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We compared the distribution of substance P (SP) and calcitonin gene-related peptide (CGRP) fibers of the superficial muscle layer (trapezius muscle), median muscle layer (rhomboideus muscle), and deep muscle layer (longissimus and spinalis muscles) of the dorsum of the rat. SP- and CGRP-immunoreactive fibers were seen along the walls of various types of blood vessels and within nerve bundles in skeletal muscles of all layers. Coexistence of SP and CGRP was evident in nerve fibers along the blood vessel walls. The total number of CGRP varicosities per millimeter square of muscle surface area was evaluated quantitatively, and CGRP varicosities were found to be significantly more numerous in the superficial muscle layer than in the deeper ones. After capsaicin treatment, most of the SP and CGRP fibers along the blood vessel walls were eliminated. These results suggest that sensory nerve fibers containing SP and CGRP are distributed more abundantly in the superficial muscle layer than in the deeper ones and that they might be involved in the regulation of local blood flow. The finding of SP- and CGRP-immunoreactive nerve fibers along the blood vessel walls connecting the trapezius muscle and the hypodermis raises the possibility that sensory stimuli to the skin affect the local blood flow of superficial muscle through collaterals of cutaneous fibers.

Neuropeptides in the motor nerves innervating the red and white muscles of the goldfish Carassius auratus were examined. In the tonic red muscles, varicose nerve endings immunoreactive for both calcitonin gene-related peptide and substance P were found spread over the surface of the muscle fibers, but in the twitch white muscles only scattered nerve endings immunoreactive for calcitonin gene-related peptide were found. At the electron microscopic observation, dense electron products immunoreactive for calcitonin gene-related peptide and for substance P (SP) were detected in the motor nerve endings making synapses on the muscle fibers of the red muscles. In the spinal cord, all of the motor neurons showed immunoreactivity to calcitonin gene-related peptide, but the motor neurons immunoreactive for substance P were restricted to the ventrolateral group that has been shown to project predominantly to the red muscles. These results suggest that the motor neurons innervating the red and white muscles of the goldfish are distinct in their neuropeptide content. The present study also raises the possibility that SP might be related to the unique physiological properties of the tonic type red muscles, probably by direct binding to the acetylcholine receptors.