Somatostatin Research Articles

A Disinhibitory Circuit for Contextual Modulation in Primary Visual Cortex

Context guides perception by influencing stimulus saliency. Accordingly, in visual cortex, responses to a stimulus are modulated by context, the visual scene surrounding the stimulus. Responses are suppressed when stimulus and surround are similar but not when they differ. The underlying mechanisms remain unclear. Here, we use optical recordings, manipulations, and computational modeling to show that disinhibitory circuits consisting of vasoactive intestinal peptide (VIP)-expressing and somatostatin (SOM)-expressing inhibitory neurons modulate responses in mouse visual cortex depending on similarity between stimulus and surround, primarily by modulating recurrent excitation. When stimulus and surround are similar, VIP neurons are inactive, and activity of SOM neurons leads to suppression of excitatory neurons. However, when stimulus and surround differ, VIP neurons are active, inhibiting SOM neurons, which leads to relief of excitatory neurons from suppression. We have identified a canonical cortical disinhibitory circuit that contributes to contextual modulation and may regulate perceptual saliency.

Вклад аналогов соматостатина в реализацию пациенториентированного подхода к лечению акромегалии

Вклад аналогов соматостатина в реализацию пациенториентированного подхода к лечению акромегалии

Somatostatin Neurons Govern Theta Oscillations Induced by Salient Visual Signals

Salient visual stimuli enhance theta oscillations and spike-phase locking in the theta band in the primary visual cortex (V1) of mice; however, the detailed mechanisms remain unknown. GABAergic neurons play a vital role in regulating these oscillations. Here, we use optogenetic recordings to tag cell-type-specific neurons in V1 of head-fixed mice and demonstrate that salient visual stimuli facilitate somatostatin (SOM)-expressing neuron responses and firing with theta band oscillations but suppress activities of parvalbumin (PV)-expressing neurons. Furthermore, inactivation of SOM neurons attenuates the enhancement of theta oscillations induced by salient visual stimuli and rhythmic activation of SOM neurons enhances theta oscillations. These results reveal a potential cortical theta oscillation mechanism governed by SOM neurons.

Somatostatin neurons in the central amygdala mediate anxiety by disinhibition of the central sublenticular extended amygdala.

Fear and anxiety are two defensive emotional states evoked by threats in the environment. Fear can be initiated by either imminent or future threats, but experimentally, it is typically studied as a phasic response initiated by imminent danger that subsides when the threats is removed. In contrast, anxiety is a sustained response, initiated by imagined or potential threats. The central amygdala (CeA) is a key structure active during both fear and anxiety but thought to engage different neural systems. Fear responses are triggered by activation of somatostatin (SOM) expressing neurons in the lateral division of the CeA (CeL), and downstream projections from the medial division. Anxiety responses engage the central extended amygdala that includes the CeA, central sublenticular extended amygdala (SLEAc) and bed nucleus of the stria terminalis, but the nature of connections between these regions is not understood. Here using a combination of tract tracing, electrophysiology, and behavioral analysis in mice, we show that a population of SOM+ neurons in the CeL project to the SLEAc where they inhibit local GABAergic interneurons. Optogenetic activation of this input to the SLEAc has no effect on movement, but is anxiogenic in both open field and elevated plus maze. Our results define the inhibitory connections between CeL and SLEAc and establish a specific CeL to SLEAc projection as a circuit element in mediating anxiety.

Effects of ethyl acetate extract of Salsola collina on brain-gut peptides and interstitial cells of gastric Cajal in rats with diabetic gastroparesis.

Objective(s):Effects of ethyl acetate extract of Salsola collina (EES) on brain-gut peptides and interstitial cells of gastric Cajal in rats with diabetic gastroparesis were explored. Materials and Methods:Rats were divided into six groups: normal control group (NC), diabetic gastroparesis model group (DGP), low, medium, and high dose of EES groups (LES, MES, and HES, respectively), and metoclopramide positive group (MPG). DGP rats were induced by streptozotocin (STZ) combined with a high-sugar-high-fat diet. The gastric emptying was measured by the phenol red labeling method. Enzyme-linked immunosorbent assay (ELISA) was employed to determine the concentrations of serum ghrelin, gastrin (GAS), somatostatin (SS), and vasoactive intestinal peptide (VIP). The expressions of c-Kit and its natural ligand stem cell factor (SCF) in gastric tissues were determined by Western blot and immunofluorescence. Results:Gastric emptying rate increased in a different degree after intervention by EES, among which MES and HES groups showed a significant effect (compared with DGP, P<0.01) and the HES group was equivalent to the MPG group; serum ghrelin and content of serum GAS increased while SS and VIP decreased (compared with the DGP group, P<0.05 or P<0.01); c-Kit and SCF protein expressions in gastric tissue increased (compared with DGP group, P<0.05 or P<0.01). Conclusion:EES significantly improved gastric emptying by regulating gastrointestinal hormone excretion and c-Kit/SCF signaling pathway. Our study provides a pharmacological basis for the use of the EES in the treatment of DGP. However, the detailed molecular mechanism remains to be clarified.

Molecular and functional characterization of somatostatin-type signalling in a deuterostome invertebrate.

Somatostatin (SS) and allatostatin-C (ASTC) are structurally and evolutionarily related neuropeptides that act as inhibitory regulators of physiological processes in mammals and insects, respectively. Here, we report the first molecular and functional characterization of SS/ASTC-type signalling in a deuterostome invertebrate—the starfish Asterias rubens (phylum Echinodermata). Two SS/ASTC-type precursors were identified in A. rubens (ArSSP1 and ArSSP2) and the structures of neuropeptides derived from these proteins (ArSS1 and ArSS2) were analysed using mass spectrometry. Pharmacological characterization of three cloned A. rubens SS/ASTC-type receptors (ArSSR1–3) revealed that ArSS2, but not ArSS1, acts as a ligand for all three receptors. Analysis of ArSS2 expression in A. rubens using mRNA in situ hybridization and immunohistochemistry revealed stained cells/fibres in the central nervous system, the digestive system (e.g. cardiac stomach) and the body wall and its appendages (e.g. tube feet). Furthermore, in vitro pharmacological tests revealed that ArSS2 causes dose-dependent relaxation of tube foot and cardiac stomach preparations, while injection of ArSS2 in vivo causes partial eversion of the cardiac stomach. Our findings provide new insights into the molecular evolution of SS/ASTC-type signalling in the animal kingdom and reveal an ancient role of SS-type neuropeptides as inhibitory regulators of muscle contractility.

Analysis of pallial/cortical interneurons in key vertebrate models of Testudines, Anurans and Polypteriform fishes.

The organization of the pallial derivatives across vertebrates follows a comparable elementary arrangement, although not all of them possess a layered cortical structure as sophisticated as the cerebral cortex of mammals. However, its expansion along evolution has only been possible by the development and coevolution of the cellular networks formed by excitatory neurons and inhibitory interneurons. Thus, the comparative analysis of interneuron types in vertebrate models of key evolutionary significance will provide important information, due to the extraordinary anatomical sophistication of their interneuron systems with simpler behavioral implications. Particularly in mammals, the main consensus for classifying interneuron types is based on non-overlapping markers, which do not form a single population, but consist of several distinct classes of inhibitory cells showing co-expression of other markers. In our study, we analyzed immunohistochemically the expression of the main markers like somatostatin (SOM), parvalbumin (PV), calretinin (CR), calbindin (CB), neuropeptide Y (NPY) and/or nitric oxide synthase (NOS) at the pallial regions of three different models of Osteichthyes. First, we selected two tetrapods, one amniote from the genus Pseudemys belonging to the order Testudine, at the base of the amniote diversification and with a three-layered simple cortex, and the Anuran Xenopus laevis, an anamniote tetrapod with a non-layered evaginated pallium, and finally the order Polypteriform, a small fish group at the base of the actinopterygian diversification with an everted telencephalon. SOM was the most conserved interneuron type in terms of its distribution and co-expression with other markers such as CR, in contrast to PV, which showed a different pattern between the models analyzed. In addition, the SOM expression supports a homological relationship between the medial pallial derivatives in all the models. CR and CB expressions in the tetrapods were observed, particularly, CR expressing cells were detected in the medial and the dorsal pallial derivatives, in contrast to CB, which appeared only in discrete scattered populations. However, the pallium of Polypteriforms fishes was almost devoid of CR cells, in contrast to the important number of CB cells observed in all the pallial regions. The NPY immunoreactivity was detected in all the pallial domains of all the models, as well as cells coexpressing CR. Finally, the pallial nitrergic expression was also conserved, which allows to postulate the homological relationships between the ventropallial and the amygdaloid derivatives. In summary, even in basal pallial models the neurochemically characterized interneurons indicate that their first appearance took place before the common ancestor of amniotes. Thus, our results suggest a shared pattern of interneuron types in the pallium of all Osteichthyes.

Study on the protective effect and mechanism of somatostatin on renal injury in paraquat intoxicated mice

Objective: To investigate the protective effect of somatostatin (SS) on acute kidney injury (AKI) of paraquat (PQ) poisoned mice and its mechanism. Methods: From December 2017 to April 2018, a total of 48 SPF male BALB/C mice were selected and randomly divided into 4 groups, with 12 mice in each group: Control group, SS group (20 mg/kg SS was injected 1 hour before and 3 hours after gavage with normal saline) , PQ group (2% PQ 60 mg/kg by gavage) and PQ+SS group (Intragastric administration was performed with 2% PQ solution of 60 mg/kg, and 20 mg/kg SS was administered 1 h before and 3 h after intragastric administration) , 12 mice in each group were observed for the general situation and behavioral effects. After 24 hours of modeling, mice were sacrificed.Then blood was extracted after eyeball was removed, and both kidneys were removed by laparotomy. Serum IL-6, TNF-α and MPO levels were determined by ELISA. The characteristic pathological changes of toxic renal tubular injury were observed under light microscope and scored accordingly. The changes of NF-κB expression were detected by Western-Blot, SOD, Caspase-3 and malondialdehyde (MDA) were detected by chemical colorimetry. Results: Mice in Control group and SS group showed normal general conditions and behaviors; Mice in PQ group were significantly worse than those in Control group, showing decreased feeding and activity, dry fur, hair shedding and listless spirit; The above symptoms in the mice of PQ+SS group were alleviated compared with the PQ group. Under the light microscope, the renal tissue structure of PQ group was obviously disordered and severely damaged, and the nephropathy score was (6.14±0.72) . The performance of PQ+SS group under light microscope was improved compared with PQ group, and nephropathy score (4.36±0.42) decreased (P<0.05) . Compared with the Control group, serum TNF-α (39.89±3.32) pg/ml, IL-6 (77.29±4.77) pg/ml, renal NF-κB (2.29±0.097) , MPO (0.31±0.017) μg/ml, MDA (0.91±0.03) mmol/mg prot, and Caspase-3 (376.51±8.24) % levels were significantly increased in the PQ group, while the level of renal SOD (2.36±0.73) U/mg prot was significantly decreased (P<0.05) . Compared with the PQ group, serum TNF-α (33.82±1.57) pg/ml, IL-6 (58.49±5.89) pg/ml, renal NF-κB (0.84±0.05) , MPO (0.22±0.01) μg/ml, MDA (0.72±0.05) mmol/mg prot, Caspase-3 (327.32±21.93) % decreased significantly, and renal SOD (4.90±0.81) U/mg prot increased significantly in the PQ+SS group (P<0.05) . Conclusion: PQ poisoning can lead to AKI in mice, while SS can reduce AKI caused by PQ poisoning, improve the general survival state of PQ poisoned mice, and play a certain protective role in kidney injury caused by PQ poisoning, which may be achieved by inhibiting oxidative stress response, inflammatory response and apoptosis caused by poisoning.

A new GABAergic somatostatin projection from the BNST onto accumbal parvalbumin neurons controls anxiety

The prevailing view is that parvalbumin (PV) interneurons play modulatory roles in emotional response through local medium spiny projection neurons (MSNs). Here, we show that PV activity within the nucleus accumbens shell (sNAc) is required for producing anxiety-like avoidance when mice are under anxiogenic situations. Firing rates of sNAcPV neurons were negatively correlated to exploration time in open arms (threatening environment). In addition, sNAcPV neurons exhibited high excitability in a chronic stress mouse model, which generated excessive maladaptive avoidance behavior in an anxiogenic context. We also discovered a novel GABAergic pathway from the anterior dorsal bed nuclei of stria terminalis (adBNST) to sNAcPV neurons. Optogenetic activation of these afferent terminals in sNAc produced an anxiolytic effect via GABA transmission. Next, we further demonstrated that chronic stressors attenuated the inhibitory synaptic transmission at adBNSTGABA → sNAcPV synapses, which in turn explains the hyperexcitability of sNAc PV neurons on stressed models. Therefore, activation of these GABAergic afferents in sNAc rescued the excessive avoidance behavior related to an anxious state. Finally, we identified that the majority GABAergic input neurons, which innervate sNAcPV cells, were expressing somatostatin (SOM), and also revealed that coordination between SOM- and PV- cells functioning in the BNST → NAc circuit has an inhibitory influence on anxiety-like responses. Our findings provide a potentially neurobiological basis for therapeutic interventions in pathological anxiety.

GABAergic Neurons in the Dorsal Raphe Nucleus that Express 5-HT3A Receptors Participate in Responses to Stress Hormones

The dorsal raphe nucleus (DRN) participates in stress responses and in mood regulation via its ascending release of serotonin (5-HT) onto neural circuits within the forebrain. Although the 5-HT DRN region is easily defined via 5-HT-expressing DRN neurons, the neuroarchitecture and microcircuitry that confer its multifunctionality have remained incompletely understood and have required further investigation. In this present study, neurochemical interactions within different subregions of the rat DRN were precisely analyzed. We found that 97.5% of GABAergic neurons in the DRN expressed ionotropic 5-HT3A receptors (5-HT3ARs), whereas there were only rare parvalbumin (PV)-positive or somatostatin (SOM)-positive GABAergic neurons. Furthermore, corticosterone administration into male rats as a rodent model of depression induced significantly higher c-Fos expression in 5-HT3AR-positive GABAergic neurons compared to that in 5-HT neurons within the DRN. Taken together, our findings suggest that 5-HT3AR-positive GABAergic neurons in the DRN participate in responses to stress hormones in a rat model of depression.

The Stroke-Induced Increase of Somatostatin-Expressing Neurons is Inhibited by Diabetes: A Potential Mechanism at the Basis of Impaired Stroke Recovery

Type 2 diabetes (T2D) hampers recovery after stroke, but the underling mechanisms are mostly unknown. In a recently published study (Pintana et al. in Clin Sci (Lond) 133(13):1367–1386, 2019), we showed that impaired recovery in T2D was associated with persistent atrophy of parvalbumin+ interneurons in the damaged striatum. In the current work, which is an extension of the abovementioned study, we investigated whether somatostatin (SOM)+ interneurons are also affected by T2D during the stroke recovery phase. C57Bl/6j mice were fed with high-fat diet or standard diet (SD) for 12 months and subjected to 30-min transient middle cerebral artery occlusion (tMCAO). SOM+ cell number/density in the striatum was assessed by immunohistochemistry 2 and 6 weeks after tMCAO in peri-infarct and infarct areas. This was possible by establishing a computer-based quantification method that compensates the post-stroke tissue deformation and the irregular cell distribution. SOM+ interneurons largely survived the stroke as seen at 2 weeks. Remarkably, 6 weeks after stroke, the number of SOM+ interneurons increased (vs. contralateral striatum) in SD-fed mice in both peri-infarct and infarct areas. However, this increase did not result from neurogenesis. T2D completely abolished this effect specifically in the in the infarct area. The results suggest that the up-regulation of SOM expression in the post-stroke phase could be related to neurological recovery and T2D could inhibit this process. We also present a new and precise method for cell counting in the stroke-damaged striatum that allows to reveal accurate, area-related effects of stroke on cell number.

Disentangling neuronal inhibition and inhibitory pathways in the lateral habenula

The lateral habenula (LHb) is hyperactive in depression, and thus potentiating inhibition of this structure makes an interesting target for future antidepressant therapies. However, the circuit mechanisms mediating inhibitory signalling within the LHb are not well-known. We addressed this issue by studying LHb neurons expressing either parvalbumin (PV) or somatostatin (SOM), two markers of particular sub-classes of neocortical inhibitory neurons. Here, we find that both PV and SOM are expressed by physiologically distinct sub-classes. Furthermore, we describe multiple sources of inhibitory input to the LHb arising from both local PV-positive neurons, from PV-positive neurons in the medial dorsal thalamic nucleus, and from SOM-positive neurons in the ventral pallidum. These findings hence provide new insight into inhibitory control within the LHb, and highlight that this structure is more neuronally diverse than previously thought.

Endometritis Changes the Neurochemical Characteristics of the Caudal Mesenteric Ganglion Neurons Supplying the Gilt Uterus.

Simple SummaryUterine inflammation is a very frequent pathology in domestic animals leading to disturbances in reproductive processes and causing significant economic losses. The uterus possesses nerves from either the autonomic or sensory part of the peripheral nervous system. Most of the uterus-projecting neurons are localized in the caudal mesenteric ganglion. These neurons synthesize and release numerous biologically active substances in the uterus, which regulate uterine functions. The effect of inflammation on uterine innervation is poorly recognized. This study showed that Escherichia coli-induced uterine inflammation in pig led to a reduction in the total population of uterine neurons in the caudal mesenteric ganglion, and in the populations of these cells in the dorsal and central areas of this ganglion. In the caudal mesenteric ganglion of gilts after intrauterine bacterial injection, the population of uterine neurons presenting positive staining for dopamine-β-hydroxylase (an enzyme participating in noradrenaline synthesis) and negative staining for galanin, as well as the population of uterine neurons presenting negative staining for dopamine-β-hydroxylase but positive staining for neuropeptide Y, were decreased. In these gilts, there were increased numbers of uterine neurons which, besides dopamine-β-hydroxylase, also expressed neuropeptide Y, galanin and vasoactive intestinal peptide. The above changes suggest that inflammation of the gilt uterus may affect the function(s) of this organ by its action on the neurons of the caudal mesenteric ganglion.This study analyzed the influence of uterine inflammation on the neurochemical characteristics of the gilt caudal mesenteric ganglion (CaMG) uterus-supplying neurons. The horns of uteri were injected with retrograde tracer Fast Blue on day 17 of the first studied estrous cycle. Twenty-eight days later (the expected day 3 of the third studied estrous cycle), either saline or Escherichia coli suspension were administered into each uterine horn. Only the laparotomy was done in the control gilts. After 8 days, the CaMGs and uteri were harvested. The infected gilts presented a severe acute endometritis. In the CaMGs, the populations of uterine perikarya possessing dopamine-β-hydroxylase (DβH) and/or neuropeptide Y (NPY), somatostatin (SOM), galanin (GAL) and vasoactive intestinal polypeptide (VIP) were analyzed using the double immunofluorescence method. In the CaMG, bacterial injection decreased the total number of the perikarya (Fast Blue-positive), the small and large perikarya populations in the dorsal and central regions, and the small and large perikarya populations coded DβH+/GAL- and DβH-/NPY+. After bacterial treatment, there was an increase in the numbers of small and large perikarya coded DβH+/NPY+, small perikarya coded DβH+/GAL+ and DβH+/SOM- and large perikarya coded DβH+/VIP+. To summarize, uterine inflammation influences the neurochemical characteristics of the CaMG uterus-supplying neurons, which may be important for pathologically changed organ functions.

SAT-127 Localization and Treatment of the Ectopic ACTH Syndrome Using Somatostatin Analogues

Ectopic ACTH syndrome (EAS) is a rare disorder with a high morbidity and mortality due to the sequelae of severe hypercortisolism and possible underlying carcinoma. It is caused by various tumors, some originating from neuroendocrine cells. Optimal management includes localization and removal of the ectopic ACTH source. About 80% of these tumors have somatostatin (SS) receptors, so that SS analogues may be useful in the localization and treatment. This case demonstrates the use of SS analogues in localization and treatment of EAS.A 49-year-old female presented with 7 months of worsening Cushingoid features including rounded plethoric face, full supraclavicular fat pads, facial hair growth, multiple bruises, violaceous abdominal striae, kyphoscoliosis, 4+ peripheral edema, weight gain, hypertension (190/110 mmHg) and thoracic compression fractures. AM plasma ACTH concentration was 333 pg/mL (nl = 6 - 58 pg/mL); serum concentration cortisol was 71.4 mcg/dL (nl = 7 - 23 mcg/dL) and serum potassium concentration was 1.2 mmol/L (nl = 3.6 - 5.1 mmol/L). A 24-hour urine cortisol could not accurately be obtained. High dose 8 mg overnight dexamethasone testing demonstrated suppression of serum cortisol concentration from 71.4 to 6.3 mcg/dL, suggesting pituitary Cushing disease. Pituitary MRI scan revealed a 2.5 mm disc shaped cystic focus suggesting a Rathke’s cleft cyst. No central-to-peripheral ACTH gradient was present on bilateral inferior petrosal sinus sampling suggesting an ectopic ACTH source. There was a 1.2 x 1.0 cm nodule in the left lower lung lobe on CT chest. A gallium-68 dotatate PET/CT scan demonstrated enhancement of this same lung lesion and mild uptake in the left inferior hilum, suggesting a neuroendocrine lung tumor with possible metastases to lymph nodes. Biopsy and resection of the lesion was deferred until after control of her hypercortisolism. 30mg of Sandostatin LAR was started every 4 weeks to control the ectopic ACTH secretion and to confirm that the SS analogue positive lung lesion was the cause of the EAS. Sandsostatin LAR therapy over the next 12 weeks resulted in a steady clinical and biochemical improvement with a decrease in serum cortisol to 9.7 mcg/dL, control of hypertension (108/64 mmHg), weight loss of 6 pounds, and resolution of supraclavicular fat pads, hypokalemia, and peripheral edema. This confirmed our impression that the neuroendocrine lung tumor was the ACTH source.In summary, this patient with EAS had severe clinical manifestations of hypercortisolism and could not safely undergo surgical intervention. This case demonstrates the value of SS analogues (gallium-68 dotatate PET/CT and Sandostatin LAR) in localizing and confirming the source of ectopic ACTH production and markedly improving the clinical and biochemical features of the EAS.

Characterization of agonist-dependent somatostatin receptor subtype 2 trafficking in neuroendocrine cells.

Somatostatin (SOM) receptor subtype 2 (SSTR2) is the major receptor subtype mediating SOM effects throughout the neuraxis. We previously demonstrated that the non-selective agonist [D-Trp8]-SOM induces intracellular sequestration of SSTR2, whereas this receptor is maintained at the cell surface after treatment with the SSTR2-selective agonist L-779,976 in cells co-expressing SSTR2 and SSTR5. In this study, we knocked-out SSTR5 in AtT20 cells endogenously expressing both SSTR2 and SSTR5 and used immuno-labeling and confocal microscopy to investigate the effect of SSTR5 on regulation of SSTR2 trafficking. Our results indicate that unlike [D-Trp8]-SOM-induced intracellular sequestration, L-779,976 stimulation results in the maintenance of SSTR2 at the cell surface regardless of whether SSTR5 is present or not. We then examined the trafficking pathways of SSTR2 upon stimulation by either agonist. We found that both [D-Trp8]-SOM and L-779,976 induce SSTR2 internalization via transferrin-positive vesicles. However, SSTR2 internalized upon L-779,976 treatment undergoes rapid recycling to the plasma membrane, whereas receptors internalized by [D-Trp8]-SOM recycle slowly after washout of the agonist. Furthermore, [D-Trp8]-SOM stimulation induces degradation of a fraction of internalized SSTR2 whereas L-779,976-dependent, rapid SSTR2 recycling appears to protect internalized SSTR2 from degradation. In addition, Octreotide which has preferential SSTR2 affinity, induced differential effects on both SSTR2 trafficking and degradation. Our results indicate that the biased agonistic property of L-779,976 protects against SSTR2 surface depletion by rapidly initiating SSTR2 recycling while SSTR5 does not regulate L-779-976-dependent SSTR2 trafficking.

Diagnostic and prognostic significance of long noncoding RNA SSTR5-AS1 in patients with gastric cancer.

Long noncoding RNAs (lncRNAs) display a functional effect on the pathogenesis of several diseases, including various tumors. Herein, we aimed to reveal the role of lncRNA somatostatin receptor 5 antisense RNA 1 (SSTR5-AS1) in gastric cancer (GC). qRT-PCR was utilized for testing the SSTR5-AS1 expression in 158 paired primary GC tissues and corresponding normal gastric specimens. Receiver operating characteristic (ROC) curves were established to determine the diagnostic values of overexpression of SSTR5-AS1 in GC. A chi-square test was performed to analyze the correlation between SSTR5-AS1 expressions and several clinicopathological features in GC patients. Kaplan-Meier survival curve was constructed to estimate the overall survival (OS) and disease-free survival (DFS). Multivariate analyses were conducted to examine the prognostic value of SSTR5-AS1. We observed that SSTR5-AS1 expression was highly expressed in GC specimens compared with adjacent non-tumor specimens (p < 0.01). High SSTR5-AS1 expression was correlated with an advanced pathologic stage. The ROC curves showed that areas under the ROC curve (AUC) for SSTR5-AS1 is 0.8419. Moreover, high expression of SSTR5-AS1 was observed to be associated with distant metastasis (p = 0.021) and TNM stage (p = 0.042). Besides, survival analysis showed that GC patients with high SSTR5-AS1 expression suffered poorer OS (p = 0.020) and DFS (p = 0.0007). Multivariate assays demonstrated that increased expressions of SSTR5-AS1 could be an independent prognostic marker of OS and DFS of GC patients. Our findings indicate that SSTR5-AS1 served as a promising novel prognostic biomarker for GC.

Effect of a novel somatostatin-14 DNA vaccine fused to tPA signal peptide and CpG adjuvant on goat lactation and milk composition

Abstract This study aimed to investigate the effect of a novel oral somatostatin-14 (SS-14) DNA vaccine fused to tissue plasminogen activator (tPA) signal peptide and CpG adjuvant on goat lactation and milk composition. Ten female goats with two newborn kids each were randomly divided into the immunized group and negative control group; The novel oral somatostatin (SS) DNA vaccine (fused to tPA signal peptide and CpG adjuvant) and empty vector bacteria were orally administered to these groups, respectively, at weeks 0, 3, and 6 of the study period. Blood samples were collected before primary immunization and at weeks 6 and 9 after primary immunization. The specific anti-SS antibody titers in the immunized group were significantly higher than that in the negative control group (P

Multiplexed Immunohistochemistry to Identify Systematically Classes of Human Colonic Myenteric Neurons

ObjectiveThe enteric nervous system (ENS) comprises multiple classes of nerve cells which can be distinguished by combinations of immunohistochemical markers (“chemical coding”) that they express. While many immunohistochemical studies have been carried out, the chemical coding of ENS of human colon has not been characterised systematically.MethodsSurgical specimens of human colon were obtained with prior informed consent (Ethics Approval SAC HREC#207.07), incubated in 83uM colchicine overnight to enhance neuropeptide immunoreactivity, then fixed. Multiplexed indirect immunohistochemistry (5 layers of quadruple labelling) included 13 common neuronal markers; HuC/D, nitric oxide synthase (NOS), choline acetyltransferase (ChAT), 5‐hydroxytryptamie (5‐HT), Substance P/tachykinins (SP), leu‐Enkephalin (ENK), calcitonin gene‐related peptide (CGRP), somatostatin (SOM), Calbindin (Calb), Calretinin (Calret), neurofilament 200 (NF200), vasoactive intestinal polypeptide (VIP) and neuropeptide Y (NPY). Antibodies were eluted between layers. The presence/absence of all 13 markers was recorded for 1303 neurons in human ascending colon (n=6) and 1087 in descending colon (n=5). HuC/D labelled all myenteric nerve cell bodies and was revealed using AMCA as a fluorophore; this staining was resistant to elution and allowed comparison between layers.ResultsChemical codes were surprisingly varied; of 4096 possible codes (212), 108 were present in ascending colon and 112 in descending colon. However, 40 codes accounted for more than 90% of neurons. There was a strong correlation between the rank order of chemical codes in ascending and descending colon (ranked by abundance; Rs=0.884[39]); p&lt;0.001) indicating that there are similar ENS populations in ascending and descending colon. In ascending colon (n=6, 1303 cells) 686 cells (52.6%) were immunoreactive for NOS; 55.5% for ChAT; 9.8% were immunoreactive for both ChAT and NOS and 22 cells (1.7%) lacked both markers. The next most abundant markers were NF200 (50.0%), VIP (31.3%) and calbindin (25.1%). Low frequency markers in myenteric nerve cell bodies included CGRP (0.9%) and SOM (1.3%). A few codes accounted for very large proportions of neurons. For example, putative inhibitory motor neurons, immunoreactive for NOS (with none of the other 11 markers), accounted for 192 of 1303 neurons (14.7%). Dogiel type II neurons (identified by NF200 morphology) were large cells (1795 +/‐ 729 μm2, 25 cells) with NF200, ChAT and SP, with either Calb or Calret (or both) or SOM. It is also possible to identify cell bodies surrounded by dense baskets of varicose axons, which represent strong synaptic inputs. For example, ChAT‐immunoreactivity was present in 55.5% of all myenteric neurons, but made up 93.6% of all myenteric neurons enclosed by dense enkephalin‐immunoreactive inputs.Conclusionsmulti‐layer multiplexed immunoreactivity reveals a wealth of information about the human colonic ENS, making possible systematic identification of classes and identification of some synaptic connectivitySupport or Funding InformationSupported by NIH SPARC grant #1OT2OD24899

Abstinence-dependent dissociable central amygdala microcircuits control drug craving

We recently reported that social choice-induced voluntary abstinence prevents incubation of methamphetamine craving in rats. This inhibitory effect was associated with activation of protein kinase-Cδ (PKCδ)-expressing neurons in central amygdala lateral division (CeL). In contrast, incubation of craving after forced abstinence was associated with activation of CeL-expressing somatostatin (SOM) neurons. Here we determined the causal role of CeL PKCδ and SOM in incubation using short-hairpin RNAs against PKCδ or SOM that we developed and validated. We injected two groups with shPKCδ or shCtrlPKCδ into CeL and trained them to lever press for social interaction (6 d) and then for methamphetamine infusions (12 d). We injected two other groups with shSOM or shCtrlSOM into CeL and trained them to lever press for methamphetamine infusions (12 d). We then assessed relapse to methamphetamine seeking after 1 and 15 abstinence days. Between tests, the rats underwent either social choice-induced abstinence (shPKCδ groups) or homecage forced abstinence (shSOM groups). After test day 15, we assessed PKCδ and SOM, Fos, and double-labeled expression in CeL and central amygdala medial division (CeM). shPKCδ CeL injections decreased Fos in CeL PKCδ-expressing neurons, increased Fos in CeM output neurons, and reversed the inhibitory effect of social choice-induced abstinence on incubated drug seeking on day 15. In contrast, shSOM CeL injections decreased Fos in CeL SOM-expressing neurons, decreased Fos in CeM output neurons, and decreased incubated drug seeking after 15 forced abstinence days. Our results identify dissociable central amygdala mechanisms of abstinence-dependent expression or inhibition of incubation of craving.

Effects of coated cysteamine on growth performance, carcass characteristics, meat quality and lipid metabolism in finishing pigs

This study was conducted to evaluate the effects of coated cysteamine (CCS) on growth performance, carcass characteristics, meat quality, and lipid metabolism in finishing pigs. A total of 160 pigs (62.12 ± 1.43 kg body weight) were randomly divided into 4 groups, with 4 pens per group and 10 pigs (5 gilts and 5 barrows) per pen. Pigs were fed a basal diet supplemented with 0, 100, 200, or 400 mg/kg CCS, corresponding to 0, 27, 54, or 108 mg/kg cysteamine, respectively. All pigs were given free access to feed and water for forty days. Average daily gain, average daily feed intake and feed to gain ratio were not affected by the CCS treatment. Supplemental CCS increased carcass lean ratio linearly (P < 0.05) and longissmus muscle area quadratically (P < 0.05), and the water-holding capacity as measured by 24 h drip loss was quadratically affected (P < 0.05) by CCS supplementation. The activity of hormone-sensitive lipase in subcutaneous adipose tissue was linearly increased (P < 0.001), while fatty acid synthase activity was linearly decreased (P < 0.05). Pigs fed CCS had a lower serum triacylglycerol (P < 0.001) level and lipase activity (P < 0.001), lower total cholesterol (P < 0.05) level, and a higher total protein (P < 0.001) and free fatty acids (P < 0.01) levels. In addition, CCS quadratically increased serum growth hormone (GH) (P < 0.001) level and quadratically affected somatostatin (SS) (P < 0.05) level. Consistently, significant effects of dietary CCS were observed on the mRNA expression of GH and SS in the anterior pituitary gland and the hypothalamus, respectively, where the intermediate CCS level showed the highest GH mRNA level and lowest SS mRNA level. The results indicate that dietary supplementation of CCS has beneficial effects on carcass characteristics and meat quality, and affects lipid metabolism in finishing pigs.