Nishino K, Kato M, Yokouchi K, Yamanouchi K, Naito K, Tojo H
Laboratory of Applied Genetics, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan.
J Exp Zool. 2000 Feb 15;286(3):320-7.
In developing mouse embryos, the Sertoli cells, Leydig cells, and seminiferous cords are differentiated in the XY gonads. The migration of mesonephric cells into the gonads is required during the developmental stage for seminiferous cord formation in the male gonads. In previous experiments, an organ coculture system has been used to examine morphologically developing gonads. However, by the process used in this system for fixing and staining the gonad/mesonephros complexes for examination, the kinetics of cell migration and the character of migrating cells cannot be observed. In the present study, we established an improved organ coculture system, using transgenic mice ubiquitously expressing Enhanced Green Fluorescent Protein (EGFP). In this system, time-dependent morphological changes in male-specific migration were observable in the gonad/mesonephros complex. The cell migration occurred at around 20 hr of coculture and began to spread at 25 hr with increases in the number of migrating cells occurring at 45 hr of coculture. No degenerative changes were detected at the end of coculture. Our results indicate that the present coculture system is very useful for investigating the mechanism of cell migration, as well as the characteristics of the migrating cells, in developing gonads. J. Exp. Zool. 286:320-327, 2000.
在发育中的小鼠胚胎中，支持细胞、间质细胞和生精索在XY性腺中分化。在发育阶段，中肾细胞迁移到性腺对于雄性性腺中生精索的形成是必需的。在先前的实验中，已使用器官共培养系统来检查形态学上发育中的性腺。然而，通过该系统用于固定和染色性腺/中肾复合体以进行检查的过程，无法观察到细胞迁移的动力学和迁移细胞的特征。在本研究中，我们使用普遍表达增强型绿色荧光蛋白（EGFP）的转基因小鼠建立了一种改进的器官共培养系统。在该系统中，在性腺/中肾复合体中可观察到雄性特异性迁移中随时间变化的形态学变化。细胞迁移发生在共培养约20小时时，并在25小时开始扩散，共培养45小时时迁移细胞数量增加。在共培养结束时未检测到退行性变化。我们的结果表明，目前的共培养系统对于研究发育中性腺中细胞迁移的机制以及迁移细胞的特征非常有用。《实验动物学杂志》286:320 - 327，2000年。
