Limits...
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.


Expression of alternative splice isoforms of Kidins220 in adult mouse and human tissues.(A-B) RT-PCR analyses for exons encoding the amino-terminus of Kidins220 and between exons 24 and 30 (24f-30r) were carried out on adult mouse (A) and human (B) tissue panels. Tissues are labelled by capital letters. N indicates PCR products obtained using primers designed to recognise exons 3 and 8 in mouse, and exon 9 and 13 in human. 24f-30r indicates samples obtained by amplification with primers recognising exons 24 and 30. Arrowheads point to samples in which a specific alternative splicing pattern was detected. (C-D) Schematics of Kidins220 splice isoforms identified in mouse heart and brain (C) and human brain (D) in the region encoded by exons 24 to 30.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4470590&req=5

pone.0129944.g001: Expression of alternative splice isoforms of Kidins220 in adult mouse and human tissues.(A-B) RT-PCR analyses for exons encoding the amino-terminus of Kidins220 and between exons 24 and 30 (24f-30r) were carried out on adult mouse (A) and human (B) tissue panels. Tissues are labelled by capital letters. N indicates PCR products obtained using primers designed to recognise exons 3 and 8 in mouse, and exon 9 and 13 in human. 24f-30r indicates samples obtained by amplification with primers recognising exons 24 and 30. Arrowheads point to samples in which a specific alternative splicing pattern was detected. (C-D) Schematics of Kidins220 splice isoforms identified in mouse heart and brain (C) and human brain (D) in the region encoded by exons 24 to 30.

Mentions: To verify the expression of alternative splice isoforms of Kidins220 between exon 24 and 29, we sequenced Kidins220 in mouse (Fig 1A) and human (Fig 1B) RNA tissue panels. We extracted RNA from wild type adult mouse tissues and compared them to a commercial RNA panel from a pool of healthy adult human tissues. The RNA from both panels was reverse-transcribed, amplified using specific Kidins220 primers (see Materials and Methods) and analysed by standard gel electrophoresis (Fig 1A and 1B). As a control (N) of the quality of our cDNA and Kidins220 expression, we designed primers specific for invariant regions of Kidins220, such as exons 3 and 8 in the case of mouse samples (Fig 1A) and for exons 9 and 13 for human samples (Fig 1B). To investigate potential alternative splicing patterns, we selected primers specific for exons 24 and 30 for both panels. As shown in Fig 1, we observed a mouse-specific and a human-specific “default pattern”, which is identical in several tissues. In the mouse tissue panel, we obtained three different bands ranging between 400 and 700 bp (samples B, E-F, H-K, M-Q and S; Fig 1A), whereas the human “default pattern” comprises only a single band migrating between 500 and 600 bp (samples A, B, D-F, I-O and P-U; Fig 1B). However, this pattern differed in brain, heart and skeletal muscle (indicated by arrowheads), both in mouse and human tissues. Additionally, a different splice pattern was detected in mouse testis (Fig 1A, arrowhead).


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)

Expression of alternative splice isoforms of Kidins220 in adult mouse and human tissues.(A-B) RT-PCR analyses for exons encoding the amino-terminus of Kidins220 and between exons 24 and 30 (24f-30r) were carried out on adult mouse (A) and human (B) tissue panels. Tissues are labelled by capital letters. N indicates PCR products obtained using primers designed to recognise exons 3 and 8 in mouse, and exon 9 and 13 in human. 24f-30r indicates samples obtained by amplification with primers recognising exons 24 and 30. Arrowheads point to samples in which a specific alternative splicing pattern was detected. (C-D) Schematics of Kidins220 splice isoforms identified in mouse heart and brain (C) and human brain (D) in the region encoded by exons 24 to 30.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0129944.g001: Expression of alternative splice isoforms of Kidins220 in adult mouse and human tissues.(A-B) RT-PCR analyses for exons encoding the amino-terminus of Kidins220 and between exons 24 and 30 (24f-30r) were carried out on adult mouse (A) and human (B) tissue panels. Tissues are labelled by capital letters. N indicates PCR products obtained using primers designed to recognise exons 3 and 8 in mouse, and exon 9 and 13 in human. 24f-30r indicates samples obtained by amplification with primers recognising exons 24 and 30. Arrowheads point to samples in which a specific alternative splicing pattern was detected. (C-D) Schematics of Kidins220 splice isoforms identified in mouse heart and brain (C) and human brain (D) in the region encoded by exons 24 to 30.
Mentions: To verify the expression of alternative splice isoforms of Kidins220 between exon 24 and 29, we sequenced Kidins220 in mouse (Fig 1A) and human (Fig 1B) RNA tissue panels. We extracted RNA from wild type adult mouse tissues and compared them to a commercial RNA panel from a pool of healthy adult human tissues. The RNA from both panels was reverse-transcribed, amplified using specific Kidins220 primers (see Materials and Methods) and analysed by standard gel electrophoresis (Fig 1A and 1B). As a control (N) of the quality of our cDNA and Kidins220 expression, we designed primers specific for invariant regions of Kidins220, such as exons 3 and 8 in the case of mouse samples (Fig 1A) and for exons 9 and 13 for human samples (Fig 1B). To investigate potential alternative splicing patterns, we selected primers specific for exons 24 and 30 for both panels. As shown in Fig 1, we observed a mouse-specific and a human-specific “default pattern”, which is identical in several tissues. In the mouse tissue panel, we obtained three different bands ranging between 400 and 700 bp (samples B, E-F, H-K, M-Q and S; Fig 1A), whereas the human “default pattern” comprises only a single band migrating between 500 and 600 bp (samples A, B, D-F, I-O and P-U; Fig 1B). However, this pattern differed in brain, heart and skeletal muscle (indicated by arrowheads), both in mouse and human tissues. Additionally, a different splice pattern was detected in mouse testis (Fig 1A, arrowhead).

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.