Ion channels in a skeletal muscle cell line from a Duchenne muscular dystrophy patient.
A cell line (RCDMD), derived from a muscle biopsy taken from a 7-year-old patient with Duchenne muscular dystrophy (DMD), was established in vitro using conditioned media from the UCHT1 thyroid cell line as described elsewhere (Biochim Biophys Acta 1992; 1134:247-255). Unlike other cell lines established by the same procedure, RCDMD cells were highly refractory to transformation and the resulting cell line grew slowly with a doubling time of approximately 72 h. Further, cells continue to grow after more than 20 doublings and 15 passages. Some of the characteristics of the cell line include lack of reaction with antidystrophin antibodies and the presence of receptors for the dihydropyridine PN200-110 (Kd) = 0.3 +/- 0.05 nmol/L and Bmax = 1.06 +/- 0.03 pmol/mg protein) and for alpha-bungarotoxin (Kd = 1.02 +/- 0.17 nmol/L and Bmax = 4.2 +/- 0.37 pmol/mg protein). Patch clamped cells in the voltage clamp configuration lack ion currents when growing in complete medium with high serum, but they can be induced to differentiate by serum deprivation and addition of hormones and trace elements. After 5 days in differentiating medium, noninactivating, delayed rectifier potassium currents are seen. At day 12, A-type, inactivating potassium currents as well as transient inward currents are seen. In conditions in which sodium and potassium currents are absent, a very fast activating and fast inactivating calcium current was evident. The cell line offers the possibility of studying cellular mechanisms in the pathophysiology of DMD.
Caviedes
R
R
Department of Physiology and Biophysics, Faculty of Medicine, University of Chile, Santiago.
Caviedes
P
P
Liberona
J L
JL
Jaimovich
E
E
D002038:Bungarotoxins / Q000378:metabolism; D015220:Calcium Channels / Q000502:physiology; D020746:Calcium Channels, L-Type; D002455:Cell Division; D002460:Cell Line; D002648:Child; D046508:Culture Techniques / Q000379:methods; D006801:Humans; D007473:Ion Channels / Q000378:metabolism / Q000502:physiology*; D017275:Isradipine / Q000378:metabolism; D007700:Kinetics; D008564:Membrane Potentials; D009124:Muscle Proteins / Q000378:metabolism; D009132:Muscles / Q000378:metabolism* / Q000503:physiopathology; D009136:Muscular Dystrophies / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology*; D011978:Receptors, Nicotinic / Q000378:metabolism; D064569:alpha7 Nicotinic Acetylcholine Receptor
D002038:Bungarotoxins; D015220:Calcium Channels; D020746:Calcium Channels, L-Type; C579167:Chrna7 protein, human; D007473:Ion Channels; D009124:Muscle Proteins; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor; D017275:Isradipine