Ammodytin L (AMDL) is a myotoxic phospholipase-like protein from the venom of Vipera ammodytes with a serine in position 49 instead of an aspartate, therefore this toxin is devoid of phospholipase activity, and the membrane-damaging effect does not involve any step of phospholipase activity. The aim of the present study was to analyze the effect of AMDL on L-6 cells from rat skeletal muscle to investigate its mechanism of action and the role of calcium ions in its muscle-damaging activity. Our data indicate that the effect of ammodytin L is strongly dependent on the degree of cell differentiation. Low doses of myotoxin gave rise to a marked release of creatine kinase in myotubes differentiated from L-6 myoblasts and the presence of calcium ions plays a role in the cytotoxic effect. The presence of EGTA in the incubation buffer reduced by 50% the release of creatine kinase. No membrane damage was observed in myoblasts, but there was a significant increase of intracellular calcium concentration measured with Fura-2. A non-specific membrane effect of AMDL was ruled out using platelets as reference cells: no platelet aggregation pattern and no increase in intracellular calcium were observed.

Incerpi, S., DE VITO, P., Luly, P., Rufini, S. (1995). Effect of ammodytin L from Vipera ammodytes on L-6 cells from rat skeletal muscle. BIOCHIMICA ET BIOPHYSICA ACTA, 1268(2), 137-142 [10.1016/0167-4889(95)00053-U].

Effect of ammodytin L from Vipera ammodytes on L-6 cells from rat skeletal muscle

INCERPI, SANDRA;DE VITO, PAOLO;LULY, PAOLO;RUFINI, STEFANO
1995-08-31

Abstract

Ammodytin L (AMDL) is a myotoxic phospholipase-like protein from the venom of Vipera ammodytes with a serine in position 49 instead of an aspartate, therefore this toxin is devoid of phospholipase activity, and the membrane-damaging effect does not involve any step of phospholipase activity. The aim of the present study was to analyze the effect of AMDL on L-6 cells from rat skeletal muscle to investigate its mechanism of action and the role of calcium ions in its muscle-damaging activity. Our data indicate that the effect of ammodytin L is strongly dependent on the degree of cell differentiation. Low doses of myotoxin gave rise to a marked release of creatine kinase in myotubes differentiated from L-6 myoblasts and the presence of calcium ions plays a role in the cytotoxic effect. The presence of EGTA in the incubation buffer reduced by 50% the release of creatine kinase. No membrane damage was observed in myoblasts, but there was a significant increase of intracellular calcium concentration measured with Fura-2. A non-specific membrane effect of AMDL was ruled out using platelets as reference cells: no platelet aggregation pattern and no increase in intracellular calcium were observed.
31-ago-1995
Pubblicato
Rilevanza internazionale
Articolo
Esperti anonimi
Settore BIO/09 - FISIOLOGIA
English
Con Impact Factor ISI
Animals; Calcium; Cell Differentiation; Cell Line; Creatine Kinase; Dose-Response Relationship, Drug; Egtazic Acid; Muscles; Rats; Time Factors; Viper Venoms
Incerpi, S., DE VITO, P., Luly, P., Rufini, S. (1995). Effect of ammodytin L from Vipera ammodytes on L-6 cells from rat skeletal muscle. BIOCHIMICA ET BIOPHYSICA ACTA, 1268(2), 137-142 [10.1016/0167-4889(95)00053-U].
Incerpi, S; DE VITO, P; Luly, P; Rufini, S
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/182447
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