Evaluation of damage in Pacific oyster (Crassostrea gigas) spermatozoa before and after cryopreservation using comet assay

Jin Chywan Gwo, Chen Yen Wu, Wu Shu Peter Chang, Hsien Yu Cheng

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51 引文 斯高帕斯(Scopus)


We examined the applicability of the comet assay (single cell gel electrophoresis assay) to estimate the quality of frozen-thawed Pacific oyster (Crassostrea gigas) spermatozoa. Comet assay was performed on semen before and after cryopreservation followed by fluorescent staining with propidium iodide to assess DNA integrity. After cryopreservation, the percentage of spermatozoa with damaged DNA significantly increased, while only about half of the cells displayed intact DNA, even when protected with 10% DMSO. All the considered parameters (head length, head area, head intensity, total length, total area, total intensity, tail length%, tail area%, and tail intensity%) were higher than the oyster sperm protected with 10%DMSO-artificial sea water after freezing and thawing. Only tail length%, tail area%, and tail intensity% were increased significantly after cryopreservation. The tail length % was found to be the most sensitive indicator of the cryopreservation-induced DNA damage. Our freeze-thawing procedure significantly affected oyster sperm DNA, as indicated by the reduced fertilization rate when frozen-thawed oyster sperm are used. Irreversible alteration of the genome may prevent fertilization or alter normal embryonic development. This study is the first to demonstrate that the comet assay is an inexpensive, rapid and sensitive method for determining DNA damage in Pacific oyster sperm quality assessments.

頁(從 - 到)171-180
出版狀態已發佈 - 5月 2003

ASJC Scopus subject areas

  • 農業與生物科學(雜項)
  • 生理學
  • 生理學(醫學)


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