SRIF Immunoreactivity Research Articles

Somatostatin Receptors Type 2 and 5 Expression and Localization During Human Pituitary Development

Somatostatin (SRIF), by acting mainly through sst2 and sst5 receptors, is a potent inhibitor of hormonal secretion by the human anterior pituitary gland. However, the pattern of protein expression of these SRIF receptors remains unknown during pituitary development. To get further insights into the physiological role of SRIF receptors in human development and pituitary function, the present study examined the developmental expression of the sst2 and sst5 receptors in the individual cell types of the anterior human pituitary. Thirteen fetal human pituitaries were investigated between 13 to 38 weeks of gestation (WG) by double-labeling immunofluorescence with antibodies raised against sst2 or sst5 receptors and GH, LH, FSH, TSH, or pro-opiomelanocortin proteins. SRIF immunoreactivity in the hypothalamus and median eminence was investigated at the same developmental ages. Immunoreactivity for the sst2 receptor was evident as early as 13 to 15 WG and onward mainly in TSH-, LH-, and FSH-expressing cells, whereas sst5 immunoreactivity was apparent at the late development stages (35-38 WG). GH-expressing cells mainly expressed sst5 immunoreactivity. SRIF-positive fibers and cells were detected as soon as 13 to 16 WG in the hypothalamus and median eminence and their densities increased with gestational age. The early appearance of hypothalamic SRIF cells and fibers suggests a physiological link between SRIF and its receptors during pituitary development. Whereas sst2 receptors might play a primary role in the differentiation and regulation of TSH, LH, and FSH cells, sst5 receptors appear to be mainly involved in GH regulation from birth onward.

Immunohistochemical detection of somatostatin receptor types 1-5 in medullary carcinoma of the thyroid.

We have analysed the distribution of the five somatostatin receptors (sst1-5) by immunohistochemistry in a large retrospective series of 51 medullary carcinoma of the thyroid (MCT) specimens and correlated the pattern of sst expression with expression of somatostatin (SRIF) peptide, tumour pathology and clinical outcome. Immunohistochemistry was performed with rabbit polyclonal antipeptide antibodies directed against the extracellular domains or cytoplasmic tail of human (h) sst1-5. SRIF immunoreactivity was investigated in parallel paraffin sections. Eighty-five percent of the tumours were positive for one or more sst, localized to both tumour cells as well as surrounding peritumoural structures, especially blood vessels. Forty-nine percent of the tumours were positive for sst1, 43% for sst2, 47% for sst3, 4% for sst4, and 57% for sst5. Fifty-one percent of tumours expressed one or two sst subtypes; 33% were positive for three or more sst isoforms. All five sst receptors were detected in only two cases. Tumours expressing octreotide sensitive subtypes (sst2,3,5) accounted for 75% of the series. 50% of the tumours co-expressed SRIF suggesting tumour cell regulation by endogenous SRIF via paracrine/autocrine circuits. There was no correlation between sst1-5 expression and age, sex, tumour size or stage, histological type or clinical outcome. Simultaneous analysis of primary tumour and lymph node metastases revealed a similar pattern of sst immunoreactivity indicating that sst expression is not modified in the course of disease progression. With the exception of sst4, medullary carcinoma of the thyroid display a rich but heterogeneous expression of sst subtypes. Immunohistochemical typing of sst receptor expression using specific antireceptor antibodies represents an ideal approach for characterizing sst subtype expression in medullary carcinoma of the thyroid for optimizing receptor targeted diagnosis and therapy with somatostatin analogs.

The over-expression of somatostatin in the gerbil entorhinal cortex induced by seizure

In present study, we investigated the immunohistochemical distribution of somatostatin (SRIF) in the hippocampal complex of the Mongolian gerbil and its association with different sequelae of spontaneous seizures, in an effort to identify the roles of SRIF in the self-recovery mechanisms in these animals. In the dentate gyrus and subiculum, SRIF immunoreactive (SRIF +) cells were similar in both the seizure resistant and the pre-seizure group of seizure sensitive gerbils. Interestingly, SRIF immunoreactivity was markedly decreased until 12 h postictal. Twenty-four hours after the on-set of seizure, the distribution of SRIF immunoreactivity in these regions had slightly increased. In contrast, in the entorhinal cortex the population of SRIF + cells and their density were significantly elevated compared to pre-seizure group 30 min postictal. Twelve hours after the on-set of seizure, however, the population of SRIF + cells and their density declined, approximately 70–80% compared to the situation at 30 min postictal. These findings suggest that the enhancement of SRIF expression in gerbil entorhinal cortex may affect tissue excitability and have a role in modulating recurrent excitation following seizures.

Granulomas in murine schistosomiasis mansoni have a somatostatin immunoregulatory circuit

The role of somatostatin (SRIF) in controlling the granulomatous inflammatory response to infection with the parasite Schistosoma mansoni was explored in mice. The murine granulomas contain SRIF-14. Immunoreactive SRIF and preproSRIF localize in the cytoplasmic granules of macrophages within the granulomas. The granulomas contain mRNA for preproSRIF and are not innervated. The production of SRIF by the inflammatory cells appears to be inducible. The granulomas contain mRNA for the SRIF receptors sst 2A and sst 2B, which are expressed mainly on CD4 + T lymphocytes and bind SRIF-14 with high affinity. Antigens from the schistosome eggs stimulate granuloma T lymphocytes to produce cytokines. Interferon-γ (IFN-γ) is one such cytokine made by CD4 + T lymphocytes. SRIF-14 suppresses antigen-induced IFN-γ production from granuloma cells, and this effect is blocked by anti-sst 2 antibody. SRIF was shown to inhibit IFN-γ-induced immunoglobulin G2a (IgG2a) synthesis in murine schistosomiasis. SRIF also blocks substance P (SP)-stimulated IFN-γ and IgG2a secretion. Schistosome-infected animals treated with the SRIF analog octreotide form smaller granulomas that secrete substantially less IFN-γ and IgG2a. Unpublished observations suggest that SRIF does not modulate schistosome egg antigen- or concanavalin A-stimulated granuloma lymphocyte proliferation in murine schistosomiasis. In conclusion, SRIF may be an important factor in the control of the granulomatous inflammatory response in murine schistosomiasis.

Somatostatin-immunoreactive neurons in the adult rabbit retina.

Somatostatin (SRIF) is a neuroactive peptide that is distributed throughout the nervous system, including the retina. This peptide has been localized to populations of amacrine cells in a variety of vertebrate species. In the rabbit retina, SRIF immunoreactivity is present in a sparse population of medium to large neurons (13.72 microns in diameter, or 147.84 mu 2) in the ganglion cell layer and in a small number of neurons in the inner nuclear layer. These cells display a preferential distribution to the inferior retina, with the highest density near the ventral and ventrolateral retinal margins (11.33 cells/mm2). SRIF-immunoreactive cells have two to five primary processes that arborize in the proximal inner plexiform layer (IPL). These give rise to a plexus of finer processes in the distal IPL. Occasional immunoreactive processes are also present in the outer plexiform layer. In the IPL, these laminar networks are present in all retinal regions. In addition, SRIF-immunoreactive cells often have a fine-caliber axonlike process that eminates from the soma or perisomal region. These processes travel for great distances across the retina in either the nerve fiber layer or in the distal IPL but are never seen to enter the optic nerve head. In addition, the number of SRIF-immunoreactive somata remains unchanged following transection of the optic nerve. Taken together, these data indicate that SRIF-immunoreactive neurons of the rabbit retina are displaced amacrine cells. Furthermore, the sparse distribution of SRIF-immunoreactive somata, the wide-ranging, asymmetric arborization of their cellular processes, and previous pharmacological studies suggest that these neurons mediate a broad modulatory role in retinal function.

Cysteamine-induced depletion of somatostatin in sheep: time course of depletion and changes in plasma metabolites, insulin, and growth hormone.

Eight crossbred wethers (51 +/- 2 kg BW), surgically fitted with abomasal cannulas, were used to determine the extent and time course of cysteamine (CSH)-induced depletion of somatostatin (SRIF) in abomasal tissue and associated changes in plasma metabolites, insulin, and growth hormone (GH). Cysteamine was administered as a single i.v. bolus (50 mg.kg BW-1 x 10 min-1) on d 0. Abomasal biopsies were obtained on d -7, -3, 0, 1, 3, and 10. On d 0, additional biopsies were taken at 2, 4, and 8 h after CSH administration. Jugular blood samples were collected over 8 h at 15-min intervals on d -2, 0, and 1. Cysteamine administration decreased (P < .05) tissue SRIF on d 0 (2, 4, and 8 h), 1, and 3; maximal depletion (42 to 55% of Pre-treatment; Pre-trt) occurred during the initial 24 h, returning to Pre-trt by d 10. Gel chromatography of pooled -7 d abomasal tissue extracts showed five peaks of SRIF immunoreactivity; the predominate peak eluted in the same position as synthetic SRIF-14. Plasma glucose, lactate, and NEFA concentrations increased (P = .001) after CSH administration and reached peak at 2 h after treatment and declined to Pre-trt concentrations by 24 h. Insulin increased (P = .001) to a maximum at h 4 and returned to Pre-trt by 24 h. Mean and baseline GH were higher (P < .07) on day of CSH administration, and pulse amplitude was lower (P < .10) on d 0 and 1. These data show that CSH rapidly reduces SRIF in abomasal tissue in a reversible manner; suggesting that CSH-treated sheep may provide a SRIF-deficient model for studying the physiological role of SRIF in ruminants.

Cellular distribution of immunoreactive somatostatin in the basal forebrain of horses and ponies

Summary In other experimental models, somatostatin (SRIF) helps coordinate central nervous system (CNS) activity associated with the response to stress. It also plays an important role in the effects of exercise, stress and bacterial infections on growth hormone (GH) release and it may interact with specific SRIF receptors found on lymphocytes. Pituitary GH is a potent immunomodulatory hor- mone which is important to normal function of the thymus and a wide range of immune defense mechanisms. This experiment mapped the distribution of immunoreactive (ir) SRIF in the basal forebrain of horses and ponies. Results are compared to those for other species in order to determine if the anatomical substrate for stress-dependant SRIF effects may be similar to that described for other animal models. Immersion fixed brain tissues were obtained from animals submitted for euthanasia and/or necropsy. Alternate sections were stained for ir-SRIF using characterized polyclonal SRIF antibody and the peroxidase-anti-peroxidase technique, as previously described. Distribution of SRIF cells and fibers was mapped on projection drawings following light microscopic examination. Tissues in this analysis included cell groups at the border between the telencephalon and the rostral diencephalon, the hypothalamus, caudate nucleus (n) subthalamus and thalamus. At rostral levels, ir-SRIF was widely dispersed throughout medial tissues. Caudally, pronounced immunoreactivity was found in ventromedial tissues. Reactive perikarya were localized in the caudate n, olfactory tuberculum, n of the lamina terminalis, rostral preoptic n, suprachiasmatic n, periventricular n, supraoptic n, paraventricular n, arcuate n and ventromedial n. Numerous SRIF fibers extended into the lateral septum, all preoptic n, paraventricular n, rostral hypothalamic n, dorsomedial n, ventromedial n and mammillary n. Reactive fibers were also found in midline thalamic n, perifomical areas, the zona incerta and the habenular n. This distribution is similar to that described in most vertebrates. It reflects the role of SRIF in multiple neurocrine interactions as well as its neuroendocrine activity. Results from this experiment suggest that equine SRIF is likely to participate in neural and endocrine pathways similar to that for other mammalian species. Similar localization patterns also suggest that the coordination of SRIF cell functions within the equine CNS may be comparable to that in other species. This suggests that studies in other species are likely to facilitate the characterization of control mechanisms for SRIF secretion in the equine species.

Developmental changes in expression of a 60-kDa somatostatin receptor immunoreactivity in the rat brain.

The neuropeptide somatostatin (SRIF) exerts several important physiological actions in the adult CNS through interactions with membrane-bound receptors. SRIF expression is developmentally regulated and this regulation is most apparent in the cerebellum, where SRIF immunoreactivity is expressed at early postnatal ages and then disappears toward adulthood. The transitory nature of SRIF expression at a time of major changes in cerebellum suggests that this peptide may have a role in cerebellar development. To further investigate the role of the SRIF transmitter system during development, we have examined the levels of expression of SRIF receptors in the developing rat brain by immunoblotting using antiserum selective for a 60-kDa brain SRIF receptor. In whole rat brain, SRIF receptor immunoreactivity first appears at embryonic day 13 (E13), is elevated at E16, increases at birth, peaks at early postnatal ages, and then gradually declines with age. No apparent changes in size of the receptor occur with age. No consistent changes in levels of SRIF receptor immunoreactivity are detected from early postnatal ages to adulthood in the hippocampus, cerebral cortex, and striatum, but levels gradually decline in the hypothalamus. In contrast, SRIF receptor immunoreactivity is expressed transiently in cerebellum. SRIF receptor immunoreactivity is detectable in cerebellum at E16, increases in levels at birth, is apparent from postnatal day 3 to postnatal day 8, and then disappears. The transitory nature of SRIF receptor expression in cerebellum is unique and parallels the expression of SRIF immunoreactivity in this brain region. These findings support the hypothesis that SRIF has a role in cerebellar development.

Transient expression of somatostatin peptide is a widespread feature of developing sensory and sympathetic neurons in the embryonic rat.

Previous studies from this and other laboratories demonstrated that many embryonic sensory ganglion cells in the rat transiently express the catecholamine synthesizing enzyme tyrosine hydroxylase (TH), a trait not expressed by most mature sensory neurons. We, therefore, sought to determine whether transient expression was uniquely associated with catecholaminergic traits, or, alternatively, whether embryonic ganglion cells transiently expressed peptidergic properties as well. Of the four peptides examined (somatostatin [somatotropin release inhibiting factor] (SRIF), galanin (Gal), calcitonin gene-related peptide (CGRP), and substance P (SP)), only SRIF was found to be transiently expressed during early stages of sensory gangliogenesis. Surprisingly, SRIF immunoreactivity was observed in virtually all cranial and spinal sensory ganglion cells on embryonic day (E) 12.5. In addition to perikaryal labeling, intense SRIF immunoreactivity was also observed in the central and peripheral processes of E12.5 sensory neurons, suggesting the peptide may be released from nerve endings. The time course of SRIF appearance in cranial ganglion cells paralleled that previously described for TH, and double-labeling studies revealed extensive co-localization of these two phenotypes. By E16.5, however, the number of neurons expressing SRIF had diminished markedly, indicating that SRIF is only transiently expressed by most sensory neurons during early stages of ganglion development. An unexpected finding was that transient expression of SRIF is also a prominent feature of sympathetic ganglion cells; however, the temporal pattern of staining in the sympathetic and sensory ganglia differed substantially. Whereas virtually no SRIF staining was observed in E12.5 sympathetics, the vast majority of cells in the E16.5 superior cervical ganglion (SCG) were labeled. This contrasted sharply with the adult SCG, in which only low levels of SRIF expression were found. These findings demonstrate that SRIF peptide is transiently expressed at high levels in peripheral sensory and sympathetic neurons during embryogenesis. The time course and widespread distribution of SRIF expression indicates that the peptide may play a role in early stages of ganglion cell growth and development. Moreover, these data, in conjunction with previous studies demonstrating SRIF immunoreactivity in developing central neurons, suggest that transient expression of this peptide is a common property of diverse neuronal cell types.

Neuroanatomical connections between corticotropin-releasing factor (CRF) and somatostatin (SRIF) nerve endings and thyrotropin-releasing hormone (TRH) neurons in the paraventricular nucleus of rat hypothalamus

In order to investigate the possible involvement of corticotropin-releasing factor (CRF) and somatostatin (SRIF) on thyrotropin-releasing hormone (TRH) neuronal cell activity in the rat hypothalamic paraventricular nucleus, we have proceeded to the simultaneous localization of CRF or SRIF and TRH. For this purpose, we used a dual immunostaining procedure that employed antibodies to CRF and SRIF and peroxidase-labeled goat anti-rabbit IgG as a first sequence, and antibodies to a cryptic fragment (Phe 178-Glu 199) of pro-TRH (to label TRH neurons) and alkaline phosphataselabeled goat anti-rabbit IgG as the second sequence. A rich innervation of the paraventricular nucleus by immunoreactive CRF and SRIF fibers was observed. A large number of CRF and SRIF nerve endings were seen intimate anatomic proximity and often appeared to surround TRH-containing cell bodies. These results strongly suggest that TRH neurons might be regulated by both CRF and SRIF. These interactions might be the neuroanatomical basis for the already observed inhibitory effects of CRF and SRIF on TRH release.

Central peptidergic neurons as targets for glucocorticoid action. Evidence for the presence of glucocorticoid receptor immunoreactivity in various types of classes of peptidergic neurons

By means of double immunolabeling procedures it has been possible to demonstrate glucocorticoid receptor (GR) immunoreactivity (IR) in large numbers of various peptidergic neurons of the brain including neurons containing gastrointestinal peptides, opioid peptides, and peptides with a hypothalamic hormone function. For each peptide system, however, marked heterogeneities exist among brain regions. Thus, in the neocortex and the hippocampal formation most of the brain peptide neurons lack GR IR, while the same types of peptide neurons in the arcuate and paraventricular nucleus [e.g. neuropeptide Y (NPY), somatostatin (SRIF) and the cholecystokinin (CCK) neurons] possess strong GR IR. Furthermore, in the arcuate, parvocellular part of the paraventricular nuclei aand the central amygdaloid nucleus practically all the peptidergic neurons are strongly GR IR, while in the lateral hypothalamus, mainly the neurotensin (NT) and galanin (GAL) IR neurons are GR IR. These marked differences among areas probably reflect functional differences dependent upon their participation in stress regulated circuits. All the paraventricular NT, corticotropin-releasing factor (CRF), growth hormone-releasing factor (GRF), thyrotropin-releasing hormone (TRH) and SRIF IR neurons appear to contain GR IR, while the luteinizing hormone-releasing hormone (LHRH) IR neurons lack GR IR, underlying the importance of glucocorticoids (GC) in controlling endocrine function. Finally, the GC may influence pain and mood control mainly via effects on enkephalin (ENK) neurons especially in the basal ganglia (mood) and on all β-endorphin (β-END) neurons of the arcuate nucleus, while most of the dynorphin neurons are not directly controlled by GC.

Cysteamine-Induced Enhancement of Growth Hormone- Releasing Factor (GRF) Immunoreactivity in Arcuate Neurons: Morphological Evidence for Putative Somatostatin/GRF Interactions within Hypothalamus*

The episodic secretion of GH is regulated through the reciprocal release into the hypophyseal portal circulation of two hypothalamic peptides, SRIF and GRF. Recent physiological evidence suggests that, in addition to exerting their well documented opposite actions at the level of the anterior pituitary, SRIF and GRF may interact within the central nervous system to modulate GH secretion. The aim of the present study was to provide morphological evidence to support the concept that SRIF exerts an influence on the GRF-containing neuronal system within the hypothalamus. To accomplish this, we used the thiol agent cysteamine (CSH; 300 mg/kg sc) to manipulate endogenous hypothalamic SRIF and examined its effects on the immunoreactivity and mRNA content of GRF-containing arcuate (ARC) neurons, using immunohistochemistry and semi-quantitative in situ hybridization, respectively. Hypothalamic SRIF content and pulsatile GH secretion were also monitored. CSH treatment reduced hypothalamic immunoreactive SRIF concentrations (to 60% of vehicle-injected control values) and severely suppressed the spontaneous surges of GH release (mean GH peak amplitude: 24.2 +/- 2.7 vs. 168.0 +/- 27.8 ng/ml in H2O-injected controls; P less than 0.001). CSH treatment also resulted in a striking increase in the number and labeling density of GRF-immunoreactive cells detected in the ARC nucleus. Cell counts revealed an overall 126% increase, over controls, in the mean number of detectable GRF-positive ARC neurons. This increase (P less than 0.01) was apparent over all rostrocaudal levels of the ARC nucleus, but was most pronounced within the caudal tier. Administration of CSH also produced a massive qualitative increase in the intensity and extent of staining of GRF-positive terminal elements in the external zone of the median eminence. Conversely, CSH treatment resulted in a decline in GRF mRNA-associated signal in cells of the ARC nucleus, with this decline being most evident in the caudal third of the nucleus. The finding of an increase in GRF immunoreactivity in ARC neurons in association with a decrease in plasma GH, as well as with a decline in relative GRF mRNA content, suggests that CSH leads to an overall decrease in GRF release from the median eminence. These results provide morphological evidence to support the concept of a SRIF-mediated central influence on the GRF-containing ARC neuronal system. Such central regulation of GRF by SRIF may be an important mechanism in the physiological control of pulsatile GH secretion.

Effects of Somatostatin on the Growth Hormone‐Releasing Factor‐Induced Growth Hormone Secretion in Rats with Electrical and Chemical Inhibitions of Endogenous Growth Hormone‐Releasing Factor and Somatostatin

Abstract The episodic pattern of growth hormone (GH) secretion of the male rat was simulated in rats exhibiting impaired GH-releasing factor (GRF) and Somatostatin (SRIF) secretion, by administering various combinations of human GRF-(1-44)NH(2) (hGRF) and SRIF. Electrical lesions of the arcuate nucleus resulted in a marked decrease in the amplitude of GH secretory bursts, while the administration of cysteamine (200 mg/kg) did not change the GH secretory profile in rats with arcuate nucleus lesions. Immunohistochemical examinations revealed a marked decrease of GRF and SRIF immunoreactivity in the median eminence of the cysteamine-treated rats with arcuate nucleus lesions. The intravenous injection of 5 mug of hGRF every 3 h caused equivalent surges of GH in the cysteamine-treated rats with arcuate nucleus lesions. The additional infusion of 4 mug/h of SRIF during the trough periods of GH secretion did not affect the amplitude of the GH surges. Hourly injection of 5 mug of hGRF caused transient desensitization to hGRF. Interestingly, the additional infusion of 4 mug/h of SRIF every 3 h enhanced the amplitude of the GH bursts induced by the fourth and the seventh hGRF injections. However, the more frequent injection of 5 mug of hGRF every 30 min caused constant desensitization to hGRF with time, and the additional infusion of 4 mug/h of SRIF every 3 h did not change the attenuated responses to hGRF. These results indicated that the simultaneous administration of hourly GRF and continuous SRIF with brief pauses was most effective for producing high GH peaks. This simulation model suggests that SRIF may play an important role not only in the production of GH troughs, but also in the maintenance of GH surges with distinct peaks in the male rat.

Depolarizing influences regulate somatostatin synthesis and processing in cultured cerebral cortical cells

There is increasing evidence that persistent depolarization plays a critical role not only in excitation-secretion coupling, but also in the mechanisms linking excitation of neuronal cells to long-term adaptative changes in biosynthesis of neuropeptides. Somatostatin (SRIF) release and synthesis are affected by numerous agents, such as high concentrations of potassium that cause depolarization of cellular membrane. In the present work, we tried to determine whether prolonged exposure to veratridine (VTD) regulates SRIF synthesis. We found that exposure to VTD (100 μM) resulted in the stimulation of total (cell content + media) immunoreactive SRIF (IR-SRIF). This effect was calcium- and sodium-dependent, since it was prevented when verapamil (VPM) 20 μM or tetrodotoxin (TTX) 1 μM were added simultaneously with VTD. Cerebral cortical cells were exposed to high potassium concentrations, and the nature of the IR-SRIF was characterized by high-pressure liquid chromatography (HPLC) or gel filtration. It was evident that chronic exposure to high potassium concentrations modified the elution profile of medium IR-SRIF on HPLC and gel filtration, causing an increase in somatostatin-28 (S-28) and a decrease in somatostatin-14 (S-14). The results indicate that chronic exposure to VTD or high potassium concentration increases immunoreactive somatostatin and augments synthesis of its high-molecular-weight forms. This suggests that chronic membrane depolarization activating sodium and calcium channels initiates the entry of calcium ions, which triggers somatostatin release and causes a depletion of its intracellular stores. The stimulation of somatostatin secretion could be coupled to synthesis of the peptide.

In situ hybridization analysis of the somatostatin-containing neuron system in developing cerebellum of rats

We used in situ hybridization histochemistry to examine the postnatal development of the somatostatin (SRIF) synthesizing system in the cerebellum of rats. There are numerous hybridizing neurons from 1 to 9 days after birth. These occur throughout the cerebellum including the developing medulla and cortex except in the external granular cell layer. The lateral cerebellar nucleus also contains SRIF gene-containing cells. The intensity of the signals for SRIF mRNA in the cerebellum decreases with age. There is a drastic decrease in SRIF mRNA in the lateral cerebellar nucleus. SRIF cells cannot be detected in the lateral cerebellar nucleus of adult rats, whereas a small, yet significant number of SRIF cells are scattered in the cerebellar medulla. However, the cerebellum of adult rats still contains a significant number of labeled cells in the granular cell layer, although the intensity for SRIF mRNA decreases from 14 days after birth to adulthood. SRIF gene-expressing cells in the cerebellar cortex are located primarily in the granular cell layer and appear to correspond to Golgi cells judging from their characteristic features. These results are consistent with our previous immunohistochemical study on the decrease of SRIF immunoreactivity in the cerebellum of adult rats. These findings, together with a recent study of transient SRIF receptor-expressing cells in the developing cerebellum suggest that SRIF acts during cerebellar development.

Somatostatin in medullary thyroid cancer.In vitro andin vivo studies

The authors evaluated the presence of somatostatin (SRIF) in the plasma and in the tumor tissue of a total of 22 patients with medullary thyroid cancer (MTC) and studied the effect of exogenous SRIF administration on basal and pentagastrin (PG)-stimulated plasma calcitonin (CT) and carcinoembryonic antigen (CEA) levels. Mean plasma SRIF concentrations were significantly higher than those found in normal controls, with five of 15 patients having plasma SRIF levels above the mean + 2 SD of normal controls. High immunoreactive SRIF concentrations were found in the extract of three tumor tissues but not in one follicular thyroid cancer or in one toxic diffuse goiter. By immunoperoxidase staining seven of 11 (63.6%) primary MTC and five of 13 (38.5%) metastases expressed SRIF antigen in a low number of cells and with a weak degree of staining. As expected, CT was expressed in almost 100% of the cases with positivity in most of the cells and strong degree of staining. Patients with positive SRIF staining in the primary tumor had longer survival than SRIF negative patients. Infusion of synthetic SRIF (11 micrograms/minute/45 minutes) produced a significant reduction of plasma CT (but not CEA) levels in 12 of the 15 patients submitted to this test. Maximal percent decrease of plasma CT ranged from 10.8% to 72.7% of the basal value and was usually observed between 30 and 45 minutes from the beginning of the infusion. When infused together with the injection of PG, SRIF was able to significantly (P less than 0.05) inhibit the PG-induced CT release in five of six patients tested. These results demonstrate the following: SRIF is present in a few cells of many primary MTC and less frequently in their metastases; tentatively, the expression of SRIF antigen in the tumor seems to be associated with longer survival; increased SRIF concentrations are found in the plasma of some patients with metastatic involvement; and treatment with exogenous SRIF reduces the basal and PG-induced CT (but not CEA) release from the tumor.

Effects of Neonatal Administration of Monosodium Glutamate on Four Neuropeptide Concentrations in the Rat Brain

The present study was designed to measure concentrations of four neuropeptides in different brain regions in monosodium glutamate(MSG)-treated rats and to assess molecular forms of each peptide with gel and high performance liquid chromatography (HPLC). MSG(4mg/kg body weight) or 10% NaCl was injected subcutaneously on postnatal days 1, 3, 5, 7 and 9 to male littermates which were subsequently used on postnatal day 100. Rats were sacrificed by decapitation, and the brains were dissected into ten discrete regions. The brain extracts were subjected to measurement of four neuropeptides; somatostatin (SRIF), neuropeptide Y (NPY), atrial natriuretic polypeptide(ANP), and a novel pituitary polypeptide 7B2 by specific radioimmunoassays. Significant increase (p less than 0.01) in midbrain SRIF content was observed in MSG-treated rats, though there was no significant change in hypothalamic SRIF content. Significant reduction (p less than 0.05) in hypothalamic NPY content was also found in MSG-treated rats. Hypothalamic ANP content was similar in both MSG-treated and control rats. A significant increase of 7B2 content was found in substantia nigra/ventral tegmentum and hypothalamus (p less than 0.05 or p less than 0.01, respectively) in MSG-treated rats. These four immunoreactivities were further characterized by gel permeation or high pressure liquid chromatography (HPLC). Chromatographic analysis of SRIF immunoreactivity revealed that there were two distinctive peaks and smaller molecular weight component corresponding to SRIF. Fractionation of NPY or 7B2 immunoreactivity by gel permeation showed a single major peak which was identical to the synthetic NPY or 7B2 immunoreactivity from porcine pituitary extract. HPLC analysis for ANP immunoreactivity also showed that the major immunoreactive component corresponded to synthetic rat ANP. MSG treatment could not produce any major alterations in proportions of molecular forms studied. These results suggest that MSG treatment in neonates might produce the alterations in SRIF, NPY and 7B2 content in the discrete brain regions including the hypothalamus.

Sexually dimorphic age-related differences in the immunocytochemical distribution of somatostatin in the platyfish

Somatostatin (SRIF) was localized by immunocytochemistry in the brains and pituitary glands of male and female platyfish ( Xiphophorus maculatus) between the ages of 8 and 30 months (average life span is 30 months). In the brain, immunoreactive (ir-) SRIF is found in the perikarya of the ventrobasal hypothalamus (nucleus lateralis tuberis, nucleus anterior tuberis), dorsomedial hypothalamus, dorsal thalamus, ventral tegmentum and rhombencephalic basal plate; in the pituitary it is localized surrounding the ir-growth hormone containing cells of the caudal pars distalis, and in older fish it is also found within certain cells of the pars intermedia. We have demonstrated that there are consistent, sexually dimorphic, age-related changes in the intensity and/or distribution of ir-SRIF. The sexual dimorphism of the changes in SRIF immunoreactivity during the aging of platyfish may be related to the different patterns of growth observed in males and females.

Immunocytochemical demonstration of growth hormone, prolactin and somatostatin-like immunoreactivities in the brain of larval, young adult and upstream migrant adult sea lamprey, Petromyzon marinus.

Growth hormone, prolactin and somatostatin-like immunoreactivities were demonstrated in the brains of larval, young adult (parasitic) and upstream migrant adult sea lampreys, Petromyzon marinus, by means of immunoperoxidase techniques. Growth hormone (GH) and prolactin (PRL) were observed within separate perikarya in the nucleus praeopticus, within fibers in the commissura praeinfundibularis, and in nerve endings within the neurohypophysis of larval and adult-stage lampreys. Cell bodies demonstrating immunoreactive growth hormone were more numerous than those reactive for prolactin. Unlike in the upstream migrant adult lamprey, no GH or PRL was demonstrated in the adenohypophysis of larval or parasitic lamprey. Somatostatin (SRIF)-like immunoreactive neurons were demonstrated in the nucleus commissurae praeinfundibularis, anterior and posterior pars ventralis hypothalami, pars dorsalis thalami, and the tegmentum motorium rhombencephali of larval, parasitic and upstream migrant adult lampreys. Many of the SRIF containing neurons within the hypothalamus were cerebrospinal fluid (CSF)-contacting cells. SRIF fibers were found throughout most of the brain predominating within the nucleus praeopticus, pars ventralis hypothalami, and the nucleus interpeduncularis. No SRIF immunoreactivity was found within the neurophyophysis. The possible functions of these peptides within the brain of the lamprey are discussed.

Effects of cysteamine administration on the in vivo incorporation of [35S]cysteine into somatostatin-14, somatostatin-28, arginine vasopressin, and oxytocin in rat hypothalamus.

The effect of cysteamine injection on the in vivo incorporation of [35S]cysteine into somatostatin-14 (SRIF-14), SRIF-28, arginine vasopressin (AVP), and oxytocin (OXT) in rat hypothalamus was studied. [35S]Cysteine was injected into the third ventricle 1 h, 4 h, or 1 week after cysteamine (300 mg/kg, sc) injection; animals were killed 4 h later. The drug was found to substantially reduce immunoreactive SRIF levels, but not OXT or AVP, 4 h after its injection. Cysteamine also caused large reductions in label incorporation into SRIF-14, SRIF-28, and OXT 1 and 4 h after drug injection. However, [35S]cysteine incorporation into AVP was increased substantially at these time points, while that into acid-precipitable protein was normal. One week after cysteamine injection, label incorporation into all hypothalamic peptides was normal. Cysteine specific activity was also measured after [35S]cysteine injection and was found to be similar in treatment and control groups. The results suggest that cysteamine inhibits the syntheses of SRIF-14, SRIF-28, and OXT and stimulates that of AVP.