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Creatine kinase transcript accumulation: effect of nerve during muscle development
C H Washabaugh1, M P Ontell, J A Kant
1Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA.
Abstract:
To determine the role of the nerve in regulating the accumulation of cytoplasmic creatine kinase (CK) mRNAs in hindleg muscles of the developing mouse, the lumbosacral spinal cords of 14-day gestation mice (E14) were laser ablated, and the accumulation of muscle CK (MCK) and brain CK (BCK) mRNAs was evaluated just prior to birth with in situ hybridization. Numbers of molecules of each of these transcripts/ng total RNA in the soleus and extensor digitorum longus (EDL) muscles were determined with competitive PCR and compared to transcripts found in innervated crural muscles. Data suggest that: 1) the level of BCK mRNA accumulation in innervated hindlimb muscles peaks at E16.5 and remains at fetal levels until the second month postnatal, when it falls to the level found in the adult. Given that MCK transcripts meet or exceed adult levels by day 28 postnatal, the "down-regulation" of the BCK gene and the "up-regulation" of the MCK gene are not tightly coupled; 2) the developmental switch from BCK to MCK, as the dominant cytoplasmic CK mRNA, occurs in innervated and aneural leg muscles between E14 and E16.5, indicating this switch is not nerve dependent; 3) the absence of innervation has no effect on BCK mRNA accumulation. MCK transcripts/ng total RNA continue to increase in aneural muscle throughout the late fetal period, but from E16.5-E19.5 the MCK transcript levels in aneural muscles become progressively lower than in age-matched innervated muscles. Thus, the accumulation of the muscle specific cytoplasmic CK, but not BCK, transcripts is affected by the absence of innervation during the fetal period. Dev Dyn 1999;215:285-296.
Insights
Nerve absence affects muscle-specific creatine kinase (CK) mRNA accumulation in developing mouse hindlegs. While the developmental switch to muscle CK (MCK) mRNA is nerve-independent, MCK mRNA levels decrease without innervation.
Area of Science:
- Developmental Biology
- Neuroscience
- Molecular Biology
Background:
- Cytoplasmic creatine kinase (CK) plays a crucial role in cellular energy homeostasis.
- The differential expression of CK isoforms, specifically muscle CK (MCK) and brain CK (BCK), is critical during muscle development.
- The influence of neural regulation on the accumulation of CK mRNAs during fetal development is not fully understood.
Purpose of the Study:
- To investigate the role of nerve supply in regulating the accumulation of cytoplasmic creatine kinase (CK) mRNAs in developing mouse hindleg muscles.
- To determine if the developmental switch from BCK to MCK mRNA is nerve-dependent.
- To assess the impact of denervation on MCK and BCK mRNA levels during fetal development.
Main Methods:
- Laser ablation of the lumbosacral spinal cord in embryonic day 14 (E14) mouse fetuses to create aneural hindleg muscles.
- In situ hybridization to evaluate the accumulation of MCK and BCK mRNAs in hindleg muscles prior to birth.
- Competitive PCR to quantify the number of MCK and BCK transcripts per nanogram of total RNA in soleus and extensor digitorum longus (EDL) muscles.
Main Results:
- The developmental switch from BCK to MCK mRNA dominance occurs between E14 and E16.5 in both innervated and aneural leg muscles, indicating it is nerve-independent.
- Absence of innervation does not affect BCK mRNA accumulation.
- While MCK mRNA levels increase in aneural muscles, they are progressively lower than in innervated muscles from E16.5 to E19.5, suggesting innervation positively influences MCK mRNA accumulation.
Conclusions:
- The developmental switch in cytoplasmic CK mRNA dominance is not regulated by nerve input.
- Innervation is crucial for the optimal accumulation of muscle-specific CK (MCK) mRNA during late fetal development.
- The absence of innervation specifically impacts the accumulation of MCK transcripts, not BCK transcripts, in developing hindleg muscles.
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