Weiss R L, Goodenough D A, Goodenough U W
J Cell Biol. 1977 Jan;72(1):144-60. doi: 10.1083/jcb.72.1.144.
Fusion of plasma membranes between Chlamydomonas reinhardtii gametes has been studied by freeze-fracture electron microscopy of unfixed cells. The putative site of cell fusion developes during gametic differentiation and is recognized in thin sections of unmated gametes as a plaque of dense material subjacent to a sector of the anterior plasma membrane (Goodenough, U.W., and R.L. Weiss. 1975.J. Cell Biol. 67:623-637). The overlying membrane proves to be readily recognized in replicas of unmated gametes as a circular region roughly 500 nm in diameter which is relatively free of "regular" plasma membrane particles on both the P and E fracture faces. The morphology of this region is different for mating-type plus (mt+) and mt- gametes: the few particles present in the center of the mt+ region are distributed asymmetrically and restricted to the P face, while the few particles present in the center of the mt- region are distributed symmetrically in the E face. Each gamete type can be activated for cell fusion by presenting to it isolated flagella of opposite mt. The activated mt+ gamete generates large expanses of particle-cleared membrane as it forms a long fertilization tubule from the mating structure region. In the activated mt- gamete, the E face of the mating structure region is transformed into a central dome of densely clustered particles surrounded by a particle-cleared zone. When mt+ and mt- gametes are mixed together, flagellar agglutination triggeeeds to fuse with an activated mt- region. The fusion lip is seen to develop within the particle-dense central dome. We conclude that these mt- particles play an active role in membrane fusion.
通过对未固定细胞进行冷冻断裂电子显微镜观察,研究了莱茵衣藻配子间质膜的融合。细胞融合的假定位点在配子分化过程中形成,在未交配配子的薄切片中,可识别为前质膜扇形区域下方的致密物质斑块(古德诺夫,U.W.,和R.L. 韦斯。1975年。《细胞生物学杂志》67:623 - 637)。在未交配配子的复制品中,覆盖其上的膜很容易被识别为一个直径约500纳米的圆形区域,在P面和E面的断裂面上相对没有“规则”的质膜颗粒。这个区域的形态在正交配型(mt +)和负交配型(mt -)配子中有所不同:mt +区域中心存在的少数颗粒不对称分布且仅限于P面,而mt -区域中心存在的少数颗粒在E面呈对称分布。通过向每种配子类型提供相反交配型的分离鞭毛,可激活其进行细胞融合。被激活的mt +配子从交配结构区域形成长长的受精管时,会产生大片颗粒清除的膜。在被激活的mt -配子中,交配结构区域的E面转化为由颗粒清除区包围的密集聚集颗粒的中央穹顶。当mt +和mt -配子混合在一起时,鞭毛凝集引发与被激活的mt -区域融合。融合唇在颗粒密集的中央穹顶内形成。我们得出结论,这些mt -颗粒在膜融合中起积极作用。