[Role of cell adhesion molecules Necl1 in synaptogenesis in primary cultured rat neurons].
OBJECTIVE
To study the role of cell adhesion molecules Necl1 in synaptogenesis in primary cultured neurons.


METHODS
Semi-quantitive reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression pattern of Necl1 in the neuronal differentiation cell model in vitro. Western blot was performed to detect the expression pattern of Necl1 in primary cultured rat neurons and in purified synaptosome. Immunofluoresence was used to detect the synapse formation in primary neurons and in 293 cells co-culture and to detect the density of synapses in primary neuron with ectopic expression of Necl1.


RESULTS
Necl1 expression increased after retinoic acid (RA) induction in SH-SY5Y and P19 cells. The increase of Necl1 expression was consistent with the days of primary neurons culture in vitro, and Necl1 partly localized in synaptosome. The overexpression of Necl1 in 293 cells induced the synapse formation between cocultured 293 cells and neurons. Ectopic expression of Necl1 in primary neurons increased the density of synapses.


CONCLUSIONS
Necl1 plays an important role in neuronal synapse formation.
Chen
Tao
T
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005, China.
Wu
Xu-Dong
XD
Gao
Jing
J
Hao
Wei
W
Yin
Bin
B
Qiang
Bo-Qin
BQ
Yuan
Jian-Gang
JG
Gong
Yan-Hua
YH
Peng
Xiao-Zhong
XZ
D000818:Animals; D015153:Blotting, Western; D015816:Cell Adhesion Molecules, Neuronal / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D006801:Humans; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013569:Synapses / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology*; D013574:Synaptosomes / Q000187:drug effects / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology
D015816:Cell Adhesion Molecules, Neuronal; C516997:nectin-like molecule 1, rat; D014212:Tretinoin