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Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Veterinary Medicine / Faculty of Veterinary Medicine >
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Simultaneous evaluation of plasma membrane integrity, acrosomal integrity, and mitochondrial membrane potential in bovine spermatozoa by flow cytometry
| Title: | Simultaneous evaluation of plasma membrane integrity, acrosomal integrity, and mitochondrial membrane potential in bovine spermatozoa by flow cytometry |
| Authors: | Kanno, Chihiro Browse this author | | Kang, Sung-Sik Browse this author | | Kitade, Yasuyuki Browse this author | | Yanagawa, Yojiro Browse this author | | Takahashi, Yoshiyuki Browse this author →KAKEN DB | | Nagano, Masashi Browse this author →KAKEN DB |
| Keywords: | Acrosomal integrity | | Bovine spermatozoa | | Flowcytometry | | Mitochondrial membrane potential | | Spermatozoon viability |
| Issue Date: | Sep-2015 |
| Publisher: | Cambridge University Press |
| Journal Title: | Zygote |
| Volume: | 24 |
| Issue: | 4 |
| Start Page: | 529 |
| End Page: | 536 |
| Publisher DOI: | 10.1017/S0967199415000490 |
| PMID: | 26369275 |
| Abstract: | 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 |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/61099 |
| Appears in Collections: | 獣医学院・獣医学研究院 (Graduate School of Veterinary Medicine / Faculty of Veterinary Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 永野 昌志
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