Oxytocin synthesis and secretion from bovine corpora lutea exposed in vitro to cycloheximide and colchicine

Two experiments were conducted to study the effects of cycloheximide and colchicine on prostaglandin F₂ (PGF₂)-induced secretion and synthesis of oxytocin in bovine luteal tissue in vitro. Corpora lutea were collected from beef heifers on Day 8 of the estrous cycle. In Experiment 1, incorporation of [¹⁴C]-leucine into oxytocin synthesized and secreted by luteal slices after exposure to PGF₂, cycloheximide and cycloheximide plus PGF₂ was examined. In Experiment 2, synthesis and secretion of oxytocin were evaluated in luteal slices incubated with colchicine and PGF₂ alone and in combination. Cycloheximide inhibited incorporation of labeled leucine into luteal proteins by more than 90% and no labeled oxytocin was detected in the media or tissue. Prostaglandin F₂ induced significant secretion of oxytocin that was not inhibited by cycloheximide. Tissue levels of oxytocin after incubation with cycloheximide and/or PGF₂ did not differ and were similar to those of the incubated control. Colchicine alone did not suppress oxytocin secretion and did not alter the ability of PGF₂ to induce significant secretion of this nonapeptide. Tissue concentrations of oxytocin after incubation with colchicine and/or PGF₂ did not differ. These studies indicate that secretion and replenishment of luteal oxytocin in vitro is not contingent upon de novo protein synthesis. Inability of colchicine to suppress oxytocin secretion and synthesis may have been due to the short duration of exposure of luteal tissue to the drug.     

Responsiveness of bovine corpora lutea to prostaglandin F2 alpha: comparison of corpora lutea anticipated to have short or normal lifespans

The objective of this study was to determine if corpora lutea anticipated to have short lifespans were more responsive to the luteolytic action of prostaglandin F2 alpha (PGF2 alpha) than corpora lutea anticipated to have normal lifespans. Sixteen cows were allotted randomly to a hysterectomized-control (HC) or hysterectomized-progestogen (norgestomet) implant (HN) group. To verify that progestogen treatment of postpartum cows prior to induction of ovulation with gonadotropin-releasing hormone (GnRH) results in an increased number of cows exhibiting normal-length luteal phases, 21 additional cows were allotted randomly to a uterine intact-control (IC) or a uterine intact-progestogen implant (IN) group. Cows allotted to the HN and IN groups received norgestomet ear implants for 9 d beginning 17 to 21 d postcalving. All cows were injected (i.m.) with 100 micrograms GnRH 28 to 32 d postcalving (48 h after implant removal in the HN and IN groups) to induce ovulation. Two or 3 d after GnRH injection (d 0), cows in the HC (n = 8) and HN (n = 8) groups were hysterectomized to remove the major endogenous source of PGF2 alpha, and on d 7 cows were injected (i.m.) with 10 mg PGF2 alpha to assess luteal sensitivity. The proportion of corpora lutea having normal lifespans was greater (P less than .1) for the IN than for the IC group. In HC and HN groups, concentration of progesterone (P) increased similarly from d 0 to 6. Injection of PGF2 alpha in HC and HN groups on d 7 decreased (P less than .01) concentration of P approximately 50% by 6 h after injection (similar for both groups). Complete luteolysis was induced by PGF2 alpha in none of eight and two of eight cows in the HC and HN groups, respectively. In remaining cows (HC and HN groups) concentration of P increased (P less than .01; similar for HC and HN groups) beginning 24 h after PGF2 alpha and remained elevated through d 30 to 34 (end of experimental-period). In summary, corpora lutea anticipated to be short-lived were not more responsive to PGF2 alpha than corpora lutea anticipated to have normal lifespans.     

Progesterone production in mares and echographic evaluation of the corpora lutea formed after follicular aspiration

Ultrasound‐guided follicular aspiration was performed in 26 Criollo crossbred mares, followed by the evaluation of ultrasonographic images of the Corpus luteum (CL) that was formed after puncture of follicles of different diameters (Group 25–29 mm; Group 30–35 mm and Group >35 mm). Serum progesterone (P₄) concentrations were measured to determine CL function. The size of the CL was measured and the CL was classified based on the following echoscore: 1– anechoic tissue; 2– poorly defined luteal structure with low echogenicity; 3– echogenicity analogous to a luteal structure. The proportion of aspirated follicles that formed a functional CL (based on P₄ concentration) 8 days after aspiration was 57.1% (4/7; CL size 25–29 mm), 75.0% (6/8; CL size 30–35 mm) and 72.7% (8/11; CL size >35 mm), respectively (p > 0.05). The echographic scores of aspirated follicles (indicating the presence or absence of a CL) were consistent with serum P₄ concentrations (p < 0.0001). Of 26 aspirations, 18 resulted in luteal function confirmed by increased progesterone concentrations ([P₄] > 1.0 ng/ml); 17 of these mares (94.4%) had an echoscore (2–3) compatible with luteinization (p = 0.0372). Eight days after aspiration, serum [P₄] > 2.0 ng/ml was associated with high (p = 0.0056) CL echoscore (3) in 15 of 17 mares (88.2%). The echoscore used in this study was valuable as a screening test to detect the presence of a functional CL after aspiration. An echoscore of 3 served as a practical and efficient method to confirm luteinization.     

Chemoattractant properties of conditioned medium from equine corpora lutea collected at various stages of the oestrous cycle

This study investigated the chemotactic activity of equine CL at different stages of the oestrous cycle. The purpose of this was to ascertain whether luteal tissue itself contributes to the massive influx of leucocytes around the time of natural and induced luteal regression. Corpora lutea were collected at different stages of dioestrus and after treatment with PGF2alpha. Culture medium harvested after incubation of luteal tissue for 20 h was chemotactic for both polymorphonuclear and mononuclear cells in late dioestrus (before functional regression) as well as after natural and induced luteal regression. By contrast, midluteal tissue showed no chemotactic activity. This is the first report of the ability of equine luteal tissue actively to recruit inflammatory cells in vitro and supports our earlier findings that this infiltration starts prior to functional luteolysis. We hypothesise that this early influx of inflammatory cells may play an active role in luteal regression. Further research is needed to identify the specific chemotactic factor(s).     

Production of heparin-binding angiogenic factor(s) by bovine corpora lutea during pregnancy.

Effects of luteal-conditioned media (LCM) on proliferation and migration of endothelial cells were used to assess angiogenic activity of corpora lutea (CL) obtained from cows on d 100 (n = 5), 150 (n = 6), 200 (n = 6), and 250 (n = 6) of gestation. Explants of CL (200 mg) were incubated for 6 h in 3 mL of serum-free media containing no hormone, LH (1 microgram/mL), prostaglandin F2 alpha (PGF2 alpha; 3 microM), or both hormones. Media from the four stages of gestation were subjected to the following procedures: 1) ultrafiltration, 2) high-salt (3.0 M NaCl) treatment and then ultrafiltration, 3) heat treatment, 4) heparin-Sepharose affinity chromatography, 5) immunoneutralization with specific antibodies against heparin-binding growth factor (HBGF)-1 and against HBGF-2, and 6) dot immunoblot assay for HBGF-2. Fractions from the first five procedures were evaluated in the endothelial cell proliferation bioassay. In addition, progesterone concentration of LCM was determined by RIA. Across all days of gestation and hormone treatments, LCM stimulated (P less than .05) proliferation and migration of endothelial cells, but activities did not differ among stages of gestation or hormone treatments. Both mitogenic and migration-stimulating fractions seemed to have Mr greater than 100,000. The mitogenic activity fraction had an apparent Mr greater than 100,000 even after treatment with high salt and was heat-labile. This endothelial mitogen was retained on heparin-Sepharose columns and was eluted with 2.0 M NaCl. Mitogenic activity was partly neutralized (P less than .05) by antibodies against HBGF-2 but not HBGF-1. Presence of HBGF-2 in LCM was detected by dot immunoblot assay.(ABSTRACT TRUNCATED AT 250 WORDS)     

Luteinizing hormone receptor number and affinity in corpora lutea from prepuberal gilts induced to ovulate and spontaneous corpora lutea of mature gilts

This study was designed to determine if luteal cell receptors for luteinizing hormone/human chorionic gonadotropin (LH/hCG) contribute to the previously demonstrated abnormal function of induced corpora lutea (CL) in gilts. Twenty-five prepuberal (P) gilts, induced to ovulate with 1,500 IU pregnant mare serum gonadotropin followed 72 h later with 500 IU hCG (d 0 = day of hCG), and 22 mature (M) gilts that had displayed two or more estrous cycles were ovariectomized (OVX) on d 10, 14, 18, 22 or 26 after the onset of estrus. All gilts except those OVX on d 10 were hysterectomized between d 6 and 9 to ensure luteal maintenance. The CL were stored at -196 degrees C until determination of LH/hCG receptor number and dissociation constant (KD) by saturation analysis. Receptor number was greater for M than for P gilts on d 14 (P less than .07) and d 18 (P less than .01). The KD was greater in M than in P gilts on d 14 (P less than .01) and d 18 (P less than .0001). The LH/hCG receptor number and KD of P gilts remained the same throughout the days studied. The LH/hCG receptor number (fmol/mg protein) of M gilts was elevated on d 10, 14, and 18 (50.8, 50.4 and 51.4, respectively) and decreased on d 22 (26.5) and d 26 (25.4) to values similar to those of P gilts. In M gilts, KD increased on d 14, remained high on d 18 and decreased on d 22. We suggest that abnormal function of induced CL in P gilts may be due to an elevated LH receptor number.     

Growth hormone, insulin and glucose concentrations in bovine fetal and maternal plasmas at several stages of gestation

For cows on d 137 (n = 6), 180 (n = 8), 226 (n = 9) and 250 (n = 5) of gestation (Exp. 1), concentrations of insulin and glucose were two- to three-fold less (P less than .01) in fetal venous plasma than in uterine arterial plasma. Concentrations of growth hormone, conversely, were 10- to 20-fold greater (P less than .01) in fetal venous than in uterine arterial plasma. Concentrations of insulin and glucose in maternal and fetal plasmas and concentrations of growth hormone in maternal plasma did not vary with stage of gestation. Concentrations of growth hormone in fetal venous plasma, however, were greater on d 226 and 250 than on d 137 and 180. For cows (n = 6) on d 198 of gestation (Exp. 2), concentrations of insulin and glucose in maternal and fetal plasmas and of growth hormone in maternal plasma remained relatively constant in samples collected every 30 min for 3 h. In contrast, growth hormone concentrations in fetal venous plasma were highly variable and appeared to be episodic, with pulses of 10 to 60 ng/ml in amplitude. No significant correlations were found among concentrations of insulin, glucose and growth hormone in fetal venous plasma. When samples were collected every 15 min for 4 h from cows (n = 5) on d 198 of gestation (Exp. 3), episodes of growth hormone in fetal venous plasma were irregular in amplitude and frequency.(ABSTRACT TRUNCATED AT 250 WORDS)     

Toll‐like receptor and related cytokine mRNA expression in bovine corpora lutea during the oestrous cycle and pregnancy

Improving our understanding of the mechanisms controlling the corpus luteum (CL) and its role in regulating the reproductive cycle should lead to improvements in the sustainability of today's global animal industry. The corpus luteum (CL) is a transient endocrine organ composed of a heterogeneous mixture steroidogenic, endothelial and immune cells, and it is becoming clear that immune mechanisms play a key role in CL regulation especially in luteolysis. Toll‐like receptors (TLR) mediate innate immune mechanisms via the production of pro‐inflammatory cytokines, especially within various tissues, although the role of TLR within CL remains unknown. Thus, the objectives of this study were to characterize TLR mRNA expression in the CL during the oestrous cycle and in pregnancy (day 30–50), and to examine the role of TLR signalling in luteal cells. Corpora lutea were collected at various stages of the cycle and pregnancy and analysed for TLR and cytokine mRNA expression. In addition, luteal cells were cultured with the TLR4 ligand (lipopolysaccharide, LPS) for 24 h to evaluate the role of TLR4 in regulating luteal function. Toll‐like receptors 1, 2, 4, 6, tumour necrosis factor alpha (TNF), interferon gamma (IFN‐G), and interleukin (IL)‐12, mRNA expressions were greatest in regressing CL compared with earlier stages (p < .05), whereas no change was observed for IL‐6 mRNA expression. Cytokine mRNA expression in cultured luteal cells was not altered by LPS. Based on these data, one or more of the TLRs found within the CL may play a role in luteolysis, perhaps via pro‐inflammatory cytokine mRNA expression.     

Microvascularization and angiogenic activity of equine corpora lutea throughout the estrous cycle

Corpus luteum growth and endocrine function are closely dependent on the formation of new capillaries. The objectives of this study were to evaluate (i) tissue growth and microvascular development in the equine cyclic luteal structures; (ii) in vitro angiogenic activity of luteal tissues in response to luteotrophic (LH, PGE₂) and luteolytic (PGF₂) hormones and (iii) to relate data to luteal endocrinological function. Our results show that microvascular density was increased in the early and mid luteal phase, followed by a fall in the late luteal phase and a further decrease in the corpus albicans. Hyperplasia of luteal tissue increased until the mid luteal phase and it was followed by tissue regression. Luteal explants were cultured with no hormone added, or with PGF₂, LH, PGE₂, LH + PGE₂ or LH + PGF₂. Media conditioned by equine luteal tissue from different stages of the luteal phase were able to stimulate mitogenesis of bovine aortic endothelial cells (BAEC), suggesting the presence of angiogenic activity. No difference was observed among luteal structures on their mitogenic capacity, for any treatment used. Nevertheless, Late-CL conditioned-media with PGF₂ showed a significant decrease in BAEC proliferation (p < 0.05) and LH + PGF₂ a tendency to reduce mitogenesis. Thus, prostaglandin F₂ may play a role on vascular regression of the CL during the late luteal phase in the mare. These data suggest that luteal angiogenesis and vascular regression in the mare are coordinated with the development of non-vascular tissue and might be regulated by many different factors.     

妊娠母猪舍的规格和设计

1引言妊娠母猪舍主要是一个能让母猪保持舒适生活环境、继续怀孕的饲养舍。因此,该设施的设计应能满足母猪维持妊娠、维护身体状况和延长生产寿命以     

Dimension and design of the gestation unit

1引言妊娠母猪舍主要是一个能让母猪保持舒适生活环境、继续怀孕的饲养舍.因此,该设施的设计应能满足母猪维持妊娠、维护身体状况和延长生产寿命以及有利于生产出大体重仔猪的要求.另外,母猪生物学上的要求决定了对技术和猪舍设计的要求.在设计猪舍时如果无法满足其生物要求,那么只能通过改善管理来消除由此造成的影响.