Fujimori K, Nakamura R M, Tonetta S A, di Zerega G S
Department of Obstetrics and Gynecology, Yamagata School of Medicine, Japan.
Biol Reprod. 1987 Nov;37(4):812-22. doi: 10.1095/biolreprod37.4.812.
The granulosa cell produces a protein inhibitor of aromatase activity (follicle-regulatory protein: FRP), which recently was purified to homogeneity. To determine the possible involvement of FRP in follicular maturation, we examined the size distribution of follicles and their morphological patterns as well as serum steroid levels after the systemic administration of FRP and/or gonadotropin to guinea pigs, which have 5-6 days between luteolysis and ovulation in a 16-day cycle. FRP was partially purified from porcine follicular fluid by ammonium sulfate precipitation (0-35%), Dye Matrex Orange A Chromatography, dialysis, and lyophylization. To investigate the effect of pregnant mare's serum (PMS) during the periovulatory period in follicular development, adult guinea pigs underwent unilateral ovariectomy on Days 10, 12, and 14 of the estrous cycle (N = 6 each). Guinea pigs were injected twice daily with vehicle or PMS (5 IU) and 2 days thereafter the remaining ovaries were removed. Another group of guinea pigs received, in addition, intraperitoneal injections of FRP (1 mg) each morning from Day 8 of estrus until they were killed. The resected ovaries were fixed, embedded in paraffin, serially sectioned (7 micron), and stained with Azan for comparative study via light microscopy. All follicles greater than 400 micron were classified by size, and the atretic pattern was determined by mural granulosa cell pyknosis and antral sloughing. The distribution of follicular size was not affected by hemicastration at Day 10, although the percentage of total atretic follicles decreased. In the PMS-treated group, there was a significant decrease in the number of viable follicles (700-899 micron) after hemicastration. Also pronounced in follicles of this size was the lack of mid-atretic follicles. After injections of FRP for 3 or 5 days, the overall number of follicles was almost doubled as compared to the number found in the normal ovary. Additionally, there was a significant increase in the percentage of follicles that were recently atretic, although the total percentage of atretic follicles was unchanged. After hemicastration at Day 10 followed by FRP treatment for 2 days, the total percentage of atretic follicles in the remaining ovary decreased to 18% compared with 35% in the normal ovary, 46% in the hemicastrated plus PMS-treated group, and 38% in the hemicastrated and PMS- and FRP-treated group (all p less than 0.01). Treating the hemicastrated animal with PMS increased the percentage of atretic follicles in all groups.(ABSTRACT TRUNCATED AT 400 WORDS)
颗粒细胞可产生一种芳香化酶活性的蛋白抑制剂（卵泡调节蛋白：FRP），该蛋白最近已被纯化至同质状态。为了确定FRP在卵泡成熟过程中可能发挥的作用，我们在豚鼠黄体溶解和排卵之间间隔5 - 6天的16天周期内，对其进行全身注射FRP和/或促性腺激素后，检查了卵泡的大小分布、形态模式以及血清类固醇水平。通过硫酸铵沉淀（0 - 35%）、染料基质橙A色谱法、透析和冻干等方法，从猪卵泡液中部分纯化得到FRP。为了研究孕马血清（PMS）在卵泡发育排卵期的作用，成年豚鼠在发情周期的第10、12和14天进行单侧卵巢切除（每组N = 6只）。豚鼠每天注射两次赋形剂或PMS（5 IU），此后2天切除剩余的卵巢。另一组豚鼠从发情期第8天开始，每天早上额外腹腔注射FRP（1 mg），直至处死。切除的卵巢经固定、石蜡包埋、连续切片（7微米），并用偶氮胭脂红染色，通过光学显微镜进行比较研究。所有大于400微米的卵泡按大小分类，闭锁模式通过壁颗粒细胞固缩和卵泡腔脱落来确定。第10天进行半侧卵巢切除虽使闭锁卵泡总数的百分比下降，但卵泡大小分布未受影响。在PMS处理组中，半侧卵巢切除后存活卵泡（700 - 899微米）数量显著减少。此大小的卵泡中也明显缺乏中期闭锁卵泡。注射FRP 3天或5天后，卵泡总数与正常卵巢相比几乎增加了一倍。此外，近期闭锁卵泡的百分比显著增加，尽管闭锁卵泡的总百分比未变。第10天进行半侧卵巢切除后再进行2天的FRP处理，剩余卵巢中闭锁卵泡的总百分比降至18%，而正常卵巢为35%，半侧卵巢切除加PMS处理组为46%，半侧卵巢切除加PMS和FRP处理组为38%（所有p均小于0.01）。用PMS处理半侧卵巢切除的动物会增加所有组中闭锁卵泡的百分比。（摘要截断于400字）
