Since the discovery of the structures of somatostatin (GIF) in 1973 by Brazeau et al, its measurement by the radioimmunoassay (RIA) methods has been reported by Arimura et al (1975), Yanaihara et al (1978) and Sawano et al (1978).As GIF does not contain tyrosine and histidine, which can be radioiodized, analogues of GIF are being used as tracers in its radioimmunoassay. In this study, two different types of trafcers (125 I tyr8of GIF and 125I-tyrosyl-GIF) were used in RIA to measure the immunoreactive GIF of fetal tissues, and their results were compared.A highly specific anti-GIF serum was generated by immunizing rabbits with GIFconjugated BSA.125 I-tyr8-GIF and 125I-tyrosyl-GIF were prepared by the lactoperoxidase method. The former was purified through a carboxymethylcellulose column and the latter through a Sephadex G-25 (fine) column. They were then incubated at 4°C in 0.01 M phosphate buffer saline containing 0.1% geratin and 0.2% BSA and used as tracers to measure the immunoreactive GIF-like substances in the pituitary gland, hypothalamus, pancreas, cerebrum, cerebellum, thyroid gland, stomach, small intestine and spinal cord of three fetuses obtained through legal abortion with gestation ages of 22 weeks, 23 weeks and 25 weeks. The free and bound forms were separated with the double antibody technique.The correlation of these GIF concentrations by the two different assay systems with results such as Y=0.905X- 87.4, r=0.992 and P<0.001 was demonstrated.From the displacement curves of the GIF-analogues (tyrosyl-GIF and tyr1 -GIF) and the standard curves of the synthetic GIF, it was found that when 125I-tyr8-GIF was used as the tracer, the anti-GIF serum could identify both the GIF-analogues and the synthetic GIF as the same substance; however, when 125I-tyrosyl-GIF was substituted as the tracer, the anti-GIF serum could only identify the tyrosyl-GIF and the synthetic GIF as being the same substance and excluded the tyr1 -GIF when its concentration reached beyond a certain level.The dilution curves of the immunoreactive-GIF extracted from the hypothalamus and pancreas of the fetuses were found to be parallel to the standard curves obtained by the synthetic GIF in both asssay systems.It is concluded that the two different assay systems which were used to measure the GIF concentrations in the fetuses can yield the same results and are useful in the application of GIF-RIA, but 125 I-tyr 8 -GIF is more stable than the 125I-tyrosyl-GIF.

With the use of the radioimmunoassay for growth hormone--releasing inhibiting factor (GIF), it was found that measurable amounts of GIF-like substance existed in the chorionic villi and decidua of pregnant women. The indirect immunofluorescent method revealed that the higher intensity of GIF-like immunofluorescence was presented in cytotrophoblasts rather than in syncytiotrophoblasts of the villi and in stromal cells of the decidua.

Since Arimura et al (1975) reported the radioimmunoassay for somatostatin (GIF), the concentration of GIF in various organ and brain regions were determined by radio-immunoassay. Dubois et al (1975) reported that immunohistochemically somatostatin was located in the discrete cells of the pancreas as well as the hypothalamus, and from this result, they presented the concept of local hormone instead of systemic hormone which was up to that time accepted in endocrinology. In this study, we developed the high specific anti-GIF serum using rabbits, and with the micro immunodiffusion method, we demonstrated that the precipitin band formed a circular fusion between the GIF and anti-GIF serum. This pattern of reaction was also seen in decidual immunodiffusion. In addition, we developed the radioimmunoassay for GIF using this anti-serum and measured immunoreactive GIF-like substances in villi and decidua of early pregnancy. The concentration of GIF-like substances with 2 N acetic acid extracted of villi and decidua were 0 to 30 pg/0.1 g dry weight. At the same time, we demonstrated the presence of GIF-like substance-containing cells in the villi and decidua by indirect immunofluorescent method. The intensity of immunofluorescence was in cytotrophoblast rather than syncytiotrophoblast, and decidual stromal cell also reacted to the immunofluorescence.

Addition of small amounts of pure or highly purified porcine GH-RH twice daily to rat anterior pituitary tissue, maintained for 5 days in Trowell's T8 medium, significantly increased the amounts of GH in the medium and tissues relative to the controls as measured by bioassay. Apparent incorporation of radioactive amino acids into the GH bands on polyacrylamide gels after electrophoresis of medium and tissue was increased also. These results support other findings from this laboratory on the identity and properties of this neurohormone.

Electron microscope studies of pituitary somatotrophs of female Sprague-Dawley rats were conducted following intracarotid injection of 2 μg of highly purified pig GRF. Animals were sacrificed by decapitation 0.5, 1, 2.5, 5 and 15 min after GRF injection. Saline-injected control animals were sacrificed according to the same schedule. Tissues were fixed in phosphate-buffered osmium tetroxide. A marked increase in the number of secretory granules undergoing extrusion into the perivascular space from the somatotroph, as compared to controls, was found to occur as early as 2.5 min after GRF injection. In mammotrophs, the number of granules undergoing extrusion also seemed somewhat larger in the GRF-injected groups. These observations corroborate the accepted morphologic identification of the somatotrophs and provide evidence that GRF acts on these cells. (Endocrinology 85: 1084, 1969)

Isolation is described on a preparative scale of growth hormone-releasing hormone (GRH) from 200,000 porcine hypothalami. Boiled, lyophilized 2N acetic acid extracts of hypothalamic tissue were re-extracted with glacial acetic acid. The GRH activity in the concentrate, as followed by in vivo and in vitro bioassays, was subjected to purification by several successive methods. Preparative gel filtration on a large column of Sephadex G-25 in batches of 45–70 g, followed by rechromatography in the same system, led to separation of GRH from other hypothalamic releasing hormones and from vasopressin. Subsequent purification steps included free-flow electrophoresis for 96 hr at pH 6.3, ion-exchange chromatography on carboxymethyl, diethylaminoethyl and triethylaminoethyl celluloses utilizing volatile ammonium acetate buffers, and finally partition chromatography using Sephadex as a support. The peptide with GRH activity, thus purified about 56,000 times, released GH in rats in vivo at doses of 1 nanogram and in ...

ABSTRACT A single subcutaneous injection of 1 mg cortisone or cortisol acetate was given to rats during the first post-natal day. As others have reported, retardation of growth, detectable a week after birth, resulted. On reaching five to six weeks of age, treated rats had no detectable growth hormone (GH)-releasing activity in their hypothalami and little GH or thyrotrophin (TSH) activity in their pituitaries. These observations suggest that the growth retardation of rats treated neonatally with these steroids may be due, at least in part, to lack of pituitary GH resulting from impaired synthesis of GH-releasing hormone (GRH**) and also to lack of TSH.

Addition of small amounts of highly purified porcine growth hormone-releasing factor (GRF) to rat anterior pituitaries incubated in vitro for 6–24 hr in Krebs-Ringer bicarbonate media significantly increased the release of growth hormone (GH). This effect was abolished by actinomycin D. Although more GH was released in vitro under the influence of GRF, the GH content of pituitaries from normal rats exposed to GRF was essentially unchanged after the incubation. When the pituitaries of donor rats were depleted of GH by a prior exposure of rats to cold in vivo, GRF was shown to stimulate the synthesis of GH in the incubated tissue as well as the release of GH. The results lend support to the thesis that one substance is responsible for both the release and synthesis of GH. (Endocrinology82: 271, 1968)

Since our group has been scheduled to deliver the concluding presentation, we would like to mention topics not discussed by previous speakers. For this reason, our presentation had to be improvised in part during the preceding talks, so that it will be composed of many topics.

ABSTRACT An attempt was made to evaluate the growth hormone (GH)-releasing activity in extracts of the stalk-median eminence region (SME) from kittens with bilateral lesions of the paraventricular nuclei, which showed a marked growth retardation, and from sham-operated animals. SME extract from the sham-operated control kittens induced a significant decrease in pituitary GH content. However, SME extract from kittens with bilateral lesions in the region of the paraventricular nuclei failed to induce the depletion of pituitary GH content. The cerebral cortical extracts from both groups elicited no effect on pituitary GH. These results show that the hypothalamus of the lesioned kittens has no GH-releasing activity, i. e. GH-releasing factor (GRF) is lacking. This suggests that, in kittens, the area of the paraventricular nuclei plays an important role in the synthesis of GRF, so that destruction of this area causes growth impairment.

Research Articles| March 18 2008 Studies on Retarded Growth of Rats with Hereditary Hypothalamic Diabetes Insipidus Subject Area: Endocrinology , Neurology and Neuroscience A. Arimura; A. Arimura Department of Medicine, Tulane University School of Medicine and Veterans Administration Hospital, New Orleans, La., USA Search for other works by this author on: This Site PubMed Google Scholar S. Sawano; S. Sawano Department of Medicine, Tulane University School of Medicine and Veterans Administration Hospital, New Orleans, La., USA Search for other works by this author on: This Site PubMed Google Scholar T.W. Redding; T.W. Redding Department of Medicine, Tulane University School of Medicine and Veterans Administration Hospital, New Orleans, La., USA Search for other works by this author on: This Site PubMed Google Scholar A.V. Schally A.V. Schally Department of Medicine, Tulane University School of Medicine and Veterans Administration Hospital, New Orleans, La., USA Search for other works by this author on: This Site PubMed Google Scholar Neuroendocrinology (1968) 3 (3): 187–192. https://doi.org/10.1159/000121707 Article history Received: January 17 1968 Accepted: February 26 1968 Published Online: March 18 2008 Content Tools Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn Email Tools Icon Tools Get Permissions Cite Icon Cite Search Site Citation A. Arimura, S. Sawano, T.W. Redding, A.V. Schally; Studies on Retarded Growth of Rats with Hereditary Hypothalamic Diabetes Insipidus. Neuroendocrinology 1 March 1968; 3 (3): 187–192. https://doi.org/10.1159/000121707 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsNeuroendocrinology Search Advanced Search Article PDF first page preview Close Modal This content is only available via PDF. 1968Copyright / Drug Dosage / DisclaimerCopyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements. You do not currently have access to this content.

Summary The effect of administration of highly purified pig GH-releasing factor (GRF) on the depletion of pituitary growth hormone (GH) content in recipient rats and levels of plasma GH-like activity were investigated using the tibia test method. It was demonstrated that GRF induced a significant increase in plasma GH-like activity which was accompanied by a simultaneous decrease in pituitary GH content. The levels of plasma GH-like activity in recipient rats treated with saline were not detectable (less than 1.25 μg/ml). These results seem to confirm that the “depletion” of pituitary GH content in rats caused by GRF indicates the “release” of GH.

Annals of the New York Academy of SciencesVolume 148, Issue 2 p. 372-388 GROWTH HORMONE-RELEASING FACTOR (GRF): PHYSIOLOGICAL AND BIOCHEMICAL STUDIES WITH GRF PREPARATIONS OF BOVINE AND PORCINE ORIGIN* A. V. Schally, A. V. Schally Merck Institute for Therapeutic Research, Rahway, N.J.Search for more papers by this authorE. E. Müller., E. E. Müller. Merck Institute for Therapeutic Research, Rahway, N.J. Endocrine and Polypeptide Laboratories, Veterans Administration Hospital and Tulane University School of Medicine, New Orleans, La.Search for more papers by this authorA. Arimura, A. Arimura Merck Institute for Therapeutic Research, Rahway, N.J.Search for more papers by this authorT. Saito, T. Saito Merck Institute for Therapeutic Research, Rahway, N.J. Present address: Dept. of Pharmacology, University of Siena, Siena, Italy.Search for more papers by this authorS. Sawano, S. Sawano Merck Institute for Therapeutic Research, Rahway, N.J.Search for more papers by this authorC. Y. Bowers, C. Y. Bowers Merck Institute for Therapeutic Research, Rahway, N.J.Search for more papers by this authorS. L. Steelman, S. L. Steelman Merck Institute for Therapeutic Research, Rahway, N.J.Search for more papers by this author A. V. Schally, A. V. Schally Merck Institute for Therapeutic Research, Rahway, N.J.Search for more papers by this authorE. E. Müller., E. E. Müller. Merck Institute for Therapeutic Research, Rahway, N.J. Endocrine and Polypeptide Laboratories, Veterans Administration Hospital and Tulane University School of Medicine, New Orleans, La.Search for more papers by this authorA. Arimura, A. Arimura Merck Institute for Therapeutic Research, Rahway, N.J.Search for more papers by this authorT. Saito, T. Saito Merck Institute for Therapeutic Research, Rahway, N.J. Present address: Dept. of Pharmacology, University of Siena, Siena, Italy.Search for more papers by this authorS. Sawano, S. Sawano Merck Institute for Therapeutic Research, Rahway, N.J.Search for more papers by this authorC. Y. Bowers, C. Y. Bowers Merck Institute for Therapeutic Research, Rahway, N.J.Search for more papers by this authorS. L. Steelman, S. L. Steelman Merck Institute for Therapeutic Research, Rahway, N.J.Search for more papers by this author First published: February 1968 https://doi.org/10.1111/j.1749-6632.1968.tb20364.xCitations: 5 * Supported in part by USPHS grant AM 08743-01, 02, 03. AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat References 1 DEUBEN, R. & J. MEITES 1964. Endocrinology 74: 408. 2 SCHALLY, A. V., S. L. STEELMAN & C. Y. BOWERS 1965. Proc. Soc. Exp. Biol. Med. 119: 208. 3 SCHALLY, A. V., S. L. STEELMAN & C. Y. BOWERS 1964. Endocrine Society Program, p. 143. 4 PECILE, A., E. E. MÜLLER and G. FALCONI and L. MARTINI 1965. Endocrinology 77: 241. 5 ISHIDA, Y. and A. KUROSHIMA, C. Y. BOWERS & A. V. SCHALLY 1965. Endocrinology 77: 759. 6 MÜLLER, E. E., S. SAWANO & A. V. SCHALLY 1967. Gen. Comp. Endocrinology In press. 7 MÜLLER, E. E. & A. PECILE 1965. Proc. Soc. Exp. Biol. Med. 119: 1191. 8 KRULICH, L., A.P.S. DHARIWAL & S. M. MCCANN 1965. Proc. Soc. Exp. Biol. Med. 120: 180. 9 MÜLLER, E. E. & A. PECILE 1966. Endocrinology 79: 448. 10 SCHALLY, A. V., E. E. MÜLLER, A. ARIMURA, C. Y. BOWERS, T. SAITO, T. W. REDDING & S. SAWANO 1967. J. Clin. Endocr. 27: 755. 11 KNOBIL, E. 1966. Physiologist 9: 25. 12 SCHALLY, A. V., A. KUROSHIMA, Y. ISHIDA, A. ARIMURA, T. SAITO, C. Y. BOWERS & S. STEELMAN 1966. Proc. Soc. Exp. Biol. Med. 122: 821. 13 DHARIWAL, A. S. P., L. KRULICH, S. H. KATZ & S. M. MCCANN 1965. Endocrinology 77: 932. 14 DEKANSKI, J. 1952. Brit. J. Pharmacol. 7: 567. 15 THOMPSON, R. E. 1944. J. Pharmacol. Exp. Ther. 80: 373. 16 SHIZUME, K., A. B. LERNER & T. B. FITZPATRICK 1954. Endocrinology 54: 533. 17 SAFFRAN, M. & A. V. SCHALLY 1955. Canad J. Biochem. Physiol. 33: 408. 18 GREENSPAN, F. S., C. H. LI, M. E. SIMPSON & H. M. EVANS 1949. Endocrinology 45: 455. 19 SCHALLY, A. V., C. Y. BOWERS & T. W. REDDING 1966. Endocrinology 78: 726. 20 SCHALLY, A. V., C. Y. BOWERS & T. W. REDDING 1966. Proc. Soc. Exp. Biol. Med. 121: 718. 21 SCHALLY, A. V. & C. Y. BOWERS 1964. Endocrinology 75: 608. 22 PORATH, J. & A. V. SCHALLY 1962. Endocrinology 70: 738. 23 HANNIG, K. 1961. Z. Anal. Chem. 181: 244. 24 LOWRY, O. H., N. J. ROSEBROUGH, A. L. FARR & R. J. RANDALL 1951. J. Biol. Chem. 193: 265. 25 SCHALLY, A. V., A. J. KASTIN, W. CARTER, C. Y. BOWERS, J. F. BARRETT & W. F. WHITE 1966. Biochemical Pharmacol. 15: 1805. 26 GESCHWIND, I. I. & C. H. LI 1957. J. Amer. Chem. Soc. 79: 615. 27 SCHALLY, A. V., R. N. ANDERSEN, J. M. LONG & R. GUILLEMIN 1960. Proc. Soc. Exp. Biol. Med. 104: 260. 28 STEELMAN, S. L., R. N. ANDERSEN & R. M. MCGREGOR 1959. Biochim. Biophys. Acta. 33: 256. 29 SCHALLY, A. V. & R. GUILLEMIN 1960. Texas Reports Biol. Med. 18: 133. 30 BOARDMAN, N. K. & S. M. PARTRIDGE 1955. Biochem. J. 59: 543. Citing Literature Volume148, Issue2Growth HormoneFebruary 1968Pages 372-388 ReferencesRelatedInformation

Hypothalamic extracts from the dog, rabbit, mouse, kitten, pigeon, and frog were found to decrease the growth hormone (GH) content of rat pituitary glands. Comparable cerebral cortical extracts were completely inactive in this test. These results broaden previous observations on the existence of a GH-releasing factor (GRF) in the hypothalami of mammals and show the presence of GRF activity in birds and amphibians.

Follicle-stimulating ho:rmone-releasing factor (FSH-RF) activity in lyophilized acetic acid extracts of hypothalamic tissue of porcine origin was concentrated by re-extraction with glacial acetic acid and then subjected to purification by several successive methods. The fractionation procedure consisted of gel filtration on Sephadex G-25, concentration and desalting by phenol extraction, chromsitography and rechromatography on carboxymethylcellulose and column electrophoresis. FSH-RF activity was measured by depletion of pituitary FSHcontent in castrated male rats pretreated with testosterone as well as in normal male rats. In these experiments highly purified FSH-RF was active in vivo at doses of the order of 10 nanograms. These fractions also released FSH from isolated rat pituitaries incubated in vitro. Purified FSH-RF was free of luteinizing hormone-releasing factor (LRF) and other known releasing factors. The response to FSH-RF was not blocked by actinomycin D nor by antihistamines. (Endocrinology81: 882, 1967)

This study was performed to investigate whether growth hormone-releasing factor (GRF) is present in the posterior pituitary lobe of the rat. The assay for GRF activity consisted of measurement of the depletion of pituitary growth hormone (GH) of recipient rats following an intracarotid injection. Since the extracts from 8 rat posterior pituitary lobes contain over 800 mU vasopressin and this large dose of vasopressin depletes the pituitary of GH, an effort was made to inactivate vasopressin without affecting GRF activity. Thioglycollate solution, which destroyed the biological activities of vasopressin, also depleted pituitary GH in the recipient rat. This effect was counteracted by adding cystine or glutathione to the thioglycollate solutions before the injection. GRF activity of a purified GRF preparation from pig hypothalami and of a rat pituitary stalk-median eminence extract (SME) was not abolished by thioglycollate treatment. Posterior pituitary extract, equivalent to 8 posterior lobes, showed GRF activity, but this activity was abolished by thioglycollate. Vasopressin treated in a similar way did not show GRF activity. The data suggest that the depletion of pituitary GH content effected by posterior pituitary extract may be ascribed to vasopressin and that GRF similar to that in the hypothalamus is lacking in the posterior lobe. (Endocrinology81: 1165, 1967)

Previous studies have shown that reserpine (1 mg/kg ip) completely blocks insulininduced growth hormone (GH) release. The possibility that the suppressive action of this drug on GH secretion might be due to its catecholaminedepleting action was investigated in the present experiments. α-Methyldopa (300 mg/kg ip), amethyl- m-tyrosine (300 mg/kg ip), or tetrabenazine (15 mg/kg ip), like reserpine, were effective in suppressing the release of GH induced by 2 U/kg of insulin. Guanethidine (30 mg/kg ip) and tyramine (15 mg/kg ip) were without effect. The blockade of GH release induced by reserpine (1 mg/kg ip) was reversed by iproniazid (150 mg/kg sc). After reserpine treatment (1 mg/kg ip), the release of growth hormone-releasing factor (GRF) from the hypothalamus induced by insulin (2 U /kg ip) was abolished. From these experiments it is concluded that: 1. The reduction of stores of brain norepinephrine impairs GH release; 2. brain amines might play a role in the release of hypothalamic neurohumoral transmitters. (Endocrinology80: 471, 1967)