title: Flotillin-2 is an acrosome-related protein involved in mouse spermiogenesis.
abstract: SPERMATOGENESIS IS A COMPLEX PROCESS OF TERMINAL DIFFERENTIATION BY WHICH MATURE SPERMS ARE GENERATED, AND IT CAN BE DIVIDED INTO THREE PHASES: mitosis, meiosis and spermiogenesis. In a previous study, we established a series of proteomic profiles for spermatogenesis to understand the regulation of male fertility and infertility. Here, we further investigated the localization and the role of flotillin-2 in spermiogenesis. Flotillin-2 expression was investigated in the testis of male CD1 mice at various developmental stages of spermatogenesis by using Western blotting, immunohistochemistry and immunofluorescence. Flotillin-2 was knocked down in vivo in three-week-old male mice using intratesticular injection of small inhibitory RNA (siRNA), and sperm abnormalities were assessed three weeks later. Flotillin-2 was expressed at high levels in male germ cells during spermatogenesis. Flotillin-2 immunoreactivity was observed in pachytene spermatocytes as a strong dot-shaped signal and in round spermatids as a sickle-shaped distribution ahead of the acrosome. Immunofluorescence confirmed flotillin-2 was localized in front of the acrosome in round spermatids, indicating that flotillin-2 was localized to the Golgi apparatus. Knockdown of flotillin-2 in vivo led to a significant increase in head sperm abnormalities isolated from the cauda epididymis, compared with control siRNA-injected testes. This study indicates that flotillin-2 is a novel Golgi-related protein involved in sperm acrosome biogenesis.
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing, Jiangsu 210029, China.
authors.forename: Yibo
authors.initials: Y
authors.lastname: Wu
authors.forename: Xin
authors.initials: X
authors.lastname: Chen
authors.forename: Shuai
authors.initials: S
authors.lastname: Wang
authors.forename: Min
authors.initials: M
authors.lastname: Jiang
authors.forename: Bo
authors.initials: B
authors.lastname: Zheng
authors.forename: Quan
authors.initials: Q
authors.lastname: Zhou
authors.forename: Ye
authors.initials: Y
authors.lastname: Bi
authors.forename: Zuomin
authors.initials: Z
authors.lastname: Zhou
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Huang
authors.forename: Jiahao
authors.initials: J
authors.lastname: Sha
keywords: Golgi apparatus; RNA interference; acrosome biogenesis; flotillin-2
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