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NOVA2-mediated RNA regulation is required for axonal pathfinding during development.

Saito Y, Miranda-Rottmann S, Ruggiu M, Park CY, Fak JJ, Zhong R, Duncan JS, Fabella BA, Junge HJ, Chen Z, Araya R, Fritzsch B, Hudspeth AJ, Darnell RB - Elife (2016)

Bottom Line: The neuron specific RNA-binding proteins NOVA1 and NOVA2 are highly homologous alternative splicing regulators.NOVA proteins regulate at least 700 alternative splicing events in vivo, yet relatively little is known about the biologic consequences of NOVA action and in particular about functional differences between NOVA1 and NOVA2.Thus we have discovered that NOVA2 uniquely regulates alternative splicing of a coordinate set of transcripts encoding key components in cortical, brainstem and spinal axon guidance/outgrowth pathways during neural differentiation, with severe functional consequences in vivo.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Neuro-Oncology, Howard Hughes Medical Institute, The Rockefeller University, New York, United States.

ABSTRACT
The neuron specific RNA-binding proteins NOVA1 and NOVA2 are highly homologous alternative splicing regulators. NOVA proteins regulate at least 700 alternative splicing events in vivo, yet relatively little is known about the biologic consequences of NOVA action and in particular about functional differences between NOVA1 and NOVA2. Transcriptome-wide searches for isoform-specific functions, using NOVA1 and NOVA2 specific HITS-CLIP and RNA-seq data from mouse cortex lacking either NOVA isoform, reveals that NOVA2 uniquely regulates alternative splicing events of a series of axon guidance related genes during cortical development. Corresponding axonal pathfinding defects were specific to NOVA2 deficiency: Nova2-/- but not Nova1-/- mice had agenesis of the corpus callosum, and axonal outgrowth defects specific to ventral motoneuron axons and efferent innervation of the cochlea. Thus we have discovered that NOVA2 uniquely regulates alternative splicing of a coordinate set of transcripts encoding key components in cortical, brainstem and spinal axon guidance/outgrowth pathways during neural differentiation, with severe functional consequences in vivo.

No MeSH data available.


Related in: MedlinePlus

DOI:http://dx.doi.org/10.7554/eLife.14371.031
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fig12: DOI:http://dx.doi.org/10.7554/eLife.14371.031

Mentions: We appreciate the reviewers’ suggestion, and have tried this challenging experiment. Since it has not been reported that Neo1 KO mice have ACC phenotype, we tried to rescue the ACC phenotype with the DCC-‐long isoform that is decreased in the cortex of Nova2-‐KO mice. We undertook in uteroelectroporation of plasmids expressing the DCC-‐long isoform. Plasmid expressing both the DCC long isoform and DsRed as a positive control were electroporated into the cingulate cortex at E14.5, the mice were sacrificed at E17.5, and subjected to immunostaining with L1 and DsRed (Author response image 1). The expression of DCC-‐long isoform in cingulate cortex at E14.5 was not sufficient to form the commissure corpus callosum, indicating that the DCC-‐long isoform was not sufficient to rescue the ACC phenotype, and suggesting the possibility that the ACC phenotype in Nova2-KO mice either does not involve the DCC-‐long isoform or is caused by the dysregulation of multiple transcriptomes. We mention this negative result in the revised Discussion part.10.7554/eLife.14371.031Author Response Image 1.


NOVA2-mediated RNA regulation is required for axonal pathfinding during development.

Saito Y, Miranda-Rottmann S, Ruggiu M, Park CY, Fak JJ, Zhong R, Duncan JS, Fabella BA, Junge HJ, Chen Z, Araya R, Fritzsch B, Hudspeth AJ, Darnell RB - Elife (2016)

DOI:http://dx.doi.org/10.7554/eLife.14371.031
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4930328&req=5

fig12: DOI:http://dx.doi.org/10.7554/eLife.14371.031
Mentions: We appreciate the reviewers’ suggestion, and have tried this challenging experiment. Since it has not been reported that Neo1 KO mice have ACC phenotype, we tried to rescue the ACC phenotype with the DCC-‐long isoform that is decreased in the cortex of Nova2-‐KO mice. We undertook in uteroelectroporation of plasmids expressing the DCC-‐long isoform. Plasmid expressing both the DCC long isoform and DsRed as a positive control were electroporated into the cingulate cortex at E14.5, the mice were sacrificed at E17.5, and subjected to immunostaining with L1 and DsRed (Author response image 1). The expression of DCC-‐long isoform in cingulate cortex at E14.5 was not sufficient to form the commissure corpus callosum, indicating that the DCC-‐long isoform was not sufficient to rescue the ACC phenotype, and suggesting the possibility that the ACC phenotype in Nova2-KO mice either does not involve the DCC-‐long isoform or is caused by the dysregulation of multiple transcriptomes. We mention this negative result in the revised Discussion part.10.7554/eLife.14371.031Author Response Image 1.

Bottom Line: The neuron specific RNA-binding proteins NOVA1 and NOVA2 are highly homologous alternative splicing regulators.NOVA proteins regulate at least 700 alternative splicing events in vivo, yet relatively little is known about the biologic consequences of NOVA action and in particular about functional differences between NOVA1 and NOVA2.Thus we have discovered that NOVA2 uniquely regulates alternative splicing of a coordinate set of transcripts encoding key components in cortical, brainstem and spinal axon guidance/outgrowth pathways during neural differentiation, with severe functional consequences in vivo.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Neuro-Oncology, Howard Hughes Medical Institute, The Rockefeller University, New York, United States.

ABSTRACT
The neuron specific RNA-binding proteins NOVA1 and NOVA2 are highly homologous alternative splicing regulators. NOVA proteins regulate at least 700 alternative splicing events in vivo, yet relatively little is known about the biologic consequences of NOVA action and in particular about functional differences between NOVA1 and NOVA2. Transcriptome-wide searches for isoform-specific functions, using NOVA1 and NOVA2 specific HITS-CLIP and RNA-seq data from mouse cortex lacking either NOVA isoform, reveals that NOVA2 uniquely regulates alternative splicing events of a series of axon guidance related genes during cortical development. Corresponding axonal pathfinding defects were specific to NOVA2 deficiency: Nova2-/- but not Nova1-/- mice had agenesis of the corpus callosum, and axonal outgrowth defects specific to ventral motoneuron axons and efferent innervation of the cochlea. Thus we have discovered that NOVA2 uniquely regulates alternative splicing of a coordinate set of transcripts encoding key components in cortical, brainstem and spinal axon guidance/outgrowth pathways during neural differentiation, with severe functional consequences in vivo.

No MeSH data available.


Related in: MedlinePlus