Somatostatin-like Immunoreactivity Research Articles

Effects of aging on quinolinic acid lesions in rat striatum

Several neurologic illnesses in which excitotoxic mechanisms may play a role increase in prevalence with age. In the present study we examined the susceptibility of rats to quinolinic acid striatal lesions at 1, 4 and 20 months of age, and susceptibility to N-methyl- d-aspartate (NMDA) at 1 and 4 months of age. The extent of the lesions was quantitated with measurements of substance P-like immunoreactivity (SPLI) and γ-aminobutyric acid (GABA). The lesions in the 4- and 20-month-old age groups showed significantly smaller depletions of SPLI and GABA than those in 1-month-old animals. Neuropeptide Y-like immunoreactivity (NPYLI) and somatostatin-like immunoreactivity (SLI) were unchanged in the lesioned striata. NMDA lesions were also attenuated in 4-month- and 12-month-old animals as compared with 1-month-old animals. Uric acid concentrations showed marked dose-dependent increases in the lesioned striatum, and to a lesser extent in the overlying cerebral cortex, in all 3 age groups. There were no changes of SLI, NPYLI or SPLI with aging in the cerebral cortex or hippocampus. Kynurenine and kynurenic acid concentrations showed significant increases with aging in frontal cortex. The present results show a reduced susceptibility of animals to striatal quinolinic acid and NMDA lesions with normal aging. The delayed onset of several neurodegenerative illnesses is therefore unlikely to be due to an increasing susceptibility to excitotoxin lesions with aging.

Somatostatin Release from Rat Cerebral Cortex Synaptosomes

Rat cerebral cortex synaptosomes were exposed in superfusion to various depolarizing stimuli and the release of somatostatin-like immunoreactivity (SRIF-LI) was measured by means of a radioimmunoassay procedure. High KCl (9-50 mM) concentration dependently evoked SRIF-LI release; the evoked overflow reached a plateau at 25 mM KCl and was completely abolished when Ca2+ ions were omitted from the superfusion medium, independently of the concentration of KCl used. The 15 mM K(+)-evoked release of SRIF-LI increased sharply as the Ca2+ concentration was raised to 0.8 mM, then leveled off and reached a plateau at 1.2 mM. The 15 mM K(+)-evoked overflow, but not the spontaneous outflow, was partially decreased (50%) by 1 microM tetrodotoxin. The presence in the superfusion fluid of a mixture of peptidase inhibitors did not improve the recovery of SRIF-LI both in the absence and in the presence of high K+. Exposure of synaptosomes to veratrine (1-50 microM) induced release of SRIF-LI in a concentration-dependent way. The effect of the alkaloid was strictly Ca2+ and tetrodotoxin sensitive. Replacement of extracellular Na+ by sucrose caused an acceleration of the spontaneous SRIF-LI outflow that was inversely correlated to the Na+ content in the superfusion medium. The release evoked by the sodium-deprived media did not exhibit any calcium dependence. HPLC analysis of the samples collected during superfusion showed that greater than 90% of the SRIF-LI released either during the spontaneous outflow or by 15 mM KCl was represented by SRIF-14 (SRIF-28(14-28]. These values reflected the ratio SRIF-14/SRIF-28 found in synaptosomes at the end of the experiments.

Somatostatin-like immunoreactivity in the region of the prepacemaker nucleus in weakly electric knifefish, Eigenmannia: a quantitative analysis

The diencephalic prepacemaker nucleus (PPn) is a bilateral cluster of neurons that controls frequency modulations of the otherwise very regular electric organ discharge cycle in weakly electric knifefish, Eigenmannia. Ultrastructural evidence had suggested that the action of excitatory and inhibitory synapses contacting PPn neurons is modulated by neuropeptides. In this investigation, we examined the distribution of somatostatin-like immunoreactive (S-IR) structures in the region of the PPn. In transverse sections, we found 5 bilateral S-IR structures in the region of the PPn: a ‘dorsohorizontal stripe’; a ‘lateral cell group’; a ‘diagonal stripe’; a ‘tubercular stripe’; and a ‘hypothalamic stripe’. The dorsohorizontal stripe, consisting of S-IR fibers, terminals, and roughly 200 cell bodies unilaterally, stretches from the edge of the third ventricle at the thalamic dorsal-posterior nucleus and the central posterior nucleus approximately 400 μm laterally to the PPn. S-IR cell bodies show a distinct pattern of distribution within this stripe. Almost half of the somata are located within 50 μm of the ventricle. These ventricular cells are small and often densely clustered. Laterally, towards the PPn, the number of labelled cells decreases, whereas their size gradually increases. The lateral cell group consists of roughly 20 somata in the medial region of the subelectrosensorius nucleus. Fibers of the diagonal stripe travel from the hypothalamus dorsalis to the PPn. Fibers of the tubercular stripe originate from S-IR cell bodies in the medial zone of the periventricular nucleus of the posterior tuberculum and merge with the diagonal stripe. Fibers of the hypothalamic stripe connect the hypothalamus lateralis and the diagonal stripe. The density of immunolabelling in the hypothalamic stripe is significantly higher in mature than in immature females.

Substance P-like immunoreactivity and somatostatin-like immunoreactivity in the ventricular fluid of patients with chronic pain syndromes

Substance P-like and somatostatin-like immunoreactivities (SPLI and SLI) were determined in ventricular fluid of patients with chronic pain syndromes and in a comparison group with multiple sclerosis, essential tremor, epilepsy and postanoxic myoclonus. Concentrations of SPLI and SLI were non-significantly decreased by 40% and 33% in chronic pain patients as compared with control patients without pain. There were no differences apparent between subgroups of pain patients (deafferentation pain, neoplasia-induced pain, thalamic pain). High pressure liquid chromatography combined with radioimmunoassay showed marked heterogeneity of SPLI and SLI.

Somatostatin and cognitive functions in Alzheimer's disease — the relationship of cerebrospinal fluid somatostatin increase with clinical response to tetrahydroaminoacridine

We studied the effect of tetrahydroaminoacridine (THA) on cerebrospinal fluid somatostatin-like immunoreactivity (CSF-SLI) in probable Alzheimer disease (AD) patients (n = 20) who took part in an open THA treatment trial. The maintenance dose (100 mg/day) was continued for 4 weeks. Samples of CSF were obtained before treatment and at the end of the treatment period. The CSF-SLI increased significantly (P = 0.01) in the responders for the treatment (increase of the Mini-Mental State Examination score greater than or equal to 3; n = 11), while the non-responders (n = 9) showed a significant decrease of CSF-SLI (P = 0.003). The change of CSF-SLI had also a significant correlation (P = 0.001) with neuropsychological performance. We conclude that the effects of of THA on the CSF-SLI may be due to presynaptic cholinergic or direct somatostatinergic stimulation.

Sucrose octasulfate stimulates gastric somatostatin release

To explore the mechanisms of the effects of sucralfate on the stomach, we investigated the action of sucrose octasulfate (SOS), a constituent of sucralfate, on the function of canine gastric parietal cells and somatostatin cells and in the isolated perfused intact rat stomach. Somatostatin cells from the canine gastric fundus were isolated by EDTA-collagenase dispersion and counterflow elutriation, and somatostatin-like immunoreactivity (SLI) release in response to SOS was measured by radioimmunoassay. Similar methods were used to isolate gastric parietal cells, in which gastric acid secretion was measured by uptake of a radiolabeled weak base, [ 14C]aminopyrine. SLI release by the intact rat stomach was examined in an isolated vascularly perfused rat stomach model. SOS, either alone or co-administered with epinephrine or gastrin heptadecapeptide (G17), dose-dependently stimulated SLI release by isolated canine fundic D-cells. At the highest doses, SOS potentiated the effect of epinephrine but not G17. Similarly, SOS potentiated the stimulating effect of dibutyryl cyclic adenosine 3′,5′-monophosphate (DBcAMP), but not 12- O-tetradecanoylphorbol 13-acetate (TPA). The effect of SOS on SLI release could be inhibited by octreotide, a somatostatin analogue. SOS did not alter acid secretion by cultured canine parietal cells either in the basal state or when coadministered with acid secretagogues. In isolated perfused rat stomach studies, SOS produced a significant (60% greater than basal) increase in SLI secretion. There was a similar effect when SOS was perfused against a background of isoproterenol. SOS stimulates SLI release from gastric somatostatin cells and from the isolated perfused stomach but has no direct effect on gastric parietal cells. These actions of SOS may mediate in part the apparent ability of sucralfate to enhance gastric mucosal defense.

“Gastrin” and “CCK” receptors on histamine-and somatostatin-containing cells from rabbit fundic mucosa—II: Characterization by means of selective antagonists (L-364, 718 and L-365, 260)

In the preceding paper, by means of selective agonists to gastrin (HG-17) and cholecystokinin (CCK-39), we evidenced the existence of “gastrin-type” receptors that could regulate histamine release and “CCK-type” receptors that could stimulate somatostatin release in isolated rabbit fundic nonparietal cells (F1 cells). Furthermore, these receptors could induce phosphoinositide breakdown. To confirm the involvement of these receptor types in these biological and biochemical processes, we used selective antagonists, L-364,718 (3-(benzoylamino)-benzodiazepine) specific to “CCK-A-type” receptor and L-365,260 (3-(acylamino)-benzodiazepine) specific to “gastrin/CCK-B-type” receptor. Neither L-364,718 nor L-365,260 alone caused any significant stimulation of [ 3H]inositol phosphate ([ 3H] InsP) production and release of histamine or somatostatin-like immunoreactivity (SLI). Each analogue inhibited in a dose-dependent manner [ 125I] HG-17 or [ 125I]CCK-39 binding to F1 cells, [ 3H]InsP accumulation and histamine and SLI release stimulated by HG-17 or CCK-39. L-365, 260 appeared to be 30–70 times more potent than L-364,718 in inhibiting [ 125I]HG-17 binding to F1 cells, as well as HG-17-induced [ 3H]InsP accumulation and HG-17- or CCK-39-enhanced histamine release ( ic 50 values: ≈5–20 nM for L-365,260 and ≈200–1500 nM for L-364,718 ). In contrast, L-364,718 was 200 to 400 times more potent than L-365,260 in inhibiting [ 125I]CCK-39 binding to F1 cells, CCK-39-induced [ 3H]-InsP accumulation and SLI release stimulated by CCK-39 or HG-17 ( ic 50 values: ≈0.3–1 nM for L-364, 718 and 100–200 nM for L-365,260 ). These results led to conclude: (i) the existence of a “gastrintype” receptor related to histamine release; (ii) the existence of a “CCK-A-type” receptor related to somatostatin release; (iii) the existence of “gastrin type” and “CCK-A-type” receptors linked to the phosphoinositide breakdown pathway.

“Gastrin” and “CCK” receptors on histamine-and somatostatin-containing cells from rabbit fundic mucosa —I: Characterization by means of agonists

A previous study has suggested the presence of two distinct binding sites for gasrin and cholecystokinin (CCK) in isolated non-parietal cells from rabbit gastric mucosa: a receptor which binds CCK-8 and CCK-39 with a high affinity and a receptor which binds gastrin and CCK-8 with the same high affinity and CCK-39 with a lower affinity. To characterize these receptors, their ability to induce phosphoinositide breakdown was investigated. Gastrin (HG-17), CCK-39 and CCK-8 induced [ 3H]-inositol phosphate ([ 3H]InsP) accumulation from [ 3H]inositol prelabelled cells with a high potency ( ec 50: 0.3–2.7 nM) but CCK-8 exhibited a higher efficacy than HG-17 or CCK-39. HG-17, CCK-8 and CCK-39 induced a rapid accumulation of [ 3H]inositol monophosphate ([ 3H]InsP 1), [ 3H]inositol bisphosphate ([ 3H]InsP 2) and [ 3H]inositol trisphosphate ([ 3H]InsP 3) but CCK-8 caused a two times higher accumulation than HG-17 or CCK-39. Histamine- and somatostatin-containing cells appeared to be located in this non-parietal cells population. HG-17, CCK-8 and CCK-39 dose-dependently induced histamine release with the following order of potency: HG-17 = CCK-8 ( ec 50 ≈ 0.2 nM) > CCK-39 ( ec 50 ≈ 4 nM) . In addition, HG-17 exhibited the highest efficacy. HG-17, CCK-8 and CCK-39 enhanced somatostatin-like immunoreactivity (SLI) release with the following order of potency: CCK-8 ( ec 50 ≈ 0.1 nM) = CCK-39 > HG-17 ( ec 50 ≈ 10 nM) ; CCK-8 and CCK-39 exhibited the highest efficacy. These results led us to the following conclusions: (i) existence of a “gastrin-type” and of a “CCK-type” receptor mediating phosphoinositide breakdown in these gastric non-parietal cells. CCK-8 interacts with both receptor-types with the same affinity; (ii) the release of histamine from histaminecontaining cells could be induced following “gastrin-type” receptors activation; (iii) somatostatin release from D-cells present in this non-parietal cells population could be induced following “CCK-type” receptors activation.

CSF monoamine metabolites and somatostatin in Alzheimer's disease and major depression

Decreased cerebrospinal fluid (CSF), somatostatinlike immunoreactivity (SLI) and alterations in the CSF monamine metabolites 3-methoxy-4-hydroxyphenylethylglycol (MHPG), 5-hydroxyindoleacetic acid (5-HIAA), and homovanillic acid (HVA) have been reported in patients with probable Alzheimer's disease (AD) and in patients with major depression. In this study, we found CSF SLI to be significantly lower in a large group of AD patients n = 60) and in a group of age-matched patients with major depression (n = 18) as compared with normal controls (n = 12). Mean CSF, MHPG, 5-HIAA, andHVA levels were not significantly different among diagnostic groups. Within a group of “depressed” AD patients, CSF levels of 5-HIAA showed a significant positive correlation (p = 0.03) with CSF SLI; a similar relationship was found within the group of patients with major depression. Further exploration of the relationship between the somatostatin and serotonin systems may provide clues as to how neuropeptides interact with monoamine neurotransmitters and what role they have in depression.

Effect of maternal exposure to nicotine in the rat on level and binding of somatostatin in brain of developing offspring

The effect of maternal exposure to nicotine on the level of somatostatin and specific binding in frontoparietal cortex and hippocampus of developing offspring was investigated. Sprague-Dawley rats were injected subcutaneously, throughout the pregnancy and the nursing period, with either: 3 mg kg nicotine base or saline vehicle. In the offspring of control rats, the level of somatostatin-like immuno-reactivity peaked at day 10 in the frontoparietal cortex, whereas the level of immunoreactivity in the hippocampus was the highest on day 30. Maternal exposure to nicotine caused enhanced levels of immunoreactivity in the frontoparietal cortex, on the day of birth and in the hippocampus, up to day 10. The maximum specific binding of somatostatin to the receptors in membranes from the frontoparietal cortex, peaked at 10 days of age in the offspring of control rats. The number of somatostatin receptors in cortical (but not in hippocampal) membranes was significantly decreased in the 0- to 10-day-old offspring of the nicotine-treated rats. Despite transient alterations in the number of somatostatin receptors, the affinity of the sites for somatostatin was consistently unchanged. The levels of somatostatin-like immunoreactivity and the number of somatostatin receptors in the frontoparietal cortex and hippocampus was comparable in the 30-day-old offspring of the control and nicotine-treated rats.

Somatostatin-like immunoreactivity in the brain of an electric fish ( Ateronotus leptorhynchus) identified with monoclonal antibodies

The immunohistochemical localization of somatostatin-like immunoreactive (SSir) cells and fibers in the brain of the gymnotiform teleost ( Apteronotus leptorhynchus) was investigated using well-characterized monoclonal antibodies directed against somatostatin-14 and -28. Large populations of SSir neurons occur in the basal forebrain, diencephalon and rhombencephalon and a dense distribution of fibers and terminal fields is found in the ventral, dorsomedial and dorsolateral telencephalon, hypothalamus, centralis posterior thalamus, subtrigeminal nucleus, the motor nucleus of vagus and in the ventrolateral medulla. Immunoreactive neurons in the forebrain are concentrated mainly in the ventral telencephalic areas, the region of the anterior commissure and entopeduncular nucleus. In a fashion similar to the large basal telencephalic cells of other species, the cells of the rostral nucleus entopeduncularis have a significant projection to the dorsal telencephalon. The preoptic region and the peri- and paraventricular hypothalamic nuclei are richly endowed with SSir cells; some of these cells contribute fibres to the pituitary stalk and gland. In the thalamus, only the n. centralis posterior stands out for the density of SSir cells and terminals; these cells appear to project to the prepacemaker nucleus, thus suggesting an SS influence on electrocommunication. In the mesencephalon most SSir cells occur in the optic tectum, torus semicircularis and interpeduncular nucleus. The rhombencephalic SSir cells have a wider distribution (central gray, raphe, sensory nuclei, reticular formation, electrosensory lateral line lobe and surrounding the central canal). The results of this study show the presence of SS in various sensory systems, electromotor system and specific hypothalamic nuclei, suggesting a modulatory role in the processing of sensory information, electrocommunication, endocrine and motor activities.

Regulation of islet somatostatin secretion and gene expression: selective effects of adenosine 3',5'-monophosphate and phorbol esters in normal islets of Langerhans and in a somatostatin-producing rat islet clonal cell line 1027 B2.

To investigate cAMP-dependent regulation of somatostatin secretion and gene expression in the islets of Langerhans, we have correlated the effects of forskolin, theophylline, and (Bu)2cAMP (dbcAMP) on the secretion of somatostatin-like immunoreactivity (SLI), cAMP generation, and somatosatin mRNA (S-mRNA) accumulation by cultured rat islet cells and a rat somatostatin-producing islet tumor cell line (1027 B2). Additionally, we have compared these effects with those of phorbol esters. Forskolin induced large acute increases in cAMP levels in islet cells, whereas theophylline produced modest sustained elevations in cAMP. During 4-h exposure to islets cells, forskolin, theophylline, and dbcAMP produced time- and dose-related increases of up to 14-fold in SLI release and up to 5-fold in S-mRNA levels. The rate of increase in S-mRNA paralleled secretion and occurred with the following order of potency: forskolin greater than dbcAMP greater than theophylline. The analog 1,9-dideoxyforskolin, which is unable to activate adenylyl cyclase, produced a small increase in SLI release without affecting S-mRNA. The effects of short term increases in islet cAMP levels and SLI release on long term changes in S-mRNA accumulation were investigated in a 48-h study with forskolin. Pretreatment of islet cells for 30 min with forskolin evoked large acute increases in cAMP levels and SLI release. S-mRNA rose in a biphasic pattern, with an acute increase at 30 min followed by a secondary increase at 12-48 h. In 1027B2 cells, forskolin and theophylline generated large increases in cAMP levels. Despite this, the two agents as well as dbcAMP produced only slight (20-35%) stimulation of SLI release and S-mRNA accumulation. Phorbol 12-myristate 13-acetate and phorbol 12,13-dibutyrate evoked dose-dependent stimulation of SLI secretion of up to 4-fold from islet cells without altering S-mRNA. Both secretion and S-mRNA were unresponsive to phorbol esters in 1027 B2 cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Interictal spikes and hippocampal somatostatin levels in temporal lobe epilepsy.

We investigated the relationship between somatostatin-like immunoreactivity (SSLI) and interictal spikes (IIS) in human temporal lobe epileptic tissue. IIS counted manually from depth electrode recordings obtained preoperatively were expressed as spike frequency in anterior, middle, and posterior portions of hippocampus. SSLI was determined by radioimmunoassay (RIA). An inverse relationship between SSLI in the entorhinal cortex (EC) and IIS frequency in hippocampus was present (r = -0.55, p = 0.06). No correlation between IIS and SSLI in CA4, CA3, CA1, or the dentate was evident. This finding suggests a role of the EC in generation, regulation, or expression of interictal paroxysmal electrical activity in temporal lobe epilepsy (TLE), for which somatostatin may be a marker.

Somatostatin modulates vascular sympathetic neurotransmission in the rabbit ear artery

Somatostatin-like immunoreactivity was localised immunohistochemically in perivascular nerves in the rabbit central ear artery. Whilst somatostatin had no direct action on this vessel, it significantly inhibited noradrenaline-induced, but not a,β-methylene ATP-induced, vasoconstriction. Somatostatin also inhibited contractions elicited by electrical field stimulation showing greater effect at low (16 Hz) compared with high (64 Hz) frequencies, and inhibited the release of tritiated noradrenaline in a concentration-dependent manner. These results confirm that somatostatin is a neuroregulatory peptide, and suggest that it is modulating vascular sympathetic cotransmission of the rabbit central ear artery by acting both prejunctionally to inhibit transmitter release, and postjunctionally to reduce the action of noradrenaline.

Quinolinic acid stimulates somatostatin gene expression in cultured rat cortical neurons.

Striatal atrophy in Huntington's disease (HD) is characterized by selective preservation of a subclass of neurons colocalizing NADPH-diaphorase (NADPH-d), somatostatin (SS), and neuropeptide Y (NPY), which have been reported to show three- to fivefold increases in SS-like immunoreactivity (SSLI) and NPY content. Since HD brain is capable of producing excessive quantities of the excitotoxin quinolinic acid (Quin), an N-methyl-D-aspartate (NMDA) receptor agonist, and since experimental Quin lesions show neuronal loss with sparing of NADPH-d/SS/NPY neurons, it has been suggested that Quin may be important in the pathogenesis of HD. In the present study we determined whether Quin stimulates SS gene function in cultured cortical cells known to be rich in NADPH-d/SS/NPY neurons. Cultures of dispersed fetal rat cortical cells were exposed to Quin (1 and 10 mM) with or without (-)-2-amino-5-phosphonovaleric acid (APV; 0.5 mM), an NMDA receptor antagonist, NMDA (0.2 and 0.5 mM), and glutamate (Glu; 0.5 mM). Medium and cellular SSLI was determined by radioimmunoassay and SS mRNA by Northern analysis with a cRNA probe. Quin induced significant (p less than 0.01) 1.6- and 2.5-4 fold increases in SSLI and SS mRNA accumulation, respectively, which were abolished by APV. Release of SSLI into the culture medium was stimulated two- to fivefold by Quin over a 2- to 20-h period. The increase in SS mRNA produced by Quin was time and dose dependent. A similar dose-dependent increase in SS mRNA comparable with that observed with Quin was induced by NMDA.(ABSTRACT TRUNCATED AT 250 WORDS)

Stimulation of somatostatin expression in developing ciliary ganglion neurons by cells of the choroid layer

An important component of neuronal development is the matching of neurotransmitter expression with the appropriate target cell. We have examined how peptide transmitter expression is controlled in a simple model system, the avian ciliary ganglion (CG). This parasympathetic ganglion contains 2 distinct types of neurons: choroid neurons, which project to vasculature in the eye's choroid layer and use somatostatin as a co-transmitter with ACh, and ciliary neurons, which innervate the ciliary body and iris and use ACh but no known peptide co-transmitter. We have found that the earliest developmental stage in which neurons with somatostatinlike immunoreactivity (SOM-IR) are consistently found in vivo is stage 30 (embryonic day 6.5), a time shortly after the extension of neurites to targets in the eye's choroid layer. In cell culture, CG neurons expressed SOM-IR in co-culture with choroid cells, but not when cultured with striated muscle myotubes or with ganglion non-neuronal cells. No significant differences in neuronal survival or in ChAT activity were observed under these different co-culture conditions, which suggests that somatostatin expression is independently regulated. The stimulation of somatostatin expression was also specific in that other neuropeptides commonly found in autonomic neurons [neuropeptide Y (NPY), substance P (SP), vasoactive intestinal polypeptide (VIP)] were not induced in the presence of choroid cells. The ability to stimulate SOM-IR was not contact dependent because a macromolecule of greater than or equal to 10 kDa in choroid-conditioned medium (ChCM) was found to stimulate somatostatin expression in a dosage-dependent fashion. The somatostatin-stimulating activity induced SOM-IR in more than 90% of CG neurons, as well as in retrogradely labeled ciliary neurons, which would not normally express SOM-IR. Thus, the expression of somatostatin in cultured CG neurons is regulated by a macromolecule produced by cells in the choroid layer, a target normally innervated in vivo by CG neurons expressing somatostatin.

Immunocytochemical evidence for the presence of somatostatin-like immunoreactive nerves in human dental pulp.

Specific antiserum to somatostatin was used for the immunocytochemical detection of this neuropeptide in human dental pulp. Immunoreactive axon varicosities were observed in the radicular as well as coronal pulp. Fibers displaying somatostatin-like immunoreactivity were detectable within radicular nerve trunks and were found to be associated mainly with blood vessels. Nevertheless, positive fibers with no apparent relation to blood vessels were also observed. No pulp cell was found to be immunoreactive. Previous physiological studies demonstrated that somatostatin may function as a regulatory peptide in feline dental pulp via a pre-synaptic inhibition of substance P release from sensory nerve terminals. It is tempting to speculate that such a mechanism may also be effective in human teeth and may be of value in the regulation of pulpal blood flow and in situations when sensory nerve fibers are activated, e.g., during pulpal inflammation.

Role of cholecystokinin in the control of gastric somatostatin in the rat: in vivo and in vitro studies

Cholecystokinin (CCK) has been shown to be a powerful stimulus for somatostatin release from isolated canine fundic D-cells in short-term culture. The influence of the CCK analogue caerulein on the secretory activity of the D-cell in the intact stomach in vitro and the effect of elevated plasma levels of endogenous CCK on gastric somatostatin stores in vivo were investigated in the rat. Basal somatostatin secretion from the isolated, vascularly perfused rat stomach preparation was not affected by various doses of caerulein. Slight stimulation of somatostatin-like immunoreactivity (SLI) release by epinephrine was significantly inhibited by caerulein, whereas caerulein did not alter half-maximal stimulation of SLI secretion by isoproterenol. Rats with chronically elevated plasma CCK levels induced by experimental exocrine pancreatic insuffiency did not show any change in tissue concentrations of SLI or in D-cell number, both in the antrum and corpus. These data suggest that CCK — in contrast to dogs — is not an important modulator of gastric somatostatin in the rat.

N-methyl- d-aspartate evokes the release of somatostatin from striatal interneurons in primary culture

Indirect immunocytochemistry of striatal neurons in primary culture, generated from the embryonic mouse brain, suggested that 2–4% of the neurons contained somatostatin-like immunoreactivity; the majority of these cells also contained neuropeptide Y immunoreactivity, characteristic of a subset of striatal interneurons. Although 10–15% of cultured striatal neurons showed moderate or intense immunoreactivity for calbindin-D 28k, the majority of neurons with somatostatin-like immunoreactivity did not contain calbindin-D 28k-like immunoreactivity; parvalbumin immunoreactivity was absent from the culture preparation. A highly sensitive radioimmunoassay was used to examine the actions of depolarizing agents and excitatory amino acids on the release of endogenous somatostatin-like immunoreactivity from striatal interneurons. During a 15 min incubation period, 47 ±10fmol of somatostatin-like immunoreactivity were released from 14 days in vitro striatal neurons, cultured in 35 mm dishes. Depolarization with 56 mM KC1 or 10 μg/ml veratrine resulted in an additional 105 ± 9 and 56 ± 5fmol, respectively, of somatostatin-like immunoreactivity released; the release evoked by veratrine was blocked by 1 μM tetrodotoxin. In the presence of 100 μM N-methyl- d-aspartate, 112 ± 21fmol of somatostatin-like immunoreactivity (above basal) were released (+238%); the N-methyl- d-aspartatevoked release was dose-dependent (EC 50, 20 μM), attenuated in the absence of added Ca 2+, potentiated in the absence of added Mg 2+ and unaffected by the presence of 1 μM tetrodotoxin. The selective antagonists 2-amino-5-phosphonovalerate (100 μM) and MK-801 (1 μM) blocked the N-methyl- d-aspartate-evoked release of somatostatin-like immunoreactivity; KCl-evoked release was unaffected. Kainate was slightly more effective, yet five-fold less potent ( ec 50, 100 μM), than N-methyl- d-aspartate in evoking somatostatin-like immunoreactivity release; quisqualate was marginally effective. The results of this study suggest that N-methyl- d-aspartate and kainate receptors are present on striatal somatostatinergic interneurons in primary culture.

Anesthetic dependence of the inhibitory effect of neurotensin on pentagastrin-stimulated acid secretion in rats. A possible role for somatostatin

The existence of possible local mediators of the inhibitory effect of neurotensin on gastric acid secretion has not been determined. We perfused rats intragastrically with warmed saline and stimulated acid secretion with intravenous pentagastrin, 32 mug/kg/hr, and found that anesthesia with pentobarbital resulted in marked inhibition of acid secretion by intravenous neurotensin; however, anesthesia with urethane prevented this inhibitory effect of neurotensin from occurring. In addition, we found a significant increase in somatostatin-like immunoreactivity in portal venous blood during neurotensin infusion in pentobarbital-anesthesized rats. Neither neurotensin nor pentagastrin infusion modified gastric luminal somatostatin-like immunoreactivity after either pentobarbital or urethane, and rats anesthesizex with urethane did not show an increase of somatostatin-like immunoreactivity in portal venous blood during neurotensin infusion. These results suggested that somatostatin-like immunoreactivity, released into the portal circulation, was necessary for exogenous neurotensin to inhibit pentagastrin-stimulated gastric acid secretion under those conditions in anesthesized rats.