Search PubMed⌕ Search

Biomedical subjects

C Kordon

Publications and source records attributed to C Kordon.

At least 127 records · Page 7Linked to original sources
PubMed1984

Mapping of LH-RH-containing projections to the mediobasal hypothalamus by differential deafferentation experiments.

Luteinizing hormone-releasing hormone (LH-RH) was immunoassayed in several hypothalamic structures of male rats after complete, anterior, posterior or lateral deafferentation of that structure performed with a rotating knife, as well as after discrete frontal or sagittal transections placed with a glass knife in various parts of the mediobasal hypothalamus (MBH). Survival time in all cases was two weeks. Taken together, the results indicate that most fibers containing LH-RH and originating in the preoptic area of the hypo-thalamus (APO) take a lateral course upon leaving that structure and travel along the medial forebrain bundle (MFB). Along that tract, they proceed caudally over some distance and enter the median eminence from the side; fibers innervating the posterior median eminence bend back towards the midline at a more posterior level than those terminating in the zona externa. A limited amount of fibers also reaches the median eminence from a midsagittal location. In addition, a few medial arcuate-median eminence connections may account for the small proportion of MBH LH-RH spared by complete, anterolateral or lateral transections. Projections to the organum vasculosum laminae terminalis, whether proceeding from the APO in an independent manner or as collaterals of fibers terminating in the median eminence, are unaffected by any of the transections tested. A partly common organization pattern of fibers supplying the median eminence in several neuropeptides (for instance somatostatin, CRF, cholecystokinin or dynorphin) is suggested.

Afferent Pathways↗
PubMed1984

Subcellular distribution of particle-bound neutral peptidases capable of hydrolyzing gonadoliberin, thyroliberin, enkephalin and substance P.

Subcellular fractions from rat anterior pituitary homogenates were obtained by differential and gradient centrifugation, identified with the help of marker enzymes and screened for peptidases capable of hydrolyzing gonadoliberin, thyroliberin, enkephalin and substance P. Since each neuropeptide is susceptible to cleavage by more than one enzyme, specific substrates or inhibitors have been used for the selective determination of the individual peptidasic activities. Among the various enzymes tested, the angiotensin-converting enzyme, the thermolysin-like metalloendopeptidase ('enkephalinase'), a thyroliberin-degrading enzyme and some aminopeptidasic activities were found to be associated with the plasma membrane. Other aminopeptidases, a gonadoliberin-degrading and a substance-P-degrading enzyme are associated with the mitochondria and thus are most likely not involved in the biological inactivation of neuropeptides.

Animals↗
PubMed1984

Subcellular distribution of somatostatin-14, somatostatin-28 and somatostatin-28 (1-12) in rat brain cortex and comparisons of their respective binding sites in brain and pituitary.

Subcellular distribution and binding characteristics of the three endogenous peptides somatostatin-14 (SRIF-14), somatostatin-28 (SRIF-28) and somatostatin-28(1-12) (SRIF-28(1-12] derived from preprosomatostatin were investigated in the rat brain cortex. The three peptides are predominantly recovered from a crude mitochondrial pellet (P2), containing the pinched off nerve endings. Specific high affinity binding sites for 125I-N-Tyr-SRIF-14 and 125I-N-Tyr-SRIF-28 are present on pituitary and brain membranes. Under the same conditions, 125I-N-Tyr-SRIF-28(1-12) binding is undetectable. Moreover, SRIF-28(1-12) does not displace 125I-N-Tyr-SRIF-14 or 125I-N-Tyr-SRIF-28 binding. SRIF-28 is more potent than SRIF-14 to displace 125I-N-Tyr-SRIF-28 binding to brain and pituitary membranes, while both peptides are equipotent to displace 125I-N-Tyr-SRIF-14 binding. Finally, the regional distribution of 125I-N-Tyr-SRIF-14 and 125I-N-Tyr-SRIF-28 binding sites in the brain is identical. In conclusion, the present results are consistent with a neurotransmitter and neurohormonal role for SRIF-14 and SRIF-28. The function of SRIF-28(1-12) in brain remains to be elucidated. Additionally, a differential role for SRIF-14 and SRIF-28 both in adenohypophysis and brain cannot be ascertained at the present time.

Animals↗
PubMed1984

Effects of 17 beta-estradiol on LH-RH release from rat mediobasal hypothalamic slices.

UNLABELLED: Mediobasal hypothalamic slices of adult ovariectomized (OVX) rats treated or not with 17 beta-estradiol (E2) were superfused in buffered (pH 7.2) Locke medium containing bacitracin. A 6-min pulse of K+ (56 m M) was less effective in releasing luteinizing hormone releasing hormone (LH-RH) from mediobasal hypothalamic slices sampled from OVX rats than from OVX animals treated subcutaneously with either E2 or stilbestrol implants for 5 days; in contrast, the basal release of the neuropeptide was identical in both cases. Direct addition to the superfusion medium of 17 beta-estradiol (10(-10) to 10(-7) M) or stilbestrol (10(-8) M) potentiated the K+-induced LH-RH release from slices of OVX animals. The K+-induced LH-RH release observed after in vivo E2 implantation was not further amplified by in vitro addition of the hormone. Tamoxifen and hydroxytamoxifen, estrogen antagonists, were ineffective by themselves, but reversed the E2 facilitation of K+-evoked LH-RH release. In contrast, 17 alpha-estradiol, progesterone, or cholesterol (10(-8) or 10(-9) M) hat no effect on either basal or stimulated release of the neurohormone. Somatostatin release measured under identical conditions was not affected by castration or by in vitro addition of the steroid. IN CONCLUSION: (1) estradiol appears selectively and specifically involved in the process coupling, nerve endings depolarization, and LH-RH release, and (2) the effect is receptor-mediated and does not appear to require nuclear translocation of the steroid or transcription processes, since it can be readily elicited upon addition of the hormone to nerve endings disconnected from their cell bodies.

Animals↗
PubMed1984

Calmodulin involvement on the Ca++-dependent release of LHRH and SRIF in vitro.

Mediobasal hypothalamic (MBH) slices of male adult rats were superfused at 37 degrees C with oxygenated Hepes-buffer Locke medium. Bacitracin (2 X 10(-5) M) was added to prevent enzymatic degradation of LHRH and SRIF. 6 min pulse of K+ (56 mM), veratridine (15 microM) or the ionophore A 23187 (10(-5) M), markedly stimulated the release of both neuropeptides. Trifluoperazine, a calmodulin inhibitor, decreased the K+-evoked LHRH and SRIF release in a dose-dependent manner; it was also effective in inhibiting the veratridine-induced neuropeptides release. Phenytoin, a calmodulin-dependent kinase inhibitor, also decreased in a dose-dependent manner the K+-induced LHRH and SRIF release; the basal release of both neuropeptides remained unaffected by either treatment. The ionophore-stimulated release of both neuropeptides was significantly inhibited as well. These data demonstrate that a Ca++-calmodulin kinase system may be involved in the mechanism of depolarization-induced LHRH and SRIF release from hypothalamic nerve terminals.

Animals↗
PubMed1984

Characterization of a neutral endopeptidase localized in the mitochondrial matrix of rat anterior pituitary tissue with GnRH as a substrate.

We have determined the subcellular localization of an endopeptidase activity able to degrade gonadotropin releasing hormone (GnRH) and present in the rat adenohypophysis. After fractionation of tissue homogenates in 0.25 M sucrose by differential centrifugation, about 25% of the total cellular GnRH degrading activity was found to be sedimentable and recovered from heavy (M) and light (L) mitochondrial fractions with a distribution pattern similar to that of the mitochondrial and lysosomal reference enzymes cytochrome oxidase and beta-galactosidase. Upon further fractionation on sucrose density gradients, the activity comigrated with mitochondria. The peptidase appears endowed with a structure-linked latency; the activity is low in a freshly prepared mitochondrial fraction and increases upon treatment with membrane disrupting agents in a manner similar to that of malate dehydrogenase, a component of the mitochondrial matrix. Determination of GnRH cleavage sites was performed by amino acid analysis of the fragments obtained after incubation of the peptidase with (3H)-GnRH labelled on the pyroglutamic acid residue, in presence of carboxypeptidase and peptidyldipeptidase inhibitors. The fragments were separated by ion-exchange chromatography on an Aminex Q-15S column and purified by chromatography on silica gel plates. Fragments 1-2, 1-3, 1-4, 1-5 and 1-6 were all present as early as 1 min after the beginning of incubation. Formation of each of them was inhibited to the same extent by EDTA, mersalyl acid, dithioerythritol and Na deoxycholate. The same fragmentation pattern was observed after partial purification of the enzyme by gel filtration. These data indicate that cleavage of several peptide bonds may result from a possibly single endopeptidase located in the mitochondrial matrix space.

Animals↗
PubMed1983

Biphasic pattern of follicle stimulating and luteinizing hormone responses to gonadotropin-releasing hormone in vitro.

Increasing concentrations of LHRH or its active analog des-gly10 (D-ala6) LHRH induced a bimodal pattern of FSH and LH release from incubated male rat pituitaries. A low amplitude response (40% increase of LH over baseline levels) was observed for concentrations of LHRH and the agonist in the range of 10(-12) and 10(-13) M respectively. After a plateau of gonadotropin stimulation, a further high amplitude response (180-240% increase over baseline levels) occurred between 3.10(-10) and 10(-8) for LHRH and 3.10(-11) and 3.10(-9) M for des-gly10 (D-ala6) LHRH. Corresponding half maximal effective concentrations (ED50) were 2 and 0.3 nM respectively. Stimulation of FSH release closely paralleled that of LH. The low amplitude, high apparent affinity response was only obtained when the peptides were diluted in low concentrations of acidic tissue extracts. A preliminary study indicated that this method of dilution minimized peptide loss by adsorption. In addition, the low amplitude, high affinity response was never observed on pituitaries sampled from castrates. These experiments suggest the presence of two distinct populations of LHRH recognition sites on pituitary gonadotrophs. Under our experimental conditions, the appearance of the higher affinity response was dependent upon prior exposure to sex steroids. This could be due to a direct action of the steroid on the expression of a high affinity LHRH receptor.

Animals↗
PubMed1983

Influence of streptozotocin-induced diabetes on growth hormone secretion in the rat.

The effects of streptozotocin-induced diabetes on pituitary growth hormone (GH) content and release from incubated pituitaries were investigated. Male rats were made diabetic by a single injection of streptozotocin (65 mg/kg) and sacrificed by decapitation 15 days later. Pituitary GH concentration was significantly reduced in streptozotocin diabetic rats as compared to that observed in control animals. The amount of GH released from hemipituitaries was also lower in diabetic rats than in controls. Kinetic characteristics of somatostatin (SRIF) inhibition of GH release were not affected by the treatment. These results suggest that the decrease in plasma GH observed by some investigators in streptozotocin diabetic rats is probably due to a deficiency in GH storage and/or synthesis rather than a change in the responsiveness of pituitary GH cells to SRIF.

Animals↗
PubMed1983

Effects of ovarian steroids on in vitro release of LHRH from mediobasal hypothalamus.

The phasic luteinizing hormone (LH) release observed in ovariectomized (OVX), estrogen-implanted rats was further amplified and advanced when progesterone (P) was given 4 h prior to the gonadotropin surge. In contrast, an inhibitory effect of P on the daily LH surge was observed when P was administered 16-36 h prior to LH peak. In order to determine whether this biphasic action of P is primarily exerted on the release of luteinizing hormone releasing hormone (LHRH), on the pituitary response to LHRH, or on both, mediobasal hypothalamic slices or pituitary fragments of adult OVX rats or of OVX rats pretreated with estrogen alone or in combination with P were tested in a perifusion system. Mediobasal hypothalamic slices were perifused in buffered (pH 7.2) oxygenated Locke's medium containing bacitracin (2 X 10(-5) M). In the absence of estrogen pretreatment, high (56 mM) concentrations of K+ were barely effective in releasing LHRH. Subcutaneous implantation of 17 beta-estradiol for 5 days markedly increased the amplitude of the LHRH secretory response to K+ depolarization. Additional administration of P (25 mg/rat s.c.) 4 h before sacrifice further amplified the K+-induced LHRH release. In contrast, the K+-evoked LHRH secretion was significantly inhibited when P was given 16 or 36 h before. Estradiol thus appears to facilitate the LHRH secretory response to depolarizing stimuli, whereas P either enhances or blocks the induced LHRH release depending upon its time of administration. At the pituitary level, the sensitivity of LHRH-induced LH release was also increased after estrogen pretreatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1983

Guanine nucleotide sensitivity of [125I]-Iodo NTyr somatostatin binding in rat adenohypophysis and cerebral cortex.

Specific [125I]-Iodo-NTyr somatostatin binding sites are present in adenohypophyseal and cerebral cortical membranes. Guanine nucleotides reduce the maximal binding capacity of adenohypophyseal binding sites without significantly affecting their apparent affinity. In pituitary as well as in cortex, GTP is the most potent nucleotide followed by GDP and guanylyl imidodiphosphate (GMP-PNP). The effect appears specific of guanine nucleotides since ATP, ADP and AMP are inactive on [125I]-Iodo-NTyr somatostatin binding. These results, showing the nucleotide sensitivity of [125I]-Iodo-NTyr somatostatin binding in pituitary and cerebral cortex, are compatible with a coupling of somatostatin receptors with adenylate cyclase.

Animals↗
PubMed1982

Somatostatin connections between the hypothalamus and the limbic system of the rat brain.

Somatostatin (SRIF) content of several brain structures was evaluated by radioimmunoassay in rats bearing various types of hypothalamic transections, as well as lesions of the amygdala. Analysis of the regional changes in SRIF concentrations after surgery suggest the following conclusions: (1) hypothalamic somatostatinergic neurons project to the limbic system, with the exception of the amygdaloid nuclei; (2) the olfactory tubercle, the lateral septal nucleus, the habenula and probably the hippocampus receive somatostatin projections from periventricular SRIF-containing cells; (3) somatostatin-containing fibers take a lateral course after leaving periventricular cells and join the medial forebrain bundle; (4) somatostatin innervation of the amygdala seems to be intrinsic.

Amygdala↗
PubMed1982

Interaction between estradiol and prolactin on vasoactive intestinal peptide concentrations in the hypothalamus and in the anterior pituitary of the female rat.

To determine whether vasoactive intestinal peptide (VIP) can be regulated by modification of plasma estradiol and prolactin levels, VIP concentrations in various structures of the rat brain and in the pituitary were measured in hyperprolactinemic female rats by means of a specific radioimmunoassay for the peptide. In ovariectomized rats treated with estradiol (E2) implants alone and with both E2 and pituitary grafts to induce an experimental hyperprolactinemia, VIP levels decreased in the anterior and mediobasal hypothalamus and increased in the pituitary as compared to ovariectomized rats. No modification of hypothalamic VIP concentrations was observed in ovariectomized rats with pituitary grafts only, whereas a significant increase was found in the pituitary. These results suggest that, in the female rats, E2 exerts an effect on hypothalamic VIP levels, probably through indirect mechanisms, and that this action can be enhanced by elevated plasma prolactin levels.

Animals↗
PubMed1982

alpha 1-adrenergic receptor involvement in the LH surge in ovariectomized estrogen-primed rats.

The circadian pattern of LH release observed in ovariectomized estrogen-implanted rats was inhibited by blockade of alpha-adrenergic receptors by phenoxybenzamine; in contrast, blockade of beta-receptors by propranolol was ineffective. Prasozin, an alpha 1-antagonist, was as effective as phenoxybenzamine, whereas yohimbine, an alpha 2-antagonist, was effective only at high doses. Neither phenoxybenzamine nor prazosin were able to modify the LHRH-induced release of LH from superfused pituitaries. These results suggest an involvement of hypothalamic alpha 1-receptors in the control of circadian LH release.

Adrenergic alpha-Antagonists↗
PubMed1982

Further evidence for the existence of opiate binding sites on neurosecretory LHRH mediobasal hypothalamic terminals.

Opiate binding sites as well as LHRH and SRIF content were evaluated in mediobasal hypothalamus (MBH) from normal male rats and animals subjected to anterolateral deafferentation of the hypothalamus. A 87 and 44% depletion of LHRH and SRIF content respectively and a 50% decrease in the number of specific opiate binding sites was observed in the deafferented MBH. Dopa-decarboxylase activity remained unchanged. These data indicate that presynaptic opiate binding sites are present on LHRH and SRIF mediobasal hypothalamic nerve endings.

Animals↗
PubMed1982

Somatostatin in catecholamine-rich nuclei of the brainstem.

Somatostatin (SRIF) concentrations in catecholamine-rich nuclei of the rat brainstem were measured by radioimmunoassay. The study was performed both in control or sham operated animals and after transecting the major projections of hypothalamic SRIF-containing neurons. Concentrations of the peptide were found to be relatively high in the locus coeruleus, the parabrachial nucleus and the nucleus of the solitary tract; they were intermediate in the lateral reticular nucleus (A1 cell group) and low in the substantia nigra. Transection of hypothalamic periventricular efferents resulted in a 58% depletion of SRIF content in the locus coeruleus, while concentrations of the peptide in other areas were unaffected. Transection of the medial forebrain bundle at the level of the lateral hypothalamus decreased SRIF content by 55% in the substantia nigra, but not in the other nuclei tested. It is concluded that the hypothalamus contributes significantly to the somatostatinergic innervation of the locus coeruleus and the substantia nigra, whereas SRIF in the other nuclei is intrinsic or originates outside the hypothalamus.

Animals↗
PubMed1982

Neurotensin stimulation of prolactin secretion in vitro.

Neurotensin stimulated prolactin (PRL) secretion from incubated rat hemipituitaries. Under the same conditions, the secretion of growth hormone, luteinizing hormone and follicle-stimulating hormone was not affected. The stimulation of PRL was dose dependent, with an apparent affinity of neurotensin of 0.56 +/- 0.12 nM and a maximal stimulation of 56.5 +/- 6.7%. The effect of neurotensin seemed to be independent of that of other PRL releasing factors. In fact, the stimulation of neurotensin and thyrotropin-releasing hormone (TRH) and also of neurotensin and vasoactive intestinal peptide were additive. The action of neurotensin on PRL cells does not appear to involve either dopamine or gamma-aminobutyric acid receptors, since antagonists to these transmitters were found ineffective on PRL stimulation by neurotensin. PRL-releasing factor activity distinct from TRH has been described in fractions of hypothalamic extracts. Neurotensin, which is present in high amounts in the median eminence and has been measured in the adenohypophysis, is a candidate as a physiological PRL-releasing factor distinct from TRH.

Animals↗