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BACKGROUND: Gap junction communication has been shown in glial and neuronal cells and it is thought they mediate inter- and intra-cellular communication. Connexin 36 (Cx36) is expressed extensively in the developing brain, with levels peaking at P14 after which its levels fall and its expression becomes entirely neuronal. These and other data have led to the hypothesis that Cx36 may direct neuronal coupling and neurogenesis during development. METHODOLOGY/PRINCIPAL FINDINGS: To investigate Cx36 function we used a neurosphere model of neuronal cell development and developed lentiviral Cx36 knockdown and overexpression strategies. Cx36 knockdown was confirmed by western blotting, immunocytochemistry and functionally by fluorescence recovery after photobleaching (FRAP). We found that knockdown of Cx36 in neurosphere neuronal precursors significantly reduced neuronal coupling and the number of differentiated neurons. Correspondingly, the lentiviral mediated overexpression of Cx36 significantly increased the number of neurons derived from the transduced neurospheres. The number of oligodendrocytes was also significantly increased following transduction with Cx36 indicating they may support neuronal differentiation. CONCLUSIONS/SIGNIFICANCE: Our data suggests that astrocytic and neuronal differentiation during development are governed by mechanisms that include the differential expression of Cx36.

Original publication

DOI

10.1371/journal.pone.0014746

Type

Journal article

Journal

PLoS One

Publication Date

09/03/2011

Volume

6

Keywords

Animals, Cell Aggregation, Cell Differentiation, Cell Lineage, Cell Proliferation, Cells, Cultured, Connexins, Gap Junctions, Gene Expression Regulation, Gene Knockdown Techniques, Genetic Vectors, In Situ Nick-End Labeling, Lentivirus, Neural Stem Cells, Neurons, RNA, Small Interfering, Rats, Reproducibility of Results