Identification of a heat-shock protein Hsp40, DjB1, as an acrosome- and a tail-associated component in rodent spermatozoa.
Iba1 is a 17-kDa EF-hand protein highly expressed in the cytoplasm of elongating spermatids in testis. Using Iba1 as a bait, we performed yeast Two-hybrid screening and isolated a heat-shock protein Hsp40, DjB1, from cDNA library of mouse testis. To characterize DjB1 that is encoded by Dnajb1 gene, we carried out immunoblot analyses, in situ hybridization, and immunohistochemistry. Immunoblot analyses showed that DjB1was constitutively expressed in mouse testis and that its expression level was not changed by heat shock. Dnajb1 mRNA was exclusively expressed in spermatocytes and round spermatids in mouse testis, and Dnajb1 protein DjB1 was predominantly expressed in the cytoplasm of spermatocytes, round spermatids, and elongating spermatids. In mature mouse spermatozoa, DjB1 was localized in the middle and the end pieces of flagella as well as in association with the head (acrosomal region). Association of DjB1 with the acrosomal region in sperm head was also observed in rat spermatozoa. These data suggested that DjB1, which was constitutively expressed in postmeiotic spermatogenic cells in testis, was integrated into spermatozoa as at least two components, that is, sperm head and tail of rodent spermatozoa.
Doiguchi
Masamichi
M
Laboratory of Zoology, Graduate School of Agriculture, Kyushu University, Higashiku Hakozaki, Fukuoka, Japan.
Kaneko
Takane
T
Urasoko
Aiko
A
Nishitani
Hideo
H
Iida
Hiroshi
H
D020101:Acrosome Reaction / Q000502:physiology*; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D050956:HSP40 Heat-Shock Proteins / Q000032:analysis* / Q000235:genetics; D017403:In Situ Hybridization; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013077:Sperm Head / Q000502:physiology; D013082:Sperm Tail / Q000502:physiology; D013094:Spermatozoa / Q000502:physiology*; D013737:Testis / Q000166:cytology; D014162:Transfection
D017931:DNA Primers; D018076:DNA, Complementary; D050956:HSP40 Heat-Shock Proteins; D012333:RNA, Messenger