Actin isoform synthesis by cultured cardiac myocytes: effects of actinomycin D and mithramycin
Cultured neonatal rat myocardial cells (CMC) were incubated with the Actinomycin D (ACT) and mithramycin (MIT) in concentrations of 1 x 10/sup -8/M to 1 x 10/sup -5/M in medium containing 35..mu..Ci of /sup 35/S-methionine to determine incorporation into myocardial contractile proteins. After 24h, cells were harvested in buffer with Triton X-100, homogenized and subjected to centrifugation. Protein content of the centrifuged extracts was determined and equivalent amounts of protein were applied to 8-15% gradient sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS PAGE). Similar aliquots were subjected to isoelectric focusing and to gel electrophoresis in the second dimension. Electrophoretic gels were autoradiographed and were quantified densitometrically. A concentration-dependent effect of ACT and MIT on protein and on actin synthesis was found in cultured myocytes with decreased intensity of the radiolabeled actin band in CMC exposed to 1 x 10/sup -8/M to 1 x 10/sup -5/M ACT of 1 x 10/sup -7/M to 1 x 10/sup -5/M MIT. Autoradiography of gels showed focusing of actin isoforms but decreased intensity of iso-actin spots in extracts from CMC exposed to 1 x 10/sup -7/M ACT. ACT and MIT decreased CMC protein and synthesis by 20% at 1 x 10/sup -8/M respectively. The effect of MIT and ACT on CMC actin synthesis and synthesis of other proteins in heart cells is not as specific as the effect of anthracycline antineoplastics on cardiac myocyte proteins and actin.
- Research Organization:
- Univ. of California School of Medicine, Los Angeles
- OSTI ID:
- 5013441
- Report Number(s):
- CONF-8604222-; TRN: 86-036853
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ACTIN
BIOSYNTHESIS
ACTINOMYCIN
BIOLOGICAL EFFECTS
AUTORADIOGRAPHY
CELL CULTURES
DOSE-RESPONSE RELATIONSHIPS
ELECTROPHORESIS
METHIONINE
MYOCARDIUM
RATS
SULFUR 35
AMINO ACIDS
ANIMALS
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
ANTIMITOTIC DRUGS
ANTINEOPLASTIC DRUGS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARBOXYLIC ACIDS
CARDIOVASCULAR SYSTEM
DAYS LIVING RADIOISOTOPES
DRUGS
EVEN-ODD NUCLEI
HEART
ISOTOPES
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
MUSCLES
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
PROTEINS
RADIOISOTOPES
RODENTS
SULFUR ISOTOPES
SYNTHESIS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques