Immunoreactive Somatostatin Research Articles

Demonstration of long‐range GABAergic connections distributed throughout the mouse neocortex

Gamma-aminobutyric acid (GABA)ergic neurons in the neocortex have been mainly regarded as interneurons and thought to provide local interactions. Recently, however, glutamate decarboxylase (GAD) immunocytochemistry combined with retrograde labeling experiments revealed the existence of GABAergic projection neurons in the neocortex. We further studied the network of GABAergic projection neurons in the neocortex by using GAD67-green fluorescent protein (GFP) knock-in mice for retrograde labeling and a novel neocortical GABAergic neuron labeling method for axon tracing. Many GFP-positive neurons were retrogradely labeled after Fast Blue injection into the primary somatosensory, motor and visual cortices. These neurons were labeled not only around the injection site, but also at a long distance from the injection site. Of the retrogradely labeled GABAergic neurons remote from the injection sites, the vast majority (91%) exhibited somatostatin immunoreactivity, and were preferentially distributed in layer II, layer VI and in the white matter. In addition, most of GABAergic projection neurons were positive for neuropeptide Y (82%) and neuronal nitric oxide synthase (71%). We confirmed the long-range projections by tracing GFP-labeled GABAergic neurons with axon branches traveled rostro-caudally and medio-laterally. Axon branches could be traced up to 2 mm. Some (n = 2 of 4) were shown to cross the areal boundaries. The GABAergic projection neurons preferentially received neocortical inputs. From these results, we conclude that GABAergic projection neurons are distributed throughout the neocortex and are part of a corticocortical network.

Somatostatin immunoreactivity in quail pterygopalatine ganglion

In the ciliary ganglion of the chicken and quail, somatostatin (SOM) is an exclusive marker for parasympathetic postganglionic neurons innervating the choroid. A second parasympathetic pathway projecting to the choroid originates from the pterygopalatine ganglion. The aim of this study was to investigate SOM immunoreactivity in the pterygopalatine ganglion of the Japanese quail (Coturnix coturnix japonica) and on neurons within the choroid, the intrinsic choroidal neurons (ICN). We did so using immunohistochemistry and subsequent light, electron and confocal laser scanning microscopy. Pterygopalatine neurons were characterized by nNOS-immunohistochemistry or NADPH-diaphorase cytochemistry. SOM immunoreactivity was absent in the perikarya, but neurons were densely surrounded by SOM-positive nerve fibres. Electron microscopy revealed that these fibres formed contacts with and without membrane specializations on pterygopalatine neurons. In the choroid, neuronal nitric-oxide synthase (nNOS)-immunoreactive ICN were likewise closely apposed by SOM-immunoreactive nerve fibres, as revealed by confocal microscopy. There was no detectable co-localization of the markers. In the absence of tracing studies, it is open to speculation whether SOM immunoreactivity originates from preganglionic fibres of the superior salivatory nucleus, postganglionic fibres of the ciliary ganglion or fibres of the brainstem via as yet unknown pathways. SOM may regulate the production of NO in pterygopalatine neurons and ICN, respectively, and is therefore involved in neuronal circuits regulating ocular homeostasis.

Age-related change of somatostatin-immunoreactive neurones in the main olfactory bulb of the rat.

Somatostatin is found in the olfactory system, including the main olfactory bulb (MOB), and is thought to be one of the neuroactive substances for olfaction. However, somatostatin immunoreactivity in the olfactory system has not been determined during ageing. Hence, we examined the age-related changes of somatostatin-immunoreactive (IR) neurones in the rat MOB over a period of 2 years, at the following various ageing stages: post-natal month 1 (PM 1), PM 3, PM 6, PM 12 and PM 24. In PM 1 group, a few somatostatin-IR neurones were detected in the granule cell layer (GCL), and had slender or oval somata and short processes. At PM 3, somatostatin-IR neurones were observed in the glomerular, external plexiform and GCL. The size of somatostatin-IR somata was larger than that at PM 1. In PM 6 group, the number and size of somatostatin-IR neurones increased, and their processes became longer while running in various directions. At PM 12, somatostatin-IR neurones increased in number, and their processes became markedly longer than those at PM 6. At this stage, somatostatin-IR neurones had multipolar somata, and were the largest in size. In PM 24 group, somatostatin-IR neurones were most numerous. However, the processes of somatostatin-IR neurones were shorter than those at PM 12. This study suggests that the increased number of somatostatin-IR neurones in the MOB of aged rats may play a role to compensate for any decrease of olfactory function.

Pituitary adenylate cyclase activating polypeptide (PACAP) decreases neuronal somatostatin immunoreactivity in cultured guinea-pig parasympathetic cardiac ganglia

Postganglionic parasympathetic neurons in guinea-pig cardiac ganglia exhibit choline acetyltransferase (ChAT)-immunoreactivity, and a large fraction (60%) of the ChAT-positive cardiac neurons co-express somatostatin-immunoreactivity. This co-expression remained when the cardiac ganglia explants were maintained in culture for 72 h (40% somatostatin-immunoreactive). The guinea-pig cardiac ganglia neurons express the high affinity pituitary adenylate cyclase activating polypeptide (PACAP)-selective PAC 1 receptor, and treatment of the ganglia explants with 20 nM PACAP27 for 72 h to evaluate PACAP regulation of somatostatin expression revealed a dramatic 85% decrease in the number of somatostatin-IR neurons (6% somatostatin-IR neurons) compared with untreated control explant preparations. The decrease in percentage of somatostatin-IR neurons by PACAP27 was time- and concentration-dependent, and selective for PACAP27; PACAP38 and vasoactive intestinal polypeptide were less effective. PACAP6-38, a PACAP antagonist, eliminated the PACAP27-induced change in somatostatin positive neurons. The PACAP-mediated decrease in somatostatin-IR neurons was eliminated in calcium-deficient solutions and by the addition of nifedipine, indicating a requirement for calcium influx through L-type calcium channels. The addition of either the calmodulin inhibitor N-(4-aminobutyl)-1-naphthalenesulfonamide or the MEK inhibitor PD98059, also eliminated the PACAP27-induced decrease in somatostatin-IR cells. The PACAP27-mediated effect on somatostatin expression was not affected by inhibitors of protein kinase A or phospholipase C, but was reduced by the adenylyl cyclase inhibitor SQ22356, suggesting cAMP involvement. Semiquantitative and quantitative reverse transcription PCR prosomatostatin transcript measurements showed that cardiac ganglia prosomatostatin mRNA levels were not diminished by chronic PACAP27 exposure despite the dramatic decrement in somatostatin-expressing neurons. Neuronal peptide-IR content represents a balance between production and secretion. These results suggested that one of the primary effects of PACAP exposure may be enhanced levels of neuropeptide release that exceeded production levels, resulting in somatostatin depletion and a decrement in the number of identifiable somatostatin-expressing cardiac neurons.

Frequent expression of immunoreactive somatostatin receptors in cervical and endometrial cancer

Objective A variety of human cancers has been shown to express somatostatin receptors which can be utilized for in vivo tumor targeting. Although the somatostatin receptor status of mammary and ovarian cancer has been studied in detail, somatostatin receptors have not been examined in other gynecological malignancies such as cervical or endometrial carcinoma. Methods We have previously developed a panel of somatostatin receptor subtype-specific antibodies that effectively stain formalin-fixed, paraffin-embedded human tumor tissue. In the present study, we have used these antibodies to determine the somatostatin receptor status of 21 randomly selected cervical carcinomas and 28 randomly selected endometrial carcinomas. Results In cervical carcinoma, the incidence of somatostatin receptor expression was sst 2A (12/21, ∼57%) > sst 3 (9/21, ∼43%) > sst 1 (8/21, ∼38%) > sst 4 (0/21, 0%) = sst 5 (0/21, 0%). In endometrial carcinoma, the incidence of somatostatin receptor expression was sst 3 (12/28, ∼43%) > sst 2A (11/28, ∼39%) > sst 1 (9/28, ∼32%) > sst 4 (1/28, ∼4%) = sst 5 (1/28, ∼4%). Somatostatin receptor-like immunoreactivity was in most cases located at the plasma membrane and completely blocked with antigenic peptide. The expression of somatostatin receptor subtypes was independent of patient age, diagnosis, tumor stage, and histological grade. Conclusion Our findings reveal a previously unappreciated high incidence of sst 1, sst 2A, and sst 3 somatostatin receptors in human cervical and endometrial cancers and suggest that a subgroup of receptor-positive uterine carcinomas may be a potential target for diagnostic and therapeutic intervention with stable somatostatin analogs.

Increase in somatostatin immunoreactivity in the suprachiasmatic nucleus of aged Wistar rats

Decreased immunoreactivity has been reported for several neuropeptides in the aged suprachiasmatic nucleus (SCN). We compared somatostatin (SS) and substance P (SP) immunoreactivity (ir) in aged (20–26 months) and young (6 months) Wistar rats. The old rat SCN revealed a significant increase in SSir (2.6-fold) and SPir. The results show that not all SCN–neuropeptidergic systems decline with age, and suggest a specific age-related role for SS in the SCN.

Comparative immunohistochemical analysis of the peptidergic innervation of human primary and permanent tooth pulp

This immunohistochemical study sought to determine whether there are any differences in the peptidergic innervation of these pulps and whether dental caries is associated with changes in neuropeptide expression. Mandibular first permanent molars and second primary molars ( n=120) were obtained from children requiring dental extractions under general anaesthesia. Extracted teeth were split longitudinally, placed in fixative, and categorized as intact, moderately carious or grossly carious. The coronal pulps were removed and 10-μm frozen sections were processed for indirect immunofluorescence. Double labelling employed combinations of the following antisera: (1) protein gene product 9.5, a general neuronal marker; (2) one of the neuropeptides calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), galanin (GAL), enkephalin (ENK) and somatostatin (SOM). Image analysis was then used to determine the percentage area of immunostaining for each label within different anatomical regions of the coronal pulp. Sparse or absent immunoreactivity for GAL, ENK and SOM made analysis impossible. Analysis of CGRP, SP and VIP revealed significant interdentition differences, with their expression being significantly greater in permanent teeth, but this was not the case for NPY, with primary and permanent teeth demonstrating a similar amount of label for this peptide. Both dentitions showed significant increases in CGRP, SP, VIP and NPY expression with caries progression. These findings could have biological and clinical importance in connection with nociception, inflammation and healing.

Somatostatin immunoreactivity in axon terminals in rat nucleus tractus solitarii arising from central nucleus of amygdala: coexistence with GABA and postsynaptic expression of sst 2A receptor

Axon terminals synapsing on neurones in the nucleus tractus solitarii (NTS) that originate from the central nucleus of the amygdala (CeA) have been shown to contain γ-aminobutyric acid (GABA) immunoreactivity. Here we investigated whether such terminals also contain somatostatin (SOM), a neuropeptide found in axons distributed throughout the NTS and in somata in the CeA, and known to modulate cardiovascular reflexes when microinjected into the NTS. With fluorescence microscopy, SOM immunoreactivity was seen in the varicosities of some axons throughout the NTS that were anterogradely labelled with biotin dextran amine injected into the CeA. Such varicosities were frequently observed in close proximity to dendrites of NTS neurones that were immunoreactive for the SOM receptor sst 2A subtype, and in many cases also for catecholamine synthesising enzymes. In the caudal, cardioregulatory zone of NTS, SOM immunoreactivity was localised by electron microscopic pre-embedding gold labelling to boutons containing dense-cored and clear pleomorphic vesicles and forming symmetrical synapses, mostly onto dendrites. Additional post-embedding gold labelling for GABA suggested that a subpopulation (29%) of GABAergic terminals sampled in this area of NTS contained SOM. Almost all boutons anterogradely labelled from the amygdala were GABA-immunoreactive (-IR) and 21% of these were SOM-IR. A similar proportion of these boutons (22%) formed synapses onto dendrites containing immunoreactivity for the SOM receptor sst 2A subtype. These observations provide evidence that some of the GABAergic projection neurones in the CeA that inhibit baroreceptor reflex responses in the NTS in response to fear or emotional stimuli could release SOM, which might modulate the activity of NTS neurones via an action on sst 2A receptors.

Ghrelin: a hypothalamic GH-releasing factor in domestic fowl (Gallus domesticus).

Ghrelin, a recently discovered peptide in the mammalian hypothalamus and gastrointestinal tract is thought to be the endogenous ligand for the GH secretagogue (GHS) receptor and it stimulates GH release in rats and humans. The possibility that ghrelin is present in birds was therefore assessed, since a GHS receptor is present in the chicken pituitary gland. Although immunoreactive ghrelin is readily detectable in the rat stomach and ileum, ghrelin immunoreactivity could not be detected in the chicken proventriculus, stomach, ileum or colon, whereas somatostatin immunoreactivity, in contrast and as expected, was readily detectable in the chicken gastrointestinal tract. Ghrelin immunoreactivity was, however, present in the chicken hypothalamus, although not in the arcuate (infundibular) nucleus, as in rats. Discrete parvocellular cells and neuronal fibers with ghrelin immunoreactivity were present in the anterior medial hypothalamus. This immunoreactivity was specific and completely abolished following the preabsorption of the antibody with an excess of human ghrelin. Ghrelin immunoreactivity was also present in clusters of large ovoid magnocellular cells in the nucleus magnocellularis preopticus pars medialis, nucleus magnocellularis preopticus supraopticus and in the chiasmaopticus. Immunoreactivity for ghrelin was restricted to the cytoplasm of the perikarya and their axonal sprouts. Immunoreactivity for ghrelin was not seen in any other hypothalamic nuclei. In a preliminary experiment, circulating GH concentrations in conscious immature chicks were promptly increased following bolus i.v. administration of human ghrelin. The increase in GH concentration (approximately three times that in the controls) was comparable with that induced by the same dose (10 microg/kg) of human GH-releasing hormone, although less than that (approximately sixfold) induced by thyrotropin-releasing hormone. These results demonstrate the presence of a ghrelin-like protein in the chicken hypothalamus and suggest that it participates in the regulation of GH secretion in birds.

Implicación de los neuropéptidos metionina­encefalina, neurotensina y somatostatina del núcleo caudal del trigémino en la migraña experimental

Primary peptidergic sensory neurons of the trigeminal ganglion that innervate the cerebral dura have been involved in the pathogenesis of headache, including the migraine. In addition, it is known that nociceptive central processes of the trigeminal neurons terminate in the caudal trigeminal nucleus. Moreover, the electrical stimulation of the trigeminal ganglion has been used as an experimental model in order to study the vascular headache, including the migraine. To study whether there is or not a decrease of the immunoreactivity for methionine enkephalin, somatostatin and neurotensin in the caudal trigeminal nucleus after electrical stimulation of the trigeminal ganglion. The trigeminal ganglia of Wistar albino rats of both sexes were electrically stimulated (frequency, 5 Hz; duration, 5 ms; intensity, 0,8 1.4 mA) and unilaterally for five minutes. Sections of the medulla oblongata containing the caudal trigeminal nucleus were obtained and processed for immunocytochemistry, in which specific antibodies were used against methionine enkephalin, neurotensin and somatostatin 28. In stimulated animals, we observed a decrease in the immunoreactivity for the three neuropeptides studied in the stimulated (ipsilateral) side, in comparison with the not stimulated side (contralateral). In control animals (not stimulated) the degree of the immunoreactivity was the same on both sides. 1. The decrease of the immunoreactivity in the ipsilateral side (stimulated) suggests that methionine enkephalin, neurotensin and somatostatin 28 are released in the caudal trigeminal nucleus after electrical stimulation of the trigeminal ganglion; 2. Methionine enkephalin and somatostatin 28 could act in the caudal trigeminal nucleus as inhibitors (with antinociceptive action) of another released exciters neuropeptides (with nociceptive action); and 3. These data will allow in the future to try new therapeutic strategies (e.g., the inhibition of the receptors implicated.), in order to alleviate certain headaches.

Absence of temporal lobe epilepsy pathology in dogs with medically intractable epilepsy.

Epilepsy is a common neurological problem in dogs. In some dogs, seizures cannot be controlled adequately with anticonvulsant medication. Temporal lobe epilepsy is the most common type of epilepsy in adult humans, it is frequently resistant to anticonvulsant therapy, and it is commonly associated with characteristic neuropathological abnormalities in the hippocampal dentate gyrus. We sought to test the hypothesis that dogs with medically intractable epilepsy have temporal lobe epilepsy. The hippocampi of 6 dogs that were euthanized because of chronic, recurrent seizures were compared with those of 8 nonepileptic controls. In control and epileptic dogs, stereological cell counting showed similar numbers of neurons in the hilus of the dentate gyrus, somatostatin immunoreactivity identified numerous immunopositive neurons in the hilus, and Timm staining showed the normal pattern of granule cell axon projections. These findings demonstrate a lack of hilar neuron loss and granule cell axon reorganization, suggesting that temporal lobe epilepsy is not a common cause of medically intractable epilepsy in dogs.

Absence of Temporal Lobe Epilepsy Pathology in Dogs with Medically Intractable Epilepsy

Epilepsy is a common neurological problem in dogs. In some dogs, seizures cannot be controlled adequately with anticonvulsant medication. Temporal lobe epilepsy is the most common type of epilepsy in adult humans, it is frequently resistant to anticonvulsant therapy, and it is commonly associated with characteristic neuropathological abnormalities in the hippocampal dentate gyrus. We sought to test the hypothesis that dogs with medically intractable epilepsy have temporal lobe epilepsy. The hippocampi of 6 dogs that were euthanized because of chronic, recurrent seizures were compared with those of 8 nonepileptic controls. In control and epileptic dogs, stereological cell counting showed similar numbers of neurons in the hilus of the dentate gyrus, somatostatin immunoreactivity identified numerous immunopositive neurons in the hilus, and Timm staining showed the normal pattern of granule cell axon projections. These findings demonstrate a lack of hilar neuron loss and granule cell axon reorganization, suggesting that temporal lobe epilepsy is not a common cause of medically intractable epilepsy in dogs.

Effects of central infusions of neuropeptide Y on the somatotropic axis in sheep fed on two levels of protein

Effects of infusions of neuropeptide Y (NPY) into 3rd ventricle of growing sheep fed on diets containing restricted (R) or elevated (E) levels of protein on the immunoreactive (ir) somatostatin neurones, ir somatotrophs, growth hormone (GH) concentration in the blood plasma were studied. The long-term restriction of protein in the diet elicited: enhancing irSS content in periventricular perikarya; diminishing irSS stores in the median eminence and elevating the number ir somatotrophs and content of irGH. NPY infusions enhanced the content of irSS in perikarya in sheep fed on E diet and diminished the number of ir somatotrophs and content of irGH of sheep fed on R diet. The R diet as well as NPY infusions caused an increase in GH mean concentrations in the blood plasma. Obtained results suggest that timulatory effect of restricted feeding and/or NPY action on GH secretion can be due to attenuated SS output. Since dietary restrictions and exogenous NPY have similar influence on the activation of GH secretion, we suggest that NPY could be a neuromodulatory link between nutritional cues and somatotropic axis in sheep.

Localization of the CB1 type cannabinoid receptor in the rat basolateral amygdala: high concentrations in a subpopulation of cholecystokinin-containing interneurons

The neuronal localization of the CB1 cannabinoid receptor in the rat basolateral amygdala was studied using peroxidase and fluorescence immunohistochemical techniques. All nuclei of the basolateral amygdala contained a large number of lightly stained pyramidal neurons and a small number of more intensely stained non-pyramidal neurons. Most of the latter cells had medium-sized to large multipolar somata and three to four aspiny dendrites, but some exhibited smaller oval somata. The axon initial segments of some of these non-pyramidal neurons exhibited large swollen varicosities in colchicine-injected animals, suggesting that much of the CB1 receptor protein is transported down the axons of these cells. Double-labeling studies using immunofluorescence histochemistry combined with confocal laser scanning microscopy revealed that the great majority of non-pyramidal neurons with CB1 receptor immunoreactivity belonged to a cholecystokinin-containing subpopulation. Whereas none of the other subpopulations of non-pyramidal neurons (exhibiting immunoreactivity for calretinin, parvalbumin, or somatostatin) expressed high levels of CB1 receptor immunoreactivity, a small percentage of these cells exhibited low levels of immunoreactivity. The results indicate that cannabinoids may modulate the activity of pyramidal projection neurons as well as a subpopulation of cholecystokinin-containing non-pyramidal neurons in the basolateral amygdala. Previous studies indicate that most of the latter are inhibitory interneurons that utilize GABA as a neurotransmitter. The intense staining of the cholecystokinin-containing interneurons and the evidence that large amounts of CB1 receptor protein are transported down the axons of these cells suggests that, as in the hippocampus, cannabinoids may inhibit the release of GABA from the axon terminals of these neurons.

Transient expression of somatostatin mRNA in developing ganglion cell layers of rat retina

Somatostatin (SOM) mRNA in developing ganglion cell layer (GCL) detected by in situ hybridization histochemistry and SOM peptide in developing optic chiasma and optic tract detected by immunocytochemistry were monitored to explore whether ganglion cells expressing SOM project to the visual center. Most of these cells in the developing GCL expressed SOM transiently from embryonic day 13 (E13) to E21. The cells expressing SOM mRNA initially followed a central-to-peripheral pattern of development. The cells expressing SOM mRNA in the retinas of fetuses became detectable at E13. From E14 to E17 the number of cells expressing SOM mRNA increased rapidly. At E17 most of the cells in the developing GCL expressed SOM mRNA. From E18 to postnatal days the positive cells became sparse except at the postnatal day 0 (PND0) the positive cells decreased dramatically in comparison with that at the E21. At PND15, the positive cells only can be found in the inner neuroblastic layer and in the ganglion cell layer. At PND20 the distribution pattern and the number of the positive cells were essentially the same as that in adult rat. SOM immunoreactivity was detectable at E16 in the developing optic chiasma and optic tract; the majority of the fibers in these area were SOM positive. From E16 to E18 the density of the immunostaining increased rapidly, whereas from E19 to E21 the density decreased. At PND0 no positive fibers were seen. The transient presence of SOM in most of the ganglion cells in the developing ganglion cell layer has prompted us to study the role of SOM in generation and differentiation of the retinal ganglion cells, and formation of the retina-visual center projections.

Distribution of somatostatin (SS) immunoreactivity using specific rabbit antibodies in preoptic-anterior hypothalamus (PO-AH) of female rat brain

Distribution of somatostatin (SS) immunoreactivity was observed in the preoptic-anterior hypothalamus (PO-AH) of the female rat brain. Six specific rabbit antibodies A (SS-14), B (SS-28), C (SS-28 complete), D (SS 14 and SS-28 both), E (SS preprohormone) and F (GHRH) were used for immunostaining using Avidin–Biotin Complex (ABC) method (Hsu et al., 1981). Immunostaining was observed with all the six antibodies, in the serial sections passing through various bregma levels (−0.3 to −3.3 mm) of preoptic-anterior hypothalamic (PO-AH) region including median eminence (ME). In conclusion, the present study suggests that immunoreactive nerve terminals for both SS-14 and SS-28 are present in internal (IZ) and external zones (EZ) of ME. High intensity of SS-14 and SS-28 containing terminals in EZ suggests that both SS fractions are involved in regulating GH secretion in anterior pituitary. This is a first report on comparative distribution of immunoreactivities of four different fractions of SS, SS-preprohormone and GHRH in PO-AH.

Helicobacter pylori infection in the cat: evaluation of gastric colonization, inflammation and function.

Further elucidation of the consequences of Helicobacter pylori infection on gastric mucosal inflammation and gastric secretory function would be facilitated by an animal model that is susceptible to infection with H. pylori, is broadly similar in gastric physiology and pathology to people, and is amenable to repeated non-invasive evaluation. The goal of this study was to examine the interrelationship of bacterial colonization, mucosal inflammation and gastric secretory function in cats with naturally acquired H. pylori infection. Twenty clinically healthy cats with naturally acquired H. pylori infection (cagA-, picB) and 19 Helicobacter-free cats were evaluated. Gastric colonization was determined by tissue urease activity, light microscopy, culture and PCR. The mucosal inflammatory response was evaluated by light microscopy, and by RT-PCR of the pro-inflammatory cytokines IL-1alpha, IL-1beta, IL-8 and TNF-alpha in gastric mucosa. Gastric secretory function was assessed by measuring pentagastrin-stimulated acid secretion, fasting plasma gastrin, and antral mucosal gastrin and somatostatin immunoreactivity. H. pylori colonized the pylorus, fundus and cardia in similar density. Bacteria were observed free in the lumen of gastric glands and were also tightly adherent to epithelial cells where they were associated with microvillus effacement. Mononuclear inflammation, lymphoid follicle hyperplasia, atrophy and fibrosis were observed primarily in H. pylori-infected cats, with the pylorus most severely affected. Neutrophilic and eosinophilic infiltrates, epithelial dysplasia, and up-regulation of mucosal IL-1beta and IL-8 were observed solely in infected cats. Fasting plasma gastrin concentrations and pentagastrin-stimulated acid output were similar in both infected and uninfected cats. There was no relationship of bacterial colonization density or gastric inflammation to plasma gastrin concentrations or gastric acid output. The pattern of colonization and the mucosal inflammatory response in cats with naturally acquired H. pylori are broadly similar to those in infected people, particularly children, and non-human primates. The upregulation of IL-8 in infected cats was independent of cagA and picB. Our findings argue against a direct acid-suppressing effect of H. pylori on the gastric secretory-axis in chronically infected cats.

PACAP peptides modulate guinea pig cardiac neuron membrane excitability and neuropeptide expression.

Morphological studies identified PACAP-immunoreactive nerve fibers in dense pericellular arrangements around virtually every cholinergic parasympathetic neuron of guinea pig cardiac ganglia; all postganglionic cardiac neurons expressed membrane-associated PAC1 receptor protein. Characterization of the alternative splice variants established predominant expression of the PAC1(very short) receptor transcript containing neither HIP nor HOP exons. PACAP depolarized cardiac neurons and increased membrane excitability; the excitability resulted from neither altered action potential properties nor inhibition of IM. Treatment of cardiac ganglia explants with PACAP significantly reduced the numbers of cholinergic neurons coexpressing somatostatin immunoreactivity, which did not appear to be correlated with prosomatostatin mRNA expression. The PACAP-mediated decrease in somatostatin immunoreactive neurons required calcium influx through L-type calcium channels and activation of adenylyl cyclase, whereas activation of phospholipase C or protein kinase A was not required. These observations indicate that PACAP through the PAC1 receptors elicits complex actions on guinea pig parasympathetic cardiac ganglia neurons, including modulation of membrane ion conductances and modulation of neuropeptide expression.

BK-Type KCaChannels in Two Parasympathetic Cell Types: Differences in Kinetic Properties and Developmental Expression

The intrinsic electrical properties of identified choroid and ciliary neurons of the chick ciliary ganglion were examined by patch-clamp recording methods. These neurons are derived from a common pool of mesencephalic neural crest precursor cells but innervate different target tissues and have markedly different action potential waveforms and intrinsic patterns of repetitive spike discharge. Therefore it is important to determine whether these cell types express different types of plasma membrane ionic channels, and to ascertain the developmental stages at which these cell types begin to diverge. This study has focused on large-conductance Ca(2+)-activated K(+) channels (K(Ca)), which are known to regulate spike waveform and repetitive firing in many cell types. Both ciliary ganglion cell types, identified on the basis of size and somatostatin immunoreactivity, express a robust macroscopic K(Ca) carried by a kinetically homogeneous population of large-conductance (BK-type) K(Ca) channels. However, the kinetic properties of these channels are different in the two cell types. Steady-state fluctuation analyses of macroscopic K(Ca) produced power spectra that could be fitted with a single Lorentzian curve in both cell types. However, the resulting corner frequency was significantly lower in choroid neurons than in ciliary neurons, suggesting that the underlying K(Ca) channels have a longer mean open-time in choroid neurons. Consistent with fluctuation analyses, significantly slower gating of K(Ca) channels in choroid neurons was also observed during macroscopic activation and deactivation at membrane potentials positive to -30 mV. Differences in the kinetic properties of K(Ca) channels could also be observed directly in single-channel recordings from identified embryonic day 13 choroid and ciliary neurons. The mean open-time of large-conductance K(Ca) channels was significantly greater in choroid neurons than in ciliary neurons in excised inside-out patches. The developmental expression of functional K(Ca) channels appears to be regulated differently in the two cell types. Although both cell types acquire functional K(Ca) at the same developmental stages (embryonic days 9-13), functional expression of these channels in ciliary neurons requires target-derived trophic factors. In contrast, expression of functional K(Ca) channels proceeds normally in choroid neurons developing in vitro in the absence of target-derived trophic factors. Consistent with this, extracts of ciliary neuron target tissues (striated muscle of the iris/ciliary body) contain K(Ca) stimulatory activity. However, K(Ca) stimulatory activity cannot be detected in extracts of the smooth muscle targets of choroid neurons.

Somatostatin-IR neurons are a major subpopulation of the cuneothalamic neurons in the rat cuneate nucleus.

This study was aimed to localize and characterize the somatostatin-immunoreactive (SOM-IR) neurons in the rat cuneate nucleus (CN). By immuno-histochemistry, the SOM-IR neurons, which were widely distributed in the nucleus, were round, spindle or multiangular in shape (mean area = 226.1 +/ -3.1 microm(2), n = 1016). By electron microscopy, the neurons shared all the ultrastructural features of the cuneothalamic neurons (CTNs) which showed a slightly indented nucleus and a fairly rich cytoplasm containing well-developed Golgi apparatuses and rough endoplasmic reticulum (rER). The SOM immunoreaction product filled the cytoplasm of the neurons extending from the soma to the proximal and distal dendrites, which were postsynaptic to unlabeled boutons. In addition to soma and dendrites, SOM-IR boutons were also identified which made axodendritic synaptic contacts with SOM-IR dendrites. The SOM-IR neurons were characterized by using anti-SOM pre-embedding immunolabeling coupled with horseradish peroxidase (HRP) retrograde method, or SOM immunolabeling along with anti-glutamate, gamma-aminobutyric acid (GABA) or glycine post-embedding immunolabeling for identification of CTNs, glutamate-IR, GABA-IR and glycine-IR neurons, respectively. It was shown that more then 80% of the CTNs contained SOM and, furthermore, they contained glutamate but not GABA or glycine. On the basis of present findings, it is suggested the majority of the SOM-IR neurons in the rat CN are CTNs and that they may be involved in modulation of somatosensory synaptic transmission.