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. 2017 Aug;369(2):395-412.
doi: 10.1007/s00441-017-2591-2. Epub 2017 Apr 22.

Calpain inhibition prevents flotillin re-ordering and Src family activation during capacitation

Deneb Maldonado-García  1 , Monica L Salgado-Lucio  1 , Ana L Roa-Espitia  1 , Tania Reyes-Miguel  1 , Enrique O Hernández-González  2
Affiliations

Calpain inhibition prevents flotillin re-ordering and Src family activation during capacitation

Deneb Maldonado-García et al. Cell Tissue Res. 2017 Aug.

Abstract

Prior to fertilization, mammalian sperm undergo several molecular, biochemical and physiological changes in a process termed capacitation. However, the mechanisms explaining the involvement of cytoskeletal remodeling and membrane re-ordering in each process prior to fertilization remain poorly understood. We found that the migration of both flotillin microdomains and Src family kinases towards the apical ridge of guinea pig sperm occurs under capacitating conditions. This re-ordering is associated with spectrin cleavage by calpain. Moreover, Src, Fyn, Lyn and Hck interact with flotillin-1; this interaction increases in a capacitation-dependent manner and the increased autophosphorylation of these kinases is linked to flotillin-1 association. The aforementioned results are prevented by the inhibition of calpain by calpeptin. Thus, spectrin cytoskeleton cleavage during capacitation seems to precede the reorganization of flotillin microdomains and Src family kinases towards the apical ridge of the sperm head in order to initiate the signaling cascade required for proper capacitation and further acrosome reaction. The significance of the Src family kinase reorganization for capacitation is demonstrated by the inhibition of calpain during capacitation also preventing the Src-family-kinase-dependent phosphorylation of FAK at Tyr576/577. Our work further highlights the scaffolding properties of flotillin microdomains and reveals the importance of their large-scale segregation during capacitation.

Keywords: Acrosome reaction; Flotillin microdomain aggregation; Role of spectrin in capacitation; Spectrin and sperm membrane domains; Src family kinases.

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