Calcitonin receptor-like receptor expression in rat skeletal muscle fibers
2011
Fernandez, Hugo L. | Smith, Andrea | Dennis, John S. | Citron, Bruce A.
The calcitonin gene related peptide (CGRP) pathway is important in many processes including several in the central and peripheral nervous systems. CGRP is present in motor neurons and in sensory tracts, with its expression likely regulated by its use. This is supported by the fact that axotomy results in increased CGRP production in the nerve cell body. The target CGRP receptor, produced in part from the calcitonin receptor-like receptor (Calcrl) gene, has been thought to be present in multiple forms based on kinetic studies; however, understanding of the regulation of the expression of the Calcrl gene remains incomplete. CALCRL is an important factor in aging and associated disorders. This study focused on the neuromuscular system where it has been unclear whether different proteins are initially translated and whether higher levels of CALCRL are localized to the endplate regions. Rat gracilis muscle neuromuscular junctions were examined by isolating endplate enriched and non-endplate regions identified by staining for acetylcholine esterase or conjugated α-bungarotoxin binding. The CALCRL protein was detected at approximately 60kDa by Western immunoblotting and, in the isolated extracts, we found that the Calcrl mRNA level was elevated 6 fold in the muscle endplate regions and that there were two distinct Calcrl messages present. Sequence analysis showed that the two different Calcrl forms were due to alternative splicing but in a non-coding region of the transcript such that only one translation product would be generated. This indicates that previously identified pharmacologic heterogeneity is most likely due to post-translational modifications and interactions.
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