Binding sites for synthetic human 1251-labeled calcitonin gene-related peptide (125I-CGRP) have been demonstrated in membranes of the human nervous system. Binding was high in the cerebellar cortex (1.35 ± 0.27 fmol/mg of tissue; mean ± SEM), spinal cord (1.06 ± 0.27 to 1.27 ± 0.23 fmol/mg), and nucleus dentatus (1.02 ± 0.15 fmol/mg), intermediate in the inferior colliculus (0.80 ± 0.14 fmol/mg) and substantia nigra (0.75 ± 0.14 fmol/mg), low in the neocortex, globus pallidus, nucleus caudatus, hippocampus, amygdala, superior colliculus, thalamus, and hypothalamus (0.15-0.32 fmol/mg), and negligible in spinal and sympathetic ganglia and pituitary (<0.04 fmol/mg). Autoradiography showed distinct 1251-CGRP binding over the molecular and Purkinje cell layers of the cerebellar cortex and over the substantia gelatinosa posterior of the spinal cord. The highest levels of CGRP-like components were recognized in the dorsal part of the spinal cord and the pituitary gland. In the ventral part of the spinal cord as well as in the pituitary and thyroid glands, CGRP values were higher when measured by radioreceptorassay as compared to RIA, indicating that at least two CGRP-like components are present. The predominant CGRPlike peak on HPLC had the retention time of synthetic human CGRP. Immunohistochemistry revealed the presence of a dense plexus of CGRP immunoreactive nerve fibers in the dorsal horn of the spinal cord.The existence of rat and human calcitonin gene-related peptide (CGRP) has been predicted by analysis of the nucleotide sequence of the calcitonin gene (1, 2). CGRP is a unique 37 amino acid peptide that shares greater structural homology with salmon calcitonin-(1-32) than with rat and human calcitonin-(1-32) (1-3). Expression of CGRP was shown immunohistochemically and by specific RIA in combination with gelpermeation chromatography and HPLC in the nervous system and in the pituitary and thyroid glands (4-6). The amino acid sequence was derived from human medullary carcinoma of the thyroid (7).Here we report distinct regional distribution of CGRP and calcitonin (CT) binding sites in the human central nervous system. The presence of endogenous CGRP-like components in the nervous system, pituitary, and thyroid was revealed by a radioreceptorassay (RRA) using human cerebellar membranes, by RIA, and by immunohistochemistry. MATERIALS AND METHODSTissues and Peptides. Tissue obtained at autopsy within 20 hr postmortem was frozen on dry ice and stored at -80°C for a maximum of 3 wk. Patients with malignant tumors, chronic renal insufficiency, or metabolic bone disease were excluded from the study.Synthetic salmon CT-( Receptor Binding Studies. Membranes were prepared and incubated as described (8, 9) with 17,000 dpm (1 dpm = 16.7 mBq) of human 125I-labeled CGRP (125I-CGRP) Ci/mmol) or salmon 125I-labeled CT (125I-CT) Ci/mmol). Specific binding was defined as total binding minus binding in the presence of 0.7 ,uM unlabeled human CGRP or salmon CT, respectively. A RRA for CGRP was developed using membranes fro...

Immunoreactive calcitonin (CT), indistinguishable from human CT-(1-32) and its sulfoxide, has been identified in extracts of the hypothalamus, the pituitary, and the thyroid obtained from human subjects at autopsy. CT concentrations were highest in a region encompassing the posterior hypothalamus, the median eminence, and the pituitary; intermediate in the substantia nigra, the anterior hypothalamus, the globus pallidus, and the inferior colliculus; and low in the caudate nucleus, the hippocampus, the amygdala, and the cerebral and cerebellar cortices. Specific CT binding measured with "MI-labeled salmon CT was high-

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A salmon calcitonin-like material indistinguishable from synthetic salmon calcitonin-(1-32) on high performance liquid chromatography (HPLC) has been recognized in thyroid extracts of normal subjects and of patients with medullary carcinoma. The same peptide was detected in extracts of the periventricular mesencephalic region which included the periventricular dorsal thalamus, the subthalamus and the hypothalamus. Human calcitonin-(1-32)- and carboxyl-terminal adjacent peptide (CCAP)-like components were also found. The content of immunoreactive salmon calcitonin of the periventricular mesencephalic region (n = 6) and of normal thyroid glands (n = 6) was comparable (mean +/- SE, 0.34 +/- 0.17 ngeq/g wet tissue and 0.39 +/- 0.22 ngeq/g, respectively); and the levels were slightly, but not significantly higher in medullary thyroid carcinoma extracts (1.95 +/- 0.69 ngeq/g) (P less than 0.1). Immunoreactive human calcitonin and CCAP occurred in roughly equimolar concentrations. They were lowest in the periventricular mesencephalic region (0.26 +/- 0.09 ngeq/g and 0.46 +/- 0.10 ngeq/g, respectively), followed by normal thyroid glands (146 +/- 26 ngeq/g and 94 +/- 19 ngeq/g, respectively), and they were highest in medullary thyroid carcinoma tissue (680 +/- 372 mu geq/g and 144 +/- 125 mu geq/g, respectively).

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