Ushiyama A, Araki T, Chiba K, Hoshi M
Department of Life Science, Tokyo Institute of Technology, Japan.
Zygote. 1993 May;1(2):121-7. doi: 10.1017/s0967199400001386.
In the starfish, spermatozoa undergo the acrosome reaction upon encountering the jelly coat of eggs. A highly sulphated glycoprotein in the jelly coat is called acrosome-reaction-inducing substance (ARIS) because it is the key signal molecule to trigger the acrosome reaction. The activity of ARIS is mainly attributed to its sulphate and saccharide residues. The extremely large molecular size and species-specific action of ARIS suggest the presence of a specific ARIS receptor on the sperm surface, but no experimental evidence for the receptor has been presented. We therefore measured specific binding of ARIS and its pronase digest (P-ARIS), which retains the full activity of ARIS, to homologous spermatozoa by using fluorescein-isothiocyanate-labelled ARIS and 125I-labelled P-ARIS, respectively. The spermatozoa had the ability to bind ARIS, as well as P-ARIS, specifically. The binding was species-specific and mostly localised to the head region of spermatozoa. Scatchard plot analysis indicated the presence of one class of ARIS receptor on the surface of acrosome-intact spermatozoa. Furthermore, the specific binding of P-ARIS to the anterior region of sperm heads was microscopically confirmed by using P-ARIS conjugated to polystyrene latex beads with intense fluorescence. It is concluded that starfish spermatozoa have a specific receptor for ARIS on the surface of the anterior region of heads.
在海星中,精子遇到卵的卵胶膜时会发生顶体反应。卵胶膜中一种高度硫酸化的糖蛋白被称为顶体反应诱导物质(ARIS),因为它是触发顶体反应的关键信号分子。ARIS的活性主要归因于其硫酸根和糖残基。ARIS极大的分子尺寸和物种特异性作用表明精子表面存在特定的ARIS受体,但尚未有该受体的实验证据。因此,我们分别使用异硫氰酸荧光素标记的ARIS和125I标记的P-ARIS(保留了ARIS的全部活性)来测量ARIS及其链霉蛋白酶消化产物(P-ARIS)与同源精子的特异性结合。精子具有特异性结合ARIS以及P-ARIS的能力。这种结合具有物种特异性,并且大多定位于精子的头部区域。Scatchard图分析表明,在顶体完整的精子表面存在一类ARIS受体。此外,通过使用与具有强荧光的聚苯乙烯乳胶珠偶联的P-ARIS,在显微镜下证实了P-ARIS与精子头部前部区域的特异性结合。结论是,海星精子在头部前部区域的表面具有ARIS的特异性受体。