Altered Ca2+ homeostasis and endoplasmic reticulum stress in myotonic dystrophy type 1 muscle cells.
The pathogenesis of Myotonic Dystrophy type 1 (DM1) is linked to unstable CTG repeats in the DMPK gene which induce the mis-splicing to fetal/neonatal isoforms of many transcripts, including those involved in cellular Ca2+ homeostasis. Here we monitored the splicing of three genes encoding for Ca2+ transporters and channels (RyR1, SERCA1 and CACN1S) during maturation of primary DM1 muscle cells in parallel with the functionality of the Excitation-Contraction (EC) coupling machinery. At 15 days of differentiation, fetal isoforms of SERCA1 and CACN1S mRNA were significantly higher in DM1 myotubes compared to controls. Parallel functional studies showed that the cytosolic Ca2+ response to depolarization in DM1 myotubes did not increase during the progression of differentiation, in contrast to control myotubes. While we observed no differences in the size of intracellular Ca2+ stores, DM1 myotubes showed significantly reduced RyR1 protein levels, uncoupling between the segregated ER/SR Ca2+ store and the voltage-induced Ca2+ release machinery, parallel with induction of endoplasmic reticulum (ER) stress markers. In conclusion, our data suggest that perturbed Ca2+ homeostasis, via activation of ER stress, contributes to muscle degeneration in DM1 muscle cells likely representing a premature senescence phenotype.
Department of Genetics, University "Tor Vergata", Roma 00133, Italy. botta@med.uniroma2.it.
Annalisa
A
Botta
Department of Neurosciences SNPSRR, University of Padova, Padova 35100, Italy. adriana.malena@unipd.it.
Adriana
A
Malena
Department of Physiology, Pennsylvania Muscle Institute, University of Pennsylvania, Philadelphia, PA 19104, USA. eloro@mail.med.upenn.edu.
Emanuele
E
Loro
Department of Neurosciences SNPSRR, University of Padova, Padova 35100, Italy. delmoro.giulia@gmail.com.
Giulia
G
Del Moro
Department of Biomedical Sciences, University of Padua and CNR Neuroscience Institute, Padua 35100, Italy. matteo.suman@gmail.com.
Matteo
M
Suman
Department of Neurosciences SNPSRR, University of Padova, Padova 35100, Italy. boris.pantic@yahoo.it.
Boris
B
Pantic
Department of Biomedical Sciences, University of Padua and CNR Neuroscience Institute, Padua 35100, Italy. g.szabadkai@ucl.ac.uk.
Gyorgy
G
Szabadkai
Department of Neurosciences SNPSRR, University of Padova, Padova 35100, Italy. lodovica.vergani@unipd.it.
Lodovica
L
Vergani