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Many neurological diseases are related to disturbances of somatostatin- (SOM-) expressing interneurons in the cingulate cortex. Therefore, their role within the circuitry of the cingulate cortex needs to be investigated. We describe here the physiological time course of SOM effects onto pyramidal cell excitability and action potential discharge pattern. Furthermore, we show that the GRK2 inhibitor Gallein had no effect on the reduced SOM-induced response following repetitive SOM applications., (Copyright © 2019 Elsevier B.V. All rights reserved.)

BRINPs (BMP/RA-inducible Neural Specific Protein-1, 2, 3) are family genes expressed mainly in both the central and peripheral nervous system. BRINP1 is abundantly expressed in many of adult brain regions including cerebral cortex and hippocampus, with expression regulated in an activity-dependent manner in the dentate gyrus. Mice with disrupted BRINP1 gene exhibit abnormal behaviors such as increased locomotive activity and poor social activity which are analogous to symptoms of human psychiatric disorders such as schizophrenia (SCZ), autism spectrum disorder (ASD) and attention-deficit hyperactivity disorder (ADHD). In the present study, to clarify the physiological roles of BRINP1 in psychiatric disorders, we examined the numbers of parvalbumin (PV)-expressing neurons and somatostatin (SST)-expressing neurons in the medial prefrontal cortex (mPFC) in BRINP1-KO mice. Immunohistochemical analysis revealed the numbers of PV-expressing neurons and SST-expressing neurons in mPFC of BRINP1-KO mice were, respectively, 50% and 20% fewer than corresponding neurons in mPFC of wild-type mice. These data suggest that the abnormal behaviors related to human psychiatric disorders in BRINP1-KO mice could be derived from the hyperexcitability of pyramidal neurons as a consequence of decreased inhibitory innervation and conceivable dysregulation of the Excitatory/Inhibitory balance in mPFC., (Copyright © 2018 Elsevier B.V. All rights reserved.)

N-Methyl-d-Aspartate receptors (NMDARs) are present in both pyramidal neurons and interneurons of the hippocampus. These receptors play a key role in the structural plasticity of excitatory neurons, but to date little is known about their influence on the remodeling of interneurons. Among hippocampal interneurons, the somatostatin expressing cells in the CA1 stratum oriens are of special interest because of their functional importance and structural characteristics: they display dendritic spines, which change their density in response to different stimuli. In order to understand the role of NMDAR activation on the structural dynamics of the spines of somatostatin expressing interneurons in the CA1 stratum oriens, we have studied entorhino-hippocampal organotypic cultures obtained from mice in which this interneuronal subpopulation expresses constitutively EGFP, and have imaged them in real-time. We have acutely infused the cultures with NMDA, a strong NMDAR agonist, and have analyzed the structural dynamics of somatostatin expressing interneurons, prior and after its administration. The appearance and disappearance rates of their dendritic spines increased 24h after the NMDA infusion and returned to baseline levels 48h afterwards. By contrast, their stability rate decreased 24h after the infusion and also returned to control levels 48h later. The relative density of the dendritic spines remained unaltered throughout the assay. Altogether, our results show that the activation of NMDARs can influence the structural dynamics of interneurons. This is especially important because of the involvement of these receptors in neuronal potentiation/depression and their putative role in the etiopathology of certain neuropsychiatric disorders, such as schizophrenia., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Dysfunction of inhibitory GABAergic interneurons is considered a major pathophysiological feature of various neurodevelopmental and neuropsychiatric disorders. The variants of SHANK3 gene, encoding a core scaffold protein of the excitatory postsynapse, have been associated with numerous brain disorders. It has been suggested that abnormalities of GABAergic interneurons could contribute to the SHANK3-related disorders, but the limitation of these studies is that they used mainly Shank3 knock-out mice. Notably, Shank3-overexpressing transgenic mice, modeling human hyperkinetic disorders, also show reduced inhibitory synaptic transmission, abnormal electroencephalography, and spontaneous seizures. However, it has not been investigated whether these phenotypes of Shank3 transgenic mice are associated with GABAergic interneuron dysfunction, or solely due to the cell-autonomous postsynaptic changes of principal neurons. To address this issue, we investigated the densities of parvalbumin- and somatostatin-positive interneurons, and the mRNA and protein levels of GAD65/67 GABA-synthesizing enzymes in the medial prefrontal cortex, striatum, and hippocampus of adult Shank3 transgenic mice. We found no significant difference in the measurements performed on wild-type versus Shank3 transgenic mice, except for the decreased GAD65 or GAD67 mRNAs in these brain regions. Interestingly, only GAD65 mRNA was decreased in the hippocampus, but not mPFC and striatum, of juvenile Shank3 transgenic mice which, unlike the adult mice, did not show behavioral hyperactivity. Together, our results suggest age-dependent decrease of GAD65/67 mRNAs but normal densities of certain GABAergic interneurons in the Shank3 transgenic mice., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Light has profound effects on mood regulation as exemplified in seasonal affective disorder (SAD) and the therapeutic benefits of light therapy. However, the underlying neural pathways through which light regulates mood are not well understood. Our previous work has developed the diurnal grass rat, Arvicanthis niloticus, as an animal model of SAD. Following housing conditions of either 12:12 h dim light:dark (DLD) or 8:16 h short photoperiod (SP), which mimic the lower light intensity or short day-length of winter, respectively, grass rats exhibit an increase in depression-like behavior compared to those housed in a 12:12 h bright light:dark (BLD) condition. Furthermore, we have shown that the orexinergic system is involved in mediating the effects of light on mood and anxiety. To explore other potential neural substrates involved in the depressive phenotype, the present study examined hypothalamic dopaminergic (DA) and somatostatin (SST) neurons in the brains of grass rats housed in DLD, SP and BLD. Using immunostaining for tyrosine hydroxylase (TH) and SST, we found that the number of TH- and SST-ir cells in the hypothalamus was significantly lower in the DLD and SP groups compared to the BLD group. We also found that treating BLD animals with a selective orexin receptor 1 (OX1R) antagonist SB-334867 significantly reduced the number of hypothalamic TH-ir cells. The present study suggests that the hypothalamic DA neurons are sensitive to daytime light deficiency and are regulated by an orexinergic pathway. The results support the hypothesis that the orexinergic pathways mediate the effects of light on other neuronal systems that collectively contribute to light-dependent changes in the affective state., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Several observations have pointed to a major pathogenic role of somatostatin depletion with respect to amyloid accumulation, which is often thought to be the crucial event in a cascade leading to Alzheimer's disease (AD). As methylation of CpG islands plays an important role in gene silencing, we studied the methylation status of the CpG islands in the promoters of somatostatin (SST) and in that of its receptor subtype in the cerebral cortex, SSTR4, in tissue samples from the middle temporal (Brodmann area 22) and superior frontal gyrus (Brodmann area 9) of 5 severely affected AD patients aged 72-94 years (Braak stages V-C or VI-C) and 5 non-demented controls aged 50-92 years. Bisulfite sequencing of DNA from cortical gray and infracortical white matter showed that the DNA methylation status at the promoters of SST and SSTR4 did not significantly differ between AD and control samples in any of the regions analyzed. We confirmed these results using deep bisulfite sequencing of PCR products from the SST promoter amplified from DNA from the cortical gray of the superior frontal gyrus of all AD patients and non-demented controls. We observed a trend toward increased DNA methylation with increasing age. In conclusion, deregulated somatostatin signaling in the AD cortices studied cannot be explained by hypermethylation of the SST or SSTR4 promoter CpG islands., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Somatostatin (SST) has been suggested to play an important role in maintaining hippocampal homeostasis by modulating excitatory neurotransmission. The putative anticonvulsant role for SST was tested in an electrical amygdala kindling model. SST was cloned into serotype 5 of the adeno-associated viral (AAV) vector and delivered bilaterally into the hippocampus of adult male Sprague Dawley rats that were subsequently electrically kindled. Behavioral severity and duration of kindled seizures was compared to uninjected and GFP-injected control rats. Results demonstrated that 70% of SST treated animals did not experience class IV or V seizures without affecting the threshold for individual stimulation-evoked seizures. This result was significantly different from control groups where 100% of animals reached class V seizures. No difference in the number of stimulations required to reach the first class I-III seizures was observed in the experimental cohort relative to age-matched controls. These preclinical results suggest a putative role for SST as an anticonvulsant therapeutic modality for epilepsy., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

The respiratory responses to bilateral microinjections (30-50 nl) of 5mM somatostatin (SOM) or 10mM cyclosomatostatin (c-SOM, a SOM antagonist) into the Bötzinger complex (BötC), the pre-Bötzinger complex (preBötC) and the rostral inspiratory portion of the ventral respiratory group (iVRG) were investigated in urethane-chloralose anesthetized, vagotomized, paralysed and artificially ventilated rabbits. SOM microinjections into the BötC decreased respiratory frequency and the rate of rise of phrenic nerve activity without obvious changes in its peak amplitude. SOM microinjected into the preBötC caused increases in respiratory frequency and decreases in peak phrenic activity associated with increases in its rate of rise. No changes in respiration were induced by SOM microinjections into the iVRG. Microinjections of c-SOM into the preBötC caused decreases in respiratory frequency as well as in peak amplitude and rate of rise of phrenic nerve activity. The results show that endogenously released SOM within the preBötC contributes to shape the pattern of baseline respiratory activity and that SOM receptors within the BötC and the preBötC have a role in the modulation of respiration in the rabbit., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

In the rat dentate gyrus, mu opioid receptor (MOR) agonists disinhibit principal cells, promoting excitation, but whether MOR protein is differentially distributed to interneuron subtypes is unknown. Here, the distribution of MOR immunoreactivity was semi-quantitatively examined in neurochemically identified interneurons using fluorescence microscopy. We find that MOR- and parvalbumin-immunoreactivities are frequently co-localized, while MOR- and somatostatin-immunoreactivities are less commonly co-localized. This suggests that MORs are most frequently on interneurons specialized to inhibit granule cell output, and are on a limited number of interneurons that inhibit granule cell distal dendrites.

This study investigated the in vivo neuronal release of somatostatin in the rat nucleus accumbens (NAc), and the effect of chronic administration of antidepressants. Microdialysis studies were performed on male Sprague-Dawley rats, in accordance with the EU guidelines (EEC Council 86/609). Somatostatin levels were quantified by radioimmunoassay (RIA) or enzyme linked immuno sorbent assay (ELISA). A high concentration of potassium ions (K(+), 100 mM) was used to ascertain the neuronal release of somatostatin. Antidepressant treatments involved the administration of citalopram (20 mg/2 ml/kg, i.p., once daily) or desipramine (DMI, 5 mg/2 ml/kg, i.p., twice daily) for 21 days. Control groups received saline (2 ml/kg for 21 days, i.p.) once or twice daily respective of the antidepressant treatment. Basal levels of somatostatin released were found to be 20.01+/-0.52 fmol/sample. K(+) (100 mM) increased somatostatin levels at 205% of basal. Chronic citalopram and desipramine treatments also increased the somatostatin levels by 83+/-32% and 40+/-6% of basal, respectively. These findings indicate that somatostatin is released neuronally in the NAc. Antidepressants influence its release in a positive manner, suggesting the necessity of further studies for the elucidation of the involvement of somatostatin in the putative therapeutic effects of these agents.

The aim of the present study was to determine the possible role of somatostatin as a modulator of dopamine release in rat retina. Basal release of dopamine, and how this is influenced by somatostatin receptor (sst) selective ligands, was examined ex vivo in rat retinal explants. Dopamine levels were quantified by high-pressure liquid chromatography (HPLC) with electrochemical detection. Basal levels of dopamine were measured over 120 min of tissue incubation and found to be 1.17+/-0.35 ng/ml. Somatostatin (10(-6), 10(-5), 10(-4)M) increased dopamine levels in a concentration-dependent manner, while the sst(2) antagonist CYN154806 (10(-4)M) reversed its actions. BIM23014 (sst(2) agonist) increased dopamine levels in a statistically significant manner only at the concentration of 10(-5)M. The sst(1) agonist L797.591 (10(-5), 10(-4)M) also increased dopamine levels, while activation of the sst(3) receptor (sst(3) agonist, L796.778, 10(-4)M) had no effect. These data substantiate a neuromodulatory role for sst(1) and sst(2) somatostatin receptors in the retina and show for the first time somatostatin's influence on dopamine release.

The aim of this study was to examine the ability of somatostatin receptor (sst(1)) to regulate the release of somatostatin in rat retina. Immunohistochemistry studies were performed to locate the somatostatin neurons, and radioligand binding to ascertain the presence of sst(1). The neuronal release of somatostatin was examined ex vivo in rat retinal explants in the presence of KCl (50 and 100 mM), and absence of Ca(++) (EGTA; 10 mM). Somatostatin levels, quantified by radioimmunoassay, were increased in the presence of KCl (100 mM, 151%) and attenuated in the absence of Ca(++) (31%). CH275 (sst(1) agonist) reduced the somatostatin levels in a concentration-dependent manner (10(-5)-10(-7) M), and this effect was reversed by NVP-SRA 880 (sst(1) antagonist;10(-5) M). MK678 (sst(2) agonist; 10(-5) M) had no effect. These data suggest an autoreceptor role for sst(1) in retina.

Most actions of anandamide (AEA) are mediated by the cannabinoid 1 (CB(1)) receptor activation, but on sensory neurones it is also an agonist on the vanilloid subtype 1 receptor (VR(1)). The aim of the present study was to analyse the effect of AEA (10(-6)-10(-4) M) on inhibitory CB(1) and excitatory VR(1) receptors by measuring sensory neuropeptide release such as somatostatin, substance P and calcitonin gene-related peptide, from isolated rat tracheae. AEA (10(-6) M) vas without significant effect, 10(-5) M inhibited neuropeptide release, which was abolished by the G protein-coupled receptor blocker pertussis toxin (100 ng/ml) and the CB(1) receptor antagonist SR141716A (5x10(-7) M). High concentrations of AEA (5x10(-5) M, 10(-4) M) increased the release of the peptides and this inhibition was prevented by the competitive VR(1) antagonist capsazepine (10(-5) M). These results indicate a dual, concentration-dependent action of AEA on CB(1) receptors and VR(1) on peripheral sensory nerve terminals., (Copyright 2002 Elsevier Science Ireland Ltd.)

We examined if axon terminals of dopaminergic neurons might make synapses upon neurokinin-1 receptor (NK1R)-expressing intrinsic neurons in the rat striatum. In a double-immunocytochemical ultrastructural study, dopaminergic terminals were labeled by the immunoperoxidase method for tyrosine hydroxylase (TH), while NK1R-immunoreactivity (-IR) was revealed by the immunogold-silver labeling method. Some TH-immunoreactive (-ir) axon terminals formed synapses of the symmetric or intermediate type on NK1R-ir neuronal profiles; usually on dendritic profiles and rarely on somatic profiles. It was further confirmed by means of triple-immunofluorescence histochemistry that NK1R-ir neurons in close association with TH-ir axon terminals showed nitric oxide synthase (NOS)- or vesicular acetylcholine transporter-IR. Since NK1R-expressing striatal neurons are segregated into cholinergic and somatostatin/NOS-containing intrinsic neurons (Brain Res. 631 (1993) 297; Neurosci. Lett. 310 (2001) 109), the present results indicate that dopaminergic neurons make synapses upon these intrinsic neurons in the striatum.

Adult teleost fish, Apteronotus leptorhynchus, exhibit an enormous regenerative capability after application of mechanical lesions to the dorsalmost subdivision of the cerebellum, the corpus cerebelli. Restoration of the neural tissue is achieved by a cascade of processes, including the guidance of migrating new neurons to the site of injury by radial glial fibers. These fibers are characterised by the expression of immunoreactive glial fibrillary acidic protein and by several morphological features. Within 12 h following the lesion, the fraction of radial glial fibers expressing the neuropeptide somatostatin (SRIF) dramatically increased from approximately 1%, as found in the intact brain, to roughly 27% 12-24 h post-lesion. Subsequently, the percentage of SRIF-expressing radial glial fibers gradually declined, until it reached background levels at about 10 days following the injury. We hypothesise that the expression of SRIF is related to the generation and/or differentiation of the new neurons produced in response to the lesion, rather than to the later guidance of these cells along their migratory pathway.

APPsw transgenic mice showing substantial features of brain Abeta amyloidosis such as senile plaques and behavioral abnormalities were examined by immunostaining to determine whether Abeta deposits could induce the subsequent disturbance of neurotransmitter systems including somatostatin, substance P and choline acetyltransferase (ChAT), which are prominent in the Alzheimer's disease brain. Somatostatin, substance P and ChAT disappeared in the areas of senile plaque and were accumulated in dystrophic neurites around the amyloid cores. These findings suggest a potential role of brain Abeta amyloidosis in disturbance of the neurotransmitter systems leading to memory disturbance of Alzheimer's disease.

The impact of hypoxia on somatostatin (SS) secretion from the median eminence (ME) of the hypothalamus and the possible glucocorticosteroid involvement in modulating secretion, were investigated in adult male rats exposed to hypoxia. SS levels were measured by specific radioimmunoassay during acute and prolonged hypoxia as well as after bilateral adrenalectomy (ADX) with or without a replacement with dexamethasone (DEX). The results were as follows: (a) acute hypoxia (5 km altitude, 10.8% O(2)) for 2 and 24 h markedly increased SS content in ME, but acute severe hypoxia (7 km, 8.2% O(2) for 24 h) markedly decreased SS level in ME. (b) Chronic hypoxia (10.8% O(2)) from 5 to 25 days exposure did not significantly affect SS content of ME. (c) ADX alone increased SS content of ME and this increase was further enhanced after 2 h exposure to hypoxia. (d) The increased SS in ME of ADX rats was blocked by replacement with DEX (500 microg/rat i.p.). The data presented suggest that acute hypoxia stress may increase or decrease SS content of ME in rats, depending on the severity and duration of the hypoxia and that the stimulatory action of hypoxia on SS content of ME be may in part mediated by the increased corticosterone levels during hypoxia.

In the present study the ability of tachykinin agonists and antagonists to modulate gamma-aminobutyric acid (GABA) release has been correlated with tachykinin receptor expression in the mouse striatum. Significant GABA release was observed when striatal slices were challenged with the NK-3 receptor agonist senktide, the selectivity of which was confirmed using the NK-3 receptor antagonist SR142801. In situ hybridisation revealed co-expression of NK-3 receptors with nitric oxide synthase (NOS)/preprosomatostatin containing GABAergic interneurones. These findings suggest that tachykinins modulate GABA release within the striatum via interaction with NK-3 receptors on somatostatin/NOS interneurones.

In rats anaesthetized with urethan and pretreated with pipecuronium bromide nocifensive reaction of blood pressure elevation evoked by intraarterial capsaicin injection was inhibited over 40 min by bilateral antidromic stimulation of the sensory fibres of the sciatic nerves. Rise in blood pressure, heart rate and respiratory frequency evoked by capsaicin were markedly diminished after smearing 1% mustard oil on the acutely denervated hindpaws indicating a release of mediators with anti-nociceptive action from cutaneous nociceptors. Intravenous injection of the putative mediator somatostatin (10 microg/kg) or its analogues RC-160 and TT-232, but not octreotide inhibited the cardiorespiratory and blood pressure responses evoked by topical cutaneous application of mustard oil or capsaicin instillation into the eye. It is concluded, that the endocrine and the anti-nociceptive effects of somatostatin are mediated through distinct receptor subtypes and therefore, TT-232, a novel heptapeptide analogue without endocrine action, is a promising analgesic compound.

Following application of mechanical lesions to the corpus cerebelli, a cerebellar subdivision, in adult individuals of the teleost fish Apteronotus leptorhynchus, the pattern of expression of the neuropeptide somatostatin was examined by employing immunohistochemical techniques. In the intact corpus cerebelli, only a very few cells displayed somatostatin-like immunoreactivity. This number dramatically increased in the area of the lesion within the granule cell layer 1 day following the injury and peaked after 2 days, when the normalized total number of somatostatin-positive cells was approximately 50 times higher than the mean number of labeled cells found after 3, 6, and 12 h of survival. Between 5 and 10 days of post-lesioning survival time, this number abruptly declined and returned to background levels at 17 and 25 days. Confocal microscopy revealed three cell types, presumably corresponding to granule cell neurons, astrocytes and microglia, which all displayed a similar temporal pattern of somatostatin expression. It is hypothesized that somatostatin is involved in regulation of the genesis and/or development of new neurons which are produced in response to injuries and which replace damaged cells at the site of the lesion.

The ontogeny of somatostatin binding sites was studied in eight fiber tracts of the human lower brainstem in 18 fetuses and infants aged from 21 weeks postconceptional to 6 months postnatal, and in two adults. The study was performed by means of quantitative autoradiography using [125I-Tyr0,DTrp8]somatostatin-14 as a radioligand. For all structures examined, the highest densities of binding sites were detected in the younger stages and the density of sites decreased during development. These results reveal the existence of a close inverse relationship between the density of somatostatin receptors and the myelination process in the fiber tracts of the human brainstem.

We measured somatostatin-like immunoreactivity, using a radioimmunoassay which does not cross react with cortistatin-like immunoreactivity, in postmortem frontal cortex (Brodmann area 9) from 32 patients, of different apolipoprotein E genotypes, and presenting with different degrees of cognitive impairment. Eleven subjects and eight patients presented with no (controls) or limited memory impairments (Borderline), respectively. Six patients with clinical criteria for possible Alzheimer's disease also presented with clinical or brain imaging of cerebrovascular disease (mixed dementia) and seven patients were classified as Alzheimer's disease (AD). In the 6 months preceeding their deaths, all subjects had been evaluated by Folstein's Mini Mental State examination (MMS). Sixty nine percent of patients with MMS >20 did not carry the epsilon 4 allele while 66% of patients with MMS <10 did. Somatostatin concentrations (ng/mg wet weight) were significantly lower in the patients carrying the epsilon 4 allele (E2/3: 0.71 +/- 0.05, n = 19 vs. E4: 0.42 +/- 0.06, n = 13; mean +/- SEM, P < 0.001). These results, which are reminiscent of those obtained on cholinergic markers, suggest that apolipoprotein E4 is involved in the somatostatinergic dysfunction early after the onset in AD.

Impairment of the basal forebrain cholinergic system is an important change in the brains of Alzheimer's disease patients. Various neurotoxins have been used to achieve this in animal models. In this study the effects of chemical lesions by ibotenic acid (IBO), a glutamate analogue and by 192 IgG-saporin, a highly specific immunotoxin against cholinergic neurons, were investigated. The toxins were delivered stereotaxically into the brains of young Sprague-Dawley rats which were later sacrificed by decapitation. Choline acetyltransferase (ChAT) activity was measured by radioenzymatic assay and substance P (SP), neuropeptide Y (NPY) and somatostatin (SOM) levels by radioimmunoassay. Decreased ChAT and SOM levels were observed in the cortex and the hippocampus in both experiments. Cortical SP levels were increased after IBO lesions but were unaffected after 192 IgG-saporin lesions. NPY levels remained unchanged in both experiments. The results indicate that there were specific changes in neuropeptide contents in the cortex and hippocampus in response to cholinergic damage in the rat brain.

To examine the possible involvement of somatostatin in growth hormone modifications induced by ethanol, we examined: (1) the effects of chronic ethanol exposure of cultured hypothalamic neurons on somatostatin content and mRNA levels; (2) the acute effect of ethanol on somatostatin release stimulated by N-methyl-D-aspartate (NMDA). The results showed that 8 days of ethanol exposure (10-100 mM) did not decrease somatostatin content or somatostatin mRNA levels. Ethanol treatment alone had no significant effect on cell morphology or on protein content. In contrast, acute application of ethanol in 8 day-old cultures significantly reduced (50 mM) or completely blocked (100 mM) somatostatin release elicited by 50 microM NMDA without modifying basal release. We conclude that chronic ethanol treatment to concentrations up to 100 mM has no effect on somatostatin biosynthesis in fetal rat hypothalamic neurons, while weaker concentrations decrease NMDA-induced somatostatin release.

The median eminence (ME) push-pull perfusion technique was used in this work and the results clearly showed that i.p. administration of MK-801 (4 mg/kg), a specific N-methyl-D-aspartate (NMDA) receptor antagonist, totally abolished dexamethasone (Dex) (300 micrograms/ 100 g i.p. injected) and immobilization stress-induced hypothalamic somatostatin release in adult male rats. We also observed that glutamate from median eminence-hypothalamic medio basal (ME-MBH) complex, measured by high performance liquid chromatography (HPLC), exhibited a conspicuous secretory pattern, with the total amount released not modified by Dex administration. This indicates that Dex and stress-induced somatostatin (SS) secretion is not mediated by endogenous glutamate variations but likely by activation of NMDA receptors.

The presence of somatostatin-like immunoreactive neurons was examined in the rabbit vestibular ganglion by using immunohistochemical techniques. About 22% (n = 153) of the total ganglion cells (n = 699) examined were somatostatin-immunoreactive and they were diffusely present throughout the ganglion. Majority of the somatostatin-immunoreactive cells were large (long diameter, 23-31 microns, 74%; short diameter, 20-25 microns, 76%) and they had oval or spherical cell bodies with well developed Nissl's body. However, about 78% (n = 546) of the total ganglion cells examined were not immunoreactive to the somatostatin. The presence of the somatostatin-immunoreactive neurons in the vestibular ganglion suggests a possibility that somatostatin may be involved in the modulation of afferent sensory transmission from the vestibular organ of the rabbit.

This work was undertaken to investigate whether the inhibitory tone exerted by GABA on somatostatin (SRIH) release operates in the control of thyrotropin (TSH) secretion in both basal and cold-stimulated conditions. In a first group of animals (G1) undergoing both carotid and third ventricle push-pull cannulation, i.vt. injection of picrotoxin (10(-5) M) induces a significant decrease in plasma TSH level under basal conditions (0.09 +/- 0.02 versus 0.27 +/- 0.4 ng/100 microliters; P < 0.03, n = 5). In a median eminence (ME) push-pull cannulated group of rats (G2), picrotoxin, peripherally administered, blocks cold-induced inhibition of SRIH release (35.0 +/- 1.8 versus 7.4 +/- 3.3 pg/15 min; P < 0.005; n = 5). In a third group of intact rats (G3), peripheral administration of picrotoxin (2 mg/kg i.p.) blunts the cold-induced TSH release (0.17 +/- 0.03 versus 0.46 +/- 0.04 ng/100 microliters; P < 0.001; n = 5). Our results strongly suggest that a decrease in SRIH release is involved in the GABAergic control of basal and cold-induced TSH secretion.

The distribution and co-localization of substance P immunoreactivity (SP-IR), somatostatin-IR (SS-IR) and vasoactive intestinal peptide-IR (VIP-IR) have been determined in canine and human jejunum by immunocytochemistry (ICC). Immunostaining with an antibody to protein gene product 9.5 revealed fewer neuronal cell bodies in the submucosal ganglion of human than in canine intestine. Double immunostaining demonstrated that SP-IR and SS-IR were always co-localized in human but never in canine submucosal neurons. In both species, VIP-IR was present in a separate population of neurons. In canine submucosal ganglia, each peptide represented approximately one-third of the neurons while the proportions of VIP and SP/SS-containing neurons in human were 40% and 42%, respectively. These results provide neuroanatomical evidence for different functions of neural SP and SS in canine and human jejunum.

The effects of acute stress on growth hormone (GH) secretion and the mechanisms involved in its changes have been investigated in sheep. An acute isolation-restraint stress induced a rapid and significant increase in jugular GH levels in 12 out of 14 rams. GH-releasing hormone (GHRH) and somatostatin secretion during the same stress were studied in 5 animals prepared for hypophysial portal blood collection. A 3.5-fold increase in portal GHRH levels was observed concomitantly with a slight elevation in portal somatostatin. Portal corticotropin-releasing hormone (CRH) and jugular cortisol plasma levels increased during the same stress. Our data suggest that an isolation-restraint stress stimulates GH secretion in the sheep and that GHRH may be responsible for GH response.

The administration of an i.p. dose of phenylephrine (2 mg/kg) increased the number of [125I]Tyr11-somatostatin ([125I]Tyr11-SS) receptors and decreased their apparent affinity in rat hippocampal membranes 7 h after its injection. Prazosin (20 mg/kg, i.p.) administered 1 h before phenylephrine reversed effects of the latter on SS binding. Prazosin alone decreased the number of SS receptors without changing the affinity. The addition of phenylephrine or prazosin (10(-5) M) to the incubation medium did not change the SS binding characteristics. The present results support the notion that the alpha 1-adrenergic system regulates the binding of SS to its specific receptors in rat hippocampus.

Interaction of beta-adrenergic and somatostatinergic systems in the hippocampus has not been investigated fully. We studied the influence of DL-isoproterenol (ISO), a beta-adrenergic agonist and DL-propranolol (PRO), a beta-adrenergic blocking agent, on the somatostatinergic system in the rat hippocampus. The short-(5h) and long-term (14 days) administration of ISO (5 mg/kg i.p.) or of PRO (10 mg/kg i.p.) did not affect somatostatin-like immunoreactivity (SSLI) content in the hippocampus of male Wistar rats. Both short- and long-term ISO administration decreased the number of specific [125I]Tyr11-somatostatin ([125I]Tyr11-SS) receptors in synaptosomes from hippocampus (29%, P < 0.05 and 34%, P < 0.05, after short- and long-term administration, respectively) without changing the affinity constant. This decrease in the number of [125I]Tyr11-SS receptors was not due to a direct effect of ISO on these receptors since no decrease in binding was produced by high concentrations of ISO (10(-5) M) when added in vitro. In addition, this decrease could be blocked by pretreatment with PRO. Short- and long-term administration of PRO alone increased the [125I]Tyr11-SS binding in hippocampus (42%, P < 0.05 and 33%, P < 0.05, after short- or long-term administration, respectively) without changing the affinity constant. Although there is no direct evidence that the regulation of SS receptors by the beta-adrenergic system has a physiological significance, this mechanism may provide a means by which the brain environment could modulate SS receptor number and, therefore, sensitivity to SS in a subset of SS-sensitive neurons.

One week after stereotaxical implantation of a push-pull cannula into the median eminence (ME), rats were stressed by immobilization for 2 h daily for 7 days. Thereafter, ME was perfused for 1 h in basal, stress and recovery conditions, respectively, and somatostatin (SRIH) was measured in perfusate fractions. Pituitaries were in vitro perifused to assess GH responsiveness to SRIH. In the stressed group, basal SRIH release was significantly higher than in the control group and stress caused a significant sharp peak in neurohormone release. GH responsiveness to SRIH was not affected in pituitaries obtained from stressed donors. High SRIH levels secreted under chronic stress thus did not impair the GH pituitary response to SRIH.

We have used sensitive and specific radioimmunoassays to examine the differential distributions of three neuropeptides, substance P (SP), calcitonin gene-related peptide (CGRP) and somatostatin (SST), in dorsal root ganglia (DRG) from levels C1 to L6 of the spinal column. SP and CGRP content both varied in proportion to ganglion wet weight, with greatest levels in cervical and lumbar DRG. By contrast, SST content showed a distinct distribution, with maximum levels in rostral, cervical DRG. We speculate that the distribution of these neuropeptides may be due to the influence of target-derived neurotrophic factors, and may underlie the particular sensitivity of the hind limbs to adjuvant arthritis in the rat.

Rats were implanted with microdialysis probes in hippocampi and striata, and somatostatin-like immunoreactivity (SS-LI) was measured in outflows obtained from awake, freely moving animals 48 and 72 h post implantation. SS-LI was measurable in all dialysates under basal conditions; concentrations were stable and within a narrow range, about 3-6 fmol/ml. Cysteamine (300 mg/kg, s.c.) markedly reduced basal SS-LI concentrations in outflows from hippocampus (P < 0.00001). KCl (100 mM, 10 min) or veratridine (50 microM, 10 min) infusion elevated hippocampal SS-LI output by 55 and 106%, respectively (P's < 0.05). EGTA (10 mM) or tetrodotoxin (2 microM) infusion inhibited the SS-LI release elicited by KCl and veratridine, respectively, without affecting the basal SS-LI outflow. Thus, our results demonstrate that SS-LI is released from rat hippocampus and striatum in vivo, and provide evidence that the peptide may be released in hippocampus by both action potential dependent and independent processes.

Optic nerve hypoplasia is seen in 50% of patients diagnosed with fetal alcohol syndrome and is due to a defect in the development of retinal ganglion cells. Somatostatin may influence the development of retinal ganglion cells, so we studied the effect of in vivo ethanol exposure on somatostatin expression in the developing retina. Somatostatin concentration is increased in retina from ethanol-exposed fetuses and pups and this increase in peptide is associated with excessive neurite formation and improper migration of the somatostatin-containing neurons at early postnatal ages. These disturbances may account for the eventual failure in retinal ganglion cell development that results in optic nerve hypoplasia.

Several lines of evidence suggest that somatostatin (SS) may interact with serotonergic neurons in the central nervous system. To assess whether SS acts presynaptically on serotonin (5-hydroxytryptamine, (5-HT)) neurons, SS receptors were measured in membranes from the hippocampus, a brain region that receives dense serotonergic innervation and has a high number of SS receptors in control and 5,7-dihydroxytryptamine (5,7-DHT)-treated rats, at 1 and 3 weeks after injection. Intracerebroventricular (i.c.v.) injection of the 5-HT-specific neurotoxin 5,7-DHT (11 micrograms (free base) dissolved in 10 microliters of isotonic saline containing 0.01% ascorbic acid) produced a 70% reduction in hippocampal 5-HT content at 3 weeks after injection but not at 1 week. This change was associated with a significant decrease in SS receptor density in rat hippocampus only at 3 weeks following the injection, without influencing the apparent affinity of the receptors at any time. Administration of 5,7-DHT did not affect somatostatin-like immunoreactivity (SSLI) levels at both times studied. These results suggest that some of the hippocampal SS receptors may be localized presynaptically on the serotonergic nerve terminals.

The dynamic release of somatostatin-like immunoreactivity (SLI) from duodenum into the lumen was studied in the isolated, vascularly perfused rat duodenum. The luminal release of duodenal SLI was stimulated by a cholinergic agonist, carbachol, and the carbachol-induced release of SLI was completely blocked by atropine, but not by hexamethonium. These data suggest that the luminal release of SLI from rat duodenum is under the control of a cholinergic muscarinic stimulation. The ratio of somatostatin-14 to somatostatin-28 in picograms was about 1 during basal release but increased to approximately 2 during carbachol stimulation.

A double-labelling immunofluorescence study of rat spinal dorsal horn was carried out with antisera to Met-enkephalin and somatostatin. Varicosities in laminae I and II were frequently immunoreactive with both antisera, and in addition some neuronal cell bodies in lamina II possessed both types of immunoreactivity. These findings suggest that enkephalin and somatostatin coexist in some axons within the rat superficial dorsal horn and that at least some of these axons are derived from local neurones.

Neuropeptides, synthesized in dorsal root ganglia (DRG), are implicated in nociception and neurogenic inflammation. Alterations in DRG neuropeptide levels have been described in polyarthritic rats, but these models are associated with widespread systemic disease. Using mild adjuvant-mediated monoarthritis of the left carpal joint we found significant increases in substance P (+69%) and calcitonin gene-related peptide (CGRP; +204+), but not somatostatin in ipsilateral C6/7 DRG. Peptide levels in contralateral DRG and other ipsilateral DRG were unaltered. Substance P and CGRP in DRG may be of importance in the pathogenesis and maintenance of adjuvant arthritis.

Previous studies have localized enkephalin and somatostatin to amacrine cell populations in the larval tiger salamander retina. Double-label immunocytochemistry was utilized to examine if enkephalin- and somatostatin-like immunoreactivities are colocalized to tiger salamander amacrine cells. Of the more than 2000 labelled cells observed in double-labelled preparations, none were found to express both enkephalin and somatostatin immunoreactivity. Therefore, these studies demonstrate that in the larval tiger salamander retina, enkephalin and somatostatin are localized to separate populations of amacrine cells.

Somatostatin (SRIF) release from fetal rat cortical cells was stimulated by exposure to 10(-5) M N-methyl-D-aspartate (NMDA) (250 +/- 20% of basal at 96 h). A similar but much less potent effect was seen with kainate (KA) but not with quisqualate (Q) which inhibited SRIF release (KA 150 +/- 13%, Q 65 +/- 18% of basal at 96 h). Similar data were obtained for intracellular levels of SRIF. Dose-dependent experiments showed that the EC50 for the stimulatory action of NMDA was 2-3 x 10(-6) M with a Bmax of around 10(-5) M. At 10(-4) M KA and Q but not NMDA reduced tissue content and release of SRIF (KA: 47 +/- 14, 67 +/- 17%; Q: 36 +/- 13, 42 +/- 6% of basal for content and release, respectively). These findings indicate that cortical SRIF content and release is enhanced by exposure to NMDA but not by KA or Q. We suggest that SRIF-containing neurones are sensitive to glutamate damage through the activation of non-NMDA rather than NMDA receptors.

Substance P and somatostatin contents were measured in the gastrointestinal tract of streptozotocin diabetic rats, 1 month after streptozotocin administration (60 mg/kg), and of age-matched controls with radioimmunoassay. Substance P and somatostatin contents were statistically increased in the extrafundus of the diabetic stomach, but not in the diabetic fundus. Substance P was significantly decreased in the diabetic ileum and caecum. Similarly, somatostatin was decreased in the diabetic caecum. Contrarily, slight increase of somatostatin contents in the diabetic duodenum, jejunum and proximal colon was not statistically significant.

To estimate the functional role of endogenous somatostatin in the production of long-term potentiation (LTP) in mossy fiber-CA3 system, an influence of depletion of somatostatin on the magnitude of it was examined in guinea-pig hippocampal slices. Administration of cysteamine (200 mg/kg, s.c.), a depletor of somatostatin, to guinea-pigs 13 h prior to preparing slices resulted in a significant decrease in the magnitude of LTP of population spikes in mossy fiber-CA3 system, being associated with a significant depletion of the content of somatostatin in the hippocampus. Furthermore, bath-applied somatostatin (1-14) at a concentration, at which the substance did not influence LTP in slices prepared from saline-treated animals, significantly augmented LTP in slices from cysteamine-treated animals. Cyclo-somatostatin (0.32 and 3.2 microM), a putative antagonist of somatostatin receptors, failed to affect the magnitude of LTP of mossy fiber-CA3 system when applied alone; however, the combined application of cyclo-somatostatin with somatostatin (0.32 microM) significantly inhibited the augmenting action of somatostatin on LTP. From these observations, it is suggested that endogenous somatostatin plays a facilitatory role in the production of LTP in mossy fiber-CA3 system in guinea-pig hippocampus.

The earliest somatostatin-immunoreactive (SS-Ir) perikarya of the human fetal frontal cortex appear in the transient subplate zone at 22 weeks of gestation. Around 32 weeks of gestation there is an increase in the number of SS-Ir neurons at the interface between the subplate zone and the cortical plate. The newborn-cortex shows decline in the overall number of SS-Ir neurons parallel to the appearance of SS-Ir neurons in the superficial layers. In conclusion, the subplate neurons are the source of the earliest peptidergic activity in the cortex. Furthermore, the distribution and density of peptidergic neurons undergo significant reorganization during the perinatal development.

As neuropeptides may play a role in the electrical kindling model of epileptogenesis, hippocampal somatostatin, Met-enkephalin and cholecystokinin were studied by immunocytochemistry in rats 24 h following full hippocampal kindling (three stage 5 seizures). As control animals we used sham-kindled rats, unoperated rats and rats subjected to a single electroshock-induced seizure. In addition, the distribution of septohippocampal, cholinergic fibers and hippocampal mossy fibers were studied by histochemistry. The important finding was that after kindling there was, as compared to unoperated control, (1) a marked increase of somatostatin immunoreactivity in cell bodies in the dentate hilus and their presumed projections area in the outer parts of the dentate molecular layer, and (2) a marked increase of Met-enkephalin immunoreactivity in hippocampal mossy fiber terminals. We found no evidence of aberrant sprouting of mossy fiber collaterals in the fascia dentata.

The gamma-Aminobutyric acid-A (GABAA) antagonist picrotoxin and bicuculline were administered to male rats to determine their effects on somatostatin (SRIF) release, measured in unanesthetized animals stereotaxically implanted with push-pull cannula in the median eminence (ME). I.p. injection (3 mg/kg) of picrotoxin (n = 5) or bicuculline (n = 6) significantly increased (35.4 +/- 10.8 vs 13.7 +/- 4.3 pg/15 min; P less than 0.03 and 38 +/- 3.5 vs 14 +/- 1.8 pg/15 min; P less than 0.001, respectively) SRIF release from the ME compared to baseline levels measured in the same animals. In contrast, with local perfusion of picrotoxin, (10(-4) to 10(-6) M) SRIF release from the ME was not affected. These data suggest a physiological endogenous inhibitory tone of GABA on SRIF release.

The messenger RNAs coding for vasopressin, oxytocin, luteinizing hormone releasing-hormone and somatostatin have been detected in tissue sections of the rat brain, especially in the hypothalamus with radioactive and biotinylated oligonucleotide probes. The results demonstrate that neuropeptide mRNAs are present in the cytoplasm of cell bodies, in processes and in punctate structures in the vicinity of the cell bodies. These results demonstrate that neuropeptide mRNAs can be transported outside the cell body most probably in proximal dendrites but also in some of their branching, and possibly at synaptic contacts. These data suggest that neuropeptide mRNA could undergo a specific compartmentation that could contribute to the targetting of the corresponding peptide inside neurons.

L-Homocysteic acid (L-HCA) is a sulfated amino acid which is present in mammalian striatum and is a putative excitatory striatal neurotransmitter. In the present study we examined the histologic and neurochemical effects of L-HCA induced striatal lesions to determine how closely changes resemble those of Huntington's disease (HD). Increasing doses of L-HCA injected into the anterior striatum resulted in dose-dependent reductions in both substance P-like immunoreactivity (SP-LI) and gamma-aminobutyric acid (GABA) while there was a relative sparing of both somatostatin-like immunoreactivity (SS-LI) and neuropeptide Y-like immunoreactivity (NPY-LI). Immunocytochemical studies showed a relative sparing of NADPH-diaphorase neurons (which colocalize with SS and NPY) within regions in which there was a significant depletion of enkephalin stained neurons. The lesions were blocked by pretreatment with MK-801, a systemically effective non-competitive antagonist of N-methyl-D-aspartate (NMDA) receptors or coinjection of equimolar concentrations of 2-amino-5-phosphonovalerate (APV). These findings are similar to those produced with the NMDA agonist quinolinic acid, and suggest that other endogenous NMDA agonists, such as L-HCA, could be potential excitotoxins in HD.

Somatostatin 28(1-14) immunoreactivity is present in dorsal root ganglion cells and in the superficial laminae of the spinal cord in the rat. The distribution of somatostatin 28(1-14) immunoreactive varicosities in the dorsal horn corresponds well to the distribution of somatostatin 14 immunoreactive elements. Some dorsal root ganglion cells exhibit both somatostatin 14 and somatostatin 28(1-14) immunoreactivities.