Gene Expression Changes in the Course of Neural Progenitor Cell Differentiation

Abstract

The molecular changes underlying neural progenitor differentiation are essentially unknown. We applied cDNA microarrays with 13,627 clones to measure dynamic gene expression changes during the in vitro differentiation of neural progenitor cells that were isolated from the subventricular zone of postnatal day 7 mice and grown in vitro as neurospheres. In two experimental series in which we withdrew epidermal growth factor and added the neurotrophins Neurotrophin-4 or BDNF, four time points were investigated: undifferentiated cells grown as neurospheres, and cells 24, 48, and 96 hr after differentiation. Expression changes of selected genes were confirmed by semiquantitative RT-PCR. Ten different groups of gene expression dynamics obtained by cluster analysis are described. To correlate selected gene expression changes to the localization of respective proteins, we performed immunostainings of cultured neurospheres and of brain sections from adult mice. Our results provide new insights into the genetic program of neural progenitor differentiation and give strong hints to as yet unknown cellular communications within the adult subventricular zone stem cell niche.

Periostin Secreted by Mesenchymal Stem Cells Supports Tendon Formation in an Ectopic Mouse Model

“…cDNA synthesis and reverse transcriptionpolymerase chain reaction Five micrograms of total RNA from cell cultures was reverse transcribed with the cDNA synthesis kit (MMLV-RT; Life Technologies) and with oligo-dT (12)(13)(14)(15)(16)(17)(18) -primer. For tendon and muscle tissue, the entire material was used for cDNA synthesis, which was carried out after DNase treatment to remove genomic DNA according to the protocol of the supplier (Revert Aid First Strand cDNA synthesis Kit; Fermentas) using random hexamer primers.…”

“…For tendon and muscle tissue, the entire material was used for cDNA synthesis, which was carried out after DNase treatment to remove genomic DNA according to the protocol of the supplier (Revert Aid First Strand cDNA synthesis Kit; Fermentas) using random hexamer primers. cDNA aliquots were subjected to polymerase chain reaction (PCR Microarrays cDNA microarrays [12] were generated using *20,000 murine cDNA clones (arrayTAG clone collection) from LION Bioscience. In this array, several genes are represented by more than one probe.…”

“…Image acquisition and data analysis was done according to Gurok et al [12]. From the hybridization data obtained we established a list of DEPs.…”

Porcine neural progenitor cells derived from tissue at different gestational ages can be distinguished by global transcriptome

“…Both culture media treatments showed a decrease in neuron-specific genes in the E45 pBPC with time, such as DCX, TUBB3, NEUROD1, and TBR1, all of which are expressed by cells committed to a neuronal fate during development. [49][50][51][52] However, unlike the UL-treated cells, the SM-treated E45 pBPCs showed an upregulation of several gliaspecific genes such as MBP and OLIG2, suggesting that the SM treatment over time induces the E45 pBPCs toward a glial cell fate, while UL treatment does not due to the lack of any change in glia-related gene expression. Over time in culture, neural stem/progenitor cells tend to lose their neurogenic potential 53 and instead become more gliogenic, 54,55 suggesting that what we observe in the E45 pBPC in SM is the beginning stages of this determination into a glia-restricted fate over time in culture.…”

Mass spectrometry based proteomic analysis of human stem cells: a brief review

“…Differentiation and maturation of stem cells comprises complex serial events, which guide the undifferentiated cells to different lineages via distinctive developmental programs (Chung et al, 2005;Hoffrogge et al, 2006;Kwak et al, 2006). Such regulatory changes can be studied systematically with transgenic technology and microarrays (Pazman et al, 2000;Böttcher et al, 2003;Gurok et al, 2004;Pahnke et al, 2004). The process of differentiation entails changes in types and amount of proteins expressed by the stem cell.…”