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Simultaneous evaluation of plasma membrane integrity, acrosomal integrity, and mitochondrial membrane potential in bovine spermatozoa by flow cytometry

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タイトル: Simultaneous evaluation of plasma membrane integrity, acrosomal integrity, and mitochondrial membrane potential in bovine spermatozoa by flow cytometry
著者: Kanno, Chihiro 著作を一覧する
Kang, Sung-Sik 著作を一覧する
Kitade, Yasuyuki 著作を一覧する
Yanagawa, Yojiro 著作を一覧する
Takahashi, Yoshiyuki 著作を一覧する
Nagano, Masashi 著作を一覧する
キーワード: Acrosomal integrity
Bovine spermatozoa
Flowcytometry
Mitochondrial membrane potential
Spermatozoon viability
発行日: 2015年 9月15日
出版者: Cambridge University Press
誌名: Zygote
開始ページ: 1
終了ページ: 8
出版社 DOI: 10.1017/S0967199415000490
抄録: The present study aimed to develop an objective evaluation procedure to estimate the plasma membrane integrity, acrosomal integrity, and mitochondrial membrane potential of bull spermatozoa simultaneously by flow cytometry. Firstly, we used frozen-thawed semen mixed with 0, 25, 50, 75 or 100% dead spermatozoa. Semen was stained using three staining solutions: SYBR-14, propidium iodide (PI), and phycoerythrin-conjugated peanut agglutinin (PE-PNA), for the evaluation of plasma membrane integrity and acrosomal integrity. Then, characteristics evaluated by flow cytometry and by fluorescence microscopy were compared. Characteristics of spermatozoa (viability and acrosomal integrity) evaluated by flow cytometry and by fluorescence microscopy were found to be similar. Secondly, we attempted to evaluate the plasma membrane integrity, acrosomal integrity, and also mitochondrial membrane potential of spermatozoa by flow cytometry using conventional staining with three dyes (SYBR-14, PI, and PE-PNA) combined with MitoTracker Deep Red (MTDR) staining (quadruple staining). The spermatozoon characteristics evaluated by flow cytometry using quadruple staining were then compared with those of staining using SYBR-14, PI, and PE-PNA and staining using SYBR-14 and MTDR. There were no significant differences in all characteristics (viability, acrosomal integrity, and mitochondrial membrane potential) evaluated by quadruple staining and the other procedures. In conclusion, quadruple staining using SYBR-14, PI, PE-PNA, and MTDR for flow cytometry can be used to evaluate the plasma membrane integrity, acrosomal integrity, and mitochondrial membrane potential of bovine spermatozoa simultaneously.
Rights: © Cambridge University Press 2015
資料タイプ: article (author version)
URI: http://hdl.handle.net/2115/61099
出現コレクション:雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

提供者: 永野 昌志

 

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