McNatty K P, Heath D A, Lun S, Hudson N L
Wallaceville Animal Research Centre, Ministry of Agriculture and Fisheries, Upper Hutt, New Zealand.
J Endocrinol. 1989 Feb;120(2):287-93. doi: 10.1677/joe.0.1200287.
Granulosa cells from ovarian follicles (greater than or equal to 1 mm diameter) in Booroola ewes which are homozygous (FF) or heterozygous (F+) for the F gene have previously been shown to produce significantly more cAMP in response to FSH or LH than those from similar sized follicles in ewes without the F gene (++). The aim of these studies was to test whether these F gene-specific differences arose because of differences in cAMP-phosphodiesterase (cAMP-PDE) activity. In the first study using 1 mumol cAMP/l as substrate, no F gene-specific effects were noted in cAMP-PDE activity in granulosa cells from small (1-2.5 mm diameter, n = 4 per genotype) or large (greater than or equal to 3 mm diameter, n = 4 per genotype) follicles from FF, F+ or ++ ewes, despite F gene-specific effects in FSH (1 microgram/ml)- and LH (0.1 microgram/ml)-induced cAMP accumulation in these same cell preparations. The overall mean levels of cAMP-PDE across all genotypes in cells from small and large follicles were 0.47 +/- 0.04 (S.E.M., n = 12) and 0.28 +/- 0.03 pmol cAMP/10(6) cells per min respectively; the mean PDE activity in cells from small follicles was significantly (P less than 0.05) higher compared with that in cells from large follicles. In a second study, granulosa cells from each genotype were pooled over all follicle sizes (greater than or equal to 1 mm diameter, one pool per genotype) and the rates of cAMP hydrolysis tested over a range of substrate concentrations (0-16 mumol/l) but no gene-specific differences with respect to the Michaelis constant and maximum velocity were noted.(ABSTRACT TRUNCATED AT 250 WORDS)
先前研究表明,携带F基因纯合子(FF)或杂合子(F+)的布鲁拉美利奴母羊,其直径大于或等于1毫米的卵泡颗粒细胞,相较于不携带F基因(++)的母羊中相似大小卵泡的颗粒细胞,对促卵泡激素(FSH)或促黄体生成素(LH)反应产生的环磷酸腺苷(cAMP)显著更多。这些研究的目的是测试这些F基因特异性差异是否源于环磷酸腺苷磷酸二酯酶(cAMP-PDE)活性的差异。在第一项研究中,以1微摩尔/升cAMP作为底物,未发现FF、F+或++母羊中小卵泡(直径1 - 2.5毫米,每种基因型n = 4)或大卵泡(直径大于或等于3毫米,每种基因型n = 4)的颗粒细胞cAMP-PDE活性存在F基因特异性影响,尽管在相同细胞制剂中,F基因对FSH(1微克/毫升)和LH(0.1微克/毫升)诱导的cAMP积累有特异性影响。大小卵泡细胞中所有基因型的cAMP-PDE总体平均水平分别为0.47±0.04(标准误,n = 12)和0.28±0.03皮摩尔cAMP/10⁶细胞每分钟;小卵泡细胞中的平均PDE活性显著高于大卵泡细胞(P < 0.05)。在第二项研究中,将每种基因型的颗粒细胞按所有卵泡大小(直径大于或等于1毫米,每种基因型一个样本池)汇集在一起,并在一系列底物浓度(0 - 16微摩尔/升)下测试cAMP水解速率,但未发现米氏常数和最大速度方面的基因特异性差异。(摘要截断于250字)