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Expression, Localization, and Binding Activity of the Ezrin/Radixin/Moesin Proteins in the Mouse Testis

Tomohiko Wakayama, Hiroki Nakata, Miho Kurobo, Yoshimichi Sai and Shoichi Iseki

Department of Histology and Embryology, Graduate School of Medical Science, Kanazawa University, Kanazawa, Japan (TW,HN,MK,SI), and Department of Pharmaceutics, Faculty of Pharmacy, Keio University, Tokyo, Japan (YS)

Correspondence to: Tomohiko Wakayama, MD, PhD, Department of Histology and Embryology, Graduate School of Medical Science, Kanazawa University, 13-1 Takara-machi, Kanazawa, Ishikawa, 920-8640, Japan. E-mail: twaka{at}basic.m.kanazawa-u.ac.jp

The ezrin, radixin, and moesin (ERM) proteins represent a family of adaptor proteins linking transmembrane proteins to the cytoskeleton. The seminiferous epithelium undergoes extensive changes in cellular composition, location, and shape, implicating roles of the membrane–cytoskeleton interaction. It remains unknown, however, whether the ERM proteins are expressed and play significant roles in the testis. In the present study, we examined the spatiotemporal expression of ERM proteins in the mouse testis by Western blotting and immunohistochemistry. Ezrin immunoreactivity was demonstrated in the cytoplasm of steps 15 and 16 spermatids from 5 weeks postpartum through adulthood, whereas radixin immunoreactivity was in the apical cytoplasm of Sertoli cells from 1 week through 2 weeks postpartum. No immunoreactivity for moesin was detected at any age. Immunoprecipitation demonstrated that ezrin was bound to the cytoskeletal component actin, whereas radixin was bound to both actin and tubulin. Of the transmembrane proteins known to interact with ERM proteins, only cystic fibrosis transmembrane conductance regulator, a chloride transporter, was bound to ezrin in elongated spermatids. These results suggest that ezrin is involved in spermiogenesis whereas radixin is involved in the maturation of Sertoli cells, through interaction with different sets of membrane proteins and cytoskeletal components. (J Histochem Cytochem 57:351–362, 2009)

Key Words: immunohistochemistry • Western blotting • immunoprecipitation • RT-PCR • mouse spermiogenesis • spermatids • Sertoli cells • ERM • CFTR • cytoskeleton

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