期刊
GENE EXPRESSION PATTERNS
卷 9, 期 2, 页码 94-108出版社
ELSEVIER
DOI: 10.1016/j.gep.2008.10.002
关键词
Embryonic stem cells; Development; Serotonin; Macaques; Affymetrix array; Gene expression; Dorsal raphe
资金
- NCRR NIH HHS [K01 RR000163, RR000163, P51 RR000163] Funding Source: Medline
- NICHD NIH HHS [U54 HD018185, P30 HD018185, HD 18185] Funding Source: Medline
- NIMH NIH HHS [MH7356, R21 MH073564, R01 MH062677-08, R21 MH073564-02, MH62677, R01 MH062677] Funding Source: Medline
The rhesus monkey embryonic stem cell line 366.4 differentiates into serotonin neurons. We examined the genetic cascade during differentiation and compared ESC-derived serotonin neurons to adult monkey serotonin neurons. RNA was extracted from ESC colonies, embryoid bodies (EBs), neurospheres in selection (N1) and proliferation stages (N2), differentiated serotonin neurons (N3) and from laser captured (LC) serotonin neurons of spayed female macaques treated with placebo, estrogen (E), progesterone (P) or E + P. The RNA was labeled and hybridized to Rhesus Monkey Affymetrix Gene Chips (n = 1 per stage and 2 per animal treatment). Gene expression was examined with GeneSifter software. 545 genes that were related to developmental processes showed a threefold or greater change between stages. TGFb, Writ, VEGF and Hedgehog signaling pathways showed the highest percent of probe set changes during differentiation. Genes in the categories (a) homeobox binding and transcription factors. (b) growth factors and receptors, (c) brain and neural specific factors and (d) serotonin specific factors are reported. Pivotal genes were confirmed with quantitative RT-PCR. In the serotonin developmental cascade, FGFR2 was robustly expressed at each stage. GATA3 was robustly expressed in EBs. Sonic hedgehog (Shh), PTCH (Shh-R) and Fev1 transcription factor expression coincided with the induction of serotonin specific marker genes during N1-selection. A majority of the examined genes were expressed in adult serotonin neurons. However, in the ESC-derived neurons, there was significant over-representation of probe sets related to cell cycle, axon guidance & dorso-ventral axis formation. This analysis suggests that the 366.4 cell line possesses Cues for serotonin differentiation at early stages of differentiation, but that ESC-derived serotonin neurons are still immature. (C) 2008 Elsevier B.V. All rights reserved.
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