In vivo investigation of luteal function in dogs: Effects of cabergoline,a dopamine agonist,and prolactin on progesterone secretion during mid-pregnancy and -diestrus

The role of prolactin on luteal function in dogs was investigated in vivo. The function of prolactin in mid-luteal phase was compared in pregnant and nonpregnant dogs. A dopamine agonist, cabergoline, known for its prolactin secretion inhibitory effects, was injected subcutaneously at a dose of 5 μg/kg body weight in five pregnant and five nonpregnant Beagle bitches. Mean plasma prolactin and progesterone were dramatically suppressed for 4 to 5 days after injection in both groups when compared with control pregnant and non-pregnant animals, whereas no effect on luteinizing hormone (LH) secretion was observed. The decline in plasma progesterone occurred after that in prolactin, suggesting plasma progesterone was impaired by inhibition of prolactin secretion. These results confirm the luteotropic importance of prolactin in pregnant bitches, and also demonstrate its importance in luteal phase of the nonpregnant dog.Second, to demonstrate that the effects of cabergoline were mediated by prolactin inhibition and not by a direct action on the corpus luteum, concomitant administration on Day 30 of cabergoline and prolactin (375 μg IV twice daily on Days 30 and 31) or cabergoline and LH (750 μg IV twice daily on Days 30 and 31) was affected in two groups of five pregnant animals each. Results showed that only prolactin was able to reverse the negative effects of cabergoline on circulating progesterone. This confirms the indirect mode of action of the dopamine agonist, cabergoline on corpus luteum function.Third, further investigation on the precise luteotropic role of prolactin was made by IV injection of 375 μg pure canine prolactin twice daily in five pregnant bitches on Days 30 and 31, and in five pregnant bitches on Days 40 and 41. No direct stimulatory effect of prolactin on plasma progesterone secretion occurred. Nor was there a noticeable effect on plasma LH secretion. These results suggest that prolactin is unable to directly stimulate progesterone secretion by the corpus luteum of pregnancy.The results of this study suggest that prolactin is an essential luteotropin in the dog from mid-luteal phase in both pregnant and nonpregnant animals. However, it appears to act by sustaining corpus luteum lifespan and function rather than by direct stimulatory effects on progesterone secretion.     

犬黄体功能调控机制研究进展

犬黄体具有异于其他家畜的生理结构和功能表现,其中最特别的是犬缺乏急性黄体溶解机制。犬黄体发生过程受催乳素、促黄体素和前列腺素E₂的复杂调控,溶解退化过程则对其他家畜主要的溶黄体激素--前列腺素F_2α不敏感。而黄体的存留对母犬的生殖活动却具有重要的意义,其自分泌或旁分泌的类固醇激素也在妊娠调控和发情周期的更迭中发挥重要作用。犬黄体的退化机制尚未明确,但国外已有包括被动退化在内的诸多假说。此外,黄体是为数不多具有完整生命周期的功能性组织（或器官）,其发生过程、功能实现和衰老溶解都在周而复始地进行,这对炎症反应、癌症发生和细胞凋亡等研究都具有重要的参考价值。作者综述了犬黄体的发生和退化过程中调控机制的研究进展,旨在为进一步深入研究犬黄体发生和退化的分子机制提供研究背景和理论依据。     

Research Progress on Regulatory Mechanisms of Canine Corpus Luteum Function

Significant differences can be found in the physiology construction and functional performance of canine corpus luteum from the other domestic animal species, and the most interesting characteristic is the lack of an acute luteolytic mechanism. It is a multifactorial process that luteinization of canine is regulated by PRL, LH and PGE₂, and the luteolysis is insensitive to PGF_2α. The remaining canine corpus luteum plays an important role in bitch reproductive activities, and the steroid hormone from it through autocrine or paracrine is the same important to pregnancy and the estrus cycle. It is still unknown about the mechanism of regression of canine corpus luteum, however, there are several hypotheses including the passive regression mechanism. Moreover, corpus luteum is one of the few tissues (organs) which includes all three parts formation, maintenance and regression of the life cycle, and it is very important for the research of inflammatory reaction, carcinogenesis and apoptosis. In this review, the advanced progress of formation and regression of canine corpus luteum were summarized, which would provide theoretical basis and research background to study the molecular mechanism in this filed.     

Temporal relationship of the pulsatile fluctuation of luteinizing hormone and progesterone in cattle: a time series cross-correlation analysis

The temporal relationship between the pulsatile patterns of plasma luteinizing hormone (LH) and progesterone (P4) was studied in mid-luteal (ML) and early-pregnant (EP) dairy cows. Blood samples were collected (via external jugular vein cannulae) at 10-min intervals for 16 h in 5 ML cows (d 10 to 12 of the cycle) and for 10 h in 5 EP cows (d 52 to 56 of gestation). Concentrations of LH and P4 were determined by radioimmunoassays and a time series cross-correlation analysis was utilized to evaluate the temporal relationship between them. A pulsatile pattern was found for both hormones in both groups, and in all animals LH peaks were uniformly followed by P4 peaks. In 80% of the cows in both groups the highest cross-correlation occurred between samples LH(n) and P4 (n + 1) (n = sample number), suggesting that a lag time of about 10 min is necessary for luteal stimulation. Results from both groups demonstrate that P4 is released from the corpus luteum in a pulsatile manner and that its release is at least partially dependent upon the pulsatile pattern of plasma LH. Correlation coefficients between LH and P4 mean level, basal levels, peak frequencies and peak amplitudes obtained in both groups indicate that the conceptus alters the hypothalamic-pituitary-ovarian relationship found in the ML cows, suggesting the existence of another factor(s) acting along with LH to release P4 in EP cows.     

The effects of prolactin and gonadotropin on luteal function and morphology in the cyclic golden hamster

The present study was conducted to evaluate the endocrinological effects of the pituitary on luteal maintenance and regression in the cyclic golden hamster (Mesocritus auratus). After hypophysectomy (Hypox) at 0900 h on day 1 of the estrous cycle (the day of ovulation), the animals received injection of prolactin (PRL) or PRL plus equine chorionic gonadotropin (eCG). They were decapitated at 1500 h on day 3 of the cycle, and trunk blood was collected for measurement of progesterone (P4). Corpora lutea (CLs) were dissected from one ovary for DNA ladder detection by electrophoresis, determination of DNA fragmentation ratio by fluorometric measurement method and measurement of P4. The other ovary was used for histological observation. After the Hypox, the daily injection of 1 mg ovine PRL restrained the DNA fragmentation ratio and number of apoptotic cell in the CLs. The PRL treatment maintained the luteal morphology and increased the luteal P4 concentration, but not in the plasma P4 concentration. In addition to PRL, injection of 2 IU eCG after the Hypox also restrained the DNA fragmentation ratio and number of apoptotic cells in the CLs to the level of a pregnant animal. The PRL plus eCG treatment maintained the luteal morphology in the same manner as the PRL only treatment and increased not only the luteal but also the plasma P4 concentration. These results suggest that PRL restrains luteal apoptosis and maintains luteal morphology and that the combination of PRL and eCG restrains not only structural but also functional luteal regression in the cyclic hamster.     

黄体期注射PGF2α对母羊血清生殖激素及相关细胞因子的影响

旨在研究黄体期不同阶段注射前列腺激素(PGF2α)对育成母羊生殖激素和生殖相关细胞因子的影响。本研究选择健康、体况良好、体重相近、发情周期正常的湖羊育成母羊60只,用“孕酮栓(MAP)+PMSG”法进行发情周期同步化处理后,选择发情正常的48只母羊随机均分为6组。发情当天记为第0天,黄体前期试验组、中期试验组和末期试验组母羊分别在第6(黄体前期)、11(黄体中期)、16天(黄体末期)注射1 mL PGF2α(0.1 mg),黄体前期对照组、中期对照组和末期对照组母羊分别在第6、11、16天注射1 mL生理盐水,每次注射后0.5、1、2、3 h采血,用于血液指标检测。结果表明,所有试验组和对照组母羊于注射后的0.5~3 h间血清中FSH、LH、PRL、P₄、E₂水平以及TNF-α、IL-1β、IL-6、IFN-β无显著变化(P＞0.05);母羊在黄体期不同阶段注射PGF2α对0.5、1、2、3 h血清中FSH、LH、PRL及IL-1β、IFN-β无显著影响(P＞0.05),前期试验组注射后3 h P₄水平显著低于前期对照组,E₂和IL-6水平显著高于前期对照组(P＜0.05),前期试验组注射后2和3 h TNF-α水平显著高于前期对照组(P＜0.05);中期试验组注射后1 h P₄水平显著低于中期对照组(P＜0.05)。黄体前期注射PGF2α后,前期试验组0.5~ 3 h内FSH、E₂、TNF-α、IL-6整体水平显著高于前期对照组(P＜0.05),P₄整体水平显著低于前期对照组(P＜0.05),对LH、PRL、IL-1β和IFN-β无显著影响(P＞0.05);黄体中期注射PGF2α后,中期试验组0.5~3 h内E₂整体水平显著高于中期对照组(P＜0.05),P₄整体水平显著低于中期对照组(P＜0.05),对FSH、LH、PRL、TNF-α、IL-1β、IL-6和IFN-β无显著影响(P＞0.05);黄体末期注射PGF2α对生殖激素与相关细胞因子没有显著性影响(P＞0.05)。本研究结果表明,PGF2α对黄体的溶解作用存在阶段性差异,母羊在黄体前期对PGF2α的短期应答反应强于黄体中、末期,且黄体前期时,卵巢可以响应PGF2α为卵泡发育营造更佳的发育环境。     

The physiological responses of anovulatory ewes to the introduction of rams — A review

If pre-conditioned by a period of isolation from rams, prepubertally, seasonally or lactationally anovulatory ewes of many breeds can be stimulated to ovulate by the reintroduction of rams. The signal from the rams is partly pheromonal and activates neural connections between the main olfactory tract and the anterior hypothalamus. This leads to an increase in the frequency of pulses of luteinizing hormone (LH), a process which is essential for the induction of ovulation and is usually completed within a few minutes. The high pulse frequency stimulates follicular growth and oestradiol secretion by the ovaries. The subsequent build-up of oestradiol in the blood has two effects: in the short term (the first 2–12 h) it reduces the levels of follicle-stimulating hormone (FSH) and the amplitude of the LH pulses; in the long term (12–48 h) it induces preovulatory surges of both LH and FSH. The LH surge induces ovulation and the formation of a corpus luteum.In some ewes, the corpus luteum is normal and a normal luteal phase follows the first ovulation. In other ewes, the first corpus luteum secretes little progesterone and regresses within 6 days. A second LH surge is then released, inducing a second ovulation and the formation of an apparently normal corpus luteum. If the ewes are treated with progestagen before the rams are introduced, all the corpora lutea formed after the first ovulation appear to be normal. This appears to be effected both through a delay in the onset of the LH surge and through a direct action of progesterone on the ovary.     

Evidence for prolactin as the main luteotrophic factor in the cyclic dog

The role of prolactin and LH in the control of the function of the corpus luteum in the dog was studied. Experiments were performed to interfere with the secretion of a) prolactin by administering a dopamine agonist and b) LH by desensitisation with a long-acting LHRH and by stimulation. Treatments with prolactin-lowering dosages of bromocriptine, (20 micrograms/kg body weight twice a day, orally; n = 8) which started between day 1-5 (n = 4) and day 20-24 (n = 4) of the luteal period resulted in a similar pattern of progesterone, concentration in peripheral blood in both groups. The progesterone release in the second half of the luteal period (13.1 +/- 1.8% (sem) of the progesterone release of the total luteal period) was significantly lower than in control dogs (24.7 +/- 2.2%). Treatment at about day 30 of the luteal period with LHRH CR (1.34 mg, intramuscularly; n = 3), which significantly suppressed the LH level, did not reduce the progesterone release in the second half of the luteal period, 21.3 +/- 4.7% compared to 24.7 +/- 2.2% in the control dogs. The endogenous LH peak resulting from treatment with LHRH had no effect on the progesterone concentration in the blood. It is concluded that prolactin is the main luteotrophic factor in the cyclic dog during the second half of the luteal period.     

Response of the bovine corpus luteum to increased secretion of luteinizing hormone induced by exogenous gonadotropin releasing hormone

Two experiments were conducted to investigate the response of the bovine corpus luteum to surges of luteinizing hormone (LH) induced by natural gonadotropin-releasing hormone (GnRH) administered twice during the same estrous cycle. In experiment 1, eight mature beef cows, each cow serving as her own control, were injected intravenously (iv) with saline on days 2 and 8 of the cycle (day of estrus = day 0 of the cycle), then with 100 micrograms GnRH on days 2 and 8 of the subsequent cycle. Jugular blood samples were taken immediately prior to an injection and at 15, 30, 45, 60, 120 and 240 min postinjection, to quantitate changes in serum luteinizing hormone. Blood was also collected on alternate days after an injection until day 16 of the cycle, to characterize changes in serum progesterone concentrations. Although exogenous GnRH caused release of LH on days 2 and 8 of the cycle, the quantity of LH released was greater on day 8 (P less than .025). Serum levels of progesterone after treatment with GnRH on day 8 of the cycle did not differ significantly from those observed during the control cycles of the heifers. Because exposure of the bovine corpus luteum to excess LH, induced by GnRH early during the estrous cycle, causes attenuated progesterone secretion during the same cycle, these data suggest that a second surge of endogenous LH may ameliorate the suppressive effect of the initial release of LH on luteal function. Duration of the estrous cycle was not altered by treatment (control, 20.4 +/- .5 vs. treated, 20.4 +/- .4 days).(ABSTRACT TRUNCATED AT 250 WORDS)     

Morphological study of the corpus luteum in the rat

The cyclic related growth and regression of the corpus luteum during four consecutive oestrous cycles of the regular four-day-cycling (ie, 16 days), virgin albino Wistar rat were followed up by light and electron microscopic investigation of the ovaries. After ovulation, follicular granulosa cells differentiated into luteal cells in the newly formed corpus luteum. Based on their specific histological characteristics, four various types of corpus luteum in each stage of the oestrous cycle could be identified. As soon as the luteal cells started to degenerate, the number of fibroblasts progressively increased and apoptotic degeneration of luteal cells was initiated and became most prominent during oestrus. Complete regression of the corpus luteum was seen after 15 days. This study shows a strictly organised pattern of luteal cell growth and degeneration in the corpus luteum of the regular four-day-cycling, virgin Wistar rat. The morphological alterations may be regulated by a hormonal fluctuation.     

Prolonged luteal activity in mares--a semantic quagmire

Prolonged luteal activity is one of the most formidable terminology challenges in mare reproductive biology. Prolonged luteal activity can be a result of persistence of an individual corpus luteum or the sequential development of luteal glands, each of which may have a normal life span. Luteal tissue can originate from an unovulated follicle or from an ovulation occurring during either follicular or luteal dominance. These complexities, together with ambiguous and inconsistent terminology, have resulted in confusion regarding those conditions which can be grouped broadly under the term prolonged luteal activity. Persistence of an individual corpus luteum can occur in association with severe damage to the endometrium resulting in loss of the uterine luteolytic mechanism. Spontaneous (no known uterine pathology) persistence of the corpus luteum from the follicular-phase ovulation has not been documented adequately as a clinical entity. The occurrence of ovulation towards the end of dioestrus may cause confusion about the origin of prolonged luteal activity. Such immature dioestrous corpora lutea may not respond to the release of uterine luteolysin, thereby leading to prolonged luteal activity even though the original corpus luteum regressed at the normal time. In the absence of critical monitoring of the corpus luteum (eg by ultrasound) the prolonged activity could be attributed erroneously to persistence of the corpus luteum from the follicular-phase ovulation. Pseudopregnancy is another confusing term that is sometimes used to describe persistence of the corpus luteum, especially when the luteal persistence is caused by embryonic loss after the embryo has blocked the uterine luteolytic mechanism.(ABSTRACT TRUNCATED AT 250 WORDS)     

Immunolocalization of steroidogenic enzymes in the corpus luteum and placenta of the Japanese Shiba goat

In this study, we performed immunohistochemistry of cholesterol side-chain cleavage cytochrome P450 (P450scc), 3beta-hydroxysteroid dehydrogenase (3betaHSD), cytochrome 17alpha-hydroxylase P450 (P450c17), and cytochrome P450 aromatase (P450arom) in the corpus luteum and placenta of Shiba goats. The aim was to clarify the steroidogenic capability of the corpus luteum and placenta of Shiba goats. Ovaries containing corpora lutea were obtained from four adult Shiba goats during the luteal phase (day10; n=2) and pregnancy (90 and 120 days of gestation). Placenta was obtained from one Shiba goat on day 120 of gestation. The sections of the ovaries and placentae were immunostained using the avidin-biotin-peroxidase complex method (ABC) with polyclonal antibodies generated against steroidogenic enzymes of mammalian origin. All luteal cells expressed P450scc, 3betaHSD, P450c17 and P450arom. The distribution of P450scc, 3betaHSD, P450c17 and P450arom were not different during the luteal phase and pregnancy. P450arom showed a weak positive staining in late pregnancy (120 days). In addition, immunoreactions for P450c17 and P450arom were observed in syncytiotrophoblast of the placenta of one Shiba goat. These results indicate that, in Shiba goats, corpus luteum is not only an important source of progesterone but also has the ability to synthesize androgen and estrogen during the luteal phase and pregnancy. Also the placenta has the ability to synthesize androgen and estrogen in late pregnancy.     

Use of milk progesterone enzyme immunoassay for differential diagnosis of follicular cyst, luteal cyst, and cystic corpus luteum in cows

In 160 cows with ovarian cysts as determined by rectal palpation, differentiation was made of follicular cyst, luteal cyst, and cystic corpus luteum on the basis of milk progesterone concentrations estimated by an enzyme immunoassay before and at 10 days after cows were treated with gonadotropin-releasing hormone. Cows having a progesterone concentration in skim milk less than 1.0 ng/ml were considered to have follicular cysts and those with concentrations of 1.0 ng/ml or higher were regarded as the cases of luteal cyst or cystic corpus luteum. Luteal cyst was characterized by progesterone values remaining high in the cows for 10 days after treatment, and cystic corpus luteum was characterized by a decrease in progesterone concentration after cows were treated. By the rectal palpation procedure it was impossible to differentiate luteal cyst and cystic corpus luteum from follicular cyst. The frequencies of follicular cyst, luteal cyst, and cystic corpus luteum were 65%, 19%, and 16%, respectively. Of 104 cows with follicular cysts as defined by milk progesterone assay result, 73 (70%) responded to the treatment with gonadotropin-releasing hormone, the milk progesterone concentration increasing from 0.7 +/- 0.2 ng/ml (mean +/- SD) to 1.8 +/- 1.1 ng/ml. The accuracy of rectal palpation 10 days after treatment for judgment of luteinization of follicular cyst confirmed by milk progesterone analysis was only 30% (48 cows of 160).     

Changes and interrelationships among luteal LH receptors, adenylate cyclase activity and phosphodiesterase activity during the bovine estrous cycle

Changes and interrelationships among the cellular mechanisms that may regulate luteal progesterone synthesis during the bovine estrous cycle were studied. Corpora lutea (CL) were enucleated from 30 cows via a transvaginal incision on d 4, 7, 10, 13, 16 and 19 following estrus (estrus = d 0). Mean corpus luteum weight, and luteal progesterone and plasma progesterone concentrations increased (P less than .05) from d 4 to 7 or 10, and declined following luteal regression (d 19). Unoccupied LH receptor (UOR) concentrations increased (P less than .05) more than fourfold from d 4 (38 fmol/mg protein) to d 16 (173 fmol/mg protein) before declining (P less than .05) by d 19 (30 fmol/mg protein). The dissociation constant for UOR concentrations increased (P less than .05) during the estrous cycle. Concentrations of occupied LH receptors (OR) were less (P less than .05) on d 7 than other days; however, the total number of OR/CL increased fourfold from d 4 (47 fmol/CL) to d 10 (221 fmol/CL) and remained similar thereafter. Basal adenylate cyclase, LH-activated adenylate cyclase, and Gpp(NH)p-activated adenylate cyclase activities were greatest (P less than .05) on d 7 to 16 as compared with d 4 and 19. Luteinizing hormone stimulated (P less than .05) adenylate cyclase activity relative to basal activity on d 7, 10, 13, and 16; whereas, Gpp(NH)p stimulated (P less than .05) adenylate cyclase activity relative to basal activity at each period. Phosphodiesterase was 46% greater on d 19 as compared with d 4.(ABSTRACT TRUNCATED AT 250 WORDS)     

乏情奶牛日粮营养水平和血浆生殖激素的变化

选取产后65～75天无发情表现的泌乳奶牛，经直检分为卵泡闭锁（Ⅰ）、卵巢囊肿（Ⅱ）、持久黄体（Ⅲ）和排卵延迟（Ⅳ）4组，每组10头奶牛。应用放射免疫测定法检测每头奶牛血浆FSH、LH、E2、P4的水平，同时分析该牛场此时期日粮中营养水平。结果表明：饲料中能量、蛋白质和磷的含量低于NRC（2001）水平。4个组奶牛血浆中的FSH含量均低于正常水平；Ⅰ组血浆中的E2含量低于同期正常水平；Ⅱ组血浆中的E2含量高于正常水平；Ⅲ组血浆中的E2含量低于正常水平，P4的含量高于正常水平；Ⅳ组血浆中的E2含量高于正常水平，P4和LH的含量低于正常水平。结论：日粮能量、蛋白和磷含量较低导致的泌乳奶牛生殖激素的紊乱是引起奶牛乏情的主要因素。     

Hormonal control of oestrous cycle in the ewe (a review)Contrôle hormonal de l'oestrus chez la brebis: revueHormonelle Kontrolle des Brunstzyklus beim Mutterschaf

During the cycle, the secretion of progesterone by the corpus luteum inhibits the positive feedback of oestrogens and thus prevents the LH discharge, and also primes the central nervous system for oestrous behaviour. Prostaglandin F_2α has been identified as the hormone produced by the uterus which causes luteal regression. The LH discharge leading to ovulation follows the demise of the CL. None of the characteristics of the LH surge (duration, maximum level, total release) can be related to ovulation rate. However, the interval from onset of oestrus to the beginning of the LH discharge is greater in highly prolific breeds than in less prolific ones.The knowledge of these physiological processes leading to oestrus and ovulation makes possible the control of ovarian activity in the ewe. In cyclic females, the control of the timing of the LH discharge and ovulation can be obtained either by inducing luteolysis with PGF_2α or its synthetic analogues after day 4–5 of the cycle, or by artificially lengthening the luteal phase with exogenous progesterone or progestagens.During the seasonal and post-partum anoestrus, PGF_2α is ineffective and progesterone or progestagens alone are generally unable to induce oestrus and ovulation. Addition at the end of progestagen treatment of inducers of follicular growth and LH release is necessary. Both PMSG and synthetic GnRH are used for this purpose.     

Hypoxia-inducible factor-1alpha and nitric oxide synthases in bovine follicles close to ovulation and early luteal angiogenesis

The objective of the study was to characterize expression patterns of hypoxia-inducible factor-1alpha (HIF1A), inducible nitric oxide synthase (iNOS) and endothelial (eNOS) isoforms in time-defined follicle classes before and after GnRH application in the cow. Ovaries containing pre-ovulatory follicles or corpora lutea were collected by transvaginal ovariectomy (n = 5 cows/group) as follow: (I) before GnRH administration; (II) 4h after GnRH; (III) 10h after GnRH; (IV) 20h after GnRH; (V) 25h after GnRH; and (VI) 60h after GnRH (early corpus luteum). The mRNA abundance of HIF1A in the follicle group before GnRH was high, followed by a significant down regulation afterwards with a minimum level 25h after GnRH (close to ovulation) and significant increase only after ovulation. The mRNA abundance of iNOS before GnRH was high, decreased significantly during LH surge, with minimum levels afterwards. In contrast, the mRNA of eNOS decreased in the follicle group 20h after GnRH, followed by a rapid and significant upregulation just after ovulation. Immunohistochemically, the granulosa cells of antral follicles and the eosinophils of the theca tissue as well of the early corpus luteum showed a strong staining for HIF1A. The location of the eosinophils could be clearly demonstrated by immunostaining with an eosinophil-specific antibody (EMBP) and transmission electron microscopy. In conclusion, the parallel and acute regulated expression patterns of HIF1A and NOS isoforms, specifically during the interval between the LH surge and ovulation, indicate that these paracrine factors are involved in the local mechanisms, regulating final follicle maturation, ovulation and early luteal angiogenesis.     

Direct effects of linoleic and linolenic acids on bovine uterine function using in vivo and in vitro studies

The aim of the present study was to evaluate the effects of continuous administration of linoleic acid or linolenic acid into the intra-uterine horn, ipsilateral to the corpus luteum, on the duration of the estrous cycle and plasma progesterone (P4) concentration. The effects of linoleic and linolenic acids on bovine uterine and luteal functions were also studied using a tissue culture system. Intra-uterine administration of linoleic or linolenic acid (5 mg/10 ml of each per day) in cows, between days 12 and 21, resulted in a prolonged estrous cycle compared to the average duration of the last one to three estrous cycles before administration in each group (P < 0.05). Moreover, plasma P4 concentration in cows treated with linoleic or linolenic acid was high between days 19 and 21 (linoleic acid), or on day 20 (linolenic acid), compared to that of the control cows (saline administration; P < 0.05 or lower). Both linoleic (500 µg/ml) and linolenic (5 and 500 µg/ml) acids stimulated prostaglandin (PG) E2 but inhibited PGF2α production by cultured endometrial tissue (P < 0.01), while P4 production by cultured luteal tissue was not affected. These findings suggest that both linoleic and linolenic acids support luteal P4 production by regulating endometrial PG production and, subsequently, prolonging the duration of the estrous cycle in cows.     

Immunohistochemical localization of steroidogenic enzymes and prolactin receptors in the corpus luteum and placenta of spotted seals (Phoca largha) during late pregnancy

To study luteal function in the late gestational period of Phocidae (seals), we analyzed the localization of steroidogenic enzymes (P450scc, 3betaHSD and P450arom) and prolactin receptors in the corpora lutea of pregnant spotted seals (Larga seal; Phoca largha) immunohistochemically. P450scc, 3betaHSD and prolactin receptors were present in all luteal cells of each corpus luteum, and most luteal cells were immunostained for P450arom. Although we analyzed only two specimens, P450scc, 3betaHSD and prolactin receptors were negatively immunostained in the placentae. P450arom was present in the syncytiotrophoblast of placentae. These findings suggest that 1) the corpus luteum of the spotted seal synthesizes pregnenolone, progesterone and estrogen during late gestational period, 2) the placenta of this species do not possess the capacity to synthesize progesterone, and 3) like other terrestrial carnivores, this species requires prolactin to maintain the corpus luteum during pregnancy. These characteristics support the recent classification of family Phocidae in the order Carnivora, and suggest a relationship between prolactin and reproductive failure during the post-implantation period in pinnipeds.