Different Transcription Factors Regulate nestin Gene Expression during P19 Cell Neural Differentiation and Central Nervous System Development

Abstract

Nestin is a molecular marker for neural progenitor cells. Rat and human nestin genes possess a central nervous system-specific enhancer within their second introns. However, the transcription factors that bind to the nestin enhancer have not been fully elucidated. Here, we show that the second intron of the mouse nestin gene is sufficient to drive reporter gene expression in the developing nervous system. The core sequence of this central nervous system-specific enhancer localizes to the 3 320-bp region. The cis-elements for Sox and POU family transcription factors and the hormone-responsive element are essential for nestin expression during embryonic carcinoma P19 cell neural differentiation and in the developing chick neural tube. Interestingly, different transcription factors bind to the nestin enhancer at different stages of P19 cell neural differentiation and central nervous system development. Sox2 and SF1 may mediate basal nestin expression in undifferentiated P19EC cells, whereas Sox2, Brn1, and Brn2 bind to the enhancer in P19 neural progenitor cells. Similarly, in vivo, Oct1 binds to the nestin enhancer in embryonic day 8.5 (E8.5) mouse embryos, and Oct1, Brn1, and Brn2 bind to this enhancer in E10.5 and E12.5 mouse embryos. Our studies therefore suggest a temporal coordination of transcription factors in determining nestin gene expression.

Combinatorial binding of transcription factors in the pluripotency control regions of the genome

“…There are examples that these factors interact to coregulate genes outside ES cells. POU2F1 and SOX2 have been shown to function synergistically in the regulation of the PAX6 and nestin genes later in neural development (Donner et al 2007;Jin et al 2009). SOX2 is widely expressed and therefore present with POU2F1 in many more tissues than POU5F1, including cell types and cell lines (e.g., HeLa) where POU2F1 is the only POU factor detectable (M Gemberling and L Ferraris, unpubl.).…”

AP2γ regulates neural and epidermal development downstream of the BMP pathway at early stages of ectodermal patterning

“…Cell culture and treatment P19 cells were cultured as previously described [75]. Mouse ESC lines R1 was used.…”

“…Reverse transcription and qRT-PCR were performed as described previously [75,84]. The primers are listed in Supplementary information, Table S3.…”

“…ChIP assay was performed according to the method described [75,84]. Mouse monoclonal anti-Smad1 antibody (10 µg per sample, CST) and normal mouse IgG in control groups were used for immunoprecipitation.…”

GATA6, SF1, NGFIB and DAX1 in the remodeled subcapsular zones in primary aldosteronism

“…These three nuclear transcription factors were also considered to play roles in the subscapular remodeling processes regardless of aldosterone overproduction. GATA6, SF1 and DAX1 are all known to play pivotal roles in developmental processes [13,22,23].…”