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Gene expression regulating epithelial intercellular junction biogenesis during human blastocyst development in vitro.

M Reza Ghassemifar1, Judith J Eckert, Franchesca D Houghton

  • 1Division of Cell Sciences, School of Biological Sciences, University of Southampton, Bassett Crescent East, Southampton SO16 7PX, UK.

Molecular Human Reproduction
|May 3, 2003
PubMed
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Investigating gene expression in human embryos reveals key junction proteins crucial for trophectoderm development and blastocyst formation. Detection levels varied, with some genes essential throughout development and others critical for later stages, impacting embryo viability.

Area of Science:

  • Developmental Biology
  • Embryology
  • Molecular Biology

Background:

  • Trophectoderm epithelial intercellular junctions, including tight junctions (TJ) and desmosomes, are vital for human embryo development.
  • These junctions facilitate vectorial transport and blastocoel cavity formation, essential processes for successful implantation.
  • Understanding the gene expression patterns of these junction proteins is critical for assessing early human embryo development and viability.

Purpose of the Study:

  • To investigate the gene expression profiles of key proteins involved in trophectoderm epithelial intercellular junction formation during human preimplantation development.
  • To correlate gene expression patterns with embryo developmental stages and morphological grade.
  • To compare human embryo junction protein expression and assembly with that of mouse blastocysts.

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Main Methods:

  • RT-PCR was employed to analyze the expression of genes encoding TJ and desmosome proteins (claudin-1, occludin, ZO-1, ZO-2, JAM, DSC2) in single human embryos.
  • Magnetic bead separation was used for polyA+ RNA isolation.
  • Immunoconfocal microscopy was utilized to assess the membrane assembly of selected proteins.

Main Results:

  • Differential gene expression was observed, with some transcripts (claudin-1, JAM, occludin TM4, ZO-1alpha-) detected throughout development, while others (ZO-2, ZO-1alpha+, DSC-2) were preferentially expressed during later cleavage stages and blastocyst formation.
  • Transcript detection sensitivity was influenced by the RT-PCR method, with solid-phase RT-PCR showing a positive correlation with embryo morphological grade.
  • Lower detection of occludin TM4-, ZO-1alpha+, and DSC2 transcripts in blastocysts suggested low expression levels potentially impacting trophectoderm differentiation competence.
  • Human embryos showed poorer membrane assembly of junction proteins compared to mouse blastocysts.

Conclusions:

  • The study elucidates stage-specific gene expression patterns of trophectoderm junction proteins in human embryos, offering insights into developmental regulation.
  • Detection method significantly impacts transcript analysis, highlighting the importance of optimized protocols for accurate assessment of human embryo development.
  • Impaired expression and assembly of junction proteins in human embryos may compromise trophectoderm differentiation and overall embryo viability, warranting further investigation.