Bohrmann J, Schill S
Institut für Biologie I (Zoologie), Universitat Freiburg, Germany.
Int J Dev Biol. 1997 Jun;41(3):499-507.
Using video-intensified fluorescence microscopy and a pseudocolor display of fluorescence intensity, we analyzed the distribution of microinjected molecules within the nurse-cell/oocyte syncytium of Drosophila ovarian follicles. We varied the composition and the osmolarity of the culture solution as well as the electrical charge and the molecular mass of the microinjected fluorescent probe. As culture solutions, we used four simple salines (IMADS) and a complex tissue-culture medium (R-14) that matched the osmolarity of adult hemolymph. Small amounts of two anionic dyes (Lucifer Yellow CH and Lucifer Yellow dextran) as well as of two cationic dyes (rhodamine 6G and tetramethylrhodamine dextran-lysine) were iontophoretically microinjected either into a nurse cell or into the oocyte of stage-10 follicles. In the tissue-culture medium, within a few seconds following microinjection, all tested dyes passed through the intercellular bridges in both the anterior direction (to the nurse cells) and the posterior direction (to the oocyte), independent of their electrical charge or molecular mass. In all simple salines, irrespective of their osmolarity, Lucifer Yellow CH was found to preferentially migrate in the posterior direction and to accumulate in the oocyte due to progressive binding to yolk spheres. Thus, with this sensitive method, no correlation was detectable between the external osmolarity, the electrical charge and the preferential direction of migration of a microinjected probe. Our results indicate that the electrical gradient described by other authors does not exert significant influence on the migration of charged molecules through intercellular bridges in situ.
利用视频增强荧光显微镜和荧光强度的伪彩色显示,我们分析了微注射分子在果蝇卵巢卵泡的滋养细胞/卵母细胞合胞体内的分布。我们改变了培养液的成分和渗透压,以及微注射荧光探针的电荷和分子量。作为培养液,我们使用了四种简单盐溶液(IMADS)和一种与成年血淋巴渗透压匹配的复杂组织培养基(R-14)。将少量两种阴离子染料(路西法黄CH和路西法黄葡聚糖)以及两种阳离子染料(罗丹明6G和四甲基罗丹明葡聚糖-赖氨酸)通过离子电渗法微注射到10期卵泡的一个滋养细胞或卵母细胞中。在组织培养基中,微注射后几秒钟内,所有测试染料都能通过细胞间桥向前(向滋养细胞)和向后(向卵母细胞)移动,与它们的电荷或分子量无关。在所有简单盐溶液中,无论其渗透压如何,发现路西法黄CH优先向后移动并由于与卵黄球的逐步结合而在卵母细胞中积累。因此,用这种灵敏的方法,未检测到外部渗透压、电荷与微注射探针优先迁移方向之间的相关性。我们的结果表明,其他作者描述的电势梯度对带电分子在原位通过细胞间桥的迁移没有显著影响。