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Na/K-ATPase β1 Subunit Expression Is Required for Blastocyst Formation and Normal Assembly of Trophectoderm Tight Junction-associated Proteins

Madan, Pavneesh ; Rose, Keeley ; Watson, Andrew J.

The Journal of biological chemistry, 2007-04, Vol.282 (16), p.12127-12134 [Periódico revisado por pares]

Elsevier Inc

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  • Título:
    Na/K-ATPase β1 Subunit Expression Is Required for Blastocyst Formation and Normal Assembly of Trophectoderm Tight Junction-associated Proteins
  • Autor: Madan, Pavneesh ; Rose, Keeley ; Watson, Andrew J.
  • É parte de: The Journal of biological chemistry, 2007-04, Vol.282 (16), p.12127-12134
  • Notas: http://www.jbc.org/
  • Descrição: Na/K-ATPase plays an important role in mediating blastocyst formation. Despite the expression of multiple Na/K-ATPase α and β isoforms during mouse preimplantation development, only the α1 and β1 isoforms have been localized to the basolateral membrane regions of the trophectoderm. The aim of the present study was to selectively down-regulate the Na/K-ATPase β1 subunit employing microinjection of mouse 1 cell zygotes with small interfering RNA (siRNA) oligos. Experiments comprised of non-injected controls and two groups microinjected with either Stealth™ Na/K-ATPase β1 subunit oligos or nonspecific Stealth™ siRNA as control. Development to the 2-, 4-, 8-, and 16-cell and morula stages did not vary between the three groups. However, only 2.3% of the embryos microinjected with Na/K-ATPase β1 subunit siRNA oligos developed to the blastocyst stage as compared with 73% for control-injected and 91% for non-injected controls. Na/K-ATPase β1 subunit down-regulation was validated by employing reverse transcription-PCR and whole-mount immunofluorescence methods to demonstrate that Na/K-ATPase β1 subunit mRNAs and protein were not detectable in β1 subunit siRNA-microinjected embryos. Aggregation chimera experiments between β1 subunit siRNA-microinjected embryos and controls demonstrated that blockade of blastocyst formation was reversible. The distribution of Na/K-ATPase α1 and tight junction-associated proteins occludin and ZO-1 were compared among the three treatment groups. No differences in protein distribution were observed between control groups; however, all three polypeptides displayed an aberrant distribution in Na/K-ATPase β1 subunit siRNA-microinjected embryos. Our results demonstrate that the β1 subunit of the Na/K-ATPase is required for blastocyst formation and that this subunit is also required to maintain a normal Na/K-ATPase distribution and localization of tight junction-associated polypeptides during preimplantation development.
  • Editor: Elsevier Inc
  • Idioma: Inglês
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  • Realização: ABCD-USP USP   Logos de Redes Sociais: Freepik

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