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Novel Kidins220/ARMS Splice Isoforms: Potential Specific Regulators of Neuronal and Cardiovascular Development.

Schmieg N, Thomas C, Yabe A, Lynch DS, Iglesias T, Chakravarty P, Schiavo G - PLoS ONE (2015)

Bottom Line: The first is located between exon 24 and exon 29, while the second site replaces exon 32 by a short alternative terminal exon 33.Neurotrophin receptor stimulation in cortical and hippocampal neurons and neuroendocrine cells induces specific Kidins220/ARMS splice isoforms and alters the appearance kinetics of the full-length transcript.Overall, this study demonstrates the existence of novel Kidins220/ARMS splice isoforms with unique properties, revealing additional complexity in the functional regulation of neurotrophin receptors, and potentially other signalling pathways involved in neuronal and cardiovascular development.

View Article: PubMed Central - PubMed

Affiliation: Molecular Neuropathobiology Laboratory, Sobell Department of Motor Neuroscience & Movement Disorders, UCL Institute of Neurology, University College London, London WC1N 3BG, United Kingdom; The Francis Crick Institute, 44 Lincoln's Inn Fields, London WC2A 3LY, United Kingdom.

ABSTRACT
Kidins220/ARMS is a transmembrane protein playing a crucial role in neuronal and cardiovascular development. Kidins220/ARMS is a downstream target of neurotrophin receptors and interacts with several signalling and trafficking factors. Through computational modelling, we found two potential sites for alternative splicing of Kidins220/ARMS. The first is located between exon 24 and exon 29, while the second site replaces exon 32 by a short alternative terminal exon 33. Here we describe the conserved occurrence of several Kidins220/ARMS splice isoforms at RNA and protein levels. Kidins220/ARMS splice isoforms display spatio-temporal regulation during development with distinct patterns in different neuronal populations. Neurotrophin receptor stimulation in cortical and hippocampal neurons and neuroendocrine cells induces specific Kidins220/ARMS splice isoforms and alters the appearance kinetics of the full-length transcript. Remarkably, alternative terminal exon splicing generates Kidins220/ARMS variants with distinct cellular localisation: Kidins220/ARMS containing exon 32 is targeted to the plasma membrane and neurite tips, whereas Kidins220/ARMS without exon 33 mainly clusters the full-length protein in a perinuclear intracellular compartment in PC12 cells and primary neurons, leading to a change in neurotrophin receptor expression. Overall, this study demonstrates the existence of novel Kidins220/ARMS splice isoforms with unique properties, revealing additional complexity in the functional regulation of neurotrophin receptors, and potentially other signalling pathways involved in neuronal and cardiovascular development.

No MeSH data available.


Related in: MedlinePlus

Different neuronal populations express specific Kidins220 splice isoforms.(A-F) RNA extracted from cortical (A, D), hippocampal (B, E) and motor neuron (C, F) primary cultures was reverse transcribed into cDNA. N indicates PCR products obtained using primers designed to recognise exons 3 and 8 of Kidins220. 24f-30r indicates samples obtained by amplification with primers recognising exons 24 and 30 (A-C). C indicates PCR products obtained using primers designed to recognise exons 31/32 and 32. 31f-33r indicates samples obtained by amplification with primers recognising exons 31 and 33 (D-F). Note the absence of ATE splicing isoforms C1 and C2 in primary motor neuron cultures.
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pone.0129944.g005: Different neuronal populations express specific Kidins220 splice isoforms.(A-F) RNA extracted from cortical (A, D), hippocampal (B, E) and motor neuron (C, F) primary cultures was reverse transcribed into cDNA. N indicates PCR products obtained using primers designed to recognise exons 3 and 8 of Kidins220. 24f-30r indicates samples obtained by amplification with primers recognising exons 24 and 30 (A-C). C indicates PCR products obtained using primers designed to recognise exons 31/32 and 32. 31f-33r indicates samples obtained by amplification with primers recognising exons 31 and 33 (D-F). Note the absence of ATE splicing isoforms C1 and C2 in primary motor neuron cultures.

Mentions: To address this point, we prepared primary cortical and hippocampal cultures (from E18.5 embryos) and primary motor neurons (from E13.5 embryos), and extracted RNA from these cells at different days in vitro (DIV). Strikingly, we found isoforms m4 and m6 from DIV 1 to DIV 7 in all neuronal populations (Fig 5A–5C). Cortical and hippocampal neurons show similar Kidins220 expression patterns between exons 24 and 30: from DIV 11 onwards, both populations express isoforms m1 and m3 in addition to isoforms m4 and m6. Motor neurons, however, show the highest variety of Kidins220 splice isoforms in the nervous system, with the expression at DIV 13 of isoform m5 in addition to those previously described for cortical and hippocampal neurons (Fig 5C).


Novel Kidins220/ARMS Splice Isoforms: Potential Specific Regulators of Neuronal and Cardiovascular Development.

Schmieg N, Thomas C, Yabe A, Lynch DS, Iglesias T, Chakravarty P, Schiavo G - PLoS ONE (2015)

Different neuronal populations express specific Kidins220 splice isoforms.(A-F) RNA extracted from cortical (A, D), hippocampal (B, E) and motor neuron (C, F) primary cultures was reverse transcribed into cDNA. N indicates PCR products obtained using primers designed to recognise exons 3 and 8 of Kidins220. 24f-30r indicates samples obtained by amplification with primers recognising exons 24 and 30 (A-C). C indicates PCR products obtained using primers designed to recognise exons 31/32 and 32. 31f-33r indicates samples obtained by amplification with primers recognising exons 31 and 33 (D-F). Note the absence of ATE splicing isoforms C1 and C2 in primary motor neuron cultures.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4470590&req=5

pone.0129944.g005: Different neuronal populations express specific Kidins220 splice isoforms.(A-F) RNA extracted from cortical (A, D), hippocampal (B, E) and motor neuron (C, F) primary cultures was reverse transcribed into cDNA. N indicates PCR products obtained using primers designed to recognise exons 3 and 8 of Kidins220. 24f-30r indicates samples obtained by amplification with primers recognising exons 24 and 30 (A-C). C indicates PCR products obtained using primers designed to recognise exons 31/32 and 32. 31f-33r indicates samples obtained by amplification with primers recognising exons 31 and 33 (D-F). Note the absence of ATE splicing isoforms C1 and C2 in primary motor neuron cultures.
Mentions: To address this point, we prepared primary cortical and hippocampal cultures (from E18.5 embryos) and primary motor neurons (from E13.5 embryos), and extracted RNA from these cells at different days in vitro (DIV). Strikingly, we found isoforms m4 and m6 from DIV 1 to DIV 7 in all neuronal populations (Fig 5A–5C). Cortical and hippocampal neurons show similar Kidins220 expression patterns between exons 24 and 30: from DIV 11 onwards, both populations express isoforms m1 and m3 in addition to isoforms m4 and m6. Motor neurons, however, show the highest variety of Kidins220 splice isoforms in the nervous system, with the expression at DIV 13 of isoform m5 in addition to those previously described for cortical and hippocampal neurons (Fig 5C).

Bottom Line: The first is located between exon 24 and exon 29, while the second site replaces exon 32 by a short alternative terminal exon 33.Neurotrophin receptor stimulation in cortical and hippocampal neurons and neuroendocrine cells induces specific Kidins220/ARMS splice isoforms and alters the appearance kinetics of the full-length transcript.Overall, this study demonstrates the existence of novel Kidins220/ARMS splice isoforms with unique properties, revealing additional complexity in the functional regulation of neurotrophin receptors, and potentially other signalling pathways involved in neuronal and cardiovascular development.

View Article: PubMed Central - PubMed

Affiliation: Molecular Neuropathobiology Laboratory, Sobell Department of Motor Neuroscience & Movement Disorders, UCL Institute of Neurology, University College London, London WC1N 3BG, United Kingdom; The Francis Crick Institute, 44 Lincoln's Inn Fields, London WC2A 3LY, United Kingdom.

ABSTRACT
Kidins220/ARMS is a transmembrane protein playing a crucial role in neuronal and cardiovascular development. Kidins220/ARMS is a downstream target of neurotrophin receptors and interacts with several signalling and trafficking factors. Through computational modelling, we found two potential sites for alternative splicing of Kidins220/ARMS. The first is located between exon 24 and exon 29, while the second site replaces exon 32 by a short alternative terminal exon 33. Here we describe the conserved occurrence of several Kidins220/ARMS splice isoforms at RNA and protein levels. Kidins220/ARMS splice isoforms display spatio-temporal regulation during development with distinct patterns in different neuronal populations. Neurotrophin receptor stimulation in cortical and hippocampal neurons and neuroendocrine cells induces specific Kidins220/ARMS splice isoforms and alters the appearance kinetics of the full-length transcript. Remarkably, alternative terminal exon splicing generates Kidins220/ARMS variants with distinct cellular localisation: Kidins220/ARMS containing exon 32 is targeted to the plasma membrane and neurite tips, whereas Kidins220/ARMS without exon 33 mainly clusters the full-length protein in a perinuclear intracellular compartment in PC12 cells and primary neurons, leading to a change in neurotrophin receptor expression. Overall, this study demonstrates the existence of novel Kidins220/ARMS splice isoforms with unique properties, revealing additional complexity in the functional regulation of neurotrophin receptors, and potentially other signalling pathways involved in neuronal and cardiovascular development.

No MeSH data available.


Related in: MedlinePlus