Analysis of neuronal gene expression with laser capture microdissection.
The brain is a heterogeneous tissue in which the numbers of neurons, glia, and other cell types vary among anatomic regions. Gene expression studies performed on brain homogenates yield results reflecting mRNA abundance in a mixture of cell types. Therefore, a method for quantifying gene expression in individual cell populations would be useful. Laser capture microdissection (LCM) is a new technique for obtaining pure populations of cells from heterogeneous tissues. Most studies thus far have used LCM to detect DNA sequences. We developed a method to quantify gene expression in hippocampal neurons from mouse brain using LCM and real-time reverse transcriptase-polymerase chain reaction (RT-PCR). This method was optimized to permit histochemical or immunocytochemical visualization of nerve cells during LCM while minimizing RNA degradation. As an example, gene expression was quantified in hippocampal neurons from the Tg2576 mouse model for Alzheimer's disease.
Vincent
Valerie A M
VA
Neuroscience Research Laboratories, Department of Psychiatry and Behavioral Sciences, Stanford University School of Medicine, Stanford, California 94305-5485, USA.
DeVoss
Jason J
JJ
Ryan
Heather S
HS
Murphy
Greer M
GM
D000165:Acridine Orange; D000544:Alzheimer Disease / Q000235:genetics; D016564:Amyloid beta-Protein Precursor / Q000235:genetics; D000818:Animals; D003851:Deoxyribonucleases / Q000378:metabolism; D004210:Dissection / Q000379:methods*; D015870:Gene Expression; D005838:Genotype; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007834:Lasers; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009154:Mutation; D016900:Neurofilament Proteins / Q000235:genetics; D009474:Neurons / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D012044:Regression Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012260:Ribonucleases / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods
D016564:Amyloid beta-Protein Precursor; D016900:Neurofilament Proteins; D012333:RNA, Messenger; C051774:neurofilament protein H; D003851:Deoxyribonucleases; D012260:Ribonucleases; D000165:Acridine Orange