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  • 2000-2004  (1)
  • 1995-1999  (1)
  • Embryonic stem cell Expressed sequence tag Neural differentiation Gene discovery  (1)
  • KN-62  (1)
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  • 2000-2004  (1)
  • 1995-1999  (1)
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  • 1
    ISSN: 1438-7948
    Keywords: Embryonic stem cell Expressed sequence tag Neural differentiation Gene discovery
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. We have used a method for synchronously differentiating murine embryonic stem (ES) cells into functional neurons and glia in culture. Using subtractive hybridization we isolated approximately 1200 cDNA clones from ES cell cultures at the neural precursor stage of neural differentiation. Pilot studies indicated that this library is a good source of novel neuro-embryonic cDNA clones. We therefore screened the entire library by single-pass sequencing. Characterization of 604 non-redundant cDNA clones by BLAST revealed 96 novel expressed sequence tags (ESTs) and an additional 197 matching uncharacterized ESTs or genomic clones derived from genome sequencing projects. With the exception of a handful of genes, whose functions are still unclear, most of the 311 known genes identified in this screen are expressed in embryonic development and/or the nervous system. At least 80 of these genes are implicated in disorders of differentiation, neural development and/or neural function. This study provides an initial snapshot of gene expression during early neural differentiation of ES cell cultures. Given the recent identification of human ES cells, further characterization of these novel and uncharacterized ESTs has the potential to identify genes that may be important in nervous system development, physiology and disease.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-7330
    Keywords: egg activation ; oocyte activation ; in vitro fertilization ; calmodulin-dependent protein kinase II ; KN-62
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Purpose: Our purpose was to determine the association of calmodulin-dependent protein kinase II (CaMK II ) with oocyte activation and to explore the network of protein kinases during mammalian fertilization. Methods: Mouse M-II oocytes were collected after superovulation induced by PMSG-hCG injection. The oocytes were inseminated or artificially activated by Ca ionophore (A23187) or 12-O-tetradecanoyl phorbol 13-acetate (TPA). The effects of KN-62, a specific and selective inhibitor of calmodulin-dependent protein kinase II, on second polar body emission (2PBE), pronuclearformation (PF), and cortical granule exocytosis (CGE) during fertilization or after artificial oocyte activation were investigated. Results: KN-62 inhibited 2PBE and PF after sperm or Ca ionophore inducing activation. Additionally, PF was inhibited by KN-62 after TPA activation, whereas KN-62 did not inhibit CGE in any case. KN-04, an inactive form of KN-62, did not inhibit significantly 2PBE, CGE, or PF. When oocytes were exposed to KN-62 after Ca ionophore or TPA activation, no inhibitory effects on 2PBE or PF were observed. Conclusions: The CaMK II activation that occurs after fertilization or artificial activation of mouse oocytes is presumably secondary to increases in the intracellular free calcium concentration. As determined by the use of inhibitor, CaMK II activity is associated with 2PBE and PF but not with CGE.
    Type of Medium: Electronic Resource
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