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Biomedical subjects

N Ling

Publications and source records attributed to N Ling.

At least 307 records · Page 17Linked to original sources
PubMed1983

Topographical study of the neurons containing hpGRF immunoreactivity in monkey hypothalamus.

Hypothalamic neurons producing growth hormone-releasing factor (GRF) have been characterized by immunohistochemistry in monkey hypothalamus, using an antiserum raised against hpGRF1-40, a peptide with GRF activity isolated from a human pancreatic tumor. Cell bodies with hpGRF immunoreactivity were found in arcuate and ventromedial nuclei. From these neurons, bundles of fibers innervate median eminence and appear to terminate in contact with portal vessels. In addition to median eminence, hpGRF immunoreactive fibers were found mostly in the anterior hypothalamus and the arcuate and ventromedial nuclei where they give perineuronal endings. These results correlate with earlier physiological data on hypothalamic control of growth hormone secretion and suggest that GRF is also involved in interneuronal relationships related or unrelated to neurohumoral control of pituitary secretions.

Animals↗
PubMed1983

A Met-enkephalin-containing-peptide, BAM 22P, as a novel substrate for glandular kallikreins.

Homogeneous preparations of two well-characterized glandular kallikreins have been examined for their ability to hydrolyze BAM 22P, a methionine-enkephalin-containing-peptide found in the adrenal medulla. Both enzymes cleaved preferentially the Arg6-Arg7 bond in this substrate. The specificity constant (kcat/Km) for this cleavage was 86 mM-1 sec-1 for horse urinary kallikrein and 566 mM-1 sec-1 for porcine pancreatic kallikrein. These results demonstrate a previously undescribed specificity for glandular kallikreins and suggest a possible role for these widely distributed enzymes in prohormone processing.

Animals↗
PubMed1983

Immunohistochemical evidence that brain enkephalins arise from a precursor similar to adrenal preproenkephalin.

We have examined rat brains by immunohistochemistry with an antiserum raised against BAM22P, a synthetic fragment of the adrenomedullary precursor of Leu- and Met-enkephalin, in order to determine whether preproenkephalin derived peptides are detectable in the central nervous system. BAM22P antiserum stained fibers, nerve endings and cell bodies in many areas of the brain, especially the striatum, the septum and the hypothalamus. Comparative topographical studies showed that neurons recognized by anti-BAM22P overlapped structures found immunoreactive for enkephalins in the present study. In addition, BAM22P antiserum stained cell bodies in cerebral cortex and in caudate putamen. The presence of BAM22P immunoreactivity in presumptive enkephalin neurons suggests that some brain enkephalins arise from a precursor identical or closely related to the preproenkephalin A found in the bovine adrenal medulla. This does not preclude the possibility of enkephalins also arising from other precursor molecules.

Adrenal Medulla↗
PubMed1983

Immunological characterization of endorphins, adrenocorticotropin, and melanotropins in frog hypothalamus.

To gain more information about the nature and regulation of opiomelanocorticotropic peptides in the frog diencephalon, radioimmunological determinations of alpha- and beta-MSH. ACTH, beta- and gamma-endorphins have been performed in hypothalamic extracts. Sephadex G-50 gel filtration revealed a single peak of alpha-MSH-like immunoreactivity (alpha-MSH-LI) comigrating with synthetic alpha-MSH. Two peaks of ACTH-LI were observed; the major one eluting slightly before human ACTH. Using a porcine beta-endorphin antiserum which exhibited 45% cross-reaction (on a molar basis) with ovine beta-LPH, one major peak of beta-endorphin-LI and two additional components were observed in the elution profile; none of these peaks coeluted with synthetic porcine beta-endorphin. No significant beta-MSH or gamma-endorphin-LI was detected. To investigate a possible role of glucocorticoids on the level of opiomelanocorticotropic peptides in frog hypothalamus, 60 male frogs were treated with dexamethasone (300 micrograms/day) during 8 days. Dexamethasone treatment did not modify the chromatographic distribution and the total amount of alpha-MSH-LI and ACTH-LI in hypothalamic extracts. A slight (15%) increase in beta-endorphin-LI was even observed after 8 days of dexamethasone treatment. From these results it is concluded that three classes of opiomelanocorticotropic peptides are present in the frog hypothalamus in the following order of concentration: beta-endorphin-LI greater than alpha-MSH greater than ACTH. In addition, circulating glucocorticoids which significantly reduce the concentration of opiomelanocorticotropic hormones in the distal lobe of the frog pituitary (S. Jégou, M. C. Tonon, F. Leboulenger, C. Delarue, J. Côté, G. Pelletier, and H. Vaudry (1981a). In "Adv. Physiol. Sci.' E. Stark, G. B. Makara, Zs. Acs, and E. Endröczi, eds., Vol. 13, pp. 129-133. Pergamon, Budapest.) do not modify the amount of these peptides in the hypothalamus.

Adrenocorticotropic Hormone↗
PubMed1983

Human growth hormone releasing factor and somatostatin from two pancreatic tumors: isolation and characterization.

Peptides with high intrinsic activity to release growth hormone from pituitary cells in tissue cultures were isolated from two different human pancreatic tumors that had caused acromegaly. Homogeneous peptides were obtained after gel filtration and two steps of reverse-phase high-performance liquid chromatography. From one tumor a 44-residue peptide (human pancreas growth hormone releasing factor, hpGRF-44) was isolated, together with two shorter fragments of reduced bioactivity having 40 and 37 amino acid residues (hpGRF-40, hpGRF-37). In contrast, the other tumor contained only one form of GRF which proved to be identical to hpGRF-40. These hpGRFs are indistinguishable from partially purified preparations of hypothalamic growth hormone releasing factor of human, porcine and murine origins with respect to biological activity and are very similar in their physicochemical properties (molecular weight, retention behavior on reverse-phase HPLC, absence of sulfhydryl groups). One of the pancreatic tumors also contained two forms of immunoreactive somatostatin. One form, after isolation and partial microsequencing, was identified as somatostatin-14 with a structure identical to that of the peptide found in other species. The second form has tentatively been identified as somatostatin-28 on the basis of chromatographic behavior.

Adenoma, Islet Cell↗
PubMed1983

Immunocytochemical localization of prosomatostatin fragments in maturing and mature secretory granules of pancreatic and gastrointestinal D cells.

Pancreatic and gastrointestinal D cells were examined by immunocytochemistry using antisera against somatostatin-28 (SS28) and its NH2-terminal fragment SS28-(1-12), followed by the staphylococcal protein A-gold (pAg) complex. In pancreatic and gastric D cells incubated with antiserum against SS28-(1-12) the gold particles produced intense staining of the mature secretory granules but weaker staining of the immature granules associated with the Golgi area, whereas after SS28 antiserum treatment the particles accumulated selectively over the population of immature secretory granules. In intestinal D cells not only SS28-(1-12) but also SS28 antiserum produced an intense gold staining over the mature delta granules. These observations show that the relative amounts of immunoreactive sites related to SS28 and its cleavage product SS28-(1-12) in maturing and mature secretory granules are different in pancreatic, gastric, and intestinal D cells.

Humans↗
PubMed1983

Cloning and sequence analysis of cDNA for the precursor of human growth hormone-releasing factor, somatocrinin.

Molecular cloning has established the primary structures of two precursors of the human pancreas growth hormone-releasing factor (hpGRF-44), somatocrinin. Both polypeptides contain the sequence of hpGRF-44 flanked by basic processing sites. Furthermore, the precursors include a putative signal sequence and a carboxyl-terminal amidation signal for hpGRF-44. The two forms of mRNA code for pre-pro-GRF-107 and pre-pro-GRF-108. Pre-pro-GRF-108 differs from pre-pro-GRF-107 by the insertion of a serine in the carboxyl-terminal portion of the precursor. In vitro translation of tumor poly(A)+ RNA followed by immunoprecipitation with GRF-specific antiserum and gel electrophoresis showed the molecular weight of preprosomatocrinin to be approximately 13,000, which is in good agreement with the molecular weight deduced from the sequences of the cDNA clones.

Amino Acid Sequence↗
PubMed1983

Somatocrinin receptor coupled with cAMP-dependent protein kinase on anterior pituitary granules.

The molecular mechanism of growth hormone release by synthetic somatocrinin was investigated on purified hog anterior pituitary secretory granules; the granules were found to contain a cAMP-dependent protein kinase that catalyzed [gamma-32P]-ATP histone phosphorylation with maximal rates ranging from 1 to 5 nmol of Pi incorporated per mg of protein per 20 min. The activity of this enzyme was further stimulated by somatocrinin. Stimulation was observed at concentrations as low as 0.3 pM, and the half-maximal effect was obtained with 35 +/- 8 pM (n = 4). Michaelis-Menten analysis of phosphorylation kinetics suggested that the peptide did not change significantly the reaction's Vmax, but produced a dramatic increase in enzyme affinity for cAMP: the apparent Km for the nucleotide decreased from 400 X 10(-9) M under unstimulated conditions to 15 X 10(-9) M in the presence of 100 pM somatocrinin. Furthermore, a Hill plot of concentration-dependence curve indicated the existence of negative cooperativity. At the concentration of 35 pM, the less potent analogs of somatocrinin [designated hpGRF-44 to indicate source (human pancreas, hp), activity (growth hormone-releasing factor, GRF), and amino acid composition], hpGRF-(1-37) and [Phe1]hpGRF-(1-40) had 20% and 7%, respectively, of the effect of somatocrinin. The biologically inactive analog hpGRF-(2-40) had no evident effect at concentrations up to 0.1 microM. Therefore, we suggest that somatocrinin stimulation of growth hormone release involves activation of exocytosis through a phosphorylation mechanism mediated by a granular receptor coupled with a cAMP-dependent protein kinase.

Animals↗
PubMed1983

Somatostatin-28 and somatostatin-14 suppression of arginine-, insulin-, and TRH-stimulated GH and PRL secretion in man.

GH and PRL responses to arginine infusion and to the combined pituitary stimulation test (0.1 u/kg of insulin, 200 micrograms of TRH, 100 micrograms of LHRH) were compared in five normal men during infusions of somatostatin-14 (SS-14) and somatostatin-28 (SS-28), and during control infusions of vehicle alone. SS-14 and SS-28 were infused at a rate of 1.8 nmol/kg over 210 min. Arginine (0.5 g/kg) was infused from 30 to 60 min and the combined pituitary stimulation test commenced at 120 min. Arginine (0.5 g/kg) infused from 30 to 60 min induced an increase in GH secretion in all subjects and this increase was completely abolished in these same subjects when infused with SS-14 and SS-28. Arginine-induced hyperglycaemia was significantly greater during infusion of SS-14 and further enhanced by infusion of SS-28. A small increase in PRL secretion occurred after arginine infusion and this was not inhibited by SS-14 or SS-28. Insulin and TRH administration induced marked increases in both GH and PRL secretion. The mean GH increase was significantly inhibited by SS-14 and SS-28 up to 165 min but not thereafter. The PRL increase was significantly inhibited by SS-28 but not by SS-14 and this greater efficacy was also indicated by administering different doses of SS-28 to one subject. Taken together with the demonstration that SS-28 is released from the median eminence, these findings indicate that SS-28 has a hormonal role in the regulation of GH and PRL secretion.

Adult↗
PubMed1983

A noninvasive functional lesion of the hypothalamo-pituitary axis for the study of growth hormone-releasing factor.

Rats were passively immunized with an antiserum against somatostatin and a monoclonal antibody against rat hypothalamic growth hormone-releasing factor (rGRF-mAb) which does not recognize the biologically active 1-40 amino acid fragment of hpGRF-44 (hpGRF-40). Using this paradigm we have observed that the pituitary possesses a resilient capacity to release growth hormone (GH) following repeated injections of hpGRF-40 and that this response follows a dose-dependent relationship.

Animals↗
PubMed1983

Immunocytochemical evidence for the presence of gamma 1-MSH-like immunoreactivity in pituitary corticotrophs and ACTH-producing tumours.

The presence of gamma 1-MSH has been demonstrated in bovine neuro-intermediate lobe by biochemical methods, thus suggesting that this peptide is cleaved from the cryptic region of pro-opiocortin. In this study we report the localisation of gamma 1-MSH-like immunoreactivity in the adenohypophysis of man, ox, pig, dog and guinea-pig using immunocytochemical procedures at both light and electron microscope levels. Antisera recognising the C-terminal Arg-Phe-amide and the C-terminal penta-peptide-amide of gamma 1-MSH have been used throughout this study. The immunostaining was found in all endocrine cells of the pars intermedia (where present) and in scattered cells of the pars distalis identified as corticotrophs. No gamma 1-MSH immunoreactivity was detected in rat adenohypophysis. In addition, 7 ACTH-producing tumours (1 pituitary adenoma and 6 ectopic) were investigated and shown to contain gamma 1-MSH immunoreactive cells.

ACTH Syndrome, Ectopic↗
PubMed1983

The absence of an age-related change in the pituitary response to growth hormone-releasing factor in rats.

The capability of the anterior pituitary gland to secrete growth hormone (GH) in response to an intravenous injection of growth hormone-releasing factor, hpGRF-44, was evaluated in male rats ranging from 22 days to 24 months of age. The increase in GH concentrations following a submaximal dose of hpGRF-44 (0.5 microgram/kg) was similar among the different aged rats suggesting there is no age-related change in pituitary sensitivity to hpGRF-44. Likewise, the response to a maximal dose of this peptide (25 micrograms/kg) was not different in young and old animals indicating there is no decrease in the readily releasable pool of GH. Thus, the age-related changes in GH secretion observed in rats do not appear to be due to a change in the pituitary response to the hypothalamic growth hormone-releasing factor.

Age Factors↗
PubMed1983

Human pancreas GH-releasing factor analog restores high-amplitude GH pulses in CNS lesion-induced GH deficiency.

Lesions of the ventromedial-arcuate (VMH-ARC) region of the hypothalamus result in impaired growth accompanied by a marked suppression in spontaneous GH secretory bursts. We studied the effects of an analog of the recently characterized human pancreas GH-releasing factor hpGRF(1-40) on GH secretory dynamics in freely-moving chronically cannulated rats bearing electrolytic lesions of the VMH-ARC. Intravenous administration of the hpGRF analog (hpGRFa) caused a dramatic surge of GH within 5 min; plasma GH levels rose to values as high as 2900 ng/ml and remained significantly elevated for 15-30 min post treatment. The simultaneous iv administration of somatostatin-14 and hpGRFa resulted in a significant inhibition of the hpGRFa-induced GH release at 5 min but not at 15 min. These results clearly demonstrate that impaired GH secretion resulting from VMH-ARC lesions can be restored by hpGRF. The findings are promising in that hpGRF and its analogs may provide valuable agents for the diagnosis and treatment of disorders of growth secondary to CNS dysfunction.

Animals↗
PubMed1983

Pituitary adenomas that caused Cushing's disease or Nelson's syndrome are not responsive to ovine corticotropin-releasing factor in vitro.

The response of pituitary adenomas obtained surgically from patients with Cushing's disease of Nelson's syndrome to synthetic ovine corticotropin-releasing factor (CRF), vasopressins, somatostatin-28, dexamethasone, 3-isobutylmethylxanthine or high [K+] was examined in vitro by measuring the amount of pro-opiomelanocortin (POMC)-derived peptides secreted into the culture medium. CRF did not stimulate the secretion of adrenocorticotropin-, beta-endorphin-, or gamma 3-melanotropin-like peptides from the pituitary adenomas at concentrations ranging from 1 x 10(-13) M to 1 x 10(-7) M whereas vasopressins, 3-isobutyrl-methylxanthine and high [K+] increased, while somatostatin-28 and dexamethasone suppressed, the secretion of these POMC-derived peptides. These findings suggest that either the pituitary ACTH-producing tumors have lost their receptors to CRF or their post-receptor mechanism to CRF is not functional.

Adenoma↗
PubMed1983

Inhibition of pancreatic hormone secretion by somatostatin-28 and somatostatin-14 in man.

The effects of a 210 min infusion of 1.8 nmol/kg somatostatin-14 (SS-14), somatostatin-28 (SS-28), and vehicle (Haemaccel) alone, on arginine- and insulin-stimulated release of pancreatic hormones were tested in 5 normal male subjects. Arginine administered at 30-60 min induced an increase in plasma glucose concentrations which was enhanced by SS-14 and further increased by SS-28. SS-28 was more effective than SS-14 in suppressing the arginine-induced secretion of insulin. Arginine-stimulated and insulin-stimulated (at 120 min) glucagon release was equally suppressed by SS-14 and SS-28, as was insulin-stimulated pancreatic polypeptide secretion. At the end of the SS-14 infusion the mean plasma somatostatin level was approximately 28% of that which occurred during the SS-28 infusion. The results are discussed in relation to similar studies in vitro and in vivo in laboratory animals and to a possible role of the two forms of SS in carbohydrate homeostasis.

Adult↗