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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: Through computational modelling, we found two potential sites for alternative splicing of Kidins220/ARMS.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.

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.


BDNF accelerates the appearance of full-length Kidins220 in cortical and hippocampal primary neurons.(A-B) Primary cortical (A) and hippocampal (B) neurons were prepared from E18.5 mouse embryos and half of each culture was plated with medium containing 100 ng/ml BDNF. RNA was extracted at different time points and reverse transcribed. N indicates PCR products obtained using primers designed to recognise exons 3 and 8. 24f-30r indicates samples obtained by amplification with primers recognising exons 24 and 30. (A) Arrowheads point to early appearance of Kidins220 isoform m1 (full-length) in BDNF treated cultures compared to non-treated cultures.
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pone.0129944.g006: BDNF accelerates the appearance of full-length Kidins220 in cortical and hippocampal primary neurons.(A-B) Primary cortical (A) and hippocampal (B) neurons were prepared from E18.5 mouse embryos and half of each culture was plated with medium containing 100 ng/ml BDNF. RNA was extracted at different time points and reverse transcribed. N indicates PCR products obtained using primers designed to recognise exons 3 and 8. 24f-30r indicates samples obtained by amplification with primers recognising exons 24 and 30. (A) Arrowheads point to early appearance of Kidins220 isoform m1 (full-length) in BDNF treated cultures compared to non-treated cultures.

Mentions: We plated primary cortical and hippocampal cultures with or without BDNF (100 ng/ml) and collected their RNA at different time points (Fig 6). In parallel, we plated PC12 cells and stimulated them with NGF (100 ng/ml) for 72 h (S5 Fig). Samples were reverse transcribed and amplified with primers recognising exons 24 and 30 and exons 31 and 33. BDNF accelerated the appearance of the m1 isoform in both primary cortical and hippocampal cultures compared to non-stimulated samples. In cortical neurons, isoform m1 appeared at DIV 7 in stimulated samples (Fig 6A, arrowhead), whilst only at DIV 11 in absence of BDNF. In hippocampal cultures, BDNF induced the robust expression of isoform 1 at DIV 11 (Fig 6B, arrowhead), whereas a band corresponding to this Kidins220 variant only appeared in significant amounts at DIV 14 in non-stimulated samples.


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)

BDNF accelerates the appearance of full-length Kidins220 in cortical and hippocampal primary neurons.(A-B) Primary cortical (A) and hippocampal (B) neurons were prepared from E18.5 mouse embryos and half of each culture was plated with medium containing 100 ng/ml BDNF. RNA was extracted at different time points and reverse transcribed. N indicates PCR products obtained using primers designed to recognise exons 3 and 8. 24f-30r indicates samples obtained by amplification with primers recognising exons 24 and 30. (A) Arrowheads point to early appearance of Kidins220 isoform m1 (full-length) in BDNF treated cultures compared to non-treated cultures.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0129944.g006: BDNF accelerates the appearance of full-length Kidins220 in cortical and hippocampal primary neurons.(A-B) Primary cortical (A) and hippocampal (B) neurons were prepared from E18.5 mouse embryos and half of each culture was plated with medium containing 100 ng/ml BDNF. RNA was extracted at different time points and reverse transcribed. N indicates PCR products obtained using primers designed to recognise exons 3 and 8. 24f-30r indicates samples obtained by amplification with primers recognising exons 24 and 30. (A) Arrowheads point to early appearance of Kidins220 isoform m1 (full-length) in BDNF treated cultures compared to non-treated cultures.
Mentions: We plated primary cortical and hippocampal cultures with or without BDNF (100 ng/ml) and collected their RNA at different time points (Fig 6). In parallel, we plated PC12 cells and stimulated them with NGF (100 ng/ml) for 72 h (S5 Fig). Samples were reverse transcribed and amplified with primers recognising exons 24 and 30 and exons 31 and 33. BDNF accelerated the appearance of the m1 isoform in both primary cortical and hippocampal cultures compared to non-stimulated samples. In cortical neurons, isoform m1 appeared at DIV 7 in stimulated samples (Fig 6A, arrowhead), whilst only at DIV 11 in absence of BDNF. In hippocampal cultures, BDNF induced the robust expression of isoform 1 at DIV 11 (Fig 6B, arrowhead), whereas a band corresponding to this Kidins220 variant only appeared in significant amounts at DIV 14 in non-stimulated samples.

Bottom Line: Through computational modelling, we found two potential sites for alternative splicing of Kidins220/ARMS.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.

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.