Premium
In Utero Electroporation(IUE) and mouse genetic model approach to assess PITP‐dependent inositol lipid signaling in NSC pool maintenance in embryonic brain
Author(s) -
Hur Seong Kwon,
Xie Zhigang,
Bankaitis Vytas
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.lb151
Subject(s) - microbiology and biotechnology , forebrain , biology , embryonic stem cell , inositol trisphosphate receptor , inositol , genetics , endocrinology , receptor , central nervous system , gene
Inositol (Ins) and phosphoinositide (PIP) signaling pathways are major intracellular regulatory systems of eukaryotic cells. Phosphatidylinositol transfer proteins (PITPs) execute novel modes of inositol lipid signal control by tightly channeling PIP production to specific biological outcomes. PITPα and PITPβ are highly expressed in mouse embryonic neocortex. We exploit an in utero electroporation approach to investigate the role of PITP‐dependent inositol lipid signaling in the embryonic neural stem cell (NSC) pool. We found that the combination of a PITPα null mouse line and PITPβ silencing evokes a dramatic depletion of NSC pools in embryonic brain. And, we finally have generated PITPα flox/flox and PITPβ flox/flox single mutants, and PITPα flox/flox , PITPβ flox/flox double mutant mouse line. In an experiment using Emx1‐Cre driver, which is forebrain specific driver, eviction of both PITPα and PITPβ in forebrain leads to a mouse that is born but has an amazing microcephaly that is due to virtual loss of the forebrain. Neither PITPα nor PITPβ eviction alone has any such effect. This experiment result is consistent with In utero electroporation data. Thus, we demonstrate that PITPα and PITPβ redundantly control NSC homeostasis in mouse brain development. Based on these data, we will observe carefully daughter cell fate during NSC division to investigate whether PITPs deficiency defeats NSC division program, asymmetric self‐renewing/differentiating cell divisions. Moreover, we will exploit our unique library of mutant PITPs with defined biochemical defects, to assess whether PtdCho‐ and PtdIns‐binding are required for PITP function in embryonic NSCs. Our research will generate completely new information regarding how lipid signaling contributes to NSC homeostasis in the developing mammalian brain. Support or Funding Information NIH 1R01GM112591‐01A1