Biochemical and Biophysical Research Communications
Ca2+-dependent splicing of neurexin IIα
Section snippets
Materials and methods
Materials. Minimal essential medium (MEM), Dulbecco’s modification of Earle’s medium (DMEM), heat-inactivated horse serum (IHS), l-glutamine, penicillin–streptomycin, and EZ- First Strand cDNA Synthesis RNA kit were purchased from Biological Industries (Beit Haemek, Israel), B-27 supplement from Invitrogen (Carlsbad, CA), Taq DNA polymerase from Bioline (Luckenwalde, Germany), actinomycin, cycloheximide, 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA),
Results
The effects of depolarization of the cortical neurons on Neurexin 2α splicing at SS#1, SS#2, SS#3, and SS#4 and of Neurexin 2β SS#4 were explored. The cortical neurons expressed neurexin 2α and β isoforms and all expected splice variants were detected. Depolarization significantly affected Neurexin 2α splicing at #SS1 and #SS3 (Supplement 1) and did not significantly affect splicing at #SS2 and #SS4. Total NRXN2α mRNA expression (sum of all transcripts at each splice site) was not significantly
Discussion
The importance of neurexin splice variants in synapse formation and maintenance is only beginning to unravel. Most studies show a role for splicing at SS#4 in the β-neurexin–neuroligin link [20], [21], [22], [23], [24]. The results of the present study show for the first time external stimulus evoked changes in NRXN2α splicing at SS#1 and SS#3 rather than SS# 2 and #4 which may preferentially increase NRXN2α forms lacking exons 2,3,4,11 and their combinations.
One plausible outcome of exon 11
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