Activated astrocyte secrete clusterin that enhances neuronal differentiation from rat hippocampal neural stem cells

doi:10.3969/j.issn.0529-1356.2013.06.006

Abstract

Abstract:

Objective To detect the secretion of clusterin in "reactive astrocytes" which are activated by fimbria-fornix transected hippocampal extracts, and to investigate the neuronal differentaition of hippocampal neural stem cells (NSCs) by clusterinin vitro. Methods The astrocytes were cultured in fimbria-fornix transected hippocampal extracts, normal hippocampal extracts and DMEM/F12 medium, the conditional medium contained normal hippocampal extracts and transected 12hours, 24hours, 48hours hippocampal extracts. The concentrations of clusterin was detected by ELISA assay. To investigate the neuronal differentaition of hippocampal neural stem cells (NSCs) by clusterin, the cells were divided into the induced group and control group. Seven days later, the neuronal differentiation of hippocampal NSCs was investigated by MAP-2 immunofluorescence and labeled by Hoechst. Results The concentrations of clusterin in normal group and control group were lower than the transected 12hours, 24hours, 48hours groups, especially the 24hours group which concentration was about 4-folds higher than normal and control group，about (0.1037±0.0119)ng/L,(0.1170±0.0078) ng/L, (0.1940±0.0364) ng/L, (0.4250±0.0724) ng/L and (0.2903±0.0472) ng/L, respectively. The number of MAP-2 positive cells in clusterin induced group was higher than the control group (P<0.05), and the percentages of the total number of cells of these two groups were (11.67± 2.57)% and (4.52±1.54)%, respectively. The processes in the induced group was longer and more than the control group. Conclusion Reactive astrocytes activated by fimbria-fornix transected hippocampal extracts secrete more clusterin. The clusterin in culture cells may promote hippocampal NSCs differentiation into mature neurons in vitro.

Key words: Astrtocyte, Clusterin, Hippocampus, Neural stem cell, Neuron, Immunofluorescence, Rat

ZOU Lin-qing JIN Guo-hua* LI Hao-ming QIN Jian-bing TIAN Mei-ling ZHU Pei-pei WANG Hai-qin. Activated astrocyte secrete clusterin that enhances neuronal differentiation from rat hippocampal neural stem cells[J]. AAS, 2013, 44(6 ): 761-766.

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