Biosynthesis of titin in cultured skeletal muscle cells.
Population
Synchronized cultures of skeletal muscle cells derived from day 12 chicken embryos
Design
Preclinical
Authors
Discussion
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Implication
Indicates stable titin integration in developing myotubes; leaves open cardiac sarcomere dynamics in vivo.
Key Points
- To examine the rate of synthesis, intracellular stability, and cytoskeletal incorporation of titin during skeletal muscle differentiation.
- Used synchronized primary skeletal muscle cultures isolated from day 12 chicken embryos.
- Tracked protein synthesis and stability via [35S]methionine pulse-labeling and immunoprecipitation with an anti-titin monoclonal antibody.
- Assessed cytoskeletal integration by evaluating titin extractability with Triton X-100 detergent during and after synthesis.
- Titin synthesis increased greater than 4-fold during the first week in culture, while muscle-specific myosin heavy chain synthesis increased greater than 12-fold.
- Newly synthesized titin exhibited high metabolic stability with an estimated half-life of approximately 70 hours.
- Titin remained insoluble and resistant to Triton X-100 extraction during and immediately following synthesis, reflecting rapid cytoskeletal association.
Structured PICO
P
Population
Synchronized cultures of skeletal muscle cells derived from day 12 chicken embryos
I
Intervention
Pulse-labeling with [35S]methionine and immunoprecipitation with an anti-titin mAb
O
Outcome
Biosynthesis rate, half-life, and extraction resistance of titinsurrogate
Newly synthesized titin molecules in developing skeletal muscle cells are stable proteins that rapidly associate with the cytoskeleton.
Cite This Study
Isaacs et al. (1989) studied this question.