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A Method for Examining the Gene Expression Profile of Developing Cortico-Spinal Projection Neuron
Qu Chunsheng1,2, Zhao Yue’e1,3, Chen Jiaqi2, Gao Ying1*
1Key Laboratory of Visual Science, National Ministry of Health, School of Optometry and Ophthalmology,Wenzhou Medical University, Wenzhou 325027, China;
2Clinical Laboratory of Lishui People’s Hospital,the Sixth Affiliated Hospital of Wenzhou Medical University, Lishui 323000, China;
3Ophthalmology Department of Lishui Central Hospital, the Fifth Affiliated Hospital of Wenzhou Medical University, Lishui 323000, China
2Clinical Laboratory of Lishui People’s Hospital,the Sixth Affiliated Hospital of Wenzhou Medical University, Lishui 323000, China;
3Ophthalmology Department of Lishui Central Hospital, the Fifth Affiliated Hospital of Wenzhou Medical University, Lishui 323000, China
Abstract: The cortico-spinal projection neurons (CSPN) are very important for the brain function.However， an efficient method to study the gene expression profile of developmental CSPN is still lacking. In the present study， mice were injected Red Retrobeads at spinal cross at postnatal day 2 (P2) for retrograde labeling of CSPN and sacrificed at P8 for laser capture micro-dissection (LCMD). Total RNAs of isolated cells were amplified linearly and verified through the marker genes of each layer by qPCR. While layer V marker gene Fezf2 (Fez family zinc finger 2) was increased， layer IV maker gene Rorβ (RAR related orphan receptor β) and layer VI maker gene Foxp2 (forkhead box P2) were decreased in amplified RNAs from LCMD， compared with those from cortical tissue. The RNA products were determined by Affymetrix Mouse GeneChip. GO category analysis found that the most enriched biological processes in the CSPN were the regulations of G-protein coupled receptor signaling pathway and motor behavior. Interestingly， genes for chromatin modification and organization are also enriched in the CSPN. Additionally， we verified the enriched genes from GeneChip by qPCR and found that 7 genes have high level of expression in the CSPN， such as Meg3 (maternally expressed gene 3) etc. Our study provides a method for examining the gene expression profile of developmental CSPN by composing several techniques such as retrograde labeling， LCMD， RNA amplification， microarray screening and qPCR.
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