蛙精子中心体的分离

中心体是动物和低等植物中构成有丝分裂器的重要结构和功能元件,是间期细胞质微管和分裂期纺锤体微管的组织中心。本文报道一种从蛙精子中分离中心体的简便方法:通过匀浆将精子尾部与头部脱离,用蔗糖离心去除脱落的尾部和杂质颗粒,从而得到纯化的精子头部。用含EGTA的低渗液使精核膨大,再以超声破碎,离心得到中心体的粗制品,以重复超声和离心去除染色质和中心体外围的线粒体,得到纯度较高的精子中心体。电镜观察和免疫荧光染色显示分离所得的中心体由一对中心粒和其外周物质组成。

ISOLATION OF CENTROSOMES FROM FROG SPERMATOZOA

Centrosome is a structurally and functio-nally important part of mitotic apparatus inanimal and lower plant cells.It serves as amicrotubule organizing center both in inter-phase cytoplasm and mitotic spindle formation.The importance of this organell has long beennoticed by cell biologists but there is still nota satisfied approach to the isolation and puri-fication of the organelle.In the present papera simple technique is reported for isolatingcentrosomes from frog spermatozoa.The majorprocedure is described as follows:The spermsuspension is homogenized in the presence ofcalcium ions to cut off the flagella from thesperm heads,then centrifuged through 2 Msucrose to collect purified sperm heads.A hy-potonic solution is then added to swell thesperm heads.The centrosome become clearlyvisible in the swelled sperm head.A suitablesonication is applied to disrupt sperm chro-matin but left the centrosome intact.Throughthree cycles of sonication and centrifugation.a pure centrosome preparation is obtained.Theisolated centrosome is structurally intact andeach contains a pair of centrioles as detectedboth under EM and with immunofluorescencemethod,and a layer of pericentriolar material.