GnRH的调控机制

ＧｎＲＨ由丘脑下部合成［１］,并以脉冲方式分泌进入丘脑下部垂体门脉血系统,到达垂体前叶发挥诱导促性腺激素释放的作用。ＧｎＲＨ除对垂体ＬＨ／ＦＳＨ分泌有强大的调控作用外,还与性腺和胎盘的生殖功能有关［２,３］。１ＧｎＲＨ信号转导途径ＧｎＲＨ调节垂体ＬＨ...     

GnRH及其类似物在牛繁殖管理中的应用（上）

促性腺激素释放激素及其类似物，能引起促黄体素和促卵泡素的大量释放，而使这两种激素在外周血液中的浓度升高持续３－５小时，ＧｎＲＨ所引起的黄体或卵泡功能的变化，是通过高节ＬＨ和ＦＳＨ的释放而间接起作用的。间情期（Ｄｉｅｓｔｒｕｓ）内多次或间情期后１次注射ＧｎＲＨ可引起血液孕酮水平急剧上升和黄体溶解推迟。这种作用的机理是ＧｎＲＨ能诱发ＬＨ水平升高。在性周期中注射ＧｎＲＨ能使卵泡发育，优势卵泡排卵或黄体化     

促性腺激素释放激素的研究进展

促性腺激素释放激素 (ＧｎＲＨ) ,又称黄体生成素释放激素(ＬＨＲＨ) ,为下丘脑神经元分泌的十肽激素 ,其化学结构为 :(焦 )谷 -组 -色 -丝 -落 -甘 -亮 -精 -脯 -甘 -ＮＨ2 。迄今还未发现哺乳类动物ＧｎＲＨ的结构有什么不同。下丘脑分泌的ＧｎＲＨ由垂体门脉系统到达垂体前叶 ,作用于垂体前叶的促性腺激素细胞 ,膜上的ＧｎＲＨ受体 ,通过磷脂酰肌醇系导细胞内钙离子浓度增加 ,促进垂体前叶分泌促卵泡素 (ＦＳＨ)和黄体生成素 (ＬＨ)。ＦＳＨ促进卵巢的卵泡生长发育 ,而在ＦＳＨ和ＬＨ共同作用下 ,使成熟的卵泡分泌雌激素和孕激素。下丘脑…     

类阿片肽对山羊生殖内分泌的调节Ⅲ．吗啡和纳洛酮对促性腺激素的影响

以周期母山羊为研究对象，在其中黄体期和早卵泡期注射阿片受体兴奋剂吗啡和阿片受体拮抗剂纳洛酮．研究了类阿片肽对垂体促性腺激素分泌的影响。结果表明，在中黄体期注射吗啡引起ＬＨ分泌波幅度和血浆平均水平显著降低（Ｐ＜０．０５），注射纳洛酮使ＬＨ血浆平均水平显著升高（Ｐ＜０．０５）。而在早卵泡期，注射吗啡和纳洛酮，只分别降低和升高血浆ＬＨ水平（Ｐ＜０．０５），对其分泌波幅度无显著影响（Ｐ＞０．０５）。在以上２种生理状态下，吗啡和纳洛酮对ＦＳＨ分泌均无显著影响（Ｐ＞０．０５）。因而认为，在山羊，体内类阿片肽参与ＬＨ分泌的调节。     

类阿片肽对山羊生殖内分泌的调节Ⅱ．吗啡和纳洛酮在体外对促性腺激素的影响

利用含早卵泡期山羊血清或中黄体期山羊血清的培养液，对山羊腺垂体细胞进行２４ｈ体外培养，并在培养液中分别加入阿片受体兴奋剂——盐酸吗啡和其拮抗剂——盐酸纳洛酮，研究了类阿片肽对山羊垂体细胞分泌促性腺激素功能的影响。结果表明，在两种培养条件下，吗啡或纳洛酮对垂体促性腺激素的分泌均无显著影响（Ｐ＞０．０５）。因而认为，在山羊，类阿片肽对垂体细胞促性腺激素的分泌无直接调节作用。     

麻醉和手术对母山羊垂体内分泌功能的影响

在给母山羊注射８４６麻醉注射液或实施外科手术前后收集颈静脉血样，监测８４６麻醉注射液或外科手术对垂体促性腺激素分泌的影响。结果表明，８４６麻醉注射液对山羊垂体ＬＨ和ＦＳＨ分泌无显著影响（Ｐ＞０．０５）；但外科手术对ＬＨ和ＦＳＨ分泌影响显著（Ｐ＜０．０５），在手术期间血浆ＬＨ和ＦＳＨ水平较手术前显著降低（Ｐ＜０．０５），而在手术结束后，血浆ＬＨ和ＦＳＨ水平又升高，与手术前差异不显著（Ｐ＞０．０５）。     

GnRH对人工孪生处理母牛下丘脑－垂体－性腺轴的调控

从农区黄牛群中选择２１头产后正常母牛，分比三组。组Ⅰ在产后发情周期第１７天注射孕马血清促性腺激素（ＰＭＳＧ），发情当天注射抗ＰＭＳＧ血清，配种时注射生理盐水。组Ⅱ和组Ⅲ的ＰＭＳＧ及其抗血清处理方法与组Ⅰ相同，但在配种时分别注射促性腺激素释放激素（ＧｎＲＨ）或其抗体。各组母牛在ＰＭＳＧ处理前安装颈静脉血管导管，每日间隔１５分钟收集血样，连续３小时。发情当天，间隔３０分钟收集血样，直至发情征兆消失。应用双重酶标免疫测定方法和酶联免疫吸咐测定方法，分别检测血清中ＧｎＲＨ、ＬＨ和Ｐ＿４（孕酮）水平。结果表明，（１）在结合应用ＰＭＳＧ及其抗体处理的母牛发情期间，外源ＧｎＲＨ可使外周血中ＧｎＲＨ和ＬＨ水平升高。用ＧｎＲＨ抗体中和内源ＧｎＲＨ，可使血中ＧｎＲＨ和ＬＨ的水平降低，并阻抑排卵。（２）在母牛排卵前，通过某种途径调控ＧｎＲＨ和ＬＨ的脉冲释放水平，可以提高母牛的超排效果，并有可能控制母牛的排卵数。（３）用ＰＭＳＧ及其抗体和ＧｎＲＨ超排处理的母牛，发情期间的ＧｎＲＨ在排卵前有多个脉冲释放峰，但ＬＨ只有一个脉冲释放峰，而且ＧｎＲＨ脉冲释放高峰出现的时间较ＬＨ峰早。（４）在配种后第８天检出的血清孕酮水平与排卵数呈强正相关?     

与GnRH分泌相关的电生理变化

下丘脑ＧｎＲＨ脉冲发生器调节ＧｎＲＨ间歇地释放到垂体门及循环中，进而调节ＬＨ的脉冲式分泌。各种内外因素可修正ＧｎＲＨ脉冲发器的活动，后者改变促性腺激素的分泌模式，并最影响生殖功能。     

LRH─A3、hCG和PMSG对体外培养条件下牦牛垂体组织LH和FSH分泌的调节

促卵泡素３号（ＬＲＨ－Ａ３）、人绒毛膜促性腺激素（ｈＣＧ）和孕马血清促性腺激素（ＰＭＳＧ）等３种外源性激素均可增加体外培养条件下牦牛垂体组织分泌ＬＨ和ＦＳＨ的能力。培养液中ＬＨ和ＦＳＨ含量与加入的ＬＲＨ－Ａ３量呈正相关，与加入的ＰＭＳＧ和ｈＣＧ的量无显著关系     

类阿片肽对山羊生殖内分泌的调节：Ⅲ.吗啡和纳洛酮对促性腺激素的…

以周期母山羊为研究对象，在其中黄体期和早卵泡期注射阿片受体兴奋剂吗啡和阿片受体拮抗剂纳洛酮，研究了类阿片肽对垂体促性腺激素分泌的影响。结果表明，在中黄体期注射吗啡引起ＬＨ分泌波幅度和血浆平均水平显著降低（Ｐ〈０．０５），注射纳洛酮使ＬＨ血浆平均水平显著升高（Ｐ〈０．０５）。而在早期卵泡期，注射吗啡和纳洛酮，只分别降低和升高血浆ＬＨ水平，对其分泌波幅度无显著影响。在以上２种生理状态下，吗啡和纳洛酮对     

山羊垂体门脉和颈静脉血样同时收集法的建立

本文描述了一种实用的山羊垂体门脉和颈静脉血样同时收集法。试验中应用该法收集了１９例山羊垂体门脉和颈静脉血样，并对其中７例血样分别进行血浆ＧｎＲＨ和ＬＨ水平的检测，然后根据两种血样红细胞压积比值曲线校正ＧｎＲＨ测值。结果表明，手术后颈静脉血浆的ＬＨ平均水平与手术前差异不显著（Ｐ＞０．０５）；两种血浆样品的激素测值分别反映了ＧｎＲＨ和ＬＨ的水平及其相应关系。因而认为，本试验所建立的采样法既能收集山羊垂体门脉血样，又能保持垂体的正常内分泌功能，对于内分泌学的研究有重要的应用价值。     

Secretion of luteinizing hormone into pituitary venous effluent of the follicular and luteal phase mare: novel acceleration of episodic release during constant infusion of gonadotropin-releasing hormone

We tested the hypothesis that continuous infusion of native GnRH into mares during the estrous cycle, at a dose of 100 μg/h, would elevate circulating concentrations of LH without disrupting the endogenous, episodic pattern of LH release. Ten cyclic mares were assigned to one of two groups (n = 5/group): (1) Control (saline) and (2) GnRH in saline (100 μg/h). On experimental day 0 (3 to 6 d after ovulation), osmotic pumps containing saline or GnRH were placed subcutaneously and connected to a jugular infusion catheter. Blood samples were collected from jugular catheters daily and at 5-min intervals from catheters placed in the intercavernous sinus (ICS) for 8 h on experimental day 4 (luteal phase; 7 to 10 d after ovulation), followed by an additional 6-h intensive sampling period 36 h after PGF(2α)-induced luteal regression (experimental day 6; follicular phase). Treatment with GnRH increased (P < 0.001) concentrations of LH by 3- to 4-fold in the peripheral circulation and 4- to 5-fold in the ICS. Continuous GnRH treatment accelerated (P < 0.01) the frequency of LH release and decreased the interepisodic interval during both luteal and follicular phases. Treatment with GnRH during the luteal phase eliminated the low-frequency, long-duration pattern of episodic LH release and converted it to a high-frequency, short-duration pattern reminiscent of the follicular phase. These observations appear to be unique to the horse. Further studies that exploit this experimental model are likely to reveal novel mechanisms regulating the control of gonadotrope function in this species.     

Effects of suckling, progestogen-impregnated pessaries or hysterectomy on ovarian function in autumn-lambing postpartum ewes

In three experiments, we examined the effects of suckling, progestogen treatment, hysterectomy or exogenous gonadotropin releasing hormone (GnRH) on ovarian function in autumn-lambing, postpartum ewes. In each experiment, GnRH was injected on approximately d 25 postpartum. Suckling reduced (P less than .01) GnRH-induced release of luteinizing hormone (LH) but not of follicle stimulating hormone (FSH), and reduced (P less than .05) the proportion of ewes that developed corpora lutea in response to GnRH. Suckling had no effect on duration (8.8 d) of GnRH-induced luteal phases. Progestogen prior to GnRH increased (P less than .01) the duration of the first luteal phase (10.1 vs 7.6 d; progestogen-treated ewes vs control ewes), but progestogen did not affect the release of LH or FSH. Progestogen treatment did not alter the interval from parturition to the first detected estrus (42.6 d). The concentration of 13,14-dihydro-15-keto-PGF2 alpha (PGFM) just after lambing was greater than 400 pg/ml of jugular plasma, but concentrations of PGFM declined thereafter. Hysterectomy the day after lambing hastened (P less than .001) the decline in concentrations of PGFM, indicating that prostaglandins from the postpartum uterus probably caused the high concentrations of PGFM in jugular plasma. Hysterectomy reduced (P less than .05) the interval from parturition to detectable luteal function (19.6 vs 25.3 d) and enhanced (P less than .001) luteal production of progesterone. This study of autumn-lambing ewes indicates that the uterus has a negative effect on ovarian function and that suckling and progestogen affect ovarian response to GnRH.     

The effect of pulsatile gonadotropin-releasing hormone and estradiol administration on luteinizing hormone and follicle-stimulating hormone concentrations in pituitary stalk-sectioned ovariectomized pony mares

Hourly pulses of gonadotropin-releasing hormone (GnRH) or bi-daily injections of estradiol (E2) can increase luteinizing hormone (LH) secretion in ovariectomized, anestrous pony mares. However, the site (pituitary versus hypothalamus) of positive feedback of estradiol on gonadotropin secretion has not been described in mares. Thus, one of our objectives involved investigating the feedback of estradiol on the pituitary. The second objective consisted of determining if hourly pulses of GnRH could re-establish physiological LH and FSH concentrations after pituitary stalk-section (PSS), and the third objective was to describe the declining time trends of LH and FSH secretion after PSS. During summer months, ovariectomized pony mares were divided into three groups: Group 1 (control, n = 2), Group 2 (pulsatile GnRH (25 μg/hr), n = 3), and Group 3 (estradiol (5 mg/12 hr), n = 3). All mares were stalk-sectioned and treatment begun immediately after stalk-section. Blood samples were collected every 30 min for 8 h on the day before surgery (DO) and 5 d post surgery (D5) to facilitate the comparison of gonadotropin levels before and after pituitary stalk-section. Additionally, jugular blood samples were collected every 12 hr beginning the evening of surgery, allowing for evaluation of the gonadotropin secretory time trends over the 10 d of treatment. On Day 10, animals were euthanized to confirm pituitary stalk-section and to submit tissue for messenger RNA analysis (parallel study). Plasma samples were assayed for LH and FSH by RIA. Mean LH secretion decreased from Day 0 to Day 5 in Groups 1 and 3, whereas LH secretion tended (P < 0.08) to decrease in Group 2 mares. On Day 5, LH was higher (P < 0.01) in Group 2 (17.26 ± 3.68 ng/ml; LSMEANS ± SEM), than either Group 1 (2.65 ± 4.64 ng/ml) or group 3 (4.28 ± 3.68 ng/ml). Group 1 did not differ from Group 3 on Day 5 (P < 0.40). Similarly, mean FSH levels decreased in all groups after surgery, yet Group 2 mares had significantly (P < 0.001) higher FSH concentrations (17.66 ± 1.53 ng/ml) than Group 1 or Group 3 (8.34 ± 1.84 and 7.69 ± 1. 63 ng/ml, respectively). Regression analysis of bi-daily LH and FSH levels indicated that the time trends were not parallel. These findings indicate: 1) Pituitary stalk-section lowered LH and FSH to undetectable levels within 5 d after surgery, 2) pulsatile administration of GnRH (25 μg/hr) maintained LH and FSH secretion, although concentrations tended to be lower than on Day 0, and 3) E2 did not stimulate LH or FSH secretion.     

Effects of sodium pentobarbital on release of gonadotropins in ewes immediately after ovariectomy and after treatment with gonadotropin-releas ing hormone

Two experiments were conducted with ewes 9 to 11 days after estrus to determine whether the secretion of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) are controlled differentially. In experiment 1, gonadotropin-releasing hormone (GnRH) was injected (100 (μg/ewe) at time = 0 min into ewes in four treatment groups. The treatment groups (9 ewes/group) were: 1) periodic iv sodium pentobarbital (NaPen) vehicle from 0 min; 2) periodic iv NaPen from 0 min; 3) vehicle iv for 120 min then iv NaPen from 120 min; 4) vehicle iv for 150 min then iv NaPen from 150 min. A surgical plane of anesthesia was maintained from the initiation of NaPen injection until the experiment ended. Jugular blood was sampled at 30-min intervals from ?30 to + 210 min for LH and FSH assays, and profiles of hormone concentrations were compared by time-trend analyses. GnRH released LH (P<.001) and FSH (P<.001), but NaPen did not affect the profiles of hormone concentrations; this indicated that NaPen did not reduce the ability of the pituitary to secrete gonadotropins in response to GnRH. Experiment 2 was a 2x2 factorial with ovariectomy (time = 0 hr) and NaPen as the main effects. One group of ovariectomized (n = 6) and one group of sham ovariectomized (n = 6) ewes were anesthetized only during surgery, while a group of ovariectomized (n = 7) and a group of sham ovariectomized (n = 6) ewes were kept at a surgical plane of anesthesia until 10 hr after surgery. Patterns of LH and FSH were compared in jugular blood collected hourly from 0 hr until 10 hr after surgery and in samples collected at 24 hr intervals from -24 to +72 hr of surgery. After ovariectomy, LH increased (P<.001) hourly and daily, but anesthesia suppressed (hourly, <.001 and daily, P<.005) these increases, which resulted in an interaction (hourly, P<.001 and daily, P<.01) of ovariectomy and anesthesia. FSH after ovariectomy increased hourly and daily (hourly, P<.02 and daily, P<.001), but the effect of anesthesia and interaction of ovariectomy and anesthesia were not significant. Because NaPen did not alter secretion of LH or FSH after exogenous GnRH in experiment 1 while it blocked the postovariectomy increase in LH but not FSH in experiment 2, we concluded that the postovariectomy increase in LH resulted from increased hypothalamic secretion of GnRH. The mechanisms responsible for the postovariectomy increase in FSH secretion are not identical to those for LH. The mechanisms that control the postovariectomy secretion of FSH might involve factors that are not suppressible by NaPen or, alternatively, the differences in LH and FSH release after ovariectomy might reflect the removal of ovarian factors that suppress FSH but not LH secretion in intact ewes.     

Responses of seasonally anovulatory mares to daily administration of thyrotropin-releasing hormone and(or) gonadotropin-releasing hormone analog

Seventeen seasonally anovulatory light horse mares were treated daily, starting January 5 (d 1), for 28 d with GnRH analog (GnRH-A; 50 ng/kg BW) and(or) thyrotropin-releasing hormone (TRH; 5 microg/kg BW) in a 2 x 2 factorial arrangement of treatments to test the hypothesis that combined treatment may stimulate follicular growth and development. Ovaries were examined via ultrasonography and jugular blood samples were collected every 3 d. Frequent blood samples were collected after treatment injections on d 1, 2, 4, 7, 11, 16, and 22; on d 29, all mares received an i.v. mixture of GnRH, TRH, sulpiride, and EP51389 (a growth hormone secretagogue) to assess pituitary responsiveness. No consistent effects (P > 0.1) of treatment were observed for plasma LH, FSH, prolactin, or thyroxine concentrations in samples collected every 3 d. The only effect on ovarian follicle numbers was a reduction in number of follicles 11 to 19 mm in diameter due to TRH treatment (P = 0.029). No mare ovulated during treatment. On the days of frequent sampling, mean LH (P = 0.0001) and FSH (P = 0.001) concentrations were higher in mares receiving GnRH-A and tended to increase from d 1 through 7. In contrast, mean prolactin (P = 0.001) and thyroid-stimulating hormone (P = 0.0001) concentrations were high in mares receiving TRH on d 1 but rapidly decreased thereafter. When mares were administered the secretagogue mixture on d 29, the LH response was greater (P = 0.0002) in mares that had previously received GnRH-A but the FSH response was not affected (P > 0.1); the prolactin response was greater (P = 0.014) and the TSH response was smaller (P = 0.0005) in mares that had previously received TRH. Surprisingly, an immediate growth hormone response to EP51389 was absent in all mares. In conclusion, daily GnRH-A treatment stimulated plasma LH and FSH concentrations immediately after injection; although no long-term elevation in preinjection concentrations was achieved, the responses gradually increased over time, indicating a stimulation of gonadotropin production and storage. Daily treatment with TRH stimulated plasma TSH and prolactin concentrations, but the response diminished rapidly and was minimal within a few days, indicating a depletion of pituitary stores and little or no stimulation of production. There was no beneficial effect of adding TRH treatment to the daily GnRH-A regimen.     

Pituitary effects of steroid hormones on secretion of follicle-stimulating hormone and luteinizing hormone

Steroid hormones have a profound influence on the secretion of the gonadotropins, follicle-stimulating hormone (FSH) and luteinizing hormone (LH). These effects can occur as a result of steroid hormones modifying the secretion of gonadotropin-releasing hormone (GnRH) from the hypothalamus, or a direct effect of steroid hormones on gonadotropin secreting cells in the anterior pituitary gland. With respect to the latter, we have shown that estradiol increases pituitary sensitivity to GnRH by stimulating an increase in expression of the gene encoding the GnRH receptor. Since an estrogen response element (ERE) has not been identified in the GnRH receptor gene, this effect appears to be mediated by estradiol stimulating production of a yet to be identified factor that in turn enhances expression of the GnRH receptor gene. However, the importance of estradiol for enhancing pituitary sensitivity to GnRH during the periovulatory period is questioned because an increase in mRNA for the GnRH receptor precedes the pre-ovulatory rise in circulating concentrations of estradiol. In fact, it appears that the enhanced pituitary sensitivity during the periovulatory period may occur as a result of a decrease in concentrations of progesterone rather than due to an increase in concentrations of estradiol. Estradiol also is capable of altering secretion of FSH and LH in the absence of GnRH. In a recent study utilizing cultured pituitary cells from anestrous ewes, we demonstrated that estradiol induced a dose-dependent increase in secretion of LH, but resulted in a dose-dependent decrease in the secretion of FSH. We hypothesized that the discordant effects on secretion of LH and FSH might arise from estradiol altering the production of some of the intrapituitary factors involved in synthesis and secretion of FSH. To examine this hypothesis, we measured amounts of mRNA for activin B (a factor known to stimulate synthesis of FSH) and follistatin (an activin-binding protein). We found no change in the mRNA for follistatin after treatment of pituitary cells with estradiol, but noted a decrease in the amount of mRNA for activin B. Thus, the inhibitory effect of estradiol on secretion of FSH appears to be mediated by its ability to suppress the expression of the gene encoding activin.     

Physiological roles of inhibin in regulation of FSH secretion and follicular development during early pregnancy in goats

The aim of the current study was to clarify the physiological role of inhibin in controlling FSH secretion and follicular development during the early pregnancy in goats. Eight goats investigated sonographically on Days 19-21 (Day 0=day of mating) for pregnancy were assigned into control (n=3) and treated (n=5) groups. The ovaries of all animals were daily scanned with ultrasound for follicles 2mm or more in diameter from 1 day before to 5 days after treatment. On Day 25 postbreeding; animals received either 10 ml, of normal goat serum or antiserum against [Tyr (30)]-inhibin alpha (1-30). Jugular blood samples were collected every 6 h starting 24 h before and until 120 h after treatment. The plasma concentration of FSH increased at 6 h and remained at significantly high levels until 120 h in treated vs. control group. The plasma concentrations of estradiol showed a marked increased at 66 h, with peak levels at 120 h after treatment of antiserum. The basal concentrations of LH and the pattern of plasma concentrations of progesterone were not significantly different between the two groups. The number of medium size (3.5-5.0 mm) follicles increased considerably from Day 2, whereas small (3.5 mm or less) and large (5 mm or more) follicles increased noticeably from Day 3, as compared with pre-treatment and controls. These results clearly indicated that inhibin is a key hormone in regulation of follicular development through regulation of endogenous FSH secretion during early pregnancy in goats.     

Bovine cystic ovarian disease: plasma hormone concentrations and treatment.

Ovarian function in 91 dairy cows with cystic ovarian disease was assessed by rectal palpation and by plasma hormone analysis before and after treatment. Plasma analysis showed that 84% of the cysts were correctly classified clinically and only these cows are considered further. Luteinised cysts occurred in 59 cows whereas only 18 had non-luteinised cysts. The mean plasma concentrations of luteinising hormone (LH), follicular stimulating hormone (FSH), progesterone, oestradiol and testosterone were not significantly different when compared with values at relevant stages of the oestrous cycle in normal cows. Success of treatment with progesterone, a synthetic prostaglandin, human gonadotrophin (HCG), or gonadotrophin releasing hormone (GnRH) was not dependent upon prior hormone concentrations, except for the prostaglandin which required active luteal tissue. LH and FSH concentrations in cows with luteinised cysts were not significantly different before and after successful treatment with GnRH or progesterone. Normal luteal function was not always established after treatment of non-luteinised cysts with GnRH.     

Characteristics of developing prolonged dominant follicles in cattle

In cattle, sub-luteal circulating progesterone induces an increase in the frequency of LH pulses, prolonged growth of the dominant follicle, increased peripheral estradiol and reduced fertility. The objective of this study was to examine the earliest stages of development of prolonged dominant follicles, to gain insight into the etiology of this aberrant condition. Heifers were treated with an intravaginal progesterone-releasing device (CIDR) from Day 4-8 post-estrus and PGF2alpha was injected on Day 6 and again 12h later (early prolonged dominant group). Follicular phase (CIDR: Day 4-6, with PGF2alpha) and luteal phase (CIDR: Day 4-8, without PGF2alpha) groups served as controls. As expected, peripheral progesterone in heifers of the early prolonged dominant group was intermediate between luteal and follicular phase groups after luteal regression (P<0.05). On Day 7, the frequency of LH pulses was higher in heifers of the follicular phase and early prolonged dominant groups than the luteal phase group (P<0.05). Dominant follicles (n = 4 per group) were collected by ovariectomy on Day 8 and were similar in size among groups (P>0.05). Estradiol and androstenedione concentrations in the follicular fluid at ovariectomy were higher in the follicular phase and early prolonged dominant groups versus the luteal phase group (P<0.01), whereas progesterone did not differ among groups (P>0.05). Granulosa cells and theca interna isolated from dominant follicles were incubated for 3h with or without gonadotropins or frozen for later analysis of mRNA for steroidogenic enzymes. Luteinizing doses (128 ng/ml) of LH and FSH increased secretion of progesterone (P<0.05) but did not affect secretion of estradiol by granulosa cells in all groups. Low (2 or 4 ng/ml) and luteinizing doses of LH increased secretion of androstenedione by theca interna to a similar extent among groups. Expression of mRNA for P450 side chain cleavage (P450scc), 3beta-hydroxysteroid dehydrogenase (3beta-HSD), P450 aromatase (aromatase) and Steroidogenic Acute Regulatory (StAR) protein by granulosa cells did not differ among groups (P>0.05). Levels of mRNA for P450scc, 3beta-HSD, 17alpha-hydroxylase (17alpha-OH) and StAR protein in theca interna were similar in the follicular phase and early prolonged dominant groups (P>0.05), but lower in the luteal phase group (P<0.05-0.1). In summary, the premature follicular luteinization observed in previous studies after prolonged periods of sub-luteal progesterone was absent in early prolonged dominant follicles, exposed to sub-luteal progesterone for 36 h, and their characteristics resembled those of control follicles during the follicular phase.