Kemper Green C N, Hawkins D A, Rocha A, Tanner J W, Harms P G, Forrest D W, Welsh T H
Department of Animal Science, Texas Agricultural Experiment Station, Texas A&M University System, College Station 77843-2471, USA.
Anim Reprod Sci. 1996 Dec 16;45(3):157-76. doi: 10.1016/s0378-4320(96)01582-5.
Ultrasonography and endocrine assay techniques were used to monitor structural and hormonal alterations made by the ovary in response to the biological actions of pituitary-derived follicle-stimulating hormone (FSH-P). Angus heifers (n = 36) were allotted to receive injections (twice per day) of either FSH-P (up to a total of 28 mg over a maximum of 4 days beginning on Day 10 of a synchronized estrous cycle) or saline in order to quantify temporal relationships among follicle growth and steroid hormone profiles. Transrectal ultrasonography was utilized at 12-h intervals to monitor and record follicle growth. Plasma was collected every 12 h for the first 48 h of the experiment and then every 6 h for the remainder of the experiment. At 48 and 60 h after the onset of treatments, prostaglandin F2 alpha (PGF2 alpha; 25 mg) was administered (i.m.). FSH-treated heifers (n = 6 at each time) were terminated at 24, 48, 72 and 96 h following the onset of treatment. Saline-treated heifers were terminated at 24 and 96 h (n = 6 at each time). After ovaries were obtained, follicular number and size were recorded and follicular fluid (FF) was collected. Plasma concentration of progesterone (P) and estradiol (E2) and FF concentration of P, E2, estrone, testosterone and androstenedione were determined by radioimmunoassays. Plasma concentration of E2 increased (P < 0.05) within 36 h of initiation of FSH treatment. Plasma P decreased (P < 0.0001) by 12 h post-PGF2 alpha. Ultrasonographic examination revealed a significant decrease in the number of small follicles by 48 h, whereas the number of medium follicles increased (P < 0.05) by 60 h after the initiation of FSH treatment. The number of large follicles (LF > or = 10 mm diameter) increased (P < 0.01) over the course of the experiment. The total number of ovarian follicles (TF) 24 h after the start of FSH treatment was correlated (r = 0.99; P < 0.0001) with the number of small follicles (SF < or = 5 mm). At 72 h after the onset of FSH treatment, the number of medium follicles (i.e. 6-9 mm) was correlated with TF (r = 0.97; P < 0.0001). Estradiol was the predominant FF steroid. Follicular fluid E2 was greatest in follicles at 72 h after FSH treatment. Follicular fluid E2 and plasma E2 were positively correlated (r = 0.66; P < 0.001). Follicular aromatase activity was estimated by evaluating the ratio of FF estrogens (E) to androgens (A). Elevated aromatase activity (E:A ratio > 1.0) was detected in 196 of 206 follicles. The estrogen to progesterone ratio was used as an estimate of follicle viability. Eighty-five percent of the follicles were estimated to be viable (E:P ratio > 1.0). The peak E:A ratio in LF preceded by 24 h the peak concentration in FF E2 and plasma E2. In MF and SF the E:A ratio increased by 72 h. Enhancement of ovarian follicular growth (i.e. increased number and size of follicles; increased steroidogenesis) by exogenous, pituitary-derived FSH is characterized by (1) increased activity of aromatase, and (2) accumulation of FF E2, events which temporally preceded the increase in plasma concentration of E2. These observations will aid efforts to incorporate recombinant bovine FSH and somatotropin in an effort to develop more predictable superstimulation and ovulation induction protocols.
采用超声检查和内分泌测定技术,监测卵巢因垂体来源的促卵泡激素(FSH-P)的生物学作用而发生的结构和激素变化。将安格斯小母牛(n = 36)分配为接受注射(每天两次)FSH-P(从同步发情周期的第10天开始,最多4天内总量达28 mg)或生理盐水,以量化卵泡生长与类固醇激素谱之间的时间关系。每隔12小时进行经直肠超声检查,以监测和记录卵泡生长情况。在实验的前48小时,每12小时采集一次血浆,实验剩余时间每6小时采集一次。在治疗开始后48和60小时,注射前列腺素F2α(PGF2α;25 mg)(肌肉注射)。FSH处理的小母牛(每次n = 6)在治疗开始后24、48、72和96小时处死。生理盐水处理的小母牛在24和96小时处死(每次n = 6)。获取卵巢后,记录卵泡数量和大小,并收集卵泡液(FF)。通过放射免疫分析法测定血浆中孕酮(P)和雌二醇(E2)的浓度以及卵泡液中P、E2、雌酮、睾酮和雄烯二酮的浓度。FSH治疗开始后36小时内,血浆E2浓度升高(P < 0.05)。PGF2α注射后12小时,血浆P浓度下降(P < 0.0001)。超声检查显示,FSH治疗开始后48小时,小卵泡数量显著减少,而中卵泡数量在60小时时增加(P < 0.05)。在实验过程中,大卵泡(直径LF≥10 mm)数量增加(P < 0.01)。FSH治疗开始后24小时,卵巢卵泡总数(TF)与小卵泡(直径SF≤5 mm)数量相关(r = 0.99;P < 0.0001)。FSH治疗开始后72小时,中卵泡(即直径6 - 9 mm)数量与TF相关(r = 0.97;P < 0.0001)。雌二醇是卵泡液中主要的类固醇激素。FSH治疗后72小时,卵泡中的卵泡液E2含量最高。卵泡液E2与血浆E2呈正相关(r = 0.66;P < 0.001)。通过评估卵泡液中雌激素(E)与雄激素(A)的比例来估计卵泡芳香化酶活性。在206个卵泡中的196个检测到芳香化酶活性升高(E:A比例> 1.0)。雌激素与孕酮的比例用于估计卵泡活力。估计85%的卵泡是有活力的(E:P比例> 1.0)。大卵泡中E:A比例峰值比卵泡液E2和血浆E2浓度峰值提前24小时出现。在中卵泡和小卵泡中,E:A比例在72小时时升高。外源性垂体来源的FSH促进卵巢卵泡生长(即卵泡数量和大小增加;类固醇生成增加)的特征为:(1)芳香化酶活性增加,(2)卵泡液E2积聚,这些事件在血浆E2浓度升高之前发生。这些观察结果将有助于将重组牛FSH和生长激素纳入其中,以努力开发更可预测的超刺激和排卵诱导方案。