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H Khachaturian

Publications and source records attributed to H Khachaturian.

At least 19 recordsLinked to original sources
PubMed1991

Pro-opiomelanocortin mRNA and peptide co-expression in the developing rat pituitary.

Pro-opiomelanocortin (POMC) is synthesized in both the pituitary gland and the brain. Various peptide products of this precursor, namely beta-endorphin, ACTH and alpha-MSH are co-localized in the anterior lobe corticotrophs, all intermediate lobe cells and in hypothalamic neurons. Messenger RNA (mRNA) for POMC has further been shown to exist in these tissues. In this study, we have shown that POMC mRNA, and peptide accumulation as detected by in situ hybridization and immunocytochemistry, respectively, occur simultaneously within the rat pituitary gland during ontogeny and that their maturation occurs in parallel during prenatal and early postnatal development.

Adrenocorticotropic Hormone↗
PubMed1987

Autoradiographic differentiation of mu, delta, and kappa opioid receptors in the rat forebrain and midbrain.

While there is an abundance of pharmacological and biochemical evidence to suggest the existence of multiple opioid receptors, their precise localization within the brain is unclear. To help clarify this issue, the present study examined the distributions of the mu, delta, and kappa opioid receptor subtypes in the rat forebrain and midbrain using in vitro autoradiography. Mu and delta receptors were labeled with the selective ligands 3H-DAGO (Tyr- D-Ala-Gly-MePhe-Gly-ol), and 3H-DPDPE (D-Pen2, D-Pen5-enkephalin), respectively, while the kappa receptors were labeled with 3H-(-)bremazocine in the presence of unlabeled DAGO and DPDPE. Based on previous findings in our laboratory, the labeling conditions were such that each ligand selectively occupied approximately 75% of each of the opioid sites. The results demonstrated that all 3 opioid receptor subtypes were differentially distributed in the rat brain. Mu binding was dense in anterior cingulate cortex, neocortex, amygdala, hippocampus, ventral dentate gyrus, presubiculum, nucleus accumbens, caudate putamen, thalamus, habenula, interpeduncular nucleus, pars compacta of the substantia nigra, superior and inferior colliculi, and raphe nuclei. In contrast, delta binding was restricted to only a few brain areas, including anterior cingulate cortex, neocortex, amygdala, olfactory tubercle, nucleus accumbens, and caudate putamen. Kappa binding, while not as widespread as observed with mu binding, was densely distributed in the amygdala, olfactory tubercle, nucleus accumbens, caudate putamen, medial preoptic area, hypothalamus, median eminence, periventricular thalamus, and interpeduncular nucleus. While all 3 opioid receptor subtypes could sometimes be localized within the same brain area, their precise distribution within the region often varied widely. For example, in the caudate putamen, mu binding had a patchy distribution, while delta and kappa sites were diffusely distributed, with delta sites being particularly dense ventrolaterally and kappa sites being concentrated ventromedially. These results support the existence of at least 3 distinct opioid receptors with possibly separate functional roles.

Animals↗
PubMed1986

Pharmacological and anatomical evidence of selective mu, delta, and kappa opioid receptor binding in rat brain.

While the distribution of opioid receptors can be differentiated in the rat central nervous system, their precise localization has remained controversial, due, in part, to the previous lack of selective ligands and insensitive assaying conditions. The present study analyzed this issue further by examining the receptor selectivity of [3H]DAGO (Tyr-D-Ala-Gly-MePhe-Gly-ol), [3H]DPDPE (2-D-penicillamine-5-D-penicillamine-enkephalin), [3H]DSLET (Tyr-D-Ser-Gly-Phe-Leu-Thr) and [3H](-)bremazocine, and their suitability in autoradiographically labelling selective subpopulations of opioid receptors in rat brain. The results from saturation, competition, and autoradiographic experiments indicated that the three opioid receptor subtypes can be differentiated in the rat brain and that [3H]DAGO and [3H]DPDPE selectively labelled mu and delta binding sites, respectively. In contrast, [3H]DSLET was found to be relatively non-selective, and labelled both mu and delta sites. [3H]Bremazocine was similarly non-selective in the absence of mu and delta ligands and labelled all three opioid receptor subtypes. However, in the presence of 100 nM DAGO and DPDPE, concentrations sufficient to saturate the mu and delta sites, [3H]bremazocine did label kappa sites selectively. The high affinity [3H]bremazocine binding sites showed a unique distribution with relatively dense kappa labelling in the hypothalamus and median eminence, areas with extremely low mu and delta binding. These results point to the selectivity, under appropriate conditions, of [3H]DAGO, [3H]DPDPE and [3H]bremazocine and provide evidence for the differential distribution of mu, delta, and kappa opioid receptors in rat brain.

Animals↗
PubMed1986

Immunocytochemical localization of pro-opiomelanocortin-derived peptides in the adult rat spinal cord.

A dispersed descending pro-opiomelanocortin (POMC) fiber system has been demonstrated by peroxidase-antiperoxidase (PAP) immunocytochemistry in the adult rat spinal cord. beta-endorphin, adrenocorticotrophic hormone (ACTH), alpha-melanocyte-stimulating hormone (alpha-MSH) and 16K immunoreactive fibers exist in the spinal cord from cervical down to sacral level. Descending fibers running parallel in the dorsolateral and lateral funiculus send collaterals ventromedially or medially to terminate in the gray matter surrounding the central canal, where nociceptive neurons have recently been located, in addition to those nociceptive cells in the dorsal horn. After spinal transection at lower thoracic level, POMC peptide immunoreactivities disappeared below the lesion. Moreover, no POMC cell bodies were found in the spinal cord. Therefore, the descending fibers are most likely of supraspinal origin.

Adrenocorticotropic Hormone↗
PubMed1986

Further characterization of the extra-arcuate alpha-melanocyte stimulating hormone-like material in hypothalamus: biochemical and anatomical studies.

Previous studies had shown the existence of an extra-arcuate cell group in lateral hypothalamus which contains alpha-melanocyte stimulating hormone (a-MSH)-like immunoreactivity, but no other pro-opiomelanocortin (POMC) immunoreactivity. The question we have attempted to address in this series of studies is whether the material is indeed a-MSH or a cross-reacting material. Chromatographic studies failed to detect any material which is different from a-MSH or des-acetyl-a-MSH, suggesting that either the material is authentic a-MSH/des-acetyl-a-MSH, or that it is not detected by our RIAs. A series of manipulations including dissections of arcuate vs. extra-arcuate hypothalamic areas, treatment with colchicine, lesions with monosodium glutamate and knife cuts were aimed at isolating the extra-arcuate region and showing that it contains an excess of a-MSH over beta-endorphin (B-END), presumably deriving from the extra-arcuate group. However, all studies showed parallel changes in a-MSH and B-END, suggesting that we were not detecting a non-POMC derived a-MSH in these studies. This led to the tentative conclusion that the material was not a-MSH and was not being detected by our RIA's. This hypothesis was tested by further characterizing the material immunohistochemically. These studies led to the conclusion that the extra-arcuate material had a carboxy-terminal homology with a-MSH but differed from it in the midregion, since antisera directed at the 4-10 region of a-MSH failed to stain this non-POMC cell group. Finally, the anatomy of this extra-arcuate group is described, particularly the projections to the striatum, hippocampus, neocortex and olfactory bulb.

Animals↗
PubMed1986

Pro-dynorphin is endogenous to the anterior pituitary and is co-localized with LH and FSH in the gonadotrophs.

Pro-dynorphin peptides have been shown to exist in the anterior lobe of the pituitary gland. The dynorphin in the anterior lobe is distinct from that which is co-localized with vasopressin in the magnocellular system in both post-translational processing and regulation of release. Here, we report on the existence of pro-dynorphin mRNA, approximately 2400 nucleotides in length, in the anterior lobe. Furthermore, we present immunocytochemical evidence for the co-existence of dynorphin, LH and FSH in a subset of gonadotrophs. These findings suggest a possible role of pro-dynorphin products in the regulation of the hypothalamic-pituitary-gonadal axis.

Animals↗
PubMed1986

Multiple opioid receptor subtypes in the pituitary-adrenal axis: a cross-species study.

This study examines the multiple opioid receptor subtypes present in the pituitary-adrenal axis of several species using in vitro autoradiography. The monkey neural lobe was found to contain all three subtypes with a preponderance of kappa sites, while the rat and bovine neurohypophyses contained only kappa sites. In the adrenal, opioid receptor sites were found exclusively in the medulla and varied with species. Monkey adrenal contained predominantly delta sites with a relatively small number of kappa sites present, while bovine adrenal medulla had predominantly kappa sites and only a relatively small proportion of delta receptor sites. No specific opioid receptor binding could be detected in the rat adrenal. These results emphasize the importance of species-differences in understanding the role of opioids in neuroendocrine regulation.

Adrenal Glands↗
PubMed1985

Time of origin of opioid peptide-containing neurons in the rat hypothalamus.

By using a combined technique of immunocytochemistry and [3H]thymidine autoradiography, we have determined the "birth date" of opioid peptide-containing neurons in several hypothalamic nuclei and regions. These include proopiomelanocortin (POMC) neurons (represented by ACTH immunoreactivity) in the arcuate nucleus; dynorphin A neurons in the supraoptic and paraventricular nuclei and the lateral hypothalamic area; and leu-enkephalin neurons in the periventricular, ventromedial, and medial mammillary nuclei, as well as in preoptic and perifornical areas. Arcuate POMC neurons were born very early in embryonic development, with peak heavy [3H]thymidine nuclear labelling occurring on embryonic day E12. Supraoptic and paraventricular dynorphin A neurons were also labelled relatively early (peak at E13). The lateral hypothalamic dynorphin A neurons showed peak heavy labelling also on day E12. By contrast, leu-enkephalin neurons in the periventricular nucleus and medial preoptic area exhibited peak heavy nuclear labelling on day E14. Furthermore, perifornical and ventromedial leu-enkephalin neurons were also born relatively early (peak on days E12 and E13, respectively). However, the leu-enkephalin neurons in the medial mammillary nucleus were born the latest of all cell groups studied (i.e., peak at E15). The results indicate a differential genesis of these opioid peptide-containing neuronal groups in different hypothalamic nuclei and regions.

Adrenocorticotropic Hormone↗
PubMed1985

Postnatal development of beta-endorphin immunoreactivity in the medulla oblongata of rat.

Beta-endorphin (B-END) like immunoreactivity (i.r.) levels were measured by radioimmunoassay in the medulla oblongata of developing rats on postnatal ages P1-P42 at 7 day intervals, and in adult rats. From P1 to P42, B-END i.r. increased from 77.0 +/- 1.3 fm to 900.0 +/- 21.6 fm per medulla region (Mean +/- S.E.M.). Adult levels of B-END i.r. were 852.0 +/- 17.0 fm per medulla region. When B-END i.r. was determined per unit protein during this developmental period, a statistically significant change in levels was noted. B-END i.r. dropped from P1 to P7, and then increased from P7 to P14 (P less than 0.01). From P14 through adult, levels did not change significantly. Despite a "drop-out" in the observed immunostaining of B-END neurons in caudal medulla (perikarya in the nucleus tractus solitarius) at P21, radioimmunoassayable levels of this peptide remained constant from P21 through adult per unit protein.

Animals↗
PubMed1985

Development of hypothalamic opioid neurons: a combined immunocytochemical and [3H]thymidine autoradiographic study.

Using a combined technique of immunocytochemistry and [3H]thymidine autoradiography, we have determined the "birth-date" of opioid peptide containing neurons in three hypothalamic nuclei. These include proopiomelanocortin neurons (indicated by ACTH immunoreactivity) in the arcuate nucleus, dynorphin A neurons in the supraoptic nucleus, and [Leu]enkephalin neurons in the periventricular nucleus. Arcuate proopiomelanocortin neurons were born very early in embryonic development, with peak heavy [3H]thymidine nuclear labelling occurring on embryonic day E12. Supraoptic dynorphin A neurons were also labelled relatively early (peak at E13). By contrast, [Leu]enkephalin neurons in the periventricular nucleus exhibited peak heavy nuclear labelling on day E14. The results indicate a differential genesis of these three opioid peptide containing neuronal groups in three different hypothalamic nuclei.

Adrenocorticotropic Hormone↗
PubMed1985

Analysis of opioid and non-opioid end products of pro-dynorphin in the substantia nigra of the rat.

The substantia nigra is among the richest pro-dynorphin terminal field regions in the rat brain. We therefore contrasted processing in this area to the known processing in the posterior pituitary. Fractionation of acid extracts of the posterior pituitary by gel filtration followed by analysis by radioimmunoassay indicated that the molar ratio of dynorphin A(1-17) to dynorphin A(1-8) averaged 1:2. The levels of dynorphin A-related end products to alpha-neo-endorphin and bridge peptide (a 2K nonopioid end product of pro-dynorphin) were approximately equimolar; however, the levels of dynorphin B-sized material were 50% lower than dynorphin A levels. Similar analyses of acid extracts of the substantia nigra also indicated that the levels of dynorphin A, alpha-neo-endorphin, and bridge peptide were approximately equimolar. In this terminal field the levels of dynorphin B-sized material were approximately 60% lower than dynorphin A. A striking feature of the nigral system was that the molar ratio of dynorphin A(1-17) to dynorphin A(1-8) averaged 1:16. Thus, in the nigra, dynorphin A(1-17) is primarily a biosynthetic intermediate rather than as an end product.

Animals↗
PubMed1985

Combined autoradiographic-immunocytochemical analysis of opioid receptors and opioid peptide neuronal systems in brain.

Using adjacent section autoradiography-immunocytochemistry, the distribution of [3H]naloxone binding sites was studied in relation to neuronal systems containing [Leu]enkephalin, dynorphin A, or beta-endorphin immunoreactivity in rat brain. Brain sections from formaldehyde-perfused rats show robust specific binding of [3H]naloxone, the pharmacological (mu-like) properties of which appear unaltered. In contrast, specific binding of the delta ligand [3H]D-Ala2,D-Leu5-enkephalin was virtually totally eliminated as a result of formaldehyde perfusion. Using adjacent section analysis, we have noted associations between [3H]naloxone binding sites and one, two, or all three opioid systems in different brain regions; however, in some areas, no apparent relationship could be observed. Within regions, the relationship was complex; for example, in caudate-putamen, patches of opioid receptors did not correspond to the distribution of enkephalin immunoreactivity, but there was a correspondence between subcallosal streaks of binding sites and enkephalin. The complexity of the association between [3H]naloxone binding sites and the multiple opioid systems, and previous reports of colocalization of mu and kappa receptors in rat brain, are inconsistent with a simple-one-to-one relationship between a given opioid precursor and opioid receptor subtype. Instead, since differential processing of the three precursors gives rise to peptides of varying receptor subtype potencies and selectivities, the multiple peptide-receptor relationships may point to a key role of post-translational processing in determining the physiological consequences of opioid neurotransmission.

Animals↗
PubMed1985

Prodynorphin peptide immunocytochemistry in rhesus monkey brain.

The present study describes the immunocytochemical distribution of peptides derived from the prodynorphin precursor in the brain of the rhesus monkey (Macaca mulatta). Animals were treated with colchicine (intracerebroventricularly) prior to perfusion to enhance the observation of perikaryal immunoreactivity. Using antisera generated against dynorphin A(1-17), dynorphin B(1-13), and prodynorphin(186-208) (or bridge peptide), the anatomical distribution of dynorphin systems was mapped. The results indicate a widespread neuronal localization of immunoreactivity from the cerebral cortex to the caudal medulla. Anti-dynorphin B and anti-bridge peptide sera proved useful for the demonstration of neuronal perikarya, while the dynorphin A antiserum was best for localizing terminal projection fields. Immunoreactive perikarya are located in numerous brain loci, including the cingulate cortex, caudate nucleus, amygdala, hypothalamus (especially the magnocellular nuclei), thalamus, substantia grisea centralis, parabrachial nucleus, nucleus tractus solitarius, and other nuclei. In addition, fiber and terminal immunoreactivity are seen in varying densities in the striatum and pallidum, substantia innominata, hypothalamus, substantia nigra pars reticulata, parabrachial nucleus, spinal trigeminal nucleus, and other areas. The distribution of prodynorphin peptides in the brain of the monkey is similar to that described for the rat brain; however, significant differences also exist. Other interspecies differences in the anatomy of prodynorphin and proenkephalin neuronal systems in the monkey and human brain are further discussed.

Animals↗
PubMed1984

Localization of neurons containing pro-opiomelanocortin-related peptides in the hypothalamus and midbrain of the lizard, Anolis carolinensis: evidence for region-specific processing of beta-endorphin.

Immunohistochemical analyses of the lizard-brain, following colchicine pretreatment, revealed two populations of POMC-producing cell bodies located in medial-basal hypothalamus and the mesencephalic tegmentum. Analyses of extracts of lizard brain regions by radioimmunoassay and gel filtration chromatography indicate that beta-endorphin-sized and alpha-MSH-sized peptides are the major POMC-related end products. Evidence is presented for region-specific processing of beta-endorphin in the lizard brain.

Animals↗
PubMed1984

Beta-endorphin/ACTH immunocytochemistry in the CNS of the lizard Anolis carolinensis: evidence for a major mesencephalic cell group.

The immunocytochemical distribution of beta-endorphin and other proopiomelanocortin (POMC) peptides in the central nervous system of the lizard Anolis carolinensis was determined. Colchicine pretreatment was used to enhance perikaryal immunoreactivity. A major finding of this study is the localization of a previously undetected mesencephalic cell group which exhibits immunoreactivity to beta-endorphin, ACTH, and alpha-MSH. The perikarya of these neurons are large, bipolar, and situated in the mesencephalic tegmental area. They appear to project to the mesencephalic central gray and other brainstem structures. In contrast, the immunoreactive parvicellular perikarya of the medial-basal hypothalamus, corresponding to the POMC perikarya of the rodent arcuate nucleus, exhibit major rostral projections to various telencephalic and diencephalic structures. The exact extent of fiber projections and innervation patterns arising from either of these two groups is not clear at this time and will require further analyses. Scattered fiber immunoreactivity was also seen in the medial cerebral cortex and the striatal complex, regions which apparently are not innervated by beta-endorphin fibers in the rodent brain. Also, no immunoreactivity was seen to an antiserum to the 16K peptide of POMC. Other similarities and differences in the brain distribution of POMC in reptiles and mammals are discussed.

Adrenocorticotropic Hormone↗