1 research outputs found
Regulation of Shootin1 Gene Expression Involves NGF-induced Alternative Splicing during Neuronal Differentiation of PC12 Cells
Shootin1 is a protein involved in neuronal polarization, and has been
shown to be a key molecule for the positive/negative feedback loop for
axon induction required during neuronal symmetry breaking. To better
understand the molecular basis of shootin1 dynamics, we analysed the
regulatory pathways and the expressional status of shootin1 gene during
NGF-induced neuronal differentiation. We demonstrated that the isoform-1
and isoform-2 of shootin1 is differentially expressed during neuronal
differentiation. By blocking individual downstream pathways of NGF
signalling, we found that PI3K/Akt pathway plays a major role in the
expression of shootin1 isoform-2. Western blot and RT-PCR results showed
that the isoform-1 of shootin1 is constitutively expressed, while the
isoform-2 is expressed in a manner that is strictly dependent on
NGF-stimulation. Isoform-specific RT-PCR results demonstrated that the
differential expression of the isoform-1 and isoform-2 of shootin1 is a
consequence of alternative splicing of shootin1 pre-mRNA, in response to
NGF-signalling. Collectively these findings provide the first
information on the molecular mechanisms regulating the expression of
shootin1 gene and represent the first example of NGF-induced alternative
splicing process that has a regulatory role in neuritogenesis