PCP4 (PEP19) overexpression induces premature neuronal differentiation associated with Ca2+/Calmodulin-Dependent kinase II- Delta d activation in mouse models of down syndrome

Pcp4/pep19 is a modulator of Ca2+-CaM, a key molecule for calcium signaling, expressed in postmitotic neuroectoderm cells during mouse embryogenesis. The PCP4 gene is located on human chromosome 21 and is present in three copies in Down syndrome (DS). To evaluate the consequences of three copies of this gene on the development of these cells in the nervous system, we constructed a transgenic (TgPCP4) mouse model, with one copy of human PCP4, and investigated the effects in this model and in the Ts1Cje, a mouse model of DS. During embryogenesis, we analyzed 1) the level of pcp4 transcript and protein in the two models; 2) the extent of colabeling for markers of neuronal differentiation ( Delta *bIII-tubulin, Map2c, calbindin, and calretinin) and pcp4 by immunofluorescence analysis and overall protein levels of these markers by Western blotting; and 3) the rate of activation of CaMKII, a Ca2+-CaM target, to evaluate the impact of pcp4 overexpression on the Ca2+-CaM signaling pathway. We showed that three copies of the pcp4 gene induced the overexpression of transcripts and proteins during embryogenesis. Pcp4 overexpression 1) induced precocious neuronal differentiation, as shown by the distribution and levels of early neuronal markers; and 2) was associated with an increase in CaMKII Delta *d activation, confirming involvement in neuronal differentiation in vivo via a Pcp4-Ca2+-CaM pathway. TgPCP4 and Ts1Cje mice developed similar modifications, demonstrating that these mechanisms may account for abnormal neuronal development in DS. J. Comp. Neurol. 519:2779-2802, 2011. ? 2011 Wiley-Liss, Inc.