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Relationship of gap junction formation to phosphorylation of connexin43 in mouse preimplantation embryos
Author(s) -
Ogawa Hidehiko,
Oyamada Masahito,
Mori Tadashi,
Mori Michio,
Shimizu Hiroshi
Publication year - 2000
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/(sici)1098-2795(200004)55:4<393::aid-mrd6>3.0.co;2-x
Subject(s) - biology , phosphorylation , western blot , embryo , microbiology and biotechnology , activator (genetics) , kinase , immunofluorescence , protein kinase a , cell , cell culture , protein kinase c , immunology , biochemistry , gene , genetics , antibody
To clarify the relationship of gap junction formation to phosphorylation of connexin43 (Cx43) in mouse preimplantation embryos, immunofluorescence and Western blot analysis were conducted. Immunofluorescence showed Cx43 positive spots first at the mid‐eight‐cell stage (6 hr postdivision to the eight‐cell stage). The number of spots increased from 6 to 15 hr postdivision to the eight‐cell stage. Western blot analysis suggested Cx43 to possibly be present in the nonphosphorylated form at the mid‐four‐cell stage (6 hr postdivision to the four‐cell stage), and phosphorylated Cx43 to increase from the mid‐eight‐cell stage (6 hr post‐division to the eight‐cell stage) onward. Dibutyryl cAMP (dbcAMP), a protein kinase A (PKA) activator, added to the culture medium increased the phosphorylation of Cx43 and Cx43 positive spots. The tumor promoter 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA), a protein kinase C (PKC) activator, increased the phosphorylation of Cx43, but decreased Cx43 positive spots. These results suggest that the phosphorylation of Cx43, induced by different protein kinase, leads to a different effect on gap junction formation in mouse preimplantation embryos. Mol. Reprod. Dev. 55:393–398, 2000. © 2000 Wiley‐Liss, Inc.