水牛附睾不同部位精子超微结构及荧光标记差异分析

doi:10.11843/j.issn.0366-6964.2021.012.012

Abstract

Abstract: In mammals, spermatozoa acquired the ability of motility and fertilization only after being matured in the epididymis. To explain the maturation process of buffalo spermatozoa in the epididymis, sexually mature buffalo (Bubalus bubalis) epididymis were selected and the spermatozoa from the caput, corpus and cauda of the epididymis were purified by Polyvinylpyrrolidone coated silica (Percoll) gradient centrifugation. Computer-assisted sperm analysis system (CASA) was used to examine sperm motility, transmission electron microscopy to observe the ultrastructure of spermatozoa in different regions of the epididymis, and after fluorescent labeling of spermatozoa, flow cytometry and fluorescence microscopy were employed to determine the plasma membrane integrity, mitochondrial membrane potential and acrosome variations in different regions of the spermatozoa. As the result showed, the purity of spermatozoa obtained from the head, body and tail of epididymis by Percoll were 95%, whereas the motility of spermatozoa in various regions was 8.35%, 20.21% and 65.60%, respectively; All the spermatozoa in different regions of epididymis existed the same abnormality pattern, and the spermatozoa in the cauda of epididymis showed the highest ratio of mitochondrial membrane potential, the sperm plasma membrane integrity rate and and the sperm acrosome integrity rate increased gradually from the caput to the cauda region of epididymis. Overall, this study demonstrated the characterization and variation of spermatozoa in distinct regions of the buffalo epididymis which provides a theoretical basis for investigating the mechanism of sperm maturation.

Key words: epididymal, sperm, ultrastructure observation, fluorescent molecular probe, acrosome

CLC Number:

HE Wengtan, ZHANG Pengfei, HUANG Yulin, XIAO Kai, HUANG Liangfeng, LU Yangqing, ZHANG Ming. Analysis of Ultrastructure and Fluorescent Markers of Sperm in Different Parts of Buffalo Epididymis[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3449-3460.

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Analysis of Ultrastructure and Fluorescent Markers of Sperm in Different Parts of Buffalo Epididymis

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