Expression of Rab3A in the cortical region in mouse metaphase II eggs.
Rab3A, a member of the small GTP-binding protein superfamily, has been implicated in regulated exocytosis at presynapses. At fertilization, sperm-egg fusion induces cytosolic calcium mobilization and cortical granule (CG) exocytosis in the egg. However, it is not yet clear whether Rab3A is involved in this process. We previously reported that Rabphilin-3A functions in calcium-dependent CG exocytosis. Rabphilin-3A is known to bind with Rab3A, Rab3B, and Rab3C. In this study, we clarified which member of the Rab3 was expressed in mouse metaphase II eggs. Messenger RNA encoding Rab3A but not Rab3B or Rab3C, was detected in unfertilized metaphase II eggs by RT-PCR. Rab3A protein was also detected in unfertilized metaphase II eggs by immunoblot analysis. Next the expression and the localization of Rab3A in eggs at various stages of development were determined by immunofluorescence analysis using confocal laser scanning microscopy. Rab3A protein was specifically distributed in the cortical region in eggs from before fertilization to the two-cell stage. However, it was not detected at the three- or four-cell stage 40 hr after fertilization. These results suggest that Rab3A may function with Rabphilin-3A in CG exocytosis.
Masumoto
N
N
Department of Obstetrics and Gynecology, Osaka University Medical School, Japan. naonao@cb3.so-net.or.jp
Ikebuchi
Y
Y
Tahara
M
M
Yokoi
T
T
Tasaka
K
K
Miyake
A
A
Murata
Y
Y
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D005089:Exocytosis / Q000502:physiology; D005260:Female; D005455:Fluorescent Antibody Technique; D019204:GTP-Binding Proteins / Q000032:analysis* / Q000235:genetics / Q000502:physiology; D005786:Gene Expression Regulation; D015151:Immunoblotting; D008677:Metaphase; D051379:Mice; D008815:Mice, Inbred Strains; D018613:Microscopy, Confocal; D010063:Ovum / Q000737:chemistry* / Q000166:cytology* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D020692:rab3 GTP-Binding Proteins
D017931:DNA Primers; D012333:RNA, Messenger; D019204:GTP-Binding Proteins; D020692:rab3 GTP-Binding Proteins