A novel avian hypothalamic peptide inhibiting gonadotropin release

The neuropeptide control of gonadotropin secretion at the level of the anterior pituitary gland is primarily through the stimulatory action of the hypothalamic decapeptide, gonadotropin-releasing hormone (GnRH), which was originally isolated from mammals and subsequently from non-mammals. To date, however, an inhibitory peptide of gonadotropin release is unknown in vertebrates. Here we show, in a bird, that the hypothalamus also contains a novel peptide which inhibits gonadotropin release. Acetic acid extracts of quail brains were passed through C-18 reversed-phase cartridges, and then the retained material was subjected to the reversed-phase and cation-exchange high-performance liquid chromatography (HPLC). The peptide was isolated from avian brain and shown to have the sequence Ser-Ile-Lys-Pro-Ser-Ala-Tyr-Leu-Pro-Leu-Arg-Phe-NH(2). Cell bodies and terminals containing this peptide were localized immunohistochemically in the paraventricular nucleus and median eminence, respectively. This peptide inhibited, in a dose-related way, gonadotropin release from cultured quail anterior pituitaries. This is the first hypothalamic peptide inhibiting gonadotropin release reported in a vertebrate. We therefore term it gonadotropin-inhibitory hormone (GnIH).     

Endotoxin administration increases hypothalamic somatostatin mRNA through nitric oxide release

Acute inflammation induced by endotoxin (LPS) administration inhibits insulin-like growth factor (IGF-I) and growth hormone (GH) secretion. The aim of this study was to elucidate the role of glucocorticoids and nitric oxide (NO) in the effect of LPS on hypothalamic somatostatin gene expression. Adult male Wistar rats were injected with different doses of LPS (5, 10 and 100 microg/kg). Rats received two i.p. injections of LPS (at 17:30 and 8:30 h the following day) and were killed 4 h after the second injection. LPS administration at the dose of 100 microg/kg increased the hypothalamic somatostatin mRNA content, as well as the serum concentrations of corticosterone. Glucocorticoids do not seem to be involved in LPS-induced increase in hypothalamic somatostatin mRNA since adrenalectomy did not prevent this effect. In order to analyze the possible effect of NO, aminoguanidine, an inducible nitric oxide synthase inhibitor, was injected (100 mg/kg s.c.) simultaneously with LPS injection. Aminoguanidine administration did not modify somatostatin mRNA in saline injected rats, but it prevented LPS-induced increase in hypothalamic somatostatin mRNA. These data suggest that the stimulatory effect of endotoxin on hypothalamic somatostatin gene expression is not mediated by glucocorticoids, but instead by the increase in NO release.     

半胱胺耗竭体内生长抑素及其机制

半胱胺,又称β巯基乙胺,相当于半胺氨酸的脱羧产物,是辅酶A分子的组成部分,在体内有重要的生理意义。临床上用于治疗放射病、胱氨酸病、扑热息痛中毒和四乙基铅中毒。Selye等发现半胱胺可诱发大鼠十二指肠溃疡。在研究其致溃疡机制过程中,Szabo     

Stimulation of immunoreactive somatostatin release from hypothalamic synaptosomes by high (K+) and dopamine

Effects of high (K+) and dopamine on the release of immunoreactive somatostatin from isolated hypothalamic synaptosomes were studied in rats. High (K+) (60 mM) and dopamine (10(-6) M) in the incubation media stimulated the release of immunoreactive somatostatin and the former effect was completely abolished by the removal of Ca++ from the media. These suggest that hypothalamic somatostatinergic synaptosomes preserved at least one of the important basic properties of secretory cells. Although it is of interest to note that dopamine stimulated the release of somatostatin. Its physiological significance awaits further studies.     

In vitro release of growth hormone-releasing factor (GRF) from the hypothalamus: somatostatin inhibits GRF release.

The manner of release of growth hormone-releasing factor (GRF) from the rat hypothalamus was studied in a perifusion system using a highly sensitive radioimmunoassay for rat GRF. The recovery of GRF in this system was 50-60%. The release of GRF from the rat hypothalamic blocks was almost stable for 20-240 min after the start of the perifusion and was stimulated by depolarization induced by high K+ concentration. The release of GRF was inhibited by somatostatin at concentrations of 10(-11) to 10(-8) M with maximum inhibition to 52.5% of the basal release at a concentration of 10(-9) M. These results suggest that this system is useful in studying the regulatory mechanism of GRF release and that, in addition to its action on the pituitary, somatostatin appears to act at the level of the hypothalamus in inhibiting GRF release in the regulation of GH secretion.     

Synthesis of a nonreducible cyclic analog of somatostatin having only growth hormone release inhibiting activity

A nonreducible cyclic analog of somatostatin (SRIF) was prepared by a combination of solid phase and solution peptide synthesis. The compound, gamma-Abu-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Asp-OH, was tested for its effect on the release of growth hormone, glucagon and insulin in rats. It significantly suppressed pentobarbital-stimulated growth hormone release but showed no effect on arginine-stimulated glucagon or insulin release. The linear form, NH2-gamma-Abu-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Asp-OH, was also prepared and tested in vivo. It was shown to have only slight activity.     

On the isolation of a prolactin inhibiting factor (hormone)

A preparation of $\text{ca.}$ < 100 ng of a prolactin inhibiting factor was isolated which could be essentially pure, because of symmetrical single peaks by high pressure liquid chromatography. The $\text{in vitro}$ activity was at $\text{ca.}$ < 5 ng which is the highest potency reported by anyone. The paucity of $\text{ca.}$ < 100 ng/80,000 hypothalami necessitates patience for definitive data on more product from $\text{ca.}$ 240,000 to 450,000 hypothalami. Weight was estimated by comparing UV absorption at 220 nm with that of synthetic peptides. This preparation is not a catecholamine by chromatography, and gives new and timely credence to the concept that prolactin secretion is mediated by complex mechanisms including a peptide inhibiting factor and a catecholamine.     

Isolation and characterization of the bovine hypothalamic corticotropin-releasing factor

A 41 amino acid peptide with high intrinsic corticotropin-releasing activity was isolated from 1000 bovine hypothalami by means of immunoaffinity chromatography, gel filtration, and two steps of reverse phase HPLC. The primary structure of the amino terminal 39 amino acids was characterized by gas phase sequence analysis. The sequence of the amidated carboxyl terminal dipeptide was established by digestion of the intact natural product with Staphylococcus aureus V8 protease, dansylation of the digest and comparative reverse phase liquid chromatography studies with the synthetic dansylated dipeptides Ile-Ala-NH2, Ile-Ala-OH, Ala-Ile-NH2 and Ala-Ile-OH. The complete structure of the bovine corticotropin-releasing factor was established as: Ser-Gln-Glu-Pro-Pro-Ile-Ser-Leu-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Glu-Val- Leu- Glu-Met-Thr-Lys-Ala-Asp-Gln-Leu-Ala-Gln-Gln-Ala-His-Asn-Asn-Arg-Lys-Leu- Leu- Asp-Ile-Ala-NH2 using approximately 650 pmol of material.     

Effect of recombinant bovine somatotropin (somidobove) in a sustained release vehicle on plasma somatotropin level and lactational performance of dairy cows.

The effects of administration of recombinantly derived bovine somatotropin (somidobove) in a sustained-release vehicle on the profiles of concentrations of bovine somatotropin (bST) in the blood plasma and on the milk yield of dairy cows of three herds were examined. Cows (36-87 days post partum) were treated subcutaneously with recombinant bST at 28-day intervals. In control animals, basal concentrations of bST averaged 1.4 ng.ml-1 in first-calf heifers and 1.5 ng.ml-1 in multiparous cows. In somidobove treated first-calf heifers, the concentration of bST was increased to 10.7, 14.5, and 27.0 ng.ml-1 at 24 h postinjection and in multiparous cows to 6.6, 11.0, and 11.7 ng.ml-1 on day 2 postinjection of 320, 640, and 960 mg of somidobove, respectively. On day 8 postinjection the average plasma bST levels of both parity groups are similar (on the average 3.4, 8.6, and 12.5 ng.ml-1 for three doses of somidobove respectively) and for the two highest doses being still significantly increased. During the 2nd week postinjection plasma bST concentration declined returning to control levels on day 15 postinjection. Somidobove-treated first-calf heifers produced 10.9, 16.7 and 17.9% and multiparous animals 25.5, 24.2 and 32.5% more milk than the controls when given 320, 640 and 960 mg somidobove, respectively. The cyclic pattern in milk yield within each 28-day injection interval was observed consistently in all herds. The milk yield increased to a maximum between day 4 to 8 postinjection and then slowly declined. Milk composition was not affected by somidobove treatment.     

Effect of pentobarbital and urethane on the release of hypothalamic somatostatin and pituitary growth hormone

Hypothalamic somatostatin release was investigated in the rat to elucidate the mechanism of anesthetic action on growth hormone (GH) release from the pituitary. Intraperitoneal injection of sodium pentobarbital (5 mg/100 gm B.W.) significantly elevated serum GH levels and increased hypothalamic somatostatin concentration from basal values of 0.98 +/- 0.01 to 1.21 +/- 0.06 ng/mg wet wt. In contrast, urethane (150 mg/100 gm B.W., IP) administration lowered serum GH levels and hypothalamic somatostatin concentration (0.64 +/- 0.04 ng/mg wet wt.). However, the mean concentration of pancreatic somatostatin showed no change in either case. In rats receiving passive immunization with 0.5 ml rabbit antiserum to somatostatin (SRIF-AS), serum GH levels were significantly increased (67.5 +/- 12.3 ng/ml) and did not differ from those in the group treated with normal rabbit serum (NRS) plus pentobarbital (101.3 +/- 18.5 ng/ml). However, serum GH levels in rats injected with SRIF-AS plus pentobarbital were increased to higher values than in rats given SRIF-AS alone. When urethane was administered to rats after passive immunization with SRIF-AS, urethane-induced suppression of serum GH levels was markedly inhibited (5.5 +/- 2.0 vs. 33.5 +/- 7.5 ng/ml). These results suggest a possibility that the changes in serum GH levels observed with pentobarbital or urethane administration may be induced at least in one part by somatostatin released from the hypothalamus.     

Isolation of a factor that reverses presynaptic inhibition of acetylcholine release

A factor (Substance B) has been isolated from brain which reverses the presynaptically-modulated inhibition of evoked ACh release from both guinea-pig myenteric plexus-longitudinal muscle synaptosomes and the intact strip. Inhibitory modulating agents whose activity is reversed by Substance B include oxotremorine, 2-chloroadenosine, clonidine, and morphine. In addition to brain, Substance B is also present in heart and ileum but not in liver or kidney. As determined by Biogel P2 chromatography, this factor appears to have a molecular weight of around 700. It is not destroyed by preincubation with periodate, amylase, adenosine deaminase, pronase, trypsin, phospholipase C or carboxypeptidase Y.