Differential expression of creatine kinase and phosphoglycerate mutase isozymes during development in aneural and innervated human muscle culture.
Several enzymes that occur in multimolecular forms undergo transitions during myogenesis. Studies of such developmentally regulated isozymes (e.g. creatine kinase) indicate that muscle cells, cultured in the absence of neural tissue never develop fully mature isozyme patterns, but continue to express large amounts of 'housekeeping' isozymes that are characteristically present in fetal muscle. We studied two developmentally controlled isozymes, creatine kinase (CK) and phosphoglycerate mutase (PGAM) in normal human muscle, both aneurally cultured and co-cultured with fetal mouse spinal cord complex. Innervated cultures attain a greater degree of maturity than non-innervated cultures, as revealed by light and electron microscopy, showing well-developed sarcomeres and motor endplates after several weeks in vitro. During early stages of muscle regeneration in co-culture, characteristic fetal isozyme patterns of CK-BB and PGAM-BB activity predominate, as in aneural cultures. The muscle-specific isozymes (CK-MM; PGAM-MM) begin to appear as the muscle differentiates, and after 2-3 months in co-culture only, virtually all enzyme activity is due to the muscle-specific forms of CK and PGAM, as is normally observed in mature skeletal muscle in vivo.
Miranda
A F
AF
Department of Pathology, Columbia University, College of Physicians and Surgeons, New York, NY 10032.
Peterson
E R
ER
Masurovsky
E B
EB
D000818:Animals; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D003402:Creatine Kinase / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D008722:Methods; D051379:Mice; D008854:Microscopy, Electron; D009132:Muscles / Q000166:cytology* / Q000201:enzymology / Q000648:ultrastructure; D009469:Neuromuscular Junction / Q000166:cytology / Q000201:enzymology; D010736:Phosphoglycerate Mutase / Q000378:metabolism*; D010770:Phosphotransferases / Q000378:metabolism*; D013116:Spinal Cord / Q000166:cytology / Q000201:enzymology / Q000648:ultrastructure
D007527:Isoenzymes; D010770:Phosphotransferases; D003402:Creatine Kinase; D010736:Phosphoglycerate Mutase