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

Kanno, Chihiro; Kang, Sung-Sik; Kitade, Yasuyuki et al.

Zygote, 2015, 24(4), 529-536

Permalink : https://hdl.handle.net/2115/61099

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アクセス権
open access
URI
https://hdl.handle.net/2115/61099
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 ; Kang, Sung-Sik ; Kitade, Yasuyuki ; Yanagawa, Yojiro ; Takahashi, Yoshiyuki ; Nagano, Masashi
著者名
Kanno, Chihiro
著者名
Kang, Sung-Sik
著者名
Kitade, Yasuyuki
著者名
Yanagawa, Yojiro
e-Rad_Researcher
70167485
著者名
Takahashi, Yoshiyuki
e-Rad_Researcher
70312402
著者名
Nagano, Masashi
言語
英語
キーワード
Acrosomal integrity
Bovine spermatozoa
Flowcytometry
Mitochondrial membrane potential
Spermatozoon viability
発行日
2015-09
出版者
Cambridge University Press
収録物名
Zygote
巻(号)
24(4)
ページ
529-536
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.
資源タイプ
学術雑誌論文
出版タイプ
Accepted Manuscript
権利情報
© Cambridge University Press 2015
PISSN
0967-1994
EISSN
1469-8730
NCID
AA10958262
関連情報
PMID
26369275
関連情報 (isVersionOf)
出版社版DOI
https://doi.org/10.1017/S0967199415000490