Deletion of a specific exon in the voltage-gated calcium channel gene cacophony disrupts locomotion in Drosophila larvae

Kayly M. Lembke, Alexander D. Law, Jasmine Ahrar, David Morton

Research output: Contribution to journalArticle

Abstract

TAR DNA-binding protein 43 (TDP-43) is an RNA-binding protein that regulates transcription, translation and alternative splicing of mRNA. We have shown previously that null mutations of the Drosophila ortholog, Tar DNA-binding homolog (tbph), causes severe locomotion defects in larvae that are mediated by a reduction in the expression of a type II voltage-gated calcium channel, cacophony (cac). We also showed that TDP-43 regulates the inclusion of alternatively spliced exons of cacophony; tbph mutants showed significantly increased expression of cacophony isoforms lacking exon 7, a particularly notable finding as only one out of the 15 predicted isoforms lacks exon 7. To investigate the function of exon 7, we generated Drosophila mutant lines with a deletion that eliminates exon 7. This deletion phenocopies many defects in tbph mutants: a reduction in cacophony protein (Dmca1A) expression, locomotion defects in male and female third instar larvae, disrupted larval motor output, and also reduced activity levels in adult male flies. All these defects were rescued by expression of cacophony transcripts containing exon 7. By contrast, expression of a cacophony cDNA lacking exon 7 resulted in reduced cacophony protein levels and failed to rescue larval locomotion.

Original languageEnglish (US)
Article numberjeb191106
JournalJournal of Experimental Biology
Volume222
Issue number1
DOIs
StatePublished - Jan 1 2019

Fingerprint

calcium channels
locomotion
Calcium Channels
Locomotion
Drosophila
exons
Larva
Exons
calcium
defect
larva
protein
gene
larvae
Genes
genes
DNA
DNA-binding proteins
alternative splicing
DNA-Binding Proteins

Keywords

  • ALS
  • Dmca1A
  • Model organism
  • Motor program
  • Neurodegeneration
  • TDP-43

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Physiology
  • Aquatic Science
  • Animal Science and Zoology
  • Molecular Biology
  • Insect Science

Cite this

Deletion of a specific exon in the voltage-gated calcium channel gene cacophony disrupts locomotion in Drosophila larvae. / Lembke, Kayly M.; Law, Alexander D.; Ahrar, Jasmine; Morton, David.

In: Journal of Experimental Biology, Vol. 222, No. 1, jeb191106, 01.01.2019.

Research output: Contribution to journalArticle

@article{cad2fe600c5e4e8b808746526c92ccca,
title = "Deletion of a specific exon in the voltage-gated calcium channel gene cacophony disrupts locomotion in Drosophila larvae",
abstract = "TAR DNA-binding protein 43 (TDP-43) is an RNA-binding protein that regulates transcription, translation and alternative splicing of mRNA. We have shown previously that null mutations of the Drosophila ortholog, Tar DNA-binding homolog (tbph), causes severe locomotion defects in larvae that are mediated by a reduction in the expression of a type II voltage-gated calcium channel, cacophony (cac). We also showed that TDP-43 regulates the inclusion of alternatively spliced exons of cacophony; tbph mutants showed significantly increased expression of cacophony isoforms lacking exon 7, a particularly notable finding as only one out of the 15 predicted isoforms lacks exon 7. To investigate the function of exon 7, we generated Drosophila mutant lines with a deletion that eliminates exon 7. This deletion phenocopies many defects in tbph mutants: a reduction in cacophony protein (Dmca1A) expression, locomotion defects in male and female third instar larvae, disrupted larval motor output, and also reduced activity levels in adult male flies. All these defects were rescued by expression of cacophony transcripts containing exon 7. By contrast, expression of a cacophony cDNA lacking exon 7 resulted in reduced cacophony protein levels and failed to rescue larval locomotion.",
keywords = "ALS, Dmca1A, Model organism, Motor program, Neurodegeneration, TDP-43",
author = "Lembke, {Kayly M.} and Law, {Alexander D.} and Jasmine Ahrar and David Morton",
year = "2019",
month = "1",
day = "1",
doi = "10.1242/jeb.191106",
language = "English (US)",
volume = "222",
journal = "Journal of Experimental Biology",
issn = "0022-0949",
publisher = "Company of Biologists Ltd",
number = "1",

}

TY - JOUR

T1 - Deletion of a specific exon in the voltage-gated calcium channel gene cacophony disrupts locomotion in Drosophila larvae

AU - Lembke, Kayly M.

AU - Law, Alexander D.

AU - Ahrar, Jasmine

AU - Morton, David

PY - 2019/1/1

Y1 - 2019/1/1

N2 - TAR DNA-binding protein 43 (TDP-43) is an RNA-binding protein that regulates transcription, translation and alternative splicing of mRNA. We have shown previously that null mutations of the Drosophila ortholog, Tar DNA-binding homolog (tbph), causes severe locomotion defects in larvae that are mediated by a reduction in the expression of a type II voltage-gated calcium channel, cacophony (cac). We also showed that TDP-43 regulates the inclusion of alternatively spliced exons of cacophony; tbph mutants showed significantly increased expression of cacophony isoforms lacking exon 7, a particularly notable finding as only one out of the 15 predicted isoforms lacks exon 7. To investigate the function of exon 7, we generated Drosophila mutant lines with a deletion that eliminates exon 7. This deletion phenocopies many defects in tbph mutants: a reduction in cacophony protein (Dmca1A) expression, locomotion defects in male and female third instar larvae, disrupted larval motor output, and also reduced activity levels in adult male flies. All these defects were rescued by expression of cacophony transcripts containing exon 7. By contrast, expression of a cacophony cDNA lacking exon 7 resulted in reduced cacophony protein levels and failed to rescue larval locomotion.

AB - TAR DNA-binding protein 43 (TDP-43) is an RNA-binding protein that regulates transcription, translation and alternative splicing of mRNA. We have shown previously that null mutations of the Drosophila ortholog, Tar DNA-binding homolog (tbph), causes severe locomotion defects in larvae that are mediated by a reduction in the expression of a type II voltage-gated calcium channel, cacophony (cac). We also showed that TDP-43 regulates the inclusion of alternatively spliced exons of cacophony; tbph mutants showed significantly increased expression of cacophony isoforms lacking exon 7, a particularly notable finding as only one out of the 15 predicted isoforms lacks exon 7. To investigate the function of exon 7, we generated Drosophila mutant lines with a deletion that eliminates exon 7. This deletion phenocopies many defects in tbph mutants: a reduction in cacophony protein (Dmca1A) expression, locomotion defects in male and female third instar larvae, disrupted larval motor output, and also reduced activity levels in adult male flies. All these defects were rescued by expression of cacophony transcripts containing exon 7. By contrast, expression of a cacophony cDNA lacking exon 7 resulted in reduced cacophony protein levels and failed to rescue larval locomotion.

KW - ALS

KW - Dmca1A

KW - Model organism

KW - Motor program

KW - Neurodegeneration

KW - TDP-43

UR - http://www.scopus.com/inward/record.url?scp=85059502309&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85059502309&partnerID=8YFLogxK

U2 - 10.1242/jeb.191106

DO - 10.1242/jeb.191106

M3 - Article

VL - 222

JO - Journal of Experimental Biology

JF - Journal of Experimental Biology

SN - 0022-0949

IS - 1

M1 - jeb191106

ER -