Contributions of Na+/H+ exchanger isoforms to preimplantation development of the mouse.

Previous work provided evidence of Na+/H+ exchanger activity in the apical domain of mouse trophectodermal plasma membranes that provides a route for entry of extracellular Na+ (Manejwala et al., 1989). This activity was hypothesized to contribute to the trans-trophectodermal Na+ flux that is required for blastocoel expansion. In the present work, we have used reverse transcriptase-polymerase chain reaction (RT-PCR) and immunocytochemistry to identify members of the Na+/H+ exchanger (NHE) family that are likely to participate in this process. When cDNA preparations from ovulated oocytes and several stages of preimplantation development were tested with PCR primers specific for the NHE-1, -2, -3, and -4 isoforms of the exchanger, only amplicons representing the NHE-1 and NHE-3 isoforms were detected. The identity of these amplicons was confirmed by direct sequencing. NHE-1 mRNA is present in oocytes and in all preimplantation stages, increasing threefold on a per embryo basis between the 4-cell and blastocyst stages. NHE-3 mRNA, on the other hand, was only detected in oocytes. Immunocytochemical analysis of blastocysts revealed that NHE-1 is localized in the basolateral domain of the trophectoderm, whereas NHE-3 is localized in the apical domain, a situation like that in epithelia of adult organs. We conclude that NHE-3, an oogenetic product that persists into the blastocyst stage, is the Na+/H+ exchanger isoform most likely to be involved in blastocoel expansion.