Connexin 36 is expressed and associated with zonula occludens-1 protein in PC-12 cells.
Connexin 36 (Cx36) is the predominant connexin isoform expressed in the mammalian neurons of the central nervous system (CNS). PC-12 cells, a neuronal-like cell line, are widely used for neuron functional studies. Many connexins have been shown to interact with zonula occludens-1 protein (ZO-1), a tight junction associated with protein. The present study is intended to investigate whether Cx36 is expressed in PC-12 cells and is associated with ZO-1. Cx36 transcripts were amplified and verified by RT-PCR. 2.9 kb Cx36 mRNA was detected in PC-12 cells through Northern blot hybridization. Western blotting showed a 36-kDa protein band in the homogenates of PC-12 cells. Immunofluorescence labeling revealed that Cx36 was present in cell-cell contacts of PC-12 cells and colocalized with ZO-1. The association of Cx36 and ZO-1 in PC-12 cells was also demonstrated by coimmunoprecipitation. In conclusion, PC-12 cells express Cx36 mRNA and Cx36 proteins that are associated with ZO-1. These results enhanced our understanding of the function of Cx36 in PC-12 cells.
Lu
S J
SJ
Department of Histology and Embryology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Li
H
H
Zhou
F H
FH
Zhang
J J
JJ
Wang
L X
LX
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D017630:Connexins / Q000378:metabolism*; D019085:Fluorescent Antibody Technique, Direct; D047468:Immunoprecipitation; D017403:In Situ Hybridization; D008565:Membrane Proteins / Q000378:metabolism*; D008970:Molecular Weight; D009474:Neurons / Q000737:chemistry / Q000378:metabolism*; D016716:PC12 Cells / Q000166:cytology / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D062826:Zonula Occludens-1 Protein; D000097004:Gap Junction delta-2 Protein
D017630:Connexins; D008565:Membrane Proteins; D010750:Phosphoproteins; D012333:RNA, Messenger; D062826:Zonula Occludens-1 Protein; D000097004:Gap Junction delta-2 Protein; C569044:Tjp1 protein, rat