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BOR - Papers in Press, published online ahead of print December 10, 2003.
Biol Reprod 2003, 10.1095/biolreprod.103.020917
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Submitted July 1, 2003
Returned for revision July 25, 2003
Accepted December 1, 2003

Embryo


Expression and Role of the Ether-à-Go-Go-Related (MERG1A) Potassium Channel Protein During Preimplantation Mouse Development

N. J. Winston , M. H. Johnson , J. M. McConnell , D. I. Cook , and M. L. Day *

* To whom correspondence should be addressed. E-mail: margotd{at}physiol.usyd.edu.au.

Abstract
Potassium channels play important roles in many cellular processes including cell cycle progression and cell differentiation. In the present studies we investigated the pattern of expression of the mouse ether-à-go-go-related (KCNH2, merg1a) potassium channel during mouse embryogenic development. Reverse transcriptase-polymerase chain reaction analysis revealed the presence of maternal merg1a transcripts until the late 2-cell stage of development, after which merg1a expression from the zygotic genome was low until the 8-cell stage, then rose in the morula, but was low in trophoblast compared to inner cell mass cells. A trophoblast stem cell line was also shown to express merg1a mRNA. Immunoblotting of oocytes, blastocysts and the trophoblast stem cell line revealed the presence of different post-translationally processed forms of MERG1A. Immunofluorescence analysis showed that the sub-cellular localization of MERG1A varied at different stages of the embryogenic cell cycle. In addition MERG1A protein levels increased following compaction at the 8-cell stage and its distribution became polarized. This re-localization of MERG1A was affected by treatment with specific inhibitors of ERG channel function and of actin polymerization. Puromycin treatment of morulae indicated that membrane-associated MERG1A had a half-life of greater than 24 h. The ERG-specific inhibitor E-4031 reduced the incidence of blastocyst formation and the number of cells per blastocyst. These results show that MERG1A is developmentally regulated and suggest that it might play a role in early mouse embryogenic development.

Key words: Conceptus • Early development • Gene regulation • Signal transduction • Trophoblast


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[Abstract] [PDF]


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