Epididymal protease inhibitor (EPPIN) is differentially expressed in the male rat reproductive tract and immunolocalized in maturing spermatozoa.
EPPIN (epididymal protease inhibitor; SPINLW1), an antimicrobial cysteine-rich protein containing both Kunitz and whey acidic protein (WAP)-type four disulfide core protease inhibitor consensus sequences, is a target for male contraception because of its critical role in sperm motility. Here, we characterized EPPIN's expression and cellular distribution in rat tissues and its in vivo regulation by androgens in the epididymis. EPPIN (mRNA and protein) was abundantly expressed in the rat testis and epididymis; we also found that the vas deferens, seminal vesicles, and brain were novel sites of EPPIN expression. PCR studies demonstrated that in addition to Sertoli cells, spermatogenic cells expressed Eppin mRNA. EPPIN was immunolocalized in Sertoli cells and spermatogenic cells (pachytene spermatocytes and round and elongated spermatids) and in epithelial cells and spermatozoa from efferent ductules and epididymis. EPPIN staining was observed on the middle and principal pieces of the flagellum of testicular spermatozoa. Epididymal spermatozoa had more intense EPPIN staining on the flagellum, and the EPPIN staining became apparent on the head and neck regions. This suggested that the EPPIN found on maturing spermatozoa was secreted primarily by the epithelial cells of the epididymis. Surgical castration down-regulated EPPIN expression levels (mRNA and protein) in the caput and cauda epididymis, an effect reversed by testosterone replacement. Altogether, our data suggested that EPPIN expression in rats is more widespread than in humans and mice, and is androgen-dependent in the epididymis. This species could be used as an experimental model to further study EPPIN's role in male fertility.
Section of Experimental Endocrinology, Department of Pharmacology, Universidade Federal de São Paulo - Escola Paulista de Medicina, SP, Brazil. ejrsilva81@gmail.com
Erick J R
EJ
Silva
Marília T C C
MT
Patrão
James K
JK
Tsuruta
Michael G
MG
O'Rand
Maria Christina W
MC
Avellar
D000728:Androgens / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000378:metabolism; D002369:Castration; D002478:Cells, Cultured; D004822:Epididymis / Q000378:metabolism; D005837:Genitalia, Male / Q000378:metabolism*; D008297:Male; D053491:Proteinase Inhibitory Proteins, Secretory / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D029607:Seminal Vesicle Secretory Proteins / Q000235:genetics / Q000378:metabolism; D012669:Seminal Vesicles / Q000378:metabolism; D013079:Sperm Maturation; D013081:Sperm Motility; D013082:Sperm Tail / Q000378:metabolism; D013094:Spermatozoa / Q000378:metabolism*; D014649:Vas Deferens / Q000378:metabolism
D000728:Androgens; D053491:Proteinase Inhibitory Proteins, Secretory; D012333:RNA, Messenger; D029607:Seminal Vesicle Secretory Proteins; C546856:eppin protein, rat