Perazzolo L M, Coward K, Davail B, Normand E, Tyler C R, Pakdel F, Schneider W J, Le Menn F
Laboratoire de Biologie de la Reproduction des Poissons, Unité Associée INRA, Université Bordeaux I, 33405 Talence, France.
Biol Reprod. 1999 May;60(5):1057-68. doi: 10.1095/biolreprod60.5.1057.
The expression and localization of vitellogenin (VTG) receptor (VTGR) mRNA were identified throughout ovarian development in the rainbow trout, Oncorhynchus mykiss. Northern blot confirmed the presence of a transcript (approximately 3.9 kilobases [kb]) that was specific to the ovary. The expression of VTGR mRNA varied throughout ovarian development and was highest in previtellogenic ovaries and in ovaries at the onset of vitellogenesis containing ovarian follicles (OF) from 35 to 600 microm in diameter. In situ hybridization using 35S riboprobes showed that the transcription of the VTGR gene was initiated in OF measuring 45-50 microm in diameter, with transcripts being exclusively localized in the ooplasm. A dramatic increase in mRNA synthesis occurred during previtellogenic growth (OF from 50 to 200 microm); this was followed by a gradual decrease during the vitellogenic growth phase. VTGR mRNA was not detected in OF greater than 1000 microm in diameter (oocytes actively sequestering VTG). Immunocytolocalization of yolk proteins derived from VTG demonstrated that oocytes started to sequester VTG when they were around 300 microm in diameter, shortly after the time of maximal density of VTGR mRNA in the ooplasm. The timing of transcription of the VTGR gene, predominantly during previtellogenesis, suggests that the VTGR is recycled to the oocyte surface during the vitellogenic growth phase.
在虹鳟鱼(Oncorhynchus mykiss)整个卵巢发育过程中,对卵黄蛋白原(VTG)受体(VTGR)mRNA的表达和定位进行了鉴定。Northern印迹证实存在一种卵巢特异性转录本(约3.9千碱基 [kb])。VTGR mRNA的表达在卵巢发育过程中有所变化,在卵黄发生前的卵巢以及卵黄发生开始时的卵巢中最高,这些卵巢中含有直径为35至600微米的卵巢卵泡(OF)。使用35S核糖探针进行原位杂交显示,VTGR基因的转录在直径为45 - 50微米的OF中开始,转录本仅定位于卵质中。在卵黄发生前的生长阶段(OF从50微米长到200微米),mRNA合成急剧增加;随后在卵黄发生生长阶段逐渐减少。在直径大于1000微米的OF中未检测到VTGR mRNA(卵母细胞正在积极摄取VTG)。对源自VTG的卵黄蛋白进行免疫细胞定位表明,卵母细胞在直径约为300微米时开始摄取VTG,这是在卵质中VTGR mRNA密度达到最大值后不久。VTGR基因转录的时间,主要发生在卵黄发生前,表明VTGR在卵黄发生生长阶段被循环到卵母细胞表面。