The Expression Pattern Of Ezrin In Mouse Testis And Ezrin’s Effect In Mice Sperm Capacitation

Ezrin, one of the membrane-cytoskeleton linker,participates in regulation of plasma membrane organization,cell migration,cellproliferation, cell signal-transduction, sustain cell form and lot of life events. Phosphorylation modification is the improtant mechanism for regulatory ezrin activation.There have been identified 7 phosphorylation sites that regulatory ezrin activation. Ezrin has been proved to expression in mouse testis, participates in mammalian spermatogenesis and sperm capacitation and so on. How about the expresson partern of ezrin in testis? What kind of phosphorylation modification happened? What relationship between those phosphorylation modification and spermatogenesis and sperm capacitation? There is no exact conclusion.Here we investigated the expression pattern of ezrin on testis of male mice at different ages and the effect of ezrin on different spermatogenesis stages,ezrin and the effect of ezrin on the development of mouse sperm capacitation by using immunofluorescence staining, immunohistochemical and Real-time PCR technology. There were three parts in this study:1. The location of ezrin in mouse testisOn the basis ofspermatogenic time, detached 1 week after birth (unmature),7 weeks (mature), and 17 weeks (adult) male mice testis, immunofluorescence staining and immunohistochemical technology analysis the location of ezrin in mouse testis, both results showed:immature male mice, ezrin mainly located in mouse testis Leydig cells,after maturity mice, ezrin not only located in Leyding cells,but within the seminiferous tubes sperm cells have partially colored, into adulthood, you can clearly see the coloring within the seminiferous tubes were deepen, broaden region.2. The expression pattern of ezrin in mouse testisDetached 1 week after birth (before maturity),7 weeks (mature), and 17 weeks (adult) male mice testis,to isolate total RNA for RT-PCR and Real-time PCR technology analysis the expression pattern of ezrin in mouse testis. The results showed:expression pattern of ezrin in immature stage testis slightly higher than the expression pattern of ezrin reach in maturity stage, adulthood ezrin expression levels gradually increased again.3. The expression and modification of ezrin in the head of epididymis sperm cellsDetched mature male mice epididymis head, releasing sperm cells and handled, take the appropriate amount of sperm cells apply on the coated with poly-lysine treatment glass slide, natural dryness, then fixed, with immunofluorescence staining technology use rabbit polyclonal anti-ezrin, phospho-ezrin (T567) antibody and phospho-ezrin (Y354) antibody for detection the expression pattern and modification of ezrin. The results showed:sperm cells in the head of epididymis have detected the expression of ezrin, between uncapacitation and capacitation group, both ezrin T567 and Y354 happened phosphorylation modification.4. The expression andmodification of ezrin in the tail of epididymis maturation sperm cellsDetched mature male mice epididymis tail, releasing sperm cells and handled, take the appropriate amount of sperm cells apply on the coated with poly-lysine treatment glass slide, natural dryness, and fixed, with its immunofluorescence staining technology for detection the phosphorylated-ezrin wild-ezrin expression level. The results showed:(1) Sperm cells in the tail of epididymis have detected the expression of ezrin, and in sperm capacitation group, the expression of ezrin have been decreased.(2) Sperm cells in the tail of epididymis, both uncapacitation and capacitation group have been detected the phosphorylation modification of ezrin,and in capacitation group, ezrin phosphorylation modification of Y354 were significant higher than uncapacitation.(3) Sperm cells in the tail of epididymis,uncapacitation or capacitation both have been detected the phosphorylation modification of ezrin, in capacitation group, ezrin phosphorylation modification of T567 were significant higher than uncapacitation group.