Laser capture microdissection and microarray analysis of dividing neural progenitor cells from the adult rat hippocampus.
Neural progenitor cells reside in the hippocampus of adult rodents and humans and generate granule neurons throughout life. Knowledge about the molecular processes regulating these neurogenic cells is fragmentary. In order to identify genes with a role in the proliferation of adult neural progenitor cells, a protocol was elaborated to enable the staining and isolation of such cells under RNA-preserving conditions with a combination of immunohistochemistry and laser capture microdissection. We increased proliferation of neural progenitor cells by electroconvulsive treatment, one of the most effective antidepressant treatments, and isolated Ki-67-positive cells using this new protocol. RNA amplification via in vitro transcription and subsequent microarray analysis revealed over 100 genes that were differentially expressed in neural progenitor cells due to electroconvulsive treatment compared to untreated control animals. Some of these genes have already been implicated in the functioning of neural progenitor cells or have been induced by electroconvulsive treatment; these include brain-derived neurotrophic factor (Bdnf), PDZ-binding kinase (Pbk) and abnormal spindle-like microcephaly-associated (Aspm). In addition, genes were identified for which no role in the proliferation of neurogenic progenitors has been described so far, such as enhancer of zeste homolog 2 (Ezh2).
Gurok
Ulf
U
Neuroscience Discovery Research, Abbott, Knollstrasse, 67061 Ludwigshafen, Germany. ulf.gurok@abbott.com
Loebbert
Ralf W
RW
Meyer
Axel H
AH
Mueller
Reinhold
R
Schoemaker
Hans
H
Gross
Gerhard
G
Behl
Berthold
B
D053687:Adult Stem Cells / Q000502:physiology* / Q000528:radiation effects; D000818:Animals; D002452:Cell Count; D049109:Cell Proliferation / Q000528:radiation effects; D004597:Electroshock / Q000379:methods; D005786:Gene Expression Regulation / Q000528:radiation effects; D006624:Hippocampus / Q000166:cytology*; D019394:Ki-67 Antigen / Q000378:metabolism; D007834:Lasers; D046228:Microarray Analysis / Q000379:methods; D042282:Microdissection / Q000379:methods; D020929:Mitogen-Activated Protein Kinase Kinases; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000502:physiology* / Q000528:radiation effects; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D019394:Ki-67 Antigen; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D017346:Protein Serine-Threonine Kinases; D020929:Mitogen-Activated Protein Kinase Kinases; C409022:PDZ-binding kinase