Live imaging and quantitative analysis of dendritic development of cultured rat hippocampal neurons.
- Author:
Wei NING
1
;
Shu-Jun XU
;
Jian-Hong LUO
Author Information
- Publication Type:Journal Article
- MeSH: Actins; genetics; metabolism; Animals; Animals, Newborn; Cells, Cultured; Dendrites; metabolism; Green Fluorescent Proteins; genetics; metabolism; Hippocampus; cytology; Microscopy, Fluorescence; Neurons; cytology; metabolism; Rats; Rats, Sprague-Dawley; Recombinant Fusion Proteins; genetics; metabolism; Transfection
- From: Journal of Zhejiang University. Medical sciences 2007;36(2):155-160
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo measure mobility of dendritic filopodia, complexity of dendritic arborization using method of live imaging in cultured rat hippocampal neurons and to analyze their morphological characters quantitatively.
METHODSVectors expressing Green Fluorescent Protein- Fibrous Actin (GFP-F-Actin) and F-GFP were co-transfected into cultured rat hippocampal neurons at 5 d in vitro (DIV 5). Neurons expressing GFP were photographed and analyzed with Metamorph software.
RESULTDendritic filopodia was observed to move actively from DIV 7 to DIV 9. The mean density of filopodia was (10.78 +/-3.78)/100 microm, (10.68 +/-2.96)/100 microm and (9.99 +/-3.67)/100 microm (P >0.05), and there were (30.18 +/-14.03)% to (87.36 +/-20.88)% filopodia were mobile (P <0.001). During DIV 7-DIV 14, the total length of dendritic branches grew from (410.74 +/-185.98) microm to (1238.21 +/-418.32)microm (P <0.001) and the number of dendritic branches increased from 18.93 +/-7.23 to 33.60 +/-10.46 (P<0.001). The density of spine was (37.17 +/-6.46)/100 microm at DIV 14.
CONCLUSIONThe combination of live imaging with quantitative analysis is a useful method to study dendritic morphological development in vitro, including indicators of dendritic filopodia, dendritic arborization and spines.
