Bertrand Bloch

Human hypothalamic growth hormone releasing factor (GRF): Evidence for two forms identical to tumor derived GRF-44-NH2 and GRF-40

Human hypothalamic growth hormone-releasing factor (GRF) was purified by gel filtration and reverse-phase HPLC. Bioassay and two radioimmunoassays of different specificity revealed the presence of two major forms of GRF-activity which coelute with human pancreas GRFs, hpGRF-44-NH2 and hpGRF-40 previously characterized in pancreas tumors. The bioactive material coeluting with hpGRF-44-NH2 is recognized by two antibodies which are directed against the amidated COOH-terminal sequence and the central portion of the GRF-44 peptide. The bioactive GRF which coelutes with hpGRF-40 reacts only with the antibody directed against the central portion of hpGRF. These data strongly suggest that the human hypothalamus contains the same major forms of GRF that were identified in pancreas tumors responsible for acromegaly in the absence of a pituitary tumor.

Topographical and ontogenetic study of the neurons producing growth hormone-releasing factor in human hypothalamus

Neurons producing growth hormone-releasing factor have been characterized and analyzed by immunohistochemistry in the hypothalami of human fetuses, neonates, infants and adults, using two antibodies against human pancreatic GRF (hpGRF). One of the antibodies recognized both the hpGRF(1-40)OH and hpGRF(1-44)NH2 in the mid portion (between the 28th and 39th amino acid), the other one specifically recognized the C-terminal end of hpGRF(1-44)NH2. These two antibodies stain a single neuronal system with cell bodies mainly located in the infundibular (arcuate) nucleus, and in the ventromedial and lateralis tuber nuclei. These neurons project to the median eminence where they give numerous endings in contact with portal vessels. These neurons are distinct from those containing LH-RH, somatostatin, CRF or pro-opiocortin. In fetuses, neurons immunoreactive with hpGRF antibodies are first detected at the 29th week. They display a neuroblastic aspect which persists after birth. Immunoreactive fibers are detectable in the median eminence after the 31st week. These results demonstrate that the infundibular nucleus plays a major role in control of GH secretion in man and that secretion of GRF appears late during fetal life; this suggests that the first stages of differentiation and development of GH producing cells in the human fetus do not depend on hypothalamic GRF secretion.

Somatocrinin, the growth hormone releasing factor

Publisher Summary This chapter discusses the isolation of tumor-derived and hypothalamic GRFs, structure-activity relationships of synthetic replicates of GRFs, in vitro studies on the mechanism of action of GRF, and antagonism between GRF and somatostatin. Clinical interest in GRF extends over its use as a diagnostic tool and a treatment of hypothalamic dwarfism. GRF helps to promote anabolism in chronic debilitating diseases, as long as the dietary intake is adequate, and to promote the healing of wounds and bone fractures. The availability of GRF with its highly specific effect in stimulating GH secretion should permit investigations of the proposed role of GH in diabetic retinopathy. Structural analogs of GRF acting as competitive antagonists may be of major clinical significance in the treatment of accidents of juvenile diabetes.

Ectopic secretion of a growth hormone-releasing factor: Report of a case of acromegaly with bronchial carcinoid tumor

Rarely, acromegaly is produced by neuroendocrine neoplasms elaborating a substance similar to or identical with growth hormone-releasing factor. This report reviews the cases described to date and presents the clinicopathologic features of a patient with acromegaly, mild sellar enlargement, and elevated growth hormone levels associated with a large bronchial carcinoid tumor. Normalization of serum growth hormone levels and regression of acromegaly followed resection of the bronchial tumor, which was shown, by bioassay and immunocytochemistry, to contain a growth hormone-releasing factor.

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.

Growth hormone-releasing factor: chemistry and physiology.

Conclusions In conclusion, the long-sought growth hormone-releasing factor has been characterized and sequenced in several species (human, porcine, bovine, and murine). The noteworthy achievement with GRF has been the rapidity with which the sum of information has been gathered. In the space of 12 months most of the pioneering studies on the mechanism of action of the peptide in vivo and in vitro were described. Immunocytochemical mapping of GRF neurons was carried out. Structure-function studies were initiated. Clinical trials were started and confirmed the potent GH-releasing activity of GRF in man. The effect of the peptide on specific GH mRNA levels was described and molecular cloning was used to establish the structures of human (pancreas) preproGRF. All the hypothalamic releasing factors which had been postulated in the early fifties as humoral regulators of the secretion of each pituitary hormone have now been characterized.

Multihormonal carcinoid tumor of the pancreas. Secreting growth hormone‐releasing factor as a cause of acromegaly

This report describes the histologic, immunocytochemical, and ultrastructural study of a multihormonal carcinoid tumor of the pancreas, secreting a growth hormone releasing factor (GRF) which provoked acromegaly. The patient presented a nonfamilial multiple endocrine neoplasia, type 1. The absence of radiologic signs of a pituitary adenoma in conjunction with elevated plasma levels of pancreatic polypeptide, glucagon, somatostatin, as well as growth hormone (GH), led to the discovery of the tumor. Its surgical excision produced a rapid disappearance of most of the clinical and biologic disorders. No immunoreactive GH was found in the tumor using radioimmunoassay and immunocyto‐chemistry. In contrast, three peptides with GH‐releasing activity were extracted and characterized. Immunocytochemistry showed that the GRF‐reactive cells, together with rare somatostatin‐storing cells, made up areas which demonstrated a medullary pattern of growth with extracellular amyloid deposits. Under electron microscopic examination, actively secreting cells were observed which carried endocrine granules of 100 to 150 nm in diameter. The other regions of the tumor presented a different type of growth and were composed of pancreatic polypeptide‐, glucagon‐, or somatostatin‐reacting cells. Cells immunostained with antisera raised against β‐endorphin were also noted. These data suggest that GRF may be a new biologic marker for pancreatic endocrine tumors.

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.

Detection of the messenger RNA coding for preproenkephalin A in bovine adrenal by in situ hybridization

The messenger RNA (mRNA) coding for the adrenal precursor of enkephalins (preproenkephalin-A) has been detected in bovine adrenal medulla cells using in situ hybridization with 32P-labelled preproenkephalin A (PPA) complementary DNA. In formaldehyde- and Carnoy-fixed tissue sections, an intense elective labelling restricted to the cells located at the periphery of the adrenal medulla can be detected after hybridization procedure, using X-ray film and classical autoradiographic procedure. Adequate controls show that this labelling is obtained only using PPA complementary DNA, inserted or not in its vector. Distribution of PPA mRNA appears identical to that of its immunoreactive end products, namely Met-enkephalin and BAM22 peptide, detected by immunohistochemistry. Norepinephrine, detectable using monoamine histofluorescence, appears restricted to the cells of the center of the gland unlabelled for PPA mRNA and its end-products. Cultured bovine adrenomedullary cells that exhibited enkephalin immunoreactivity also contain PPA mRNA located in their cytoplasm.

Pituitary response to growth hormone-releasing factor in rats with functional or anatomical lesions of the central nervous system that inhibit endogenous growth hormone secretion

The pituitary growth hormone (GH) response to the growth hormone-releasing factor, hpGRF-44, was evaluated in male rats with various lesions of the central nervous system. These included an electrical lesion of the ventromedial hypothalamus, a chemical lesion of the arcuate nucleus induced by neonatal treatment with monosodium glutamate, a functional lesion of catecholamine synthesis with alpha-methyl-p-tyrosine or a functional lesion of catecholamine storage with reserpine. The first three lesions appear to partially inhibit normal somatostatin secretion since in every instance hpGRF-44 administration induced a significant increase in plasma GH concentrations. In contrast, reserpine blocked the GH response to hpGRF-44, presumably by stimulating somatostatin secretion. The pituitary GH response to hpGRF-44 in the above described models was enhanced by pretreatment of the rats with antibodies against somatostatin. The pituitary GH response to repeated injections of hpGRF-44 was also evaluated in rats with an anatomical lesion of the arcuate nucleus or a functional lesion of catecholamine synthesis. The maximum GH response did not vary over time to the repeated injections of hpGRF-44 in rats with lesions of the arcuate nucleus; however, interruption of catecholamine synthesis resulted in a significant decrease in the GH response to hpGRF-44 over time.