Kanungo J, Petrino T R, Wallace R A
Whitney Laboratory, University of Florida, St. Augustine 32096.
J Exp Zool. 1990 Jun;254(3):313-21. doi: 10.1002/jez.1402540310.
A procedure was developed for studying vitellogenin (VTG) incorporation by vitellogenic oocytes of Fundulus heteroclitus in vitro. Since homologous VTG can be obtained from this animal only with great difficulty, the use of [32P]VTG from Xenopus laevis was explored as an alternative. Vitellogenic as well as maturational-stage oocytes were found to sequester X. laevis [32P]VTG from the medium, and incorporation was found to be linear with time for at least up to 12 hr. Once incorporated into the oocyte, [32P]VTG did not appear to undergo turnover. The effect of different [32P]VTG concentrations on incorporation indicated that the uptake mechanism was saturable. Unlabeled F. heteroclitus VTG and X. laevis VTG were also found to compete effectively with X. laevis [32P]VTG, whereas bovine serum albumin did not. These results represent the first documentation of a successful culture system for receptor-mediated VTG incorporation by teleost oocytes.
开发了一种体外研究异育银汉鱼卵黄生成期卵母细胞摄取卵黄蛋白原(VTG)的方法。由于从这种动物中获取同源VTG非常困难,因此探索使用来自非洲爪蟾的[32P]VTG作为替代方法。发现卵黄生成期以及成熟阶段的卵母细胞能从培养基中摄取非洲爪蟾的[32P]VTG,并且摄取量至少在长达12小时内与时间呈线性关系。一旦[32P]VTG被摄取到卵母细胞中,似乎不会发生周转。不同浓度的[32P]VTG对摄取的影响表明摄取机制是可饱和的。还发现未标记的异育银汉鱼VTG和非洲爪蟾VTG能与非洲爪蟾的[32P]VTG有效竞争,而牛血清白蛋白则不能。这些结果首次证明了一种成功的培养系统,用于硬骨鱼卵母细胞通过受体介导摄取VTG。