Parrott J A, Vigne J L, Chu B Z, Skinner M K
Reproductive Endocrinology Center, University of California, San Francisco 94143-0556.
Endocrinology. 1994 Aug;135(2):569-75. doi: 10.1210/endo.135.2.8033804.
Mesenchymal-epithelial cell interactions between thecal and granulosa cells in bovine ovarian follicles were investigated. Experiments were designed to examine the local production and action of two mesenchymal (stromal)-derived growth factors, keratinocyte and hepatocyte growth factors (KGF and HGF). Using reverse transcription-polymerase chain reaction, gene expression for KGF and HGF was detected in the mesenchymal-derived thecal cells, but not in the epithelial granulosa cells. The bovine polymerase chain reaction products for KGF and HGF were sequenced and found to be similar to known mouse, rat, and human sequences. The bovine KGF sequence was found to have a high degree of identity (86-95%) with the other species, whereas bovine HGF has a lesser degree of identity (60-63%). Immunoprecipitation of radiolabeled thecal cell secreted proteins with a KGF antibody demonstrated production of the 28-kilodalton (kDa) KGF protein. An immunoblot of thecal cell secreted proteins with HGF antibodies detected the 87-kDa HGF as well as relevant 69- and 34-kDa subunits. Therefore, thecal cells were found to express the genes and secrete the proteins for KGF and HGF. Granulosa cells had no detectable KGF or HGF expression. Treatment with recombinant KGF or HGF stimulated the proliferation of granulosa cells, but not thecal cells. Therefore, the actions of KGF and HGF in the ovarian follicle appear to be restricted to granulosa cells. The combined results indicate that KGF and HGF are produced locally in the bovine ovarian follicle by thecal cells, and that both of these growth factors can act on granulosa cells to influence cell proliferation. These observations demonstrate that KGF and HGF can mediate mesenchymal-epithelial cell interactions between thecal and granulosa cells. The potential importance that the mesenchymal derived thecal cells may have in ovarian follicle development is discussed.
研究了牛卵巢卵泡中膜细胞与颗粒细胞之间的间充质 - 上皮细胞相互作用。实验旨在检测两种间充质（基质）来源的生长因子，即角质形成细胞生长因子和肝细胞生长因子（KGF和HGF）的局部产生和作用。利用逆转录 - 聚合酶链反应，在间充质来源的膜细胞中检测到KGF和HGF的基因表达，但在上皮颗粒细胞中未检测到。对牛KGF和HGF的聚合酶链反应产物进行测序，发现其与已知的小鼠、大鼠和人类序列相似。发现牛KGF序列与其他物种具有高度同源性（86 - 95%），而牛HGF的同源性较低（60 - 63%）。用KGF抗体对放射性标记的膜细胞分泌蛋白进行免疫沉淀，证明产生了28千道尔顿（kDa）的KGF蛋白。用HGF抗体对膜细胞分泌蛋白进行免疫印迹，检测到87 kDa的HGF以及相关的69 kDa和34 kDa亚基。因此，发现膜细胞表达KGF和HGF的基因并分泌其蛋白。颗粒细胞未检测到KGF或HGF表达。用重组KGF或HGF处理可刺激颗粒细胞增殖，但不刺激膜细胞增殖。因此，KGF和HGF在卵巢卵泡中的作用似乎仅限于颗粒细胞。综合结果表明，KGF和HGF由牛卵巢卵泡中的膜细胞在局部产生，并且这两种生长因子均可作用于颗粒细胞以影响细胞增殖。这些观察结果表明，KGF和HGF可介导膜细胞与颗粒细胞之间的间充质 - 上皮细胞相互作用。讨论了间充质来源的膜细胞在卵巢卵泡发育中可能具有的潜在重要性。
