Formation of triads without the dihydropyridine receptor alpha subunits in cell lines from dysgenic skeletal muscle

doi:10.1083/jcb.134.2.375

This Article

	Full Text (PDF, 5385K)
	Alert me when this article is cited
	Citation Map

Services

	Email this article
	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new content in the JCB
	Download to citation manager

Citing Articles

	Citing Articles via HighWire
	Citing Articles via CrossRef
	Citing Articles via Google Scholar

Google Scholar

	Articles by Powell, J. A.
	Articles by Flucher, B. E.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Powell, J. A.
	Articles by Flucher, B. E.


Social Bookmarking

	What's this?

ARTICLES

Formation of triads without the dihydropyridine receptor alpha subunits in cell lines from dysgenic skeletal muscle

JA Powell, L Petherbridge and BE Flucher
Department of Biological Sciences, Smith College, Northampton, Massachusetts 01063, USA.

Muscular dysgenesis (mdg/mdg), a mutation of the skeletal muscle dihydropyridine receptor (DHPR) alpha 1 subunit, has served as a model to study the functions of the DHPR in excitation-contraction coupling and its role in triad formation. We have investigated the question of whether the lack of the DHPR in dysgenic skeletal muscle results in a failure of triad formation, using cell lines (GLT and NLT) derived from dysgenic (mdg/mdg) and normal (+/+) muscle, respectively. The lines were generated by transfection of myoblasts with a plasmid encoding a Large T antigen. Both cell lines express muscle-specific proteins and begin organization of sarcomeres as demonstrated by immunocytochemistry. Similar to primary cultures, dysgenic (GLT) myoblasts show a higher incidence of cell fusion than their normal counterparts (NLT). NLT myotubes develop spontaneous contractile activity, and fluorescent Ca2+ recordings show Ca2+ release in response to depolarization. In contrast, GLTs show neither spontaneous nor depolarization-induced Ca2+ transients, but do release Ca2+ from the sarcoplasmic reticulum (SR) in response to caffeine. Despite normal transverse tubule (T-tubule) formation, GLT myotubes lack the alpha 1 subunit of the skeletal muscle DHPR, and the alpha 2 subunit is mistargeted. Nevertheless, the ryanodine receptor (RyR) frequently develops its normal, clustered organization in the absence of both DHPR alpha subunits in the T-tubules. In EM, these RyR clusters correspond to T-tubule/SR junctions with regularly spaced feet. These findings provide conclusive evidence that interactions between the DHPR and RyR are not involved in the formation of triad junctions or in the normal organization of the RyR in the junctional SR.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Facebook Add to Reddit Reddit Add to Technorati Technorati Twitter What's this?

This article has been cited by other articles:

Fuller-Bicer, G. A., Varadi, G., Koch, S. E., Ishii, M., Bodi, I., Kadeer, N., Muth, J. N., Mikala, G., Petrashevskaya, N. N., Jordan, M. A., Zhang, S.-P., Qin, N., Flores, C. M., Isaacsohn, I., Varadi, M., Mori, Y., Jones, W. K., Schwartz, A. (2009). Targeted disruption of the voltage-dependent calcium channel {alpha}2/{delta}-1-subunit. Am. J. Physiol. Heart Circ. Physiol. 297: H117-H124 [Abstract] [Full Text]
Jorquera, G., Juretic, N., Jaimovich, E., Riveros, N. (2009). Membrane depolarization induces calcium-dependent upregulation of Hsp70 and Hmox-1 in skeletal muscle cells. Am. J. Physiol. Cell Physiol. 297: C581-C590 [Abstract] [Full Text]
Tuluc, P., Kern, G., Obermair, G. J., Flucher, B. E. (2007). Computer modeling of siRNA knockdown effects indicates an essential role of the Ca2+ channel {alpha}2{delta}-1 subunit in cardiac excitation-contraction coupling. Proc. Natl. Acad. Sci. USA 104: 11091-11096 [Abstract] [Full Text]
Zhou, J., Yi, J., Royer, L., Launikonis, B. S., Gonzalez, A., Garcia, J., Rios, E. (2006). A probable role of dihydropyridine receptors in repression of Ca2+ sparks demonstrated in cultured mammalian muscle. Am. J. Physiol. Cell Physiol. 290: C539-C553 [Abstract] [Full Text]
Cardenas, C., Liberona, J. L., Molgo, J., Colasante, C., Mignery, G. A., Jaimovich, E. (2005). Nuclear inositol 1,4,5-trisphosphate receptors regulate local Ca2+ transients and modulate cAMP response element binding protein phosphorylation. J. Cell Sci. 118: 3131-3140 [Abstract] [Full Text]
Obermair, G. J., Kugler, G., Baumgartner, S., Tuluc, P., Grabner, M., Flucher, B. E. (2005). The Ca2+ Channel {alpha}2{delta}-1 Subunit Determines Ca2+ Current Kinetics in Skeletal Muscle but Not Targeting of {alpha}1S or Excitation-Contraction Coupling. J. Biol. Chem. 280: 2229-2237 [Abstract] [Full Text]
Ursu, D., Schuhmeier, R. P., Melzer, W. (2005). Voltage-controlled Ca2+ release and entry flux in isolated adult muscle fibres of the mouse. J. Physiol. 562: 347-365 [Abstract] [Full Text]
Takekura, H., Paolini, C., Franzini-Armstrong, C., Kugler, G., Grabner, M., Flucher, B. E. (2004). Differential Contribution of Skeletal and Cardiac II-III Loop Sequences to the Assembly of Dihydropyridine-Receptor Arrays in Skeletal Muscle. Mol. Biol. Cell 15: 5408-5419 [Abstract] [Full Text]
Weiss, R. G., O'Connell, K. M. S., Flucher, B. E., Allen, P. D., Grabner, M., Dirksen, R. T. (2004). Functional analysis of the R1086H malignant hyperthermia mutation in the DHPR reveals an unexpected influence of the III-IV loop on skeletal muscle EC coupling. Am. J. Physiol. Cell Physiol. 287: C1094-C1102 [Abstract] [Full Text]
Cardenas, C., Muller, M., Jaimovich, E., Perez, F., Buchuk, D., Quest, A. F. G., Carrasco, M. A. (2004). Depolarization of Skeletal Muscle Cells induces Phosphorylation of cAMP Response Element Binding Protein via Calcium and Protein Kinase C{alpha}. J. Biol. Chem. 279: 39122-39131 [Abstract] [Full Text]
Kugler, G., Weiss, R. G., Flucher, B. E., Grabner, M. (2004). Structural Requirements of the Dihydropyridine Receptor {alpha}1S II-III Loop for Skeletal-type Excitation-Contraction Coupling. J. Biol. Chem. 279: 4721-4728 [Abstract] [Full Text]
Araya, R., Liberona, J. L., Cardenas, J. C., Riveros, N., Estrada, M., Powell, J. A., Carrasco, M. A., Jaimovich, E. (2002). Dihydropyridine Receptors as Voltage Sensors for a Depolarization-evoked, IP3R-mediated, Slow Calcium Signal in Skeletal Muscle Cells. JGP 121: 3-16 [Abstract] [Full Text]
Flucher, B. E., Weiss, R. G., Grabner, M. (2002). Cooperation of two-domain Ca2+ channel fragments in triad targeting and restoration of excitation- contraction coupling in skeletal muscle. Proc. Natl. Acad. Sci. USA 99: 10167-10172 [Abstract] [Full Text]
Powell, J. A., Carrasco, M. A., Adams, D. S., Drouet, B., Rios, J., Muller, M., Estrada, M., Jaimovich, E. (2002). IP3 receptor function and localization in myotubes: an unexplored Ca2+ signaling pathway in skeletal muscle. J. Cell Sci. 114: 3673-3683 [Abstract] [Full Text]
Flucher, B. E., Kasielke, N., Grabner, M. (2000). The Triad Targeting Signal of the Skeletal Muscle Calcium Channel Is Localized in the Cooh Terminus of the {alpha}1S Subunit. JCB 151: 467-478 [Abstract] [Full Text]
Jaimovich, E., Reyes, R., Liberona, J. L., Powell, J. A. (2000). IP3 receptors, IP3 transients, and nucleus-associated Ca2+ signals in cultured skeletal muscle. Am. J. Physiol. Cell Physiol. 278: C998-C1010 [Abstract] [Full Text]
Menard, C., Pupier, S., Mornet, D., Kitzmann, M., Nargeot, J., Lory, P. (1999). Modulation of L-type Calcium Channel Expression during Retinoic Acid-induced Differentiation of H9C2 Cardiac Cells. J. Biol. Chem. 274: 29063-29070 [Abstract] [Full Text]
Flucher, B. E., Conti, A., Takeshima, H., Sorrentino, V. (1999). Type 3 and Type 1 Ryanodine Receptors Are Localized in Triads of the Same Mammalian Skeletal Muscle Fibers. JCB 146: 621-630 [Abstract] [Full Text]
Neuhuber, B., Gerster, U., Doring, F., Glossmann, H., Tanabe, T., Flucher, B. E. (1998). Association of calcium channel alpha 1S and beta 1a subunits is required for the targeting of beta 1a but not of alpha 1S into skeletal muscle triads. Proc. Natl. Acad. Sci. USA 95: 5015-5020 [Abstract] [Full Text]
Protasi, F., Franzini-Armstrong, C., Allen, P. D. (1998). Role of Ryanodine Receptors in the Assembly of Calcium Release Units in Skeletal Muscle. JCB 140: 831-842 [Abstract] [Full Text]
Protasi, F., Franzini-Armstrong, C., Flucher, B. E. (1997). Coordinated Incorporation of Skeletal Muscle Dihydropyridine Receptors and Ryanodine Receptors in Peripheral Couplings of BC3H1 Cells. JCB 137: 859-870 [Abstract] [Full Text]
Gregg, R.�G., Messing, A., Strube, C., Beurg, M., Moss, R., Behan, M., Sukhareva, M., Haynes, S., Powell, J.�A., Coronado, R., Powers, P.�A. (1996). Absence of the beta �subunit (cchb1) of the skeletal muscle dihydropyridine receptor alters expression of the alpha 1 subunit and eliminates excitation-contraction�coupling. Proc. Natl. Acad. Sci. USA 93: 13961-13966 [Abstract] [Full Text]
Proenza, C., Wilkens, C., Lorenzon, N. M., Beam, K. G. (2000). A Carboxyl-terminal Region Important for the Expression and Targeting of the Skeletal Muscle Dihydropyridine Receptor. J. Biol. Chem. 275: 23169-23174 [Abstract] [Full Text]
Wilkens, C. M., Kasielke, N., Flucher, B. E., Beam, K. G., Grabner, M. (2001). Excitation-contraction coupling is unaffected by drastic alteration of the sequence surrounding residues L720-L764 of the alpha 1S II-III loop. Proc. Natl. Acad. Sci. USA 98: 5892-5897 [Abstract] [Full Text]