Peptide neuroanatomy of adjuvant-induced arthritic inflammation in rat.

The influence of adjuvant-induced arthritis of the rat on central and peripheral peptide neuroanatomy was investigated by immunohistochemistry. The most striking feature of arthritic rats was the differential intensification of neuronal proenkephalin- and prodynorphin-related staining in dorsal horn. Changes were ipsilateral in monoarthritic and bilateral in polyarthritic rats as compared to controls. Opioid responsive neurons were target of substance P (SP) and calcitonin gene-related peptide (CGRP) fibers. Changes of SP and CGRP predominated in peripheral inflamed tissue and consisted of intensified immunostaining and an apparent sprouting of sensory fibers particularly around venules, in the epidermis and in areas infiltrated by immunocompetent cells. Opioid staining was absent from primary afferents but present in some immune cells of inflamed tissue. Endogenous antinociceptive opioids and pro-nociceptive/pro-inflammatory SP and CGRP may be crucial in the concerted response of the neuroimmune system to chronic inflammatory pain.

Isolation and structure of a C-terminally amidated nonopioid peptide, amidorphin-(8-26), from bovine striatum: a major product of proenkephalin in brain but not in adrenal medulla.

We have isolated and sequenced a C-terminally amidated peptide from bovine striatum. The peptide was purified to homogeneity by adsorption to XAD-2 resins and four different HPLC steps. Amino acid composition analysis and gas-phase sequence analysis revealed identity of this peptide with residues 8-26 of the proenkephalin-derived opioid peptide amidorphin, which we have recently isolated from bovine adrenal medulla. C-terminal amidation of amidorphin-(8-26) from bovine striatum was demonstrated by its stability to carboxypeptidase A digestion and full crossreactivity in a radioimmunoassay that required the C-terminal amide group as part of the recognition site. The nonopioid peptide amidorphin-(8-26), which lacks the N-terminal [Met]enkephalin sequence of amidorphin, is a major product of the opioid peptide precursor proenkephalin in the brain. In the adrenal medulla, however, where amidorphin occurs in remarkably high concentrations, amidorphin-(8-26) could not be detected. This is indicative of differential post-translational processing of proenkephalin in different tissues. In the brain, as opposed to the adrenal medulla, amidorphin is further processed at the typical cleavage signals of two basic residues, giving rise to the nonopioid peptide amidorphin-(8-26) and, possibly, to the opioid peptide [Met]enkephalin. Thus, proenkephalin in the brain might be considered as a precursor in which an opioid peptide is linked with a nonopioid peptide of possibly different biological function.

Activation of hypothalamic beta-endorphin pools by reward induced by highly palatable food.

Experiments were performed to find biochemical evidence of an activation of endogenous opiate peptides in the brain by incentive reward. A method used to estimate specific in vivo opiate binding in rats using the labelled opiate agonist, 3H-etorphine, indicated a considerable reduction in opiate binding exclusively in the hypothalamus of non-deprived animals given a highly palatable food to eat for 20 min. Radioimmunoassay of the hypothalamus of rats under similar conditions found a pronounced drop in the concentration of beta-endorphin, but not in dynorphin, in the hypothalamus, indicating a release and breakdown of beta-endorphin. Therefore, the reduction in opiate binding in the hypothalamus may at least be partially explained by an occupation of opiate receptors by beta-endorphin, causing a reduced availability of receptors to etorphine. A possible role of hypothalamic beta-endorphin in the facilitation of reward pathways in the brain is discussed.

Stress-induced release of brain and pituitary beta-endorphin: major role of endorphins in generation of hyperthermia, not analgesia.

The present paper examines the conjectured causal relationship between the alterations in brain, pituitary and plasma levels of endorphins and the antinociception (analgesia) and hyperthermia elicited by acute stress. A 5-min foot-shock instigated a significant depression in the levels of beta-endorphin immunoreactivity (beta-EI) in both the hypothalamus and periventricular beta-endorphinergic fibre-containing tissue. A large elevation in plasma levels of beta-EI, consisting of about 70% beta-endorphin (beta-EP), and 30% beta-lipotropin (beta-LPH) was associated with a significant reduction in the beta-EI content of both the anterior (AL) and neurointermediate (NIL) lobes of the pituitary. No concomitant changes in the levels of Met-enkephalin immunoreactivity (M-EI) in discrete areas of brain and pituitary were detectable. Application of a high (10 mg/kg) but not a low (1 mg/kg) dose of naloxone, prior to foot-shock, slightly reduced the increase in tail-flick latency evoked by this stress. In contrast, both of these doses strongly and dose-dependently attenuated the accompanying rise in core temperature (Tc). Chronic (approximately 30 day) morphine treatment resulted in a 45% decrease in the NIL content of beta-EI and a clear depression in its basal plasma levels, although a substantial post-stress rise in plasma beta-EI was still found: stress-induced analgesia (SIA) was enhanced, but the concurrent stress-induced hyperthermia (SIH), reduced in morphinized animals. These data demonstrate that stress produces a generalized mobilization of both central and pituitary pools of beta-EI, and indicate that endorphins may play a more important role in the mediation of changes in Tc than in the generation of the concomitant increase in nociceptive threshold, upon activation by stress.

Extrahypothalamic corticotropin and alpha-melanotropin in human brain.

The distribution of corticotropin (ACTH) and alpha-melanotropin (alpha-MSH) in human brain was investigated by radioimmunoassay using an antiserum which recognized h-ACTH1-39 and alpha-MSH to an equal degree on a molar basis. Significant amounts of material, which migrated on calibrated Sephadex G-50 columns as synthetic h-ACTH1-39 and synthetic alpha-MSH, were detected in distinct brain areas. The highest concentrations of ACTH and alpha-MSH were found in the diencephalon (hypothalamus 4.2 and 12.5 pmol/g wet weight, respectively) and in midbrain (periaqueductal gray 0.5 and 1.7 pmol/g, respectively) and smaller quantities in the rhombencephalon and telencephalon. The ACTH concentration in human pituitary (adenohypophysis) was 10(4)-fold greater than that in the hypothalamus. alpha-MSH was confined to the midpart and/or stalk region of the pituitary.