Electrogenesis of embryonic chick skeletal muscle cells differentiated <i>in vitro</i>

Kano, Masaakira; Shimada, Yutaka; Ishikawa, Koichi

doi:10.1002/jcp.1040790306

Cited by 39 publications

(19 citation statements)

References 14 publications

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“…A prolonged AP outlasting the stimulating pulse by several seconds has been described in embryonic chick myotubes in culture (18,24) and in situ (17). During development in situ this response appears before the fast AP and disappears after hatching (17 (25).…”

Section: Discussionmentioning

confidence: 96%

“…The expression of these properties is autonomously regulated with no requirement for neuronal intervention. Although a rise in RP during differentiation of myotubes has been reported in culture (13)(14)(15) and in situ (16,17), low synchrony in differentiation produced large variability in electrical responses (17,18) and has hampered determination of the time of appearance and sequence of development of membrane excitability. Fischbach et al (13) The development of contractile activity is indicated by the early appearance of local twitches, which spread through the entire myotube upon maturation.…”

Section: Discussionmentioning

confidence: 99%

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Development of electrophysiological and biochemical membrane properties during differentiation of embryonic skeletal muscle in culture.

Spector¹,

Prives²

1977

Proc. Natl. Acad. Sci. U.S.A.

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Newly fused chick myotubes undergo simultaneous and rapid changes in cell membrane properties during synchronous differentiation in culture. These changes are coordinately regulated and include increases in acetylcholine receptor, acetylcholinesterase, and resting potential, as well as the appearance of action potentials in discrete membrane areas upon stimulation. Subsequently, the acetylcholine receptor reaches maximal levels, whereas the development of electrical properties is marked by a further increase in resting potential, changes in the characteristics of the elicited action potential, and the recruitment of additional membrane areas for action potential generation. Maturation of electrical excitability, marked by the acquisition of the ability to fire repetitively and to conduct action potentials along the membrane, occurs well after resting potential has reached a maximum. During postmaturational development, myotubes exhibit spontaneous electrical and contractile activity, and levels of acetylcholine receptor aecessible to externally applied 125I-labeled a-bungarotoxin decrease markedly. It is suggested that electrophysiological membrane maturation is autonomously regulated with no requirement for neuronal intervention and involves the coordinated biosynthesis of discrete membrane components and their subsequent organization in the myotube membrane.Skeletal muscle cells differentiating in culture are highly suitable for studying the development of the distinctive biochemical and electrophysiological properties of excitable membranes. Fusion of myoblasts in culture to form multinucleated myotubes is accompanied by biosynthesis of contractile proteins, cytoplasmic enzymes, and specialized membrane components and by changes in electrophysiological properties (1-4). Like adult skeletal muscle, mature cultured myotubes are cross-striated, maintain a large resting membrane potential (RP), generate action potentials (APs), and contract rapidly (5). The sequence of development of these characteristics has been obscured by the lack of sufficient synchrony in fusion and myotube maturation in previous electrophysiological studies. Furthermore, while the expression of individual properties has been studied extensively, the sizeable variations in rate and extent of differentiation under different conditions make it difficult to correlate electrophysiological and biochemical aspects of membrane differentiation.In order to determine the developmental sequence of electrophysiological properties and to relate this development to changes in specialized membrane components assayed by biochemical methods in the same cultures, we have used chick embryonic muscle cultures showing rapid and synchronous differentiation kinetics. Our findings show that electrophysiological membrane differentiation is initiated shortly afterThe costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 sole...

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Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

Development of electrophysiological and biochemical membrane properties during differentiation of embryonic skeletal muscle in culture.

Spector¹,

Prives²

1977

Proc. Natl. Acad. Sci. U.S.A.

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“…Earlier work has demonstrated a relationship between the mechanical activity of myotubes and the accumulation of muscle protein (10,(19)(20)(21) . In chick myotubes cultured in the presence of 1 p,M tetrodotoxin (TTX), early development proceeds normally, but by 5-6 d, when normal cultures are exhibiting spontaneous contraction, TTX cultures exhibit little or no mechanical activity and contain only 50% the myosin of control cultures (21).…”

mentioning

confidence: 99%

“…electrical properties of myotube membranes (8,10) . Newly formed myotubes exhibit a resting potential of -15 mV which increases to -60 mV as the myotube matures (8,14).…”

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confidence: 99%

Increased K+ inhibits spontaneous contractions reduces myosin accumulation in cultured chick myotubes.

Bandman¹,

Strohman²

1982

The Journal of Cell Biology

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Increasing the K+ from 5.4 mM to 12 mM in the culture medium of developing chick myotubes causes an immediate cessation of spontaneous contractions and leads to an inhibition of myosin accumulation. The synthesis of myosin continues at the same rate in 12 mM K+ as in 5.4 mM K+ as measured by [3H]leucine incorporation into myosin corrected for differences in pool specific activity. Total protein synthesis and total protein accumulation are unaffected by growth in 12 mM K+. In addition, growth in 12 mM K+ did not alter the type of myosin heavy-chain isoform expression nor did it alter the pattern of myosin light-chain synthesis. However, the rate of myosin turnover increased threefold in cultures grown in 12 mM K+ compared to cultures grown in 5.4 mM K+, while total protein turnover was only marginally increased. We conclude that suppressed electrical or contractile activity of myotubes leads to an increased rate of myofibrillar protein turnover and that spontaneous mechanical and or electrical activity is required for continued myotube maturation in culture.

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“…et a!., 1971; KANO et al, 1972KANO et al, , 1977RITCHIE and FAMBROUGH, 1975;SPECTOR and PRIvEs, 1977) and the culture of an established myogenic cell line (LAND et al, 1973;KIDOKORO, 1973KIDOKORO, , 1975. Of these, the clonal cell line would offer an advantageous system for these studies because of its homogeneous cell population and synchronism of differentiation.…”

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confidence: 99%

Development of excitability during the in vitro differentiation of a newly established myogenic cell line.

Amagai¹,

Iijima²,

Kasai³

1983

Jpn.J.Physiol

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Developmental changes in the membrane electrical properties during the differentiation of a newly established clonal myogenic cell line MC3T3-Al/M13 (M13) derived from newborn mouse calvaria were studied using the conventional intracellular recording method. M13 cells proliferated in vitro with a population doubling time of about 20 hr when they were cultured in a-MEM containing 10% newborn bovine serum at 37°C. After they had achieved confluence and stopped growing, myotube formation by fusion of individual postmitotic mononucleated cells took place within 48 hr, and it advanced until 70-80% of the total number of nuclei were incorporated into such myotubes. Mononucleated M13 cells had a resting membrane potential (Em) of -22.5+1.7 mV (mean+S. D.) and responded passively to current stimuli, indicating that they are nonexcitable. On the contrary, multinucleated myotubes had Ems ranging from -25 to -70 mV, depending on the stage of their development. Newly fused myotubes had relatively less negative Ems and showed no response, whereas myotubes later in development showed delayed rectification against depolarizing current pulses, proving the development of a voltage-sensitive outward current system. Further, mature myotubes had an Em of -58.5+3.2 mV and generated fast action potentials having a maximum rate of rise of 315+11 V/sec and a duration of 3.0+0.5 msec (measured at half height). These action potentials were identified as tetrodotoxin-insensitive Na+ spikes. These results indicate that the membrane excitability of the M13 myogenic cell line develops well after the formation of myotubes, with an increase in Em as maturation proceeds.

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Electrogenesis of embryonic chick skeletal muscle cells differentiated in vitro

Cited by 39 publications

References 14 publications

Development of electrophysiological and biochemical membrane properties during differentiation of embryonic skeletal muscle in culture.

Development of electrophysiological and biochemical membrane properties during differentiation of embryonic skeletal muscle in culture.

Increased K+ inhibits spontaneous contractions reduces myosin accumulation in cultured chick myotubes.

Development of excitability during the in vitro differentiation of a newly established myogenic cell line.

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