Expression of muscle-gene-specific isozymes of phosphorylase and creatine kinase in innervated cultured human muscle (original) (raw)
Related papers
Neuroscience Letters, 1988
Expression of muscle specific isozymes (MSIs) of creatine kinase (CK, EC 2.7.3.2), glycogen phosphorylase (GP, EC 2.4.1.1)~ lactate dehydrogenase (LDH, EC 1.1.1.27) and phosphoglycerate mutase (PGAM, EC 2.7.5.3) was studied both in cultured human muscle fibers which had been innervated (InnCHMFs) for 2# 83 days, and in their non-innervated (non-lnnCHMFs) sister control. In non-InnCHMFs, the MSI of PGAM was never detected, and there was no change in the expression of the MSI of CK during the entire period examined; the expression of MSIs of LDH and GP showed linear increase during the entire period of growth. The expression of MSIs of all 4 enzymes was significantly enhanced in InnCHMFs as compared to non-innervated control. The expression of MSIs of GP and PGAM, and to a lesser degree of LDH increased significantly in correlation with the duration of innervation; the MSI of CK increased linearly only up to 54 days of innervation and plateaued afterward. This study demonstrates: (1) innervation of cultured human muscle fibers by fetal rat spinal cord exerts a time-related maturational influence on their cellular isoenzymatic pattern; (2) to achieve induction and characteristic time-related expression of various MSIs, the requirements for neuronal influences seem to differ. Creatine kinase (CK), glycogen phoshorylase (GP), phosphoglycerate mutase (PGAM) and lactate dehydrogenase (LDH) are present in the adult normal, innervated human skeletal muscle virtually only in the form of their muscle-specific isozyme (MSI) [18]. In the fetus, 'fetal' (in respect to muscle) isozymes of these enzymes predominate, while their MSIs are either weakly or not at all expressed [18]. Fetal
Histochemistry of Cultured, Embryonic and Regenerating Rat Muscle
Journal of Histochemistry & Cytochemistry, 1973
Histochemical features of cultured rat muscle were compared to those of cultured chicken muscle, as well as of young embryonic and regenerating rat muscle. Primary cultures were established from trypsin-dissociated myoblasts of thigh muscle from 17- to 19-day-old rat embryos and of breast muscle from 10- to 12-day-old chicken embryos. Histochemical reactions for myofibrillar adenosine triphosphatase (ATPase) performed at pH 9.4 and for ATPase after acid pH 4.35 preincubation, phosphorylase, succinic dehydrogenase, reduced diphosphopyridine nucleotide dehydrogenase and myoglobin were applied to examine cultured muscle, gastrocnemius muscle of 15-17 day-old embryos and regenerating muscle fibers in cold-injured gastrocnemius muscle of adult rats. Cultured rat and chicken muscles could be best distinguished by the ATPase reaction following acid preincubation (positive in cultured rat muscle, negative in cultured chicken muscle) and by a different pattern of phosphorylase staining. Howe...
Role of innervation on the embryonic development of skeletal muscle
Cell and Tissue Research, 1980
The extent to which the motor innervation regulates the embryonic development of skeletal muscle was investigated by comparing changes in normal, aneural, and paralyzed superior oblique muscle of the duck embryo. The muscle was made aneural by permanently destroying the trochlear motor neurons with electrocautery on day 7 i.e., three days prior to innervation. Embryos were paralyzed by daily application of α-bungarotoxin onto the chorioallantoic membrane from day 10 onwards. The differentiation of myoblasts and myotubes in the aneural muscle was severely affected and did not progress to the myofiber stage. A mass of dead cells in the aneural muscle was replaced by connective tissue. Although the differentiation of myoblasts and myotubes was also retarded in the paralyzed muscle, numerous muscle cells progressed to the myofiber stage. Neuromuscular junctions of normal ultrastructure were seen in all paralyzed muscles. Degeneration of some cells in the paralyzed muscle occurred but there was no evidence of a massive wave of cell death similar to that observed in the aneural muscle. These observations suggest that both the trophic factors from the nerve and the nerve-evoked muscle activity are essential for the execution of the developmental program of the muscle. Trophic factors may play a larger role in differentiation, and maintenance of the muscle than muscle activity.
Activity of creatine kinase in a contracting mammalian muscle of uniform fiber type
Biophysical Journal, 1994
We investigated whether the creatine kinase-catalyzed phosphate exchange between PCr and-yATP in vivo equilibrated with cellular substrates and products as predicted by in vitro kinetic properties of the enzyme, or was a function of ATPase activity as predicted by obligatory "creatine phosphate shuttle" concepts. A transient NMR spin-transfer method was developed, tested, and applied to resting and stimulated ex vivo muscle, the soleus, which is a cellularly homogeneous slowtwitch mammalian muscle, to measure creatine kinase kinetics. The forward and reverse unidirectional CK fluxes were equal, being 1.6 mMs-1 in unstimulated muscle at 220C, and 2.7 mMs-1 at 300C. The CK fluxes did not differ during steady-state stimulation conditions giving a 10-fold range of ATPase rates in which the ATP/PCr ratio increased from approximately 0.3 to 1.6. The observed kinetic behavior of CK activity in the muscle was that expected from the enzyme in vitro in a homogeneous solution only if account was taken of inhibition by an anion-stabilized quaternary dead-end enzyme complex: E-Cr-MgADP-anion. The CK fluxes in soleus were not a function of ATPase activity as predicted by obligatory phosphocreatine shuttle models for cellular energetics.
In vitro development of skeletal muscle fiber
2006
The present review will discuss new insights of myogenesis that begins when embryonic mononucleated progenitor cells become committed to myogenic lineage and subsequently proliferate and fuse to form multinucleated contracting skeletal muscle fibers, responsible for generation of force and movement. This review will focus mostly on the influence of specific myogenic transcription factors on skeletal muscle differentiation and on the compartmentalized expression of nicotinic acetylcholine receptors and acetylcholinesterase at specific muscle fiber microdomains. Experiments using cultured muscle model obtained from embryonic or adult animals, have provided detailed information on myogenesis. Differentiated cultured skeletal muscle fibers contract spontaneously and preserve several properties of in vivo multinucleated muscle fiber, including the expression of specific myogenic transcription factors as well as the compartmentalized synthesis and expression of neuromuscular synaptic proteins around individual nuclei. Besides, cultured muscle cells express multiple receptors coupled to G protein, including muscarinic acetylcholine receptors. Considering that many aspects of the present knowledge about the development and differentiation of muscle fiber and formation of the neuromuscular synapse were established in studies using muscles cultures, protocols of primary tissue-cultured skeletal muscle obtained either from embryonic myoblasts or adult satellite cells will be presented.
European journal of biochemistry / FEBS, 1981
Purified, homodimeric creatine kinases from chicken were subjected to two-dimensional gel analysis under dissociating conditions. Each of the subunits M-creatine kinase and B-creatine kinase was resolved into a basic and an acidic subspecies with very similar mobilities in the sodium dodecylsulfate dimension. The M-creatine kinase subspecies were found in myogenic cells, fast muscle, slow muscle and the B-creatine kinase subspecies were present in heart, gizzard and brain. The creatine kinase subunits were identified in these tissues by a variety of methods like immunoreplicas of two-dimensional gels, immunoprecipitations, or coelectrophoresis with purified creatine kinase and all gave the same results. In the course of myogenic development in vitro the subspecies were synthesized coordinately and no indication was found for a differential regulation of any of the subspecies of the creatine kinase subunits. No radioactive phosphorus was incorporated into either one of the subspecies...