Ultrastructural localization of myoglobin mRNA in human skeletal muscle

Abstract

The intracellular localization of myoglobin mRNA in the skeletal muscles of normal subjects was examined by in situ hybridization using a biotin-labeled cDNA probe. By phase-contrast microscopy, myoglobin mRNA signals were demonstrated to be located preferentially on the A-band. Two different methods of tissue preparation, i.e., pre-embedding method and post-embedding method, were used for the electron microscopic study. With the pre-embedding method, only a few gold particles were found to be associated with cytoskeletal filaments in the intermyofibrillar space. With the post-embedding method, superior preservation of sections and higher signal intensities were obtained. Although most of the gold particles were localized on the A-band, some were seen in other regions; i.e., in the intermyofibrillar space, perinuclear space, or the I-band, where myoglobin is localized. These findings suggest that myoglobin is primarily synthesized on the A-band, where ribosomes predominantly exist, although myoglobin is also localized on the I-band. The predominant localization of myoglobin mRNA on the A-band may aid in the mRNA transcription and may be related to the regulation of myoglobin synthesis in skeletal muscle cells.