Green T, Dockray G J
Physiological Laboratory, University of Liverpool, U.K.
Neuroscience. 1988 Apr;25(1):181-93. doi: 10.1016/0306-4522(88)90017-6.
Retrograde tracing of the fluorescent marker, True Blue, has been used together with immunohistochemistry employing antibodies to substance P, calcitonin gene-related peptide, somatostatin, vasoactive intestinal polypeptide and morphine-modulating peptide to study the afferent innervation of the stomach in rat, mouse and guinea-pig. Up to 85% of spinal afferents to the stomach in all three species contained immunoreactive calcitonin gene-related peptide, and up to 50% contained substance P. In all three species less than 10% of vagal afferents to the stomach reacted with antibodies to calcitonin gene-related peptide, or substance P. Cacitonin gene-related peptide-immunoreactive fibres were found in the myenteric plexus, circular muscle and around submucosal blood vessels in the stomach. In the rat, removal of the coeliac ganglion, splanchnic nerve section, or capsaicin treatment virtually abolished calcitonin gene-related peptide immunoreactivity in the stomach. Capsaicin and splanchnic section also abolished the staining of immunoreactive calcitonin gene-related peptide fibres in the coeliac ganglion. The same treatments abolished substance P staining of fibres around submucosal blood vessels, but in the myenteric plexus and circular smooth muscle there were still abundant immunoreactive fibres, presumably arising from intrinsic cell bodies. No somatostatin-containing visceral afferents could be found, although somatostatin was localized to cell bodies in rat dorsal root ganglia. Immunoreactive vasoactive intestinal polypeptide-containing dorsal root ganglia neurons were not found; although antibodies to morphine-modulatory peptide revealed immunoreactive nerve cell bodies, we were unable to exclude the possibility that this result is attributable to cross reactivity with calcitonin gene-related peptide. These results provide direct evidence that calcitonin gene-related peptide is a marker for a major subset of visceral primary afferent neurons and suggest that this population of spinal afferents makes a major contribution to the total gastric content of calcitonin gene-related peptide.
荧光标记物真蓝逆行追踪技术已与免疫组织化学相结合，使用针对P物质、降钙素基因相关肽、生长抑素、血管活性肠肽和吗啡调节肽的抗体，来研究大鼠、小鼠和豚鼠胃的传入神经支配。在所有这三个物种中，高达85%的胃脊髓传入神经含有免疫反应性降钙素基因相关肽，高达50%含有P物质。在所有这三个物种中，不到10%的胃迷走传入神经与降钙素基因相关肽或P物质的抗体发生反应。在胃的肌间神经丛、环形肌和黏膜下血管周围发现了降钙素基因相关肽免疫反应性纤维。在大鼠中，切除腹腔神经节、切断内脏神经或用辣椒素处理几乎消除了胃中的降钙素基因相关肽免疫反应性。辣椒素处理和切断内脏神经也消除了腹腔神经节中降钙素基因相关肽免疫反应性纤维的染色。相同的处理消除了黏膜下血管周围纤维的P物质染色，但在肌间神经丛和环形平滑肌中仍有大量免疫反应性纤维，推测这些纤维来自内在细胞体。尽管生长抑素定位于大鼠背根神经节的细胞体，但未发现含生长抑素的内脏传入神经。未发现含血管活性肠肽的背根神经节神经元具有免疫反应性；尽管吗啡调节肽抗体显示出免疫反应性神经细胞体，但我们无法排除这一结果可能归因于与降钙素基因相关肽交叉反应的可能性。这些结果提供了直接证据，表明降钙素基因相关肽是内脏初级传入神经元主要亚群的标志物，并表明这群脊髓传入神经对胃中降钙素基因相关肽的总量有主要贡献。
