Domestic animal endocrinology最新文献

The aim of this study was to determine if intranasal administration of oxytocin modifies sexual behaviour and the stress response in young rams during sexual tests with ewes in oestrus. Ten rams were used in a cross-over design. At Day 0, the control group (CG, n = 5) received isotonic saline spray intranasally, and the treated group (OTG, n = 5) received oxytocin (24 IU) intranasally, 40 min before the sexual test. At Day 15, the groups were reversed. In each sexual test (20 min) with an oestrous-induced ewe, the sexual behaviour of the young rams was recorded. Serum cortisol concentrations were determined before and after the test. Less flehmen was observed in the OTG, but mounts with ejaculation were increased. The OTG presented lower serum cortisol concentration than the CG. In conclusion, intranasal administration of oxytocin modified the sexual behaviour of rams, evidenced by a decrease in flehmen behaviour and an increase in mounts with ejaculation, making sexual activity more efficacious. In addition, the treatment decreased the stress response of the rams in the sexual tests. Therefore, intranasal administration of oxytocin could be used to increase sexual activity in rams, and with less stress, providing better welfare conditions.

Feline diabetes mellitus is a common endocrine disease with increasing prevalence. It shows similarities with human type 2 diabetes and is characterized by insulin resistance and deficient insulin secretion. Moreover, cats and humans belong to the very few species that form amyloid depositions in the pancreatic islets. However, little is known about cat islet function and no studies have addressed insulin secretion from isolated islets ex vivo. The aim of this study was to establish a protocol for isolation of islets of Langerhans from pancreata of cats euthanized due to disease, and to evaluate insulin secretion responses to various physiological and pharmacological stimuli. Collagenase digestion of pancreatic tissue from 13 non-diabetic cats and two cats with diabetic ketoacidosis yielded individual islets surrounded by a layer of exocrine tissue that was reduced after two days in culture. Histological examination showed islet amyloid in pancreatic biopsies from most non-diabetic and in one diabetic cat. Islets from non-diabetic cats cultured at 5.5 mM glucose responded with increased insulin secretion to 16.7 mM glucose, 30 mM K⁺ and 20 µM of the sulfonylurea glipizide (2-3 times basal secretion at 3 mM glucose). The glucagon-like peptide-1 receptor agonist exendin-4 (100 nM) had no effect under basal conditions but potentiated glucose-triggered insulin release. Only one of nine islet batches from diabetic cats released detectable amounts of insulin, which was enhanced by exendin-4. Culture of islets from non-diabetic cats at 25 mM glucose impaired secretion both in response to glucose and K⁺ depolarization. In conclusion, we describe a procedure for isolation of islets from cat pancreas biopsies and demonstrate that isolated cat islets secrete insulin in response to glucose and antidiabetic drugs. The study provides a basis for future ex vivo studies of islet function relevant to the understanding of the pathophysiology and treatment of feline diabetes.

Laparoscopic ovum pick-up (LOPU) combined with in vitro embryo production (IVEP) is a technology platform that improves the utilization rate of the elite ewe's ovarian oocytes and increases the number of obtained offspring. This study aimed to evaluate the effects of FSH pre-stimulation, serial oocyte collection, and breed on LOPU-IVEP under field conditions. Donors were randomly assigned to five groups (group A: decreasing doses of pituitary FSH (p-FSH); group B: constant doses of p-FSH; group C: two doses of long-acting recombinant ovine FSH (ro-FSH); group D: single administration of a long-acting ro-FSH in; group E: no FSH stimulation). Oocyte yield following LOPU (average recovered oocytes: 20.9 ± 0.5; average viable oocytes: 17.2 ± 0.4) and oocyte developmental competence (average blastocysts: 7.0 ± 0.2) in group C were significantly better than these of group D and group E, and similar to these of groups A and B. Meanwhile, there were no differences in oocyte yield and developmental capacity using repeated LOPU session at 1-, 2-, and 3-month intervals (p > 0.05). Finally, we compared LOPU-IVEP outcomes among five sheep breeds. The results indicated that East Friesian × Chinese Mongolian crossbred sheep and purebred East Friesian sheep had the more recovered oocytes and viable oocytes compared with the Suffolk, Dorper, and Texel breeds, and average number of blastocysts in East Friesian × Chinese Mongolian sheep group was also highest among the groups (8.1 ±0.3, p < 0.05). In summary, the results of this study indicate long-acting ro-FSH pre-stimulation combined with 12 times LOPU sessions over one year maximizes embryo production of elite donor ewes under field conditions.

This study aims to evaluate the effects of melatonin and its mechanisms of action on preantral follicle activation and survival, stromal cell density and collagen distribution in extracellular matrix (ECM). The involvement of melatonin receptors and mTORC1 pathway in these procedures were also investigated. To this end, ovarian fragments were cultured for six days in α‐MEM⁺ alone or supplemented with 1000 pM melatonin, 1000 pM melatonin with 1000 pM luzindole (inhibitor of melatonin receptors), or 1000 pM melatonin with 0.16 µg/ml rapamycin (mTORC1 inhibitor). At the end of culture period, tissues were processed for classical histology, and the follicles were classified as normal or degenerated, as well as in primordial or growing follicles. The ovarian stromal cell density and ECM collagen distribution were also evaluated. Samples of ovarian tissues were also destined to measure the levels of thiol and mRNA for CAT, SOD, GPX1 and PRDX1, as well as the activity of antioxidant enzymes CAT, SOD, and GPX1. The results demonstrated that ovarian tissues cultured with melatonin, melatonin with luzindole or melatonin with rapamycin had significantly higher percentage of morphologically normal follicles than those cultured in control medium (α‐MEM⁺). However, the presence of either luzindole or rapamycin, did not block the positive effects of melatonin on follicle survival (P > 0.05). Although the presence of melatonin in culture medium reduced the percentage of primordial follicles and increased the percentage of development follicles, these positive effects of melatonin were blocked by either luzindole or rapamycin (P < 0.05). Melatonin, melatonin with luzindole or melatonin with rapamycin did not influence the number of ovarian stromal cells. In contrast, melatonin significantly increased the percentages of collagen in ovarian tissues, but the positive effects of melatonin were blocked by either luzindole or rapamycin. Tissues cultured with melatonin and rapamycin had higher levels of mRNA for CAT and lower GPx activity when compared to those cultured in control medium. In conclusion, melatonin promotes primordial follicle activation, increases collagen fiber in ECM of in vitro cultured bovine ovarian tissue through its membrane-coupled receptors and mTORC1. Oppositely, melatonin increase follicles survival by acting through other pathways, since it can pass through cell membranes and directly regulate oxidative stress.

In this study, changes in salivary and serum proteome of dogs with hypothyroidism were studied using tandem mass tags (TMT) labelling and liquid chromatography-mass spectrometry (LC-MS/MS). Saliva and serum proteome from 10 dogs with hypothyroidism were compared with 10 healthy dogs. In saliva, a total of seven proteins showed significant changes between the two groups, being six downregulated and one upregulated, meanwhile, in serum, a total of six proteins showed significant changes, being five downregulated and one upregulated. The altered proteins reflected metabolic and immunologic changes, as well as, skin and coagulation alterations, and these proteins were not affected by gender. One of the proteins that were downregulated in saliva, lactate dehydrognease (LDH), was measured by a spectrophotometric assay in saliva samples from 42 dogs with hypothyroidism, 42 dogs with non-thyroid diseases and 46 healthy dogs. The activity of LDH was lower in the saliva of hypothyroid dogs when compared to non-thyroid diseased dogs and healthy controls.

This study indicates that canine hypothyroidism can produce changes in the proteome of saliva and serum. These two sample types showed different variations in their proteins reflecting physiopathological changes that occur in this disease, mainly related to the immune system, metabolism, skin and coagulation. In addition, some of the proteins identified in this study, specially LDH in saliva, should be further explored as potential biomarkers of canine hypothyroidism.

The underlying molecular mechanisms leading to insulin dysregulation are poorly understood in horses. Therefore, this study aimed to determine if insulin dysregulation is associated with an altered basal expression and extent of phosphorylation of key proteins of the insulin signaling cascade in liver (LT), muscle (MT), and subcutaneous adipose tissue (AT) under basal and stimulated conditions. Twelve Icelandic horses were subjected (1) to an oral glucose (Gluc PO) challenge and (2) to an intravenous (Ins IV) insulin challenge in a crossover study. Biopsies of LT, MT, and AT were taken in vivo under basal conditions and after Gluc PO and Ins IV stimulation. Corresponding insulin levels were measured by an equine optimized ELISA (Mercodia AB, Uppsala). Insulin levels ≥ 110 µIU/mL at 120 min indicated that six horses were insulin dysregulated (HI), while six were not (NI). Gluc PO stimulation resulted in a more pronounced hyperinsulinemia and hyperglycemia in HI horses compared to NI horses. Western blot analysis of key proteins of the insulin signaling cascade revealed an enhanced phosphorylation of the insulin receptor (InsR) under Gluc PO (P = 0.001) and Ins IV stimulation (P = 0.017) within LT, but not in MT and AT. Phosphorylation of protein kinase B was enhanced under Gluc PO stimulation in all tissues and under Ins IV stimulation in MT and AT, while phosphorylation of adenosine monophosphate protein kinase α was reduced after glucose administration (P = 0.005) in all horses. Interestingly, HI horses had significantly higher amounts of phosphorylated mechanistic target of rapamycin (mTOR) in MT (P = 0.049), irrespective of any stimulation. In LT, the amount of phosphorylated mTOR decreased under Gluc PO conditions in HI horses, while an increase was observed in NI horses (P = 0.015). A major limitation was the inclusion of only Icelandic horses of advanced age since insulin dysregulation could be related to both the equine metabolic syndrome and/or pituitary pars intermedia dysfunction. In summary, insulin signaling appeared to be maintained in both HI and NI Icelandic horses, although post-receptor alterations were observed. Thus, ID might be an equine-specific metabolic condition, in which alterations of the mTOR signaling pathway may play a crucial role, as emphasized by higher mTOR phosphorylation in HI horses.

The study aimed to evaluate the role of vitamin D on redox balance, insulin resistance and its predicting value for subclinical pregnancy toxemia (SPT) in pregnant ewes. At four weeks pre-lambing, fifteen healthy pregnant ewes were divided into two groups, ewes with sufficient vitamin D (25-hydroxy-vitamin D (25VitD) (SVD, n = 9) and ewes with insufficient 25VitD (ISVD, n = 6). Blood samples were collected at 4 weeks pre-lambing using modified frequently sampled intravenous glucose tolerance test for the estimation of various metabolites. The baseline glucose, insulin, non-esterified fatty acid (NEFA), fructosamine, beta-hydroxy butyric acid (β-BHA), calcium, phosphorus concentration and total oxidant status (TOS) did not differ significantly between the two groups, however, total antioxidant capacity (TAC) was significantly (p = 0.031) low in ISVD ewes. Area under the curve for glucose, insulin, elimination rate of glucose and peak insulin also did not differ significantly between the two groups. Correlation analysis revealed, positive association of 25VitD with fructosamine, calcium and TAC, and negative correlation with NEFA and TOS. Subsequent blood sampling at 2 weeks pre-lambing and at lambing showed significant difference in NEFA (p = 0.001), β-HBA (p = 0.001), and fructosamine(p = 0.012) between the two groups. A significant time x group interaction was observed in NEFA (p = 0.019), β-HBA (p = 0.031), and fructosamine (p = 0.026) concentration. The NEFA concentrations were increased and fructosamine decreased at 2 weeks pre-lambing and at lambing along with significantly increased β-HBA at 2 weeks pre-lambing in ISVD compared to SVD. Taking 0.8 mmol/L β-HBA as the cut off limit for SPT, ISVD ewes had higher odds of developing SPT two weeks prior to lambing (OD 16.00; p = 0.042) and at lambing (OD 10; p = 0.077). This study concludes that 25VitD significantly influence redox balance and energy profile and serves as a valuable predictor for SPT in pregnant sheep.

Synchronized cyclicity of replacement gilts is crucial to optimize breeding herd management, however, protocols with oral progestogen are expensive and require daily administration. This study tested two synchronization protocols without progestogens during the luteal phase in gilts. In Experiment I, on the day of the expression of the third estrus (D0), gilts were assigned to three groups (n = 6, each): control, with no treatment; PGF25: in which gilts received two doses of hCG (1,500 IU each) on D12 and D15 and two doses of a prostaglandin F2α (PGF) analogue (sodium cloprostenol; 250 µg) 6-h apart, on D25; and PGF30: in which gilts received two doses of hCG (1,500 IU each) on D12 and D15 and two doses of the PGF analogue (sodium cloprostenol; 250 µg) 6-h apart, on D30. The interval between PGF treatment and estrus expression was shorter in PGF30 than in PGF25 (P < 0.01). The PGF treatment failed to decrease serum progesterone (P4) for gilts from the PGF25 group (P > 0.05), but it was effective for gilts in the PGF30 group (P = 0.01). In Experiment II, gilts were assigned to three groups (n = 12, each): control (no treatment); eCG+hCG (400 IU eCG on D10 plus 500 IU hCG on D12); and hCG2 (two hCG doses, 1,500 IU each on D12 and D15). On D30, gilts from eCG+hCG and hCG2 that did not express estrus received two doses of the PGF analogue (250 µg each, 6-h apart). All gilts were inseminated when estrus was detected. Serum P4 concentrations were similar for all groups on D10 (P > 0.05) and greater on D20 and D25 for gilts in eCG+hCG and hCG2 (P < 0.01) than for those in the control, whereas P4 concentration was greater in hCG2 than in eCG+hCG, on both moments. The inter-estrus interval (IEI) was shorter for control gilts and intermediate for gilts in eCG+hCG, while the longest IEI was observed for gilts in hCG2 (P < 0.01). Total litter size was larger for gilts in the control (P = 0.02) compared to those in hCG2 and did not differ from the other groups for gilts in eCG+hCG (P > 0.05). In conclusion, Experiment I showed that PGF treatment did not induce luteolysis 10 days after the second hCG treatment but it was effective 15 days after the second hCG application. Additionally, Experiment II showed that both eCG+hCG and hCG2 were efficient in prolonging the luteal phase; however, number of piglets born alive and total litter size were negatively affected by the hCG2 protocol. In this sense, treatment with eCG+hCG or hCG2 may represent a steroid-free approach to prolong the luteal phase in gilts, although the doses and number of treatments must be further investigated.

The widespread use of silver nanoparticles (AgNPs) in consumer products and animal husbandry raises the need to study their impact on living organisms. This study was conducted on Hy-Line Brown hens at the age of 25 weeks with an average weight of 1.58 kg. Hens for 2 weeks received a solution of 50 nm AgNPs at a concentration of 100 pm (experimental group; n = 6) or a solution in which the nanoparticles were suspended (control group; n = 6). Thyroid hormones (thyroxine – T4, triiodothyronine – T3) were evaluated in the blood plasma and expression profiles of genes involved in thyroid hormone (TH) synthesis (TSHR, NIS, TPO, TG), metabolism (DIO1, DIO2, DIO3) and transport (MCT8, MCT10, LAT1) were determined in the chicken thyroid gland. Furthermore, iodothyronine deiodinase, TH transporter and TH receptor (THRA, THRB) mRNA expressions were evaluated in the livers isolated from the same chickens. AgNPs did not affect serum T₄ levels but elevated serum T₃ concentration. The results showed that AgNPs increased DIO3 mRNA in the thyroid gland. In turn, in the liver AgNPs administration significantly upregulated DIO2 and downregulated MCT10 mRNA levels. These results indicate that exposure to AgNPs leads to a tissue-specific alternative expression of genes engaged in TH metabolism. Moreover, the mRNA expression of DIO2 in the liver showed a positive correlation with plasma T₃ levels. In conclusion, AgNPs may have an impact on TH metabolism by affecting deiodinases and TH transporter MCT10 mRNA expression.