Clark D E, Tisdall D J, Fidler A E, McNatty K P
AgResearch, Wallaceville Animal Research Centre, Upper Hutt, New Zealand.
J Reprod Fertil. 1996 Mar;106(2):329-35. doi: 10.1530/jrf.0.1060329.
The c-kit protein is a transmembrane tyrosine kinase receptor that binds the growth factor stem cell factor. Mutant alleles of the genes coding for both the receptor (c-kit) and its ligand (stem cell factor) affect gametogenesis, development of melanoblasts and some aspects of haematopoiesis. The aim of this study was to examine expression of the c-kit gene during folliculogenesis in fetal sheep ovaries using in situ hybridization. A 422 bp cDNA encoding the extracellular domain of the c-kit protein was amplified from sheep ovarian RNA using reverse-transcription-polymerase chain reaction (RT-PCR), cloned and sequenced. Riboprobes transcribed from the ovine cDNA encoding c-kit were used to detect the presence of mRNA encoding c-kit within the ovaries of fetal sheep on days 90, 100, 120 and 135 of gestation (term = 147 days). In day 90 and 100 fetal ovaries, mRNA encoding c-kit was not detected in association with oogonia during the period of meiosis (to prophase I) but was present in some of the isolated oocytes. In ovaries from day 90 to day 135, mRNA encoding c-kit was detected in the oocytes at every stage of follicular growth--primordial through to antral follicles. This pattern of localization is consistent with that demonstrated in mice.
c-kit蛋白是一种跨膜酪氨酸激酶受体,可结合生长因子干细胞因子。编码该受体(c-kit)及其配体(干细胞因子)的基因突变等位基因会影响配子发生、成黑素细胞的发育以及造血的某些方面。本研究的目的是使用原位杂交技术检测胎儿绵羊卵巢卵泡发生过程中c-kit基因的表达。使用逆转录-聚合酶链反应(RT-PCR)从绵羊卵巢RNA中扩增出编码c-kit蛋白细胞外结构域的422 bp cDNA,进行克隆和测序。从编码c-kit的绵羊cDNA转录而来的核糖探针用于检测妊娠第90、100、120和135天(足月为147天)胎儿绵羊卵巢内编码c-kit的mRNA的存在情况。在妊娠第90天和100天的胎儿卵巢中,在减数分裂期(至前期I)与卵原细胞相关的编码c-kit的mRNA未被检测到,但在一些分离的卵母细胞中存在。在妊娠第90天至135天的卵巢中,在卵泡生长的每个阶段——从原始卵泡到窦状卵泡的卵母细胞中都检测到了编码c-kit的mRNA。这种定位模式与在小鼠中显示的一致。