近江蛏精子超微形态结构观察及与缢蛏精子的比较
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福建省海洋与渔业厅重点项目(闽海渔合同[2008]2-4号)


Comparative ultrastructure of spermatozoa of Sinonovacula rivularis sp. nov. and Sinonovacula constricta
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    摘要:

    利用扫描电镜和透射电镜分别观察了近江蛏和缢蛏成熟精子的超微形态结构。发现近江蛏精子与缢蛏精子在超微形态结构上差异很大。近江蛏精子为典型的原生型,成熟精子由头部、中段和尾部组成,头部包括顶体和细胞核。近江蛏精子与缢蛏精子顶体均为保龄球状,但近江蛏精子顶体全长比缢蛏短。近江蛏精子细胞核略扁圆形,外缘具8~9个圆弧形凸起;缢蛏精子细胞核则呈圆球状。近江蛏精子中段由线粒体和中心粒复合体构成,线粒体一般5个,个别4~6个;缢蛏线粒体5个或6个。近江蛏精子尾部为细长的鞭毛,轴丝为“9 2”结构,与缢蛏的相同。从近江蛏与缢蛏的精子形态差异看,两者应属于种间差异。

    Abstract:

    Sinonovacula rivularis sp. nov., found in Minjiang Estuary Waters of Fujian Province,was identified as a new species after study,and it belonged to Sinonovacula,Pharellidae,Bivalvia.The outer shape for the shell of S. rivularis sp. nov. was similar to that of S.constricta (Lamarck,1818), and it was likely that S. rivularis sp. nov. and S. constricta had a relatively closer affinity.Because the morphological structure of spermatozoon had the specificity of species,the differences between the species could be shown by studying and comparing the ultrastructure of spermatozoa with relatively closer affinity and could be used as the auxiliary evidence for related classification.This article observed the ultrastructure of mature sperms of S.rivularis sp. nov. and S.constricta respectively by scanning electron microscope and transmission electron microscopy aiming at providing a certain basic data and related figures for the reproductive biology research of S.rivularis sp.nov. and also providing strong evidences for differentiating S.rivularis sp.nov. and S.constricta. The samples of S.rivularis sp.nov. and S.constricta were collected in mating season(in November).Samples of S.rivularis sp. nov. were collected from coastal waters of Meihua town,Changle city,Fujian,while samples of S.constricta were collected from Xiamen Seafood Market.The sample preparation method of scanning electron microscope:dissecting the mature brood stock,extracting the sperms by suction tube and fixating by glutaric dialdehyde of 2.5%(V/V);taking 1-5 mL of the fixated and concentrated sample,and filtering it into the nuclear porous filter membrane at the size of 1.8 μm;then washing them three times with sterilized sea water and distilled water respectively,each time 10 minutes,dehydrate it once using 0%,50%,70%,90% and 95% graded ethanol respectively; and then dehydrate it twice using absolute ethyl alcohol,10 minutes for each grade.The sample is to be dried in CO2 Critical Point Dryer; the sample after being dried is to be sprayed with metal for 25 s;and using LEO1530 field emission scanning electron microscope to observe and take photographs.The sample preparation method of transmission electron microscopy:selecting male S.rivularis sp. nov. with full and well-developed gonad,absorbing spermary by the suction tube,and quickly placing it into the fixative solution(2% paraformaldehyde 2.5% glutaraldehyde 0.05% CaCl2,pH 7.6,0.1 MPBS),and then fixating it in an environment of 4 ℃,after dehydration and embedding,and then cutting it into slices by NOVA ultramicrotome; dyeing it with 3% uranyl acetate-lead citrate; using JEM-2100HC High contrast TEM to observe and take photographs.The ultrastructures of spermatozoa of Sinonovacula rivularis sp. nov. and Sinonovacula constricta were examined by transmission electron microscopy(TEM)and scanning electron microscopy(SEM).The results showed that there were a great deal of differences between S.rivularis sp. nov. and S.constricta. The mature sperm of S.rivularis sp. nov. was a typical primitive type,consisting of head,middle piece and tail. S.rivularis sp. nov. and S.constricta both have ellipse-shaped acrosome,but the length of acrosome for S.rivularis sp. nov. was shorter than S.constricta. The nucleus of S.rivularis sp. nov. was oblate with 8-9 arc-tubes in outer margin,whereas the nucleus of S.constricta was spherical.The middle piece of S.rivularis sp. nov. contained five mitochondria,and typically a proximal and a distal centriole.Few mature sperm of S.rivularis sp. nov. contained 4 or 6 mitochondria.There were 5 or 6 mitochondria of S.constricta based on published information.The flagellum exhibited the typical 9 2 microtubule structure in both species.

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黄瑞,黄标武,李林春,张云飞.近江蛏精子超微形态结构观察及与缢蛏精子的比较[J].水产学报,2011,35(1):58~65

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  • 收稿日期:2010-09-25
  • 最后修改日期:2010-11-09
  • 录用日期:2010-12-01
  • 在线发布日期: 2011-01-14
  • 出版日期: 2011-01-15