Abstract: Publication date: Available online 18 May 2013
Source:Domestic Animal Endocrinology
Author(s): C. Gilor , S. Gilor , T.K. Graves , L.B. Borst , P. Labelle , T.K. Ridge , D. Santoro , O. Dossin
Glucose-dependent insulinotropic peptide (GIP), glucagon-like peptide (GLP)-1 and GLP-2 are hormones secreted from specialized K cells (GIP) and L cells (GLP-1, GLP-2) in the intestinal mucosa. These hormones play major roles in health and disease by modulating insulin secretion, satiety, and multiple intestinal functions. The aim of this study was to describe the distribution of K cells and L cells in the intestines of healthy cats. Samples of duodenum, mid-jejunum, ileum, cecum, and colon were collected from 5 cats that were euthanized for reasons unrelated to this study and had no gross or histologic evidence of gastrointestinal disease. Samples stained with rabbit-anti-porcine GIP, mouse-anti-(all mammals) GLP-1, or rabbit-anti-(all mammals) GLP-2 antibodies were used to determine the number of cells in 15 randomly selected 400× microscopic fields. In contrast to other mammals (eg, dogs) in which K cells are not present in the ileum and aborally, GIP-expressing cells are abundant throughout the intestines in cats (>6/high-power field in the ileum). Cells expressing GLP-1 or GLP-2 were most abundant in the ileum (>9/high-power field) as in other mammals, but, although GLP-1–expressing cells were abundant throughout the intestines, GLP-2–expressing cells were rarely found in the duodenum. In conclusion, the distribution of GIP-secreting K cells in cats is different from the distribution of K cells that is described in other mammals. The difference in distribution of GLP-2– and GLP-1–expressing cells suggests that more than 1 distinct population of L cells is present in cats.
Abstract: Publication date: Available online 17 May 2013
Source:Domestic Animal Endocrinology
Author(s): A. Cools , D. Maes , R. Decaluwé , J. Buyse , T.A. van Kempen , G.P. Janssens
Highly prolific sows often experience peripartum hypophagia, resulting in decreased production rate. Leptin, ghrelin, and resistin are known as feed intake–regulating hormones in many species, but it is yet unknown how feeding strategy and body condition will affect these hormones around parturition in sows. In the present study, a total of 63 sows, parity 2 to 7 were divided over 2 treatment groups which were fed either restricted (RESTRICT) or ad libitum (ADLIB) during the peripartum period (day 106 of gestation until day 7 of lactation). Within each treatment group, sows were assigned to 1 of 3 body condition groups based on back fat thickness at day 106 of gestation: <18 mm (LEAN), between 18 and 22 mm (MODERATE), and >22 mm (FAT). Postprandial blood samples were taken on days 107, 109, and 112 of gestation and on days 1, 3, and 5 of lactation. With RIA, leptin, ghrelin, and resistin of each sample were analyzed. For both leptin and resistin, the hormonal profile gradually increased throughout the peripartum period (P < 0.001), whereas ghrelin peaked on day 109 of gestation compared with day 107 of gestation and day 1 of lactation. Other time points were intermediate between those two (P < 0.001). The peripartum profile of leptin was significantly higher for FAT sows than for the 2 other condition groups. No effect of body condition on ghrelin and resistin concentrations was observed. None of the 3 measured hormones were affected by feeding strategy. In conclusion, during the peripartum period feed intake of sows did not affect leptin, ghrelin, or resistin profiles. Leptin was the only hormone investigated that reflected body condition. Although body condition and late gestation feed intake have been previously described as risk factors for peripartum hypophagia, they did not induce hypophagia in any of the sows or affect the profile of the observed feed intake–regulating hormones during the peripartum period.
Abstract: Publication date: Available online 7 May 2013
Source:Domestic Animal Endocrinology
Author(s): B. Mishra , K. Koshi , K. Kizaki , K. Ushizawa , T. Takahashi , M. Hosoe , T. Sato , A. Ito , K. Hashizume
A disintegrin and metalloproteinase with thrombospondin motifs 1 (ADAMTS1) is a secreted protease. Through the regulation of extracellular matrix remodeling or developmental processes or both, ADAMTS1 is involved in several biological functions, including ovulation and embryo receptivity. However, the expression and possible role of ADAMTS1 in bovine endometrium is unknown. In this study, we analyzed ADAMTS1 mRNA expression in bovine endometrium during the estrous cycle, peri-implantation period, and at different stages of gestation by using quantitative real-time RT-PCR (qPCR) and in situ hybridization. The qPCR results indicated that the expression of ADAMTS1 mRNA was not affected by the day of the estrous cycle and was similar to cyclic levels on day 35 of gestation; however, the expression was more abundant in cotyledonary tissues of the placenta during late gestation. The in situ hybridization study showed that ADAMTS1 mRNA was detected mainly in uterine luminal epithelia and stromal cells during the estrous cycle and peri-implantation period. A disintegrin and metalloproteinase with thrombospondin motifs 1 mRNA was also expressed in the peri-implantation conceptus as well as in trophoblast cells, which include binucleate cells, and increased during late gestation. Furthermore, treatment of stromal cell with progesterone (300 nM) stimulated the expression of ADAMTS1 mRNA. This study indicates that ADAMTS1 participates in bovine endometrial remodeling, which is required for implantation and placental development in coordination with ovarian steroids.
Abstract: Publication date: Available online 4 May 2013
Source:Domestic Animal Endocrinology
Author(s): Z. Bohák , F. Szabó , J.-F. Beckers , N. Melo de Sousa , O. Kutasi , K. Nagy , O. Szenci
Daily fluctuations of cortisol concentration in the blood or saliva have been repeatedly reported. However, several contradictions in the existing literature appear on this subject. The present study was performed to definitively establish options for testing adrenocortical function. To the best of our knowledge, this is the first study to evaluate parallel circadian rhythms in salivary and serum cortisol concentrations during a 24-h period. Twenty horses were examined under the same conditions. Blood and saliva samples were taken every 2 h for 24 h to determine the daily changes in cortisol concentrations of saliva and serum at rest and to determine the relationship between salivary and serum cortisol levels. Cosinor analysis of group mean data confirmed a significant circadian component for both serum and salivary cortisol concentrations (P < 0.001 in both cases). The serum cortisol circadian rhythm had an acrophase at 10:50 AM (95% CI, 10:00 AM–11:40 AM), a MESOR of 22.67 ng/mL, and an amplitude of 11.93 ng/mL. The salivary cortisol circadian rhythm had an acrophase at 10:00 AM (95% CI, 9:00 AM–11:00 AM), a MESOR of 0.52 ng/mL, and an amplitude of 0.12 ng/mL. We found a significant but weak association between salivary and serum cortisol concentrations; the Pearson correlation coefficient was 0.32 (P < 0.001). The use of salivary cortisol level as an indicator of hypothalamic-pituitary-adrenal axis activity may be warranted. However, the salivary cortisol levels are more likely to be correlated with free plasma cortisol than with the total plasma cortisol concentration.
Abstract: Publication date: Available online 12 April 2013
Source:Domestic Animal Endocrinology
Author(s): S. Yonekura , Y. Tokutake , S. Hirota , M.T. Rose , K. Katoh , H. Aso
Leptin is thought to be not only a satiety factor but also a stimulator of angiogenesis. We examined leptin, PPARγ2, and vascular endothelial growth factor (VEGF) expression in bovine intramuscular preadipocyte (BIP) cells during proliferation. The cells were seeded at 0.85 × 104 cells/cm2 and collected every day until the fifth day after passage. Leptin mRNA was present in the cells between days 2 and 4, as indicated by RT-PCR analysis. Western blot analysis showed a band for leptin at approximately 16 kDa on all of the days during growth, and the cytoplasmic concentration of leptin was highest on day 2 and decreased gradually thereafter. A PPARγ2 band at approximately 54 kDa was also observed on all days. The concentration was highest on day 2 and decreased thereafter, which is similar to the expression pattern of leptin. In constant, the expression level of VEGF protein did not change while in culture. We have demonstrated that BIP cells can synthesize both leptin and PPARγ2, with maximal synthesis occurring during maximal proliferation. Given the role of leptin in angiogenesis, we speculate that leptin is involved in the neovascularization of adipose tissue, because new organization of adipose tissue requires the growth of new blood vessels.
Abstract: Publication date: Available online 11 April 2013
Source:Domestic Animal Endocrinology
Adipogenesis, the development from preadipocytes or mesenchymal stem cells (MSCs) to mature adipocytes, is regulated by a network of signaling pathways and transcription factors. The involvement of Notch signaling and its effector HES1 in adipogenesis has been investigated in several studies with conflicting results. The underlying mechanisms remain unclear because of the lack of information about HES1 target genes during adipocyte differentiation. As a novel gene transiently up-regulated in early adipogenesis, FAD24 functions as a positive regulator of adipocyte differentiation in both preadipocytes and MSCs. In the present study, we report that the expression level of FAD24 is inversely associated with that of HES1 in porcine MSCs after adipogenic induction. Enforced overexpression of HES1 in MSCs during the early stage of adipogenesis significantly repressed the transcription of FAD24 (P < 0.01) and the other pro-adipogenic genes (P < 0.05), resulting in reduced intracellular lipid accumulation. Sequence analysis showed that porcine FAD24 harbors an evolutionarily conserved HES1 binding site in its proximal promoter region. Functional HES1, but not its dominant-negative mutant, markedly reduced the promoter activity of FAD24 (P < 0.01). Site-directed mutation and chromatin immunoprecipitation further confirmed that HES1 inhibits FAD24 transcription by direct binding to the promoter. Taken together, we identified FAD24 as a novel downstream target of HES1 during adipogenesis. Our data suggest that HES1-mediated repression of FAD24 transcription at the early stage of adipocyte differentiation may contribute to the impaired adipogenesis induced by the Notch-HES1 signaling pathway.
Abstract: Available online 6 April 2013 Publication year: 2013
Source:Domestic Animal Endocrinology
To determine the relationships among vasculature, mitotic activity, and expression of endothelial nitric oxide synthase (eNOS) of antral follicles in Bos indicus, bovine ovaries were obtained on day 6 of the estrous cycle from 10 crossbred (Brahman to Thai native cows) after a synchronized estrus with prostaglandin F2α analogue. Ovaries were fixed, paraffin-embedded, and used for immunofluorescence detection of factor VIII (a marker of endothelial cells). Immunostaining of eNOS and proliferating cell nuclear antigen (PCNA) were performed with specific monoclonal antibodies. Vasculature and positive staining of eNOS and PCNA were quantitatively evaluated with the image analysis. Follicles were classified by size (small, medium, and large) and by structure as healthy and atretic follicles (n = 82). The expression of factor VIII and eNOS were detected greater in the blood vessels of the theca layers of the healthy follicles than those in atretic follicles. The labeling indices (LIs) in granulosa and theca cells were greater (P < 0.05) in the healthy small and medium follicles than in the healthy large follicles. Vasculature, capillary area density, and capillary number density were positively correlated with eNOS expression and the LIs of granulosa and theca cells but were negatively correlated with the healthy follicle size. During the growing phase of antral follicle in Bos indicus, relationships among vasculature, mitotic activity, and eNOS were observed predominantly in healthy antral follicles. Thus, these data highlight the importance of vasculature, cell proliferation, and eNOS expression of growing and atretic follicles in the first follicular wave.
Abstract: Available online 16 March 2013 Publication year: 2013
Source:Domestic Animal Endocrinology
A preadipocyte primary cell culture was established to gain knowledge about adipose tissue development in gilthead sea bream (Sparus aurata), one of the most extensively produced marine aquaculture species in the Mediterranean. The preadipocytes obtained from the stromal-vascular cell fraction of adipose tissue proliferated in culture, reaching confluence around day 8. At that time, the addition of an adipogenic medium promoted differentiation of the cells into mature adipocytes, which showed an enlarged cytoplasm filled with lipid droplets. First, cell proliferation and differentiation were analyzed under control and adipogenic conditions during culture development. Next, the effects of insulin, GH, and IGF-I on cell proliferation were evaluated at day 8. All peptides significantly stimulated proliferation of the cells after 48 h of incubation (P < 0.002 for GH and IGF-I and P < 0.05 for insulin), despite no differences observed between the different doses tested. Subsequently, the effects of insulin and IGF-I maintaining differentiation when added to growth medium were studied at day 11, after 3 d of induction with adipogenic medium. The results showed that IGF-I is more potent than insulin-enhancing differentiation (P < 0.01 for IGF-I compared with the control). In summary, a primary culture of gilthead sea bream preadipocytes has been characterized and the effects of several regulators of growth and development have been evaluated. This cellular system can be a good model to study the process of adipogenesis in fish, which may help improve the quality of the product in aquaculture.
Abstract: Available online 16 March 2013 Publication year: 2013
Source:Domestic Animal Endocrinology
Recent reports have indicated a role of cell-to-cell interactions during gonadal development and functions. Numerous reports indicate that fetal hormonal disruption induces abnormalities in the developing reproductive system and, therefore, may interfere with reproductive functions later in adult life. Hence, this study investigated the effect of androgen deficiency during late prenatal periods on the gap junction-associated connexin 43 (Cx43) and the adherens junction-associated β-catenin expression in the fetal porcine gonads. Thus, pregnant gilts were injected with anti-androgen flutamide (for 7 d, 50 mg/kg BW per day) or corn oil (control groups) starting at 83 (GD90) or 101 (GD108) gestational day. On GD90 and GD108 the fetuses were excised and fetal gonads were obtained. To assess Cx43 and β-catenin expression real-time PCR and immunohistochemistry were performed. In fetal testes, Cx43 was localized between Leydig cells, whereas β-catenin was observed mainly within the seminiferous tubules. In fetal ovaries, Cx43 was detected between interstitial cells and between granulosa cells of forming follicles, whereas β-catenin was found within egg nests, in oocytes' membrane, and in granulosa cells of forming follicles. Immunohistochemistry showed decreased Cx43 and β-catenin expression in fetal gonads from flutamide-treated pigs compared with respective controls. However, the ovaries from animals treated with flutamide on GD108 showed increased Cx43 expression. The changes of Cx43 and β-catenin expression after prenatal flutamide treatment were confirmed at the mRNA level. These findings suggest that androgen deficiency during late gestation may lead to disturbed intercellular interactions in fetal porcine testes affecting testicular functions, as well as impaired follicular formation in fetal ovaries. Our results further signify the role of androgens in the regulation of cell-to-cell interactions within fetal porcine gonads.
Abstract: Available online 16 March 2013 Publication year: 2013
Source:Domestic Animal Endocrinology
Six insulin-sensitive and 6 insulin-insensitive mares were used in a replicated 3 by 3 Latin square design to determine the pituitary hormonal responses (compared with vehicle) to sulpiride and thyrotropin-releasing hormone (TRH), 2 compounds commonly used to diagnose pituitary pars intermedia dysfunction (PPID) in horses. Mares were classified as insulin sensitive or insensitive by their previous glucose responses to direct injection of human recombinant insulin. Treatment days were February 25, 2012, and March 10 and 24, 2012. Treatments were sulpiride (racemic mixture, 0.01 mg/kg BW), TRH (0.002 mg/kg BW), and vehicle (saline, 0.01 mL/kg BW) administered intravenously. Blood samples were collected via jugular catheters at −10, 0, 5, 10, 20, 30, 45, 60, 90, and 120 min relative to treatment injection. Plasma ACTH concentrations were variable and were not affected by treatment or insulin sensitivity category. Plasma melanocyte-stimulating hormone (MSH) concentrations responded (P < 0.01) to both sulpiride and TRH injection and were greater (P < 0.05) in insulin-insensitive mares than in sensitive mares. Plasma prolactin concentrations responded (P < 0.01) to both sulpiride and TRH injection, and the response was greater (P < 0.05) for sulpiride; no effect of insulin sensitivity was observed. Plasma thyroid-stimulating hormone (TSH) concentrations responded (P < 0.01) to TRH injection only and were higher (P < 0.05) in insulin-sensitive mares in almost all time periods. Plasma LH and FSH concentrations varied with time (P < 0.05), particularly in the first week of the experiment, but were not affected by treatment or insulin sensitivity category. Plasma GH concentrations were affected (P < 0.05) only by day of treatment. The greater MSH responses to sulpiride and TRH in insulin-insensitive mares were similar to, but not as exaggerated as, those observed by others for PPID horses. In addition, the reduced TSH concentrations in insulin-insensitive mares are consistent with our previous observation of elevated plasma triiodothyronine concentrations in hyperleptinemic horses (later shown to be insulin insensitive as well).
Abstract: Available online 16 March 2013 Publication year: 2013
Source:Domestic Animal Endocrinology
A mutation in the ryanodine receptor gene (RYR1) of the calcium release channel is responsible for increased stress susceptibility in pigs. In the present study, the relation of a mutation in RYR1 with the neuroendocrine (stress-related hormone) response and the immune defense represented by natural killer cell cytotoxicity (NKCC) during a 4-h restraint and recovery phase in 60 male pigs was investigated. Blood samples were collected from pigs previously divided into RYR1 genotypes (nn, Nn, NN), based on PCR amplification and restriction analyses. The blood samples collected during the restraint and recovery phases of the experiment were used to determine NKCC (51Cr-release assay), large granular lymphocyte number (hematologic method), and plasma concentrations of prolactin (PRL), GH, ACTH, and cortisol (COR) (by specific RIA). The greatest degree of NKCC response (P < 0.05) to restraint stress relative to controls was observed for the stress-susceptible homozygote group (nn). Measures of stress-related hormones were positively correlated with NKCC during the entire experimental period (P < 0.001 for all investigated hormones) in the nn group. Immunostimulatory effects in the early (0–60 min) phase of restraint were associated with increased hormone responses, especially PRL and GH. In the late (180–240 min) phase of stress and the recovery phase (480 min), a decrease in immune response was accompanied by an elevated COR response in all RYR1 genotypes. Moreover, divergent responses of both PRL (greatest in nn, P < 0.001) and GH (greatest in NN, P < 0.001) to the 4-h restraint were observed. Our results suggest that stress-susceptible RYR1-mutated homozygotes develop a greater level of immune defense, including cytotoxic activity of NK cells, and accompanied by more pronounced stress-induced changes in neuroendocrine response than stress-resistant heterozygous (Nn) and homozygous (NN) pigs.
Abstract: Available online 15 February 2013 Publication year: 2013
Source:Domestic Animal Endocrinology
An increasing demand for calcium during pregnancy and lactation can result in both clinical and subclinical hypocalcemia during the early lactation period in several mammalian species, in particular the dairy cow. Serotonin (5-HT) was recently identified as a regulator of lactation and bone turnover. The purpose of this study was to determine whether supplementation of the maternal diet with a 5-HT precursor would increase maternal bone turnover and calcium mobilization to maintain appropriate circulating maternal concentrations of ionized calcium during lactation. Female Sprague-Dawley rats (n = 30) were fed either a control diet (n = 15) or a diet supplemented with the 5-HT precursor 5-hydroxytryptophan (5-HTP, 0.2%; n = 15) from day 13 of pregnancy through day 9 of lactation. Maternal serum and plasma (day 1 and day 9 of lactation), milk and pup weight (daily), mammary gland and bone tissue (day 9 of lactation) were collected for analysis. The 5-HTP diet elevated circulating maternal concentrations of 5-HT on day 1 and day 9 of lactation and parathyroid hormone related-protein (PTHrP) on day 9 of lactation (P < 0.033). In addition, 5-HTP supplementation increased total serum calcium concentrations on day 1 of lactation and total milk calcium concentration on day 9 of lactation (P < 0.032). Supplemental 5-HTP did not alter milk yield, maternal body weight, mammary gland structure, or pup litter weights (P > 0.05). Supplemental 5-HTP also resulted in increased concentrations of mammary 5-HT and PTHrP, as well as increased mRNA expression of rate-limiting enzyme in 5-HT synthesis, tryptophan hydroxylase 1, and PTHrP mRNA on day 9 of lactation (P < 0.028). In addition, supplementation of 5-HTP resulted in increased mRNA expression of maternal mammary calcium transporters and resorption of bone in the femur, indicated by increase osteoclast number and diameter as well as mRNA expression of classical markers of bone resorption on day 9 of lactation (P < 0.048). These results show that increasing 5-HT biosynthesis during the transition from pregnancy to lactation could be a potential therapeutic target to explore for prevention of subclinical and clinical hypocalcemia.
Abstract: Available online 15 February 2013 Publication year: 2013
Source:Domestic Animal Endocrinology
Type 1 diabetes mellitus is one of the most frequently diagnosed endocrinopathies in dogs, and prevalence continues to increase. Pancreatic islet transplantation is a noninvasive and potentially curative treatment for type 1 diabetes mellitus. Institution of this treatment in dogs will require a readily available source of canine islets. We hypothesized that clinically acceptable islet yield and purity could be achieved by using deceased canine donors and standard centrifugation equipment. Pancreata were procured from dogs euthanized for reasons unrelated to this study. Initial anatomic studies were performed to evaluate efficacy of pancreatic perfusion. Infusion into the accessory pancreatic duct resulted in perfusion of approximately 75% of the pancreas. Additional cannulation of the distal right limb of the pancreas allowed complete perfusion. Collagenase digestion was performed with a Ricordi chamber and temperature-controlled perfusion circuit. Islets were separated from the exocrine tissue with the use of a discontinuous density gradient and a standard laboratory centrifuge. After isolation, islet yield was calculated and viability was assessed with dual fluorescent staining techniques. Islet isolation was completed in 6 dogs. Median (interquartile range) islet yield was 36,756 (28,527) islet equivalents per pancreas. A high degree of islet purity (percentage of endocrine tissue; 87.5% [10%]) and viability (87.4% [12.4%]) were achieved. The islet yield achieved with this technique would require approximately 1 pancreas per 5 kg body weight of the recipient dog. Purity and viability of the isolated islets were comparable with those achieved in human islet transplantation program. According to initial results, clinically relevant islet yield and quality can be obtained from deceased canine donors with the use of standard laboratory equipment.
Abstract: Available online 9 February 2013 Publication year: 2013
Source:Domestic Animal Endocrinology
Growth hormone is a key component of the somatotropic axis and is critical for the interplay between nutrition, regulation of metabolic functions, and subsequent processes of growth. The objective of this study was to investigate potential relations between meal feeding concentrates differing in the glycemic responses they elicit and GH secretory patterns in young growing horses. Twelve quarter horse weanlings (5.4 ± 0.4 mo of age) were used in a crossover design, consisting of two 21-d periods and two treatments, a high-glycemic (HG) or low-glycemic (LG) concentrate meal, fed twice daily. Horses were individually housed and fed hay ad libitum. On the final day of each period, quarter-hourly blood samples were drawn for 24 h to measure plasma glucose, insulin, nonesterified fatty acids, and GH. Growth hormone secretory characteristics were estimated with deconvolution analysis. After a meal, HG-fed horses exhibited a longer inhibition until the first pulse of GH secretion (P = 0.012). During late night hours (1:00 AM to 6:45 AM), HG horses secreted a greater amount of pulsatile GH than LG horses (P = 0.002). These differences highlight the potential relations between glycemic and insulinemic responses to meals and GH secretion. Dietary energy source and metabolic perturbations associated with feeding HG meals to young, growing horses have the potential to alter GH secretory patterns compared with LG meals. This may potentially affect the developmental pattern of various tissues in the young growing horse.
Abstract: Available online 8 February 2013 Publication year: 2013
Source:Domestic Animal Endocrinology
German black headed mutton (GBM) ewes are recognized as being highly susceptible to ovine pregnancy toxemia (OPT). The present trial was performed to evaluate whether a breed-dependent gestational diabetes mellitus-like insulin resistance during late pregnancy might be responsible for the high incidence of OPT in the GBM breed. Modified frequently sampling intravenous glucose tolerance tests (300 mg glucose and 0.03 mIU insulin per kg of BW) were performed during mid and late pregnancy, the periparturient, and the dry period in polytocous 3.5-y-old GBM and Finnish Landrace (FL) ewes fed according to their requirements. The corresponding blood samples were analyzed for plasma concentrations of glucose, insulin, nonesterified fatty acids (NEFAs) and β-hydroxybutyrate (β-HB). In addition, the baseline plasma cortisol concentrations were determined during late pregnancy. The BW gain during pregnancy and the rearing success did not differ between the GBM and FL ewes. In both breeds, late pregnancy was associated with decreased basal plasma glucose concentrations and enhanced glucose disposal, as well as elevated baseline β-HB values. Only in the GBM ewes did the plasma NEFA concentrations increase significantly during advancing pregnancy. Moreover, significantly higher baseline plasma NEFA concentrations as well as lower (P < 0.05) basal plasma glucose values were recorded during late pregnancy in the GBM than in the FL ewes. The first-phase insulin secretion, the peripheral insulin sensitivity, and the baseline plasma cortisol values did not differ between both breeds during late pregnancy. It is concluded that increased lipolysis during late pregnancy is a characteristic of the GBM breed. Moreover, elevated plasma NEFA concentrations may contribute to impaired pancreatic insulin response and peripheral insulin resistance in GBM ewes and thus promote OPT.
Abstract: Available online 25 January 2013 Publication year: 2013
Source:Domestic Animal Endocrinology
Fertility of domestic roosters decreases at ∼50 wk of age. In a previous study on aging white leghorn roosters, low fertility was accompanied by low levels of both hypothalamic vasoactive intestinal peptide (VIP) and pituitary prolactin (PRL) mRNA expression; however, their role in aging broiler breeder rooster reproduction is still unclear. In this study we compared reproductive activities of young (35-wk-old) and aging (73-wk-old) broiler breeder roosters. Weekly semen volume; concentration and ejaculation grade; and concentrations of plasma testosterone, estradiol, and PRL were examined. Every other week, 10 roosters from each group were euthanized, their testes weighed, and hypothalamus and pituitary removed to determine mRNA expression of hypothalamic GnRH-I, pituitary FSH, pituitary LH, hypothalamic VIP, and pituitary PRL. Aging roosters had significantly lower testis weight and semen volume, sperm concentration, ejaculation grade and plasma testosterone and low hypothalamic GnRH-I, pituitary FSH, and pituitary LH mRNA expression than young roosters (P ≤ 0.05). Aging roosters had higher concentrations of plasma estradiol and PRL and higher hypothalamic VIP and pituitary PRL mRNA expression than young roosters (P ≤ 0.05). We suggest that PRL, which is known to inhibit the gonadal axis, and its releasing factor, VIP, play an important role in the reproductive failure associated with age in broiler breeder roosters.
Abstract: Available online 24 January 2013 Publication year: 2013
Source:Domestic Animal Endocrinology
Glucose transport across the enterocyte brush border membrane by sodium/glucose cotransporter-1 (SGLT1, coded by SLC5A1) is the rate-limiting step for intestinal glucose transport. The relevance of SGLT1 expression in predisposition to diabetes mellitus and to obesity was investigated in dogs. Cultured Caco-2/TC7 cells were shown to express SGLT1 in vitro. A 2-kbp fragment of the SLC5A1 5′ flanking region was cloned from canine genomic DNA, ligated into reporter gene plasmids, and shown to drive reporter gene expression in these cells above control (P < 0.001). To determine the effect of the 3 known SNPs in this region on promoter function, new promoter/reporter constructs (all permutations of these 3 SNPs) were created by site-directed mutagenesis. No significant differences in promoter function were seen, suggesting that these SNPs do not have a significant effect on the constitutive transcription of SGLT1 mRNA in dogs. A search for novel SNPs in this region in dogs was made in 2 breeds predisposed to diabetes mellitus (Samoyed and cairn terrier), 2 breeds that rarely develop diabetes (boxer and German shepherd), and 2 breeds predisposed to obesity (Labrador retriever and cocker spaniel). The SLC5A1 5′ flanking region was amplified from 10 healthy individuals of each of these breeds by high-fidelity PCR with the use of breed-labeled primers and sequenced by pyrosequencing. The sequence of the SLC5A1 5′ flanking region in all individuals of all breeds tested was identical. On this evidence, variations in SLC5A1 promoter sequence between dogs do not influence the pathogenesis of diabetes mellitus or obesity in these breeds.
Abstract: Available online 24 January 2013 Publication year: 2013
Source:Domestic Animal Endocrinology
Photoperiod is the most important “noise-free” seasonal environmental cue for synchronizing physiological states (such as reproductive activity) in birds. However, in photoperiodic birds such as Japanese quail, the effect of photoperiod on adrenocortical activity remains unclear, particularly in males with differences in cloacal gland photoresponsiveness. At 8 wk of age, birds (n = 55) were either assigned to a short photoperiod (8L:16D; SD) or maintained under long photoperiod (16L:8D; LD; control). After 5 wk of SD exposure, males were classified as nonresponsive (SD-NR; with foam production) or responsive (SD-R; with no foam production) to short days, depending on the cloacal gland volume was above or below 1,000 mm3. At 14 wk of age, droppings were collected during 3 consecutive days to determine corticosterone (CM) and androgen metabolites (AMs) by enzyme immunoassays. Male Japanese quail under LD showed significantly higher concentrations of CM (300 ± 10 ng/g) and AMs (1,257 ± 115 ng/g) than birds kept under SD. Under short days, SD-NR and SD-R showed differences (P < 0.0001), both in CM (153 ± 8 ng/g and 98 ± 6 ng/g, respectively) and AM concentrations (1,294 ± 309 ng/g and 275 ± 53 ng/g, respectively). Interestingly, although SD-NR males exhibited no cloacal gland arrest (according to cloacal gland volume and foam production) and similar testicular activity (AM values) to LD males, they showed lower concentrations of CM compared with males that remained on LD (P < 0.05). Our findings suggest length of photoperiod affected hypothalamic-pituitary-adrenal activity; however, that was not the only factor involved, because birds subjected to shorter days but whose hypothalamic-pituitary-gonadal axis failed to respond had intermediate CM values. Further research is required to elucidate the underlying mechanisms of this interesting finding.
Abstract: Available online 29 September 2012 Publication year: 2012
Source:Domestic Animal Endocrinology
Oxyntomodulin (OXM), glucagon, glucagon-like peptide-1 (GLP-1), and exendin-4 (Ex-4) are peptide hormones that regulate glucose homeostasis in monogastric and ruminant animals. Recently, we reported that the insulin-releasing effects of OXM and glucagon in cattle are mediated through both GLP-1 and glucagon receptors. The purpose of this study was to examine the mechanisms of the glucoregulatory actions induced by Ex-4, GLP-1, OXM, and glucagon and the interrelationships among these hormones in cattle. Two experiments were performed in Holstein cattle. In Experiment 1, we initially assessed the effects of intravenous (iv) bolus injection of 0, 0.25, 1, and 2 μg/kg body weight (BW) of Ex-4, GLP-1, and OXM on insulin and glucose concentrations in 3-mo-old intact male Holstein calves. In Experiment 2, we studied insulin and glucose responses to iv coinjection of 0.25 μg of Ex-4 or GLP-1/kg BW with 2 μg of OXM or glucagon/kg BW in 4-mo-old Holstein steers. Administration of peptides and blood sampling were done via a jugular catheter. Plasma was separated and the concentrations of peptides and glucose in plasma were analyzed using radioimmunoassay and enzymatic methods, respectively. Results showed that the potent glucoregulatory action of Ex-4 in 4-mo-old steers was delayed and attenuated when Ex-4 was coinjected with OXM. The decline in plasma glucose concentrations began at 5 min in the Ex-4-injected group (P < 0.05) vs 15 min in the Ex-4 + OXM–injected group (P < 0.05). Plasma concentrations of glucose at 30 min were reduced 26% from basal concentrations in the Ex-4-injected group and 13% in the Ex-4 + OXM–injected group (P < 0.001). Results also showed that the glucose concentrations initially increased in the Ex-4 + glucagon–treated group, but declined to a relatively hypoglycemic condition by 90 to 120 min. In contrast, the glucose concentrations at specific time points between the GLP-1 + OXM–injected group and the OXM-injected group did not differ. Similarly, the glucose concentrations in the GLP-1 + glucagon–injected group did not differ from those in the glucagon-injected group. Because OXM and glucagon mediate glucose concentrations via the glucagon receptor, it is suggested that the potent glucose-lowering action of Ex-4 might include the glucagon receptor antagonistic action of Ex-4.
Abstract: Available online 29 September 2012 Publication year: 2012
Source:Domestic Animal Endocrinology
During the onset of lactation, there is a dramatic increase in the expression of glucose transporters (GLUT) and a group of enzymes involved in milk fat synthesis in the bovine mammary gland. The objective of this study was to investigate whether the lactogenic hormones mediate both of these increases. Bovine mammary explants were cultured for 48, 72, or 96 h with the following hormone treatments: no hormone (control), IGF-I, insulin (Ins), Ins + hydrocortisone + ovine prolactin (InsHPrl), or Ins + hydrocortisone + prolactin + 17β-estradiol (InsHPrlE). The relative expression of β-casein, α-lactalbumin, sterol regulatory element binding factor 1 (SREBF1), fatty acid synthase (FASN), acetyl-CoA carboxylase α (ACACA), stearyol-CoA desaturase (SCD), GLUT1, GLUT8, and GLUT12 were measured by real-time PCR. Exposure to the lactogenic hormone combinations InsHPrl and InsHPrlE for 96 h stimulated expression of β-casein and α-lactalbumin mRNA by several hundred-fold and also increased the expression of SREBF1, FASN, ACACA, and SCD genes in mammary explants (P < 0.01). However, those hormone combinations had no effect on GLUT1 or GLUT8 expression and inhibited GLUT12 expression by 50% after 72 h of treatment (P < 0.05). In separate experiments, the expression of GLUTs in the mouse mammary epithelial cell line HC11 or in bovine primary mammary epithelial cells was not increased by lactogenic hormone treatments. Moreover, treatment of dairy cows with bovine prolactin had no effect on GLUT expression in the mammary gland. In conclusion, lactogenic hormones clearly stimulate expression of milk protein and lipogenic genes, but they do not appear to mediate the marked up-regulation of GLUT expression in the mammary gland during the onset of lactation.
Abstract: Available online 1 October 2012 Publication year: 2012
Source:Domestic Animal Endocrinology
The hypothalamic-pituitary-adrenal (HPA) axis exerts a large range of effects on metabolism, the immune system, inflammatory processes, and brain functions. Together with the sympathetic nervous system, it is also the most important stress-responsive neuroendocrine system. Both systems influence production traits, carcass composition, and meat quality. The HPA axis may be a critical target for genetic selection of more robust animals. Indeed, numerous studies in various species have demonstrated the importance of genetic factors in shaping the individual HPA axis phenotype, and genetic polymorphism can be found at each level of the axis, including hormone production by the adrenal cortices under stimulation by adrenocorticotropic hormone (ACTH), hormone bioavailability, or receptor and postreceptor mechanisms. The aim of the present experiment was to extend these findings to the brain neurochemical systems involved in stress responses. To this end, a number of candidate genes were sequenced for molecular polymorphisms and their association was studied with stress neuroendocrine and production traits in a genetically diverse population consisting of 100 female pigs from an advanced intercross (F10-F12) between 2 highly divergent breeds, Large White (LW) and Meishan (MS). The LW breed has a high production potential for lean meat and a low HPA axis activity, and the MS breed has low growth rate, fat carcasses—but large litters of highly viable piglets—and a high HPA axis activity. Candidate genes were chosen in the catecholaminergic and serotonergic pathways, in the pituitary control of cortisol production, among genes previously demonstrated to be differentially expressed in ACTH-stimulated adrenal glands from LW and MS pigs, and in cortisol receptors. Sixty new polymorphisms were found. The association study with carcass and meat quality traits and with endocrine traits showed a number of significant results, such as monoamine oxidase (MAOA) polymorphisms with growth rate (P = 0.01); lean content and intramuscular fat (P < 0.01), which are the most important traits for carcass value; dopamine receptor D3 (DRD3) with carcass composition (P < 0.05); and vasopressin receptor 1B (AVPR1B) with meat quality traits (P ≤ 0.05). The effect of these polymorphisms on neuroendocrine parameters (eg DRD3 and HPA axis or AVPR1B and catecholamines) indicates information regarding their biological mechanism of action.
Cosinor analysis was used to evaluate whether pituitary and adrenal hormones exhibit circadian rhythmicity in horses. The effect of season and animal age on their respective rhythms was also determined. In addition, the usefulness of evaluating cortisol rhythmicity for the diagnosis of pituitary pars intermedia dysfunction (PPID) was assessed. Serum cortisol concentrations (P < 0.01), but not plasma ACTH or α-melanocyte-stimulating hormone (α-MSH), showed a significant circadian periodicity in horses. An effect of season on hormone concentration was observed with plasma ACTH and α-MSH concentration greater in the fall and cortisol concentration greater in the spring (P < 0.001). Age did not affect cortisol rhythm, but it did blunt the variation in cortisol concentration in horses, similar to what has been previously reported to occur in aged people and dogs. In addition, our results suggest that clinically and diagnostically normal, non–PPID-affected horses commonly have a loss of cortisol diurnal rhythm. Therefore, measurement of circadian rhythm is not an appropriate diagnostic test for PPID.
Abstract: Available online 8 October 2012 Publication year: 2012
Source:Domestic Animal Endocrinology
Porcine corpora lutea (CL) develop sensitivity to regression by prostaglandin F2α (PGF2α), termed luteolytic capacity, about 13 d after estrus. We postulated that PGF2α regulation of activating protein 1 (AP-1) transcriptional factor expression underlies acquisition of luteolytic capacity. CL were collected from gilts on day 9 (estrous cycle) or day 17 (pseudopregnancy) before or after PGF2α treatment with mRNA measured for FOS, FOSB, FOSL1, FOSL2, JUN, JUNB, and JUND and the AP-1 target genes CCL2 and SERPINE1. At 0.5 h after PGF2α, both day-9 and day-17 CL had increased (P < 0.01) mRNA for FOS (2,225% and 1,817%), JUNB (237% and 358%), and FOSB (1,060% and 925%). Intriguingly, at 0.5 h after PGF2α there was increased (P < 0.01) mRNA encoding JUN (1,099%) and JUND (300%) in day-17 but not day-9 CL. At 10 h after PGF2α there was elevated FOSB mRNA in day-17 (771%) but not day-9 CL and no PGF2α-induced change in FOS, JUN, JUND, and JUNB mRNA in day-9 or day-17 CL. Treatment with PGF2α increased mRNA for AP-1-responsive genes, CCL2 at 0.5 h (202%) and CCL2 and SERPINE1 at 10 h (719% and 1,515%), only in day-17 CL. Thus, many of the fos family of transcription factors are dramatically induced by PGF2α in CL with or without luteolytic capacity. However, PGF only induced JUN and JUND expression in CL with luteolytic capacity, a finding that may be key for understanding the acquisition of luteolytic capacity, given that JUN is the only AP-1 family member with strong N-terminal trans-activation activity.
Negative energy balance at the onset of lactation is unfavorably associated with fitness traits in high-producing dairy cows. Angiopoietin-like protein 4 (ANGPTL4) is an adipokine that has been associated with the regulation of lipid metabolism through the inhibition of lipoprotein lipase activity and regulation of lipolysis. Expression of ANGPTL4 messenger RNA (mRNA) increases during early lactation, but its regulation with changing energy status is currently unknown. Accordingly, the objective of this study was to determine whether ANGPTL4 mRNA abundance is responsive to declining energy balance induced by the transition from pregnancy to lactation, feed restriction, and GH administration in lactating dairy cows. The mRNA abundance of leptin, adiponectin, and adiponectin receptor 2 were also measured to compare adipokine mRNA profiles during changes in energy metabolism. Repeated adipose tissue biopsies were taken from different cows during transition from late pregnancy to lactation (n = 26), feed restriction (n = 19), and GH administration (n = 20). As expected, milk yield increased with the onset of lactation and GH administration (P < 0.01) but declined during feed restriction. Energy balance declined in each experiment, resulting in negative energy balance at the onset of lactation and after feed restriction. Abundance of ANGPTL4 mRNA expression increased 2- to 6-fold with declining energy balance in each experiment. Leptin mRNA declined with feed restriction, and adiponectin mRNA decreased with the onset of lactation. The consistency and magnitude of the increase in ANGPTL4 mRNA across multiple models of altered energy balance identifies it as an adipokine that is uniquely responsive to changes in energy balance in the lactating dairy cow.
Glucagon-like peptide-1 (7-36) amide (GLP-1), secreted by the small intestine, has insulinotropic and glucose-lowering action. Basal plasma GLP-1 concentrations were measured in calves around the weaning period, the effect of short-chain fatty acids (SCFA) on plasma GLP-1 concentrations was examined, and the effects of GLP-1 administration on plasma insulin, glucagon, and glucose concentrations were measured. Thirteen Holstein bull calves were fed whole milk and solid feed and weaned at 7 wk of age. Preprandial plasma samples were obtained from 5 calves once a week from week 0 to 13 to measure basal concentrations of plasma GLP-1 and insulin (experiment 1). Four calves were intravenously administered with a mixed solution of SCFA (2.4 mmol/kg body weight [BW]) in week 2 and 11 to measure plasma GLP-1 concentrations (experiment 2). Another 4 calves were intravenously injected with GLP-1 (1.0 μg/kg BW) to elucidate the response of plasma insulin, glucagon, and glucose concentrations in week 1, 2, 4, 6, 7, 9, 11, and 13 (experiment 3). In experiment 1, age and weaning did not affect preprandial basal concentrations of plasma GLP-1 throughout the experimental period. Preprandial insulin concentrations increased after weaning (P < 0.05), and GLP-1 and insulin were more strongly correlated postweaning than preweaning. In experiment 2, intravenous treatment with SCFA increased plasma GLP-1 concentrations in both week 2 and 11 (P < 0.05.) In experiment 3, intravenous GLP-1 treatment decreased plasma glucose concentrations throughout the experiment (P < 0.05), but increased plasma insulin concentrations only after weaning (P < 0.05). Treatment with GLP-1 did not affect plasma glucagon concentrations, regardless of age. These results indicate that preprandial basal concentrations of plasma GLP-1 in calves are not changed by weaning, but SCFA stimulate GLP-1 secretion. The insulinotropic action of GLP-1 is detected only after weaning, but the glucose-lowering action of GLP-1 is not affected by weaning.
To determine effects of maternal diet on in vitro fertilization (IVF) and early embryonic development, ewes (n = 48) were divided into control, overfed (ad libitum feeding), and underfed (60% of control) nutritional planes for 8 wk before oocyte collection. Follicular development was induced by twice-daily injections of FSH on days 13 and 14 of the estrous cycle, and ovaries and blood samples were collected on day 15 of the estrous cycle. During the 8-wk experiment, for control ewes BW and BCS did not change, but for overfed ewes mean (± SEM) BW and BCS increased (11.8 ± 1.1 kg and 2.0 ± 0.1, respectively) and for underfed ewes decreased (14.2 ± 0.9 kg and 0.7 ± 0.1, respectively). The number of follicles was determined; oocytes were collected and subjected to in vitro maturation and fertilization. After IVF, developing embryos were evaluated throughout the 8-d culture period. The proportion of cleaved oocytes after IVF and developing morula and blastocyst were less (P < 0.0001) in overfed and underfed ewes than in control ewes. However, number of visible follicles, total number of oocytes, number of healthy oocytes, and percentage of healthy oocytes were similar for control, overfed, and underfed ewes. Serum insulin concentration was greater (P < 0.05) in overfed ewes than in underfed ewes, estradiol 17-β (E2) concentration was greater (P < 0.05) in underfed ewes than in overfed ewes, but triiodothyronine (T3) and thyroxine (T4) concentrations were similar in all treatment groups. These data show that inadequate feeding has a negative effect on oocyte quality which results in lower oocyte cleavage after IVF and morula and blastocyst formation; overfeeding increased serum insulin and underfeeding increased serum E2 but not T3 or T4. These data emphasize the importance of diet for reproductive and metabolic functions. Furthermore, the mechanisms through which enhanced or decreased energy in diet affect oocyte quality and serum insulin and E2 concentrations remain to be elucidated.
The aims of this study were to determine the effects of lipopolysaccharides (LPS), tumor necrosis factor (TNF), interleukin 1 alpha (IL-1α), nitric oxide donor (NONOate), or the combination of TNF + IL-1α + NONOate on the following: (i) secretion of prostaglandin (PG)-F2α, PGE2, leukotriene (LT)-B4, and LTC4 by epithelial cells of the teat cavity and lactiferous sinus of bovine mammary gland; (ii) messenger RNA (mRNA) transcription of enzymes responsible for arachidonic acid (AA) metabolism (prostaglandin-endoperoxide synthase 2 [PTGS2], prostaglandin E synthase [PTGES], prostaglandin F synthase [PGFS], and arachidonate 5-lipooxygenase [ALOX5]); and (iii) proliferation of the cells. The cells were stimulated for 24 h. Prostaglandins and LT were measured by enzyme immunoassay, mRNA transcription of enzymes was determined by real-time reverse transcription polymerase chain reaction, and the cell viability was measured by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide. All factors increased PG secretion, but the highest stimulation was observed after TNF and IL-1α (P < 0.001). Tumor necrosis factor, NONOate, and TNF + IL-1α + NONOate increased LTB4 production (P < 0.01), whereas LTC4 was increased by LPS, TNF, and IL-1α (P < 0.01). Lipopolysaccharides, TNF, IL-1α, and the reagents combination increased PTGS2, PTGES, and PGFS mRNA transcription (P < 0.01), whereas ALOX5 mRNA transcription was increased only by TNF (P < 0.001). Lipopolysaccharides, TNF, IL-1α, NONOate, and the combination of reagents increased the cell number (P < 0.001). Mediators of acute-clinical Escherichia coli mastitis locally modulate PG and LT secretion by the epithelial cells of the teat cavity and lactiferous sinus, which might be a useful first line of defense for the bovine mammary gland. Moreover, the modulation of PG and LT secretion and the changing ratio of luteotropic (PGE2, LTB4) to luteolytic (PGF2α, LTC4) metabolites may contribute to disorders in reproductive functions.
The objective of this study was to investigate how weight-loss program would alter the proteome of the serum of Beagle dogs. For this purpose, serum samples from 5 Beagle dogs, before and after weight loss, were analyzed using 2-dimensional electrophoresis. Protein profiles of all samples were obtained, divided into 2 classes (obese and lean), and compared using specific 2-dimensional software, giving a total of 144 spot matches. Statistical analysis revealed 3 spot matches whose expressions were modulated in response to weight loss: 2 protein spots were upregulated and 1 protein spot was downregulated in the obese state in comparison with the lean state of the dogs. Mass spectrometric identification of differentially regulated spots revealed that these protein spots corresponded to retinol-binding protein 4, clusterin precursor, and α-1 antitrypsin, respectively, which could be considered potential markers of obesity and obesity-related disease processes in dogs.
Abstract: Available online 15 November 2012 Publication year: 2012
Source:Domestic Animal Endocrinology
The present study was designed to determine the effect of subcutaneous rivastigmine treatment on IL-1β expression and IL-1 type I receptor (IL-1R1) gene expression in the hypothalamic structures (preoptic area [POA], anterior hypothalamus [AHA], and medial basal hypothalamus [MBH]) of ewes after lipopolysaccharide (LPS) treatment. Endotoxin treatment increased (P ≤ 0.01) both IL-1β and IL-1R1 gene expression in the POA, AHA, and MBH compared with the control group, whereas concomitant rivastigmine and LPS injection abolished this stimulatory effect. It was also found that LPS elevated (P ≤ 0.01) IL-1β concentration in the hypothalamus (71.0 ± 2.3 pg/mg) compared with controls (16.1 ± 3.6 pg/mg). The simultaneous injection of LPS and rivastigmine did not increase IL-1β concentration in the hypothalamus (24.6 ± 13.0 pg/mg). This central change in IL-1β synthesis seems to be an effect of acetylcholinesterase (AChE) inhibition by rivastigmine, which decreases (P ≤ 0.01) the activity of this enzyme from 78.5 ± 15.0 μmol · min−1 · g−1 of total protein in the control and 68.8 ± 9.8 μmol · min−1 · g−1 of total protein in LPS-treated animals to 45.2 ± 5.6 μmol · min−1 · g−1 of total protein in the rivastigmine and LPS-treated group. Our study showed that rivastigmine could effectively reverse the stimulatory effect of immune stress induced by LPS injection on IL-1β synthesis through a decrease in AChE activity in the hypothalamic area of sheep. Our results also proved that the cholinergic anti-inflammatory pathway could directly modulate the central response to endotoxin.
Abstract: Available online 15 November 2012 Publication year: 2012
Source:Domestic Animal Endocrinology
Members of the bone morphogenetic protein (BMP) family regulate follicular development and granulosa cell function. However, changes in expression of BMP2 and its receptors during follicular waves in cattle and ability of BMP2 to modulate bovine granulosa cell estradiol production are not well understood. The objectives of this study were to determine temporal regulation of mRNA for BMP2 and its type I and II receptors (BMPR1A and BMPR2) in bovine follicles collected at specific stages of a follicular wave (predeviation, early dominance, mid dominance, preovulatory), ability of BMP2 to modulate bovine granulosa cell steroidogenesis, and whether effects of BMP2 on granulosa cell estradiol production are influenced by cotreatment with cocaine- and amphetamine-regulated transcript (CART), an intrafollicular regulatory peptide shown to inhibit estradiol production in response to other trophic hormones (FSH and IGF1). Relative abundance of mRNAs for BMP2 and BMPR2 was elevated at the mid dominance stage relative to earlier stages of the follicular wave and further increased at the preovulatory stage. Abundance of mRNA for BMPR1A was lowest at early dominance stage and highest at preovulatory stage relative to other stages of the follicular wave examined. Treatment of bovine granulosa cells in vitro with BMP2 increased estradiol but decreased progesterone concentrations. Co-incubation with CART reduced the BMP2-stimulated increase in granulosa cell estradiol production. Results suggest that BMP2 may play a regulatory role in development of bovine follicles to the preovulatory stage and that CART can inhibit granulosa cell estradiol production in response to multiple hormones/growth factors, including BMP2.
Abstract: Available online 15 November 2012 Publication year: 2012
Source:Domestic Animal Endocrinology
This study investigated the effect of an acute stress on food intake and on the expression of neuropeptide Y (NPY), corticotropin-releasing hormone (CRH), and ghrelin and its receptors, growth hormone secretagogue receptors (GHSRs) in the tilapia (Oreochromis mossambicus). Food intake was significantly (P < 0.01) reduced in fish after a 30-min crowding and handling stress. In a second group of animals exposed to the same 30-min stressor, tissue samples were collected immediately after the stressor to determine changes in the neuroendocrine regulators of food intake. Although CRH and NPY are considered the major mediators of appetite during stress, both mRNA levels were unaltered in the telencephalon/pre-optic area and in the hypothalamic/optic tectum. Interestingly, there was an elevation in the ghrelin transcript (P < 0.05) in the telencephalon/pre-optic area and elevation of its functional receptor (GHSR1a-LR) (P < 0.001) in the hypothalamic/optic tectum. Elevation of GHSR-LR heteronuclear RNA (P < 0.01) in the telencephalon/pre-optic area and suppression in the hypothalamic/optic tectum (P PubDate: 2012-12-16T03:02:12Z
Abstract: Available online 29 November 2012 Publication year: 2012
Source:Domestic Animal Endocrinology
Adiponectin is an adipose tissue-derived glycoprotein circulating as highly abundant multimers. It regulates glucose metabolism and insulin sensitivity. In ruminants, valid data about serum concentrations and tissue-specific protein expression are lacking, and we, therefore, aimed to generate a polyclonal antibody against bovine adiponectin to apply it in immunodetection. The specificity of the purified anti-adiponectin antibody was established by Western blot analysis with the use of reducing and denaturing conditions applied to both the purified protein and the bovine serum samples. Besides bovine serum, the applicability of the antibody for immunodetection of adiponectin was confirmed for the supernatant fluid of in vitro–differentiated bovine adipocytes, for protein extracts from bovine adipose tissue, and also in a multispecies comparison: bands comparable in size with monomeric bovine adiponectin were obtained under completely denaturing conditions in serum of camel, horse, human, mouse, pig, roe deer, and sheep. In addition, when used in immunohistochemistry on bovine adipose tissue sections, a characteristic adipocyte-specific staining pattern was obtained with this antibody. The antibody was used for establishing a semiquantitative Western blot procedure and the development of an ELISA. Both methods were extensively validated and were first applied to characterize the serum adiponectin concentrations in multiparous dairy cows during the transition from pregnancy to lactation, that is, 3 wk before until 5 wk after calving. With both assays a time effect (P = 0.017, P = 0.026, respectively) with lowest values at the day of parturition was observed. We thus established 2 useful tools to validly assess bovine adiponectin at the protein level.
Abstract: Available online 3 December 2012 Publication year: 2012
Source:Domestic Animal Endocrinology
Adiponectin's beneficial effects are mediated by the AdipoR1 and AdipoR2 receptors (AdipoRs). The pig is a good model to study complex disorders such as obesity. We analyzed the expression of adiponectin, AdipoRs and some key molecules of energy metabolism (AMP-activated protein kinase α [AMPKα], p38 mitogen-activated protein kinase [p38 MAPK], and PPARα) in 2 pig breeds that displayed an opposite genetic behavior for energy metabolism: Casertana (CE), a fat-type animal, and Large White (LW), a lean-type animal. Muscle, liver, visceral and subcutaneous adipose tissues, and brain tissues were examined. The AdipoRs cDNA sequences were identical in the 2 breeds. AdipoRs mRNA expression, measured in all tissues, was significantly lower only in the 2 adipose tissues of CE pigs (P < 0.05). The muscle expression of AdipoRs, AMPKα, p38 MAPK, and PPARα was lower in CE than in LW animals (P < 0.01, P < 0.05, P < 0.01, P < 0.01, respectively). In liver, no molecule differed between breeds. The expression of both AdipoRs in visceral and subcutaneous adipose tissues was lower in CE pigs (P < 0.01). In brain, AdipoR1 and AMPKα expression was lower in CE pigs (P < 0.01), whereas AdipoR2 tended to be lower in CE than LW pigs (P = 0.05). In conclusion, our results suggest that tissue-specific downregulation of adiponectin, AdipoRs, and of the key molecules of energy metabolism may be associated with the tendency of CE pigs to accumulate fat.
Increased secretion of prostaglandin F2α (PGF2α) within the uterus because of uterine inflammation can cause luteolysis and result in early embryonic loss. Supplementation with polyunsaturated fatty acids (PUFAs) has been shown to influence PG production in many species, although the effects on the mare remain unknown. The present study aimed to determine fatty acid uptake in equine endometrial explants and evaluate their influence on PG secretion and expression of enzymes involved in PG synthesis in vitro. Equine endometrial explants were treated with 100 μM arachidonic acid, eicosapentaenoic acid, or docosahexaenoic acid and then challenged with oxytocin (250 nM) or lipopolysaccharide (LPS; 1 μg/mL). Production of PGF2α and PG E2 (PGE2) was measured, and mRNA expression of enzymes involved in PG synthesis was determined with quantitative real-time PCR. Media concentrations of PGF2α and PGE2 were higher (P < 0.0001) from endometrial explants challenged with oxytocin or LPS compared with controls despite which fatty acid was added. Only DHA lowered (P < 0.0001) media concentrations of PGF2α and PGE2 from explants. Endometrial explants stimulated with oxytocin had increased expression of PG-endoperoxide synthase 1 (PTGS1; P < 0.02), PG-endoperoxide synthase 2 (PTGS2; P < 0.001), PG F2α synthase (PGFS; P < 0.01), PG E2 synthase (PGES; P < 0.01), and phospholipase A2 (PLA2; P < 0.005) compared with controls and regardless of fatty acid treatment; whereas stimulation with LPS increased expression of PTGS2 (P < 0.004), PGFS (P < 0.03), PGES (P < 0.01), and PLA2 (P < 0.01) compared with controls and regardless of fatty acid treatment. Treatment with PUFAs, specifically DHA, can influence PG secretion in vitro through mechanisms other than enzyme expression.
The bovine mammary alveolar cell-T (MAC-T) cell line is able to uniformly differentiate and secrete casein proteins in response to dexamethasone, insulin, and prolactin and is extensively used to study bovine mammary epithelial cell (MEC) function. Somatotropin, or growth hormone (GH), has been shown to increase milk protein synthesis both in vivo and in mammary cell models and to induce cytoskeletal rearrangement in a 3T3 fibroblast cell line and a Chinese hamster ovary cell line. To identify the nature of the effects of GH in MECs cultured with lactogenic hormones, changes in global protein expression were assessed in the MAC-T cell line with the use of two-dimensional gel electrophoresis and matrix-assisted laser desorption ionization tandem time of flight mass spectrometry. Forty proteins were differentially expressed in response to GH (P < 0.05) and were related to metabolism, the cytoskeleton, protein folding, RNA and DNA processing, and oxidant stress. These widespread changes in protein expression are indicative of a global role of GH in overall cellular differentiation that may underlie the direct modulation of milk component synthesis in MEC models that have been described to date.
Although the exact mechanism(s) by which estradiol (E2) enhances muscle growth in a number of species, including humans and cattle, is not known, E2 treatment has been shown to stimulate proliferation of cultured bovine satellite cells (BSCs). This is particularly significant because satellite cells are the source of nuclei needed to support postnatal muscle fiber hypertrophy and are thus crucial in determining the rate and extent of muscle growth. The objective of this study was to assess the role of estrogen receptor-α (ESR1) and the type 1 insulin-like growth factor receptor (IGFR1) in E2-stimulated proliferation of cultured BSCs. To accomplish this, we have used small interfering RNA (siRNA) to silence expression of ESR1 or IGFR1 and assessed the effects on E2-stimulated proliferation in BSC cultures. In BSCs treated with nonspecific siRNA, E2 significantly (P < 0.05) stimulates proliferation under conditions in which neither IGF-1 nor IGF-2 expression is increased; however, treatment of ESR1- or IGFR1-silenced cells with E2 does not significantly stimulate proliferation. These results indicate that both ESR1 and IGFR1 are required for E2 to stimulate proliferation in BSC cultures. The fact that this occurs under culture conditions in which neither IGF-1 nor IGF-2 mRNA expression is increased strongly suggests that E2 activates IGFR1 via a mechanism that does not involve increased IGF-1 or IGF-2 binding to the receptor.
Equine insulin resistance is important because of its association with laminitis. The insulin-response test is described to diagnose insulin resistance in clinical settings. Practitioners may be reluctant to perform this test because of the time needed for the test and the fear of inducing hypoglycemia. The objective of the study was to compare a 2-step insulin-response test with a complete insulin-response test. A complete insulin-response test was performed on 6 insulin-resistant horses and 6 controls. A 2-step insulin-response test consisting of an intravenous injection of 0.1 IU/kg human insulin and blood glucose determination at 0 and 30 min after injection was performed on the same horses. Times to reach a 50% reduction of glucose baseline were compared between tests and horses. All the horses tolerated both tests well. No significant difference was observed between baseline glucose concentrations of insulin-resistant horses and controls (P = 0.09). Time to reach 50% reduction of glucose baseline for controls was not significantly different with the use of the complete insulin-response test or the 2-step test (P = 0.98). For insulin-resistant horses, the time to reach 50% reduction of glucose baseline with the use of the 2-step test was significantly longer than for controls (P = 0.004). With a cut-off time of 30 min, the 2-step test had the same characteristics as the complete test. The 2-step test provided a safe, rapid, and low-cost method to diagnose insulin resistance in horses in a clinical setting.
The use of soy-based products in pig diets had raised concerns regarding the reproductive toxicity of genistein, the predominant isoflavone in soybeans. Genistein was reported to exhibit weak estrogenic activity but its mechanism of action is not fully recognized. The aim of the study was to examine the in vitro effects of genistein on (1) progesterone (P4) and estradiol (E2) secretion by porcine granulosa cells harvested from medium follicles, (2) the viability of cultured granulosa cells, and (3) the mRNA and protein expression of estrogen receptors α and β (ERα and ERβ) in these cells. In addition, to verify the role of protein tyrosine kinase (PTK)–dependent mechanisms possibly involved in genistein biological action, we tested the effects of lavendustin C, the nonsteroidal PTK inhibitor, on granulosa cell steroidogenesis. We found that genistein inhibited (P < 0.05) basal P4 secretion by granulosa cells harvested from medium follicles of pigs. In contrast, lavendustin C did not affect basal P4 secretion by the cells. Moreover, genistein increased (P < 0.05) basal granulosal secretion of E2. In contrast, lavendustin C did not alter basal E2 secretion by porcine granulosa cells. In addition, we demonstrated that genistein increased mRNA and protein expression of ERβ (P < 0.05) in the examined cells. The expression of ERα mRNA was not affected by genistein and ERα protein was not detected in the cultured granulosa cells of pigs. In summary, the genistein action on follicular steroidogenesis in pigs involved changes in the granulosal expression of ERβ. However, the genistein action on P4 and E2 production by granulosa cells harvested from medium follicles did not seem to be associated with PTK.
The aim of this study was to investigate the effects of melatonin and follicle-stimulating hormone (FSH) on the in vitro culture of goat preantral follicles. Ovarian fragments were cultured for 7 d in α-minimum essential medium (α-MEM+) containing melatonin (100, 250, 500, or 1,000 pM), FSH (50 ng/mL), or a combination of the 2 hormones and further analyzed by histology and transmission electron and fluorescent microscopy. The results showed that after 7 d of culture, tissues cultured in α-MEM+ alone or supplemented with FSH alone, melatonin (500 and 1,000 pM), or the combination of FSH and melatonin (1,000 pM) maintained percentages of normal preantral follicles similar to the fresh control. In contrast to the noncultured tissues, the percentage of developing follicles was increased under all culture conditions after 7 d (P < 0.05). The addition of 1,000 pM melatonin associated with FSH to the culture medium increased follicular and oocyte diameters compared with α-MEM+ alone after 7 d of culture (P < 0.05). Ultrastructural and fluorescent analyses confirmed the integrity of follicles cultured with 1,000 pM of melatonin plus FSH for 7 d. In conclusion, this study demonstrated that the interaction between melatonin and FSH maintains ultrastructural integrity and stimulates further growth of cultured caprine preantral follicles.
PubDate: 2012-12-16T03:02:12Z
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