Serotonin-immunoreactive Cells Research Articles

Clarifying the Functional Role of Serotonin in Meloidogyne graminicola Host Plant Parasitism by Immunolocalization and RNA Interference.

Serotonin (5-hydroxytryptamine) is an essential neurotransmitter involved in regulating various behaviors in plant-parasitic nematodes, including locomotion, egg laying, feeding, and mating. However, the functional role of serotonin in root-knot nematode invasion of host plants and the molecular mechanisms underlying feeding behavior remain poorly understood. In this study, we tested the effects of exogenous serotonin and the pharmacological compounds fluoxetine and methiothepin on the feeding behaviors of Meloidogyne graminicola. Our results suggested that M. graminicola possesses an endogenous serotonin signaling pathway and that serotonin plays a crucial role in modulating feeding behaviors in M. graminicola second-stage juveniles. We also identified and cloned the serotonin synthesis enzyme tryptophan hydroxylase (Mg-tph-1) in M. graminicola and investigated the role of endogenous serotonin by generating RNA interference nematodes in Mg-tph-1. Silencing Mg-tph-1 substantially reduced nematode invasion, development, and reproduction. According to the immunostaining results, we speculated that these serotonin immunoreactive cells near the nerve ring in M. graminicola are likely homologous to Caenorhabditis elegans ADFs, NSMs, and RIH serotonergic neurons. Furthermore, we investigated the impact of phytoserotonin on nematode invasion and development in rice by overexpressing OsTDC-3 or supplementing rice plants with tryptamine and found that an increase in phytoserotonin increases nematode pathogenicity. Overall, our study provides insights into the essential role of serotonin in M. graminicola host plant parasitism and proposes that the serotonergic signaling pathway could be a potential target for controlling plant-parasitic nematodes.

Serotonin Immunoreactive Cells in Extrahepatic Bile Ducts, Major Duodenal Papilla and Gallbladder in the Domestic Pig

Abstract The main part of serotonin in the body is synthesized and released by a certain type of enteroendocrine cells in the intestinal mucosa called enterochromaffin cells. The scarce qualitative and quantitative data on enterochromaffin and serotonin-positive mast cells in porcine extrahepatic bile ducts and gallbladder, motivated us to undertake the present study. The aim of this study was to determine the localization and density of serotonin-positive cells in the wall of the extrahepatic bile ducts and gallbladder in pigs. An immunohistochemical method was used to identify enterochromaffin cells and determine their percentage relative to the total number of endocrine cells labeled with chromogranin A. Serotonin-positive mast cells were identified after tryptase staining of serial sections. The endocrine function of mast cells was demonstrated by chromogranin A immunolabeling. The highest number of enterochromaffin cells were found in the intramural part of the ductus choledochus, followed by the papilla duodeni major, extramural part of the ductus choledochus, ductus hepaticus comunis, ductus cysticus, and gallbladder. In all parts of the extrahepatic bile ducts, the highest number of mast cells was found in the muscle layer, followed by the serosal layer and the propria. The expression of serotonin in the enterochromaffin cells of the biliary glands and in the mast cells of the analyzed organs suggests a possible synthesis of serotonin, which probably regulates physiological and pathological processes.

Moderate intestinal immunopathology after acute oral infection with Toxoplasma gondii oocysts is associated with expressive levels of serotonin

BackgroundInvasion of the intestinal mucosa by T. gondii elicits a local immune response of variable intensity. These reactions can be lethal in C57BL/6 mice. The tissue damage caused by inflammation and the functional effects depend on the host immunity, strain, and developmental form of the parasite. We investigated the effects of acute oral infection with T. gondii on histoarchitecture, enteric nervous system (ENS), and inflammatory markers in the jejunum and ileum of mice. MethodsFemale C57BL/6 mice were divided into a control group and a group orally infected with 1000 sporulated T. gondii oocysts (ME-49 strain). After 5 days, jejunum and ileum were collected and processed for analyzes (e.g., histological and histopathological examinations, ENS, cytokine dosage, myeloperoxidase, nitric oxide activity). Main resultsIn infected mice, we observed a significant increase in serotonin-immunoreactive cells (5-HT IR) in the intestinal mucosa, as well as cellular infiltrates in the lamina propria, periganglionitis, and ganglionitis in the myenteric plexus. We also noted decreased neuron density in the jejunum, increased population of enteric glial cells in the ileum, histomorphometric changes in the intestinal wall, villi, and epithelial cells, remodeling of collagen fibers, and increased myeloperoxidase activity, cytokines, and nitric oxide in the intestine. Conclusions and inferencesAcute infection of female mice with T. gondii oocysts resulted in changes in ENS and a marked increase in 5-HT. These changes are consistent with its modulatory role in the development of moderate acute inflammation. The use of this experimental model may lend itself to studies aimed at understanding the pathophysiological mechanisms of intestinal inflammation in humans involving ENS.

Neuroanatomical Distribution of the Serotonergic System in the Brain and Retina of Holostean Fishes, The Sister Group to Teleosts

Among actinopterygian fishes, holosteans are the phylogenetically closest group to teleosts but they have been much less studied, particularly regarding the neurochemical features of their central nervous system. The serotonergic system is one of the most important and conserved systems of neurotransmission in all vertebrates. By means of immunohistochemistry against serotonin (5-hydroxytryptamine), we have conducted a comprehensive and complete description of this system in the brain and retina of representative species of the 3 genera of holostean fishes, belonging to the only 2 extant orders, Amiiformes and Lepisosteiformes. Serotonin-immunoreactive cell groups were detected in the preoptic area, the hypothalamic paraventricular organ, the epiphysis, the pretectal region, the long and continuous column of the raphe, the spinal cord, and the inner nuclear layer of the retina. Specifically, the serotonergic cell groups in the preoptic area, the epiphysis, the pretectum, and the retina had never been identified in previous studies in this group of fishes. Widespread serotonergic innervation was observed in all main brain regions, but more abundantly in the subpallium, the hypothalamus, the habenula, the optic tectum, the so-called cerebellar nucleus, and the area postrema. The comparative analysis of these results with those in other groups of vertebrates reveals some extremely conserved features, such as the presence of serotonergic cells in the retina, the pineal organ, and the raphe column, while other characteristics, like the serotonergic populations in the preoptic area, the paraventricular organ, the pretectum, and the spinal cord are generally present in all fish groups, but have been lost in most amniotes.

The distribution of serotonergic nerve on the hippocampus of the fruit bats (Rousettus amplexicaudatus).

Background:Rousettus amplexicaudatus is one of the old world fruit bats, commonly known in Javanese word as codot belongs to Order Chiroptera and suborder Megachiroptera. R. amplexicaudatus possessed an outstanding visual ability, which helps in the process of searching for food. Those activities process and save within the brain involving the learning and memory activities. Learning and memory activities are centered around the hippocampus with the help of serotonergic nerve.Aim:This study conducted to identify the morphology and distribution of serotonergic nerve (5-hydroxytryptamine) in the hippocampus of R. amplexicaudatus, which related to the function of serotonin within the learning and memory process.Materials and Methods:Five R. amplexicaudatus bats were brought from Gunung Kidul cave, Yogyakarta, Java Island, Indonesia. The bats were anesthetized using ketamine and xylazine. R. amplexicaudatus then perfused intracardially. The brain samples were collected processed into paraffin block, and a further cut in 8 µm thickness serially. The tissue slides were stained using hematoxylin-eosin, cresyl echt violet, and immunohistochemistry with rabbit’s anti-serotonin (1:200). The results observed and analyzed quantitatively and qualitatively using image J (https://imagej.nih.gov/ij/) (Bethesda, Maryland, USA) and GraphPad Prism 7 (La Jolla, CA, USA).Results:The hippocampus of R. amplexicaudatus composed of the dentate gyrus (DG), hippocampus proper (cornu ammonis 3 [CA3], and CA1 as the main area for learning memory), and subiculum. On the DG serotonin immunoreactive cells found within the granular layer (132±35.03 cells/mm2), polymorphic stratum (86.33±11.23 cells/mm2), and molecular layer (93±1 cells/mm2). Meanwhile, on CA3 area, the immunoreactive cells for serotonin found in each stratum. The number of immunoreactive cells on each stratum from highest to the lowest are stratum pyramidal 123.33±15.88 cell/mm2, stratum molecular 63±13.11 cell/mm2, stratum lucidum 62.67±8.08 cell/mm2, stratum radiatum 55.33±510.21 cell/mm2, stratum oriens 48±3.46 cell/mm2, and stratum alveus 28.67±2,52 cell/mm2. In addition, in CA1 also hampers the immunoreactive cells in the pyramidal stratum, molecular, lucidum, oriens, radiatum, and alveus layer, respectively, of each 91±27.40 cell/mm2, 60.33±20.65 cell/mm2, 53.67±4.51 cell/mm244.33±10.40 cell/mm2, 41.33±5.51 cell/mm2, and 27±4 cell/mm2.Conclusion:Taking together the distribution of serotonin-immunoreactive cells in the hippocampus of R. amplexicaudatus mostly found on CA3 followed by CA 1 and DG.

Cognitive and anxiety-like impairments accompanied by serotonergic ultrastructural and immunohistochemical alterations in early stages of parkinsonism

Parkinson’s disease (PD) is mostly known as a dopamine deficiency syndrome due the structural and functional changes in striatal projection neurons. However, studies have considered this pathology as a multi-systemic disease in which the neurodegenerative process extends beyond the dopaminergic system. Therefore, the purpose of the present study was to investigate the morphological and immunohistochemical changes associated with behavioral and cognitive alterations in a model of parkinsonism induced by low dose of reserpine. Animals showed anxiety–like behavior and deficits in short-term recognition memory. Besides, Tyrosine Hydroxylase (TH) immunoreactive cells decreased in reserpine (RES) group in CA1 and serotonin (5-HT) immunoreactive cells decreased in RES group in CA1, CA3 and medial prefrontal cortex (mPFC). Moreover, an increase in the area (μm2) of 5 H T labeled ultrastructure (axon terminal) was observed in RES group only in CA1 and mPFC. The evidence of alterations in 5-HT immunoreactive in the premotor phase of model of parkinsonism highlights the importance of looking beyond the nigrostriatal system to elucidate the underling mechanisms and deficits in other neurotransmitters systems. This provides vital information regarding novel interventions for the management of non-motor symptoms. Additionally, the low-dose reserpine treatment has an early effect on axonal ultrastructure. As the axonopathy in PD has been increasingly recognized, the focus on axonal neurobiology is noteworthy for both neuroprotective and restorative therapeutics, and the progressive reserpine rat model can be a useful tool in this search.

On the larva and the zooid of the pterobranch Rhabdopleura recondita Beli, Cameron and Piraino, 2018 (Hemichordata, Graptolithina)

Hemichordates (Enteropneusta and Pterobranchia) belong to a small deuterostome invertebrate group that may offer insights on the origin and evolution of the chordate nervous system. Among them, the colonial pterobranch Rhabdopleuridae are recognized as living representatives of Graptolithina, a taxon with a rich fossil record. New information is provided here on the substrate selection and the life cycle of Rhabdopleura recondita Beli, Cameron and Piraino, 2018, and for the first time, we describe the nervous system organization of the larva and the adult zooid, as well as the morphological, neuroanatomical and behavioural changes occurring throughout metamorphosis. Immunohistochemical analyses disclosed a centralized nervous system in the sessile adult zooid, characterized by different neuronal subsets with three distinct neurotransmitters, i.e. serotonin, dopamine and RFamide. The peripheral nervous system comprises GABA-, serotonin-, and dopamine-immunoreactive cells. These observations support and integrate previous neuroanatomical findings on the pterobranch zooid of Cephalodiscus gracilis. Indeed, this is the first evidence of dopamine, RFamide and GABA neurotransmitters in hemichordates pterobranchs. In contrast, the lecithotrophic larva is characterized by a diffuse basiepidermal plexus of GABAergic cells, coupled with a small group of serotonin-immunoreactive cells localized in the characteristic ventral depression. It is envisaged the use of R. recondita as a novel and easily accessible hemichordate model organism to shed light on the evolution of hemichordates and more generally on the origin of deuterostome developmental mechanisms.

Neuroendocrine structures of the small intestine of the capybara Hydrochoerus hydrochaeris (Mammalia, Rodentia)

Abstract A complex network of nerve fibers of the enteric nervous system and enteroendocrine cells is known to regulate the gastrointestinal tract. The distribution and frequency of the argyrophil, argentaffin and serotonin immunoreactive endocrine cells and of the submucosal and myenteric nervous ganglia were studied in the small intestine of the capybara Hydrochoerus hydrochaeris, aiming to verify the existence of possible numerical correlations between endocrine cells and nervous ganglia. Fragments of the duodenum, jejunum and ileum of adult animals were collected and processed according to routine histological techniques. To study the nervous ganglia, hematoxylin and eosin staining was used, while specific staining techniques were used to study the argyrophil, argentaffin and serotonin immunoreactive endocrine cells: Grimelius, modified Masson-Fontana and peroxidase anti-peroxidase, respectively. Endocrine cells were more abundant in the area of the crypts and, in relation to their morphology, ‘open type’ endocrine cells prevailed. The population of argyrophil cells was larger than that of argentaffin cells, and these cells were larger than serotonin immunoreactive cells. The frequency of endocrine cells was apparently greater in the duodenum, indicating the importance of this intestinal segment in digestive and absorptive functions. Prominent nervous ganglia were observed in the submucosal and myenteric plexi, and were larger and more frequent in the myenteric plexus. A numerical correlation was found among the endocrine cells (argentaffin and serotonin immunoreactive cells) and the myenteric nervous ganglia, suggesting the presence of physiological interactions among the endocrine and nervous systems for the control of intestinal activities. The findings in this study contribute to the understanding of the digestive processes of this species, which may also help in its conservation and future survival.

Immunohistochemical Study of Endocrine Cells in the Stomach and Small Intestine of the Lion (Panthera leo)

Abstract. The distribution and relative frequency of gastrointestinal endocrine cells were investigated in the stomach and small intestine (duodenum and jejunum) of a lion (Panthera leo). The present study was an immunohistochemical investigation of endocrine cells showing immunoreactivity for serotonin, gastrin, or cholecystokinin (CCK). Serotonin-immunoreactive (IR) cells were detected throughout the gastrointestinal tract with the highest frequency in the duodenum. Gastrin-IR cells were numerous in the pyloric region of the stomach and moderate numbers were also seen in the duodenum and jejunum. Moderate numbers of CCK-IR cells were only detected in the duodenum and jejunum.

Dietary guidance and ileal enteroendocrine cells in patients with irritable bowel syndrome.

The enteroendocrine cells of the ileum are stimulated by the luminal contents to release specific hormones that regulate its functions. The density of ileal enteroendocrine cells is abnormal in patients with irritable bowel syndrome (IBS), and the majority of patients with IBS associate their symptoms to the consumption of certain foodstuffs. The present study investigated the effect of dietary guidance on the enteroendocrine cells of the ileum in 11 patients with IBS. A total of 10 control subjects were also included. Each patient received three sessions of dietary guidance. Colonoscopies were performed on both controls and patients with IBS (at baseline and 3–9 months after the patients had received dietary guidance). Biopsy samples from the ileum were immunostained for all enteroendocrine cells and quantified by computerized image analysis. The densities of serotonin-immunoreactive cells in controls and in patients with IBS prior to and following dietary guidance were 35.5±5.7, 38.7±7.1 and 22.3±2.6 cells/mm2, respectively (mean ± standard error of the mean; P=0.046); the corresponding values for PYY-immunoreactive cells were 16.7±2.8, 20.2±5.1 and 21.3±2.7 cells/mm2 (P=0.86). These results suggest that changes in enteroendocrine cell densities in the ileum along with changes in enteroendocrine cells throughout the gastrointestinal tract may contribute to the improvement in IBS symptoms following dietary guidance.

시스플라틴으로 유발된 랫트의 위장관 운동장애에 대한 태음조위탕의 효과

Objectives This study aimed to observe the effect of Taeumjowi-tang on the cisplatin-induced gastrointestinal dysfunctions in rats. Methods Four groups, each of 8 rats per group, were used in this study. Saline and distilled water treated control rats were intact vehicle control group. Delayed gastrointestinal motility was induced by intraperitoneal treatment of cisplatin 2mg/kg, once a week for 5 weeks(Cisplatin control group). Taeumjowi-tang aqueous extracts(TJ) were orally administered in a volume of 5ml/kg, once a day for 14 days from 4th cisplatin treatment(TJ group). Ondansetron 1mg/kg was subcutaneously treated, in a volume of 1ml/kg, as same as TJ(ondansetron group). We measured the body weights, intestinal charcoal transit ratio, fecal parameters, fundus MDA(malondialdehyde), GSH(glutathione) contents and SOD(superoxide dismutase), CAT(catalase) activities, TPH(tryptophanhydroxylase) and MAO(monoamine oxidase) activities, pyloric gastrin and serotonin contents with their immunoreactive cells, colonic serotonin-immunoreactive cells, the histopathology of pylorus, fundus mucosa and colon. Results 1) The body weight gains, the small intestinal charcoal transfer rates, the fecal parameters(numbers, weights and water contents) were increased in TJ, ondansetron group. 2) The inhibit of fundus antioxidant defense systems by cisplatin were decreased in TJ, ondansetron group. 3) The pyloric TPH activities were increased and the pyloric MAO activities were decreased in TJ group. 4) The pyloric gastric contents and the gastrin-immunoreactive cells were increased and the pyloric serotonin contents and the pyloric and colonic serotonin-immunoreactive cells were decreased in TJ group. 5) The pylorus atrophic changes and the gastric surface erosive damage regions by cisplatin were favorably inhibited by treatment of TJ group. Conclusions The results obtained in this study suggest that TJ favorably retarded the cisplatin related GI(gastrointestinal) dysfunctions and constipation through modulations of GI enterochromaffin cells, serotonin and gastrin-producing cells and antioxidative systems.

Fine taxonomic sampling of nervous systems within Naididae (Annelida: Clitellata) reveals evolutionary lability and revised homologies of annelid neural components.

IntroductionAn important goal for understanding how animals have evolved is to reconstruct the ancestral features and evolution of the nervous system. Many inferences about nervous system evolution are weak because of sparse taxonomic sampling and deep phylogenetic distances among species compared. Increasing sampling within clades can strengthen inferences by revealing which features are conserved and which are variable within them. Among the Annelida, the segmented worms, the Clitellata are typically considered as having a largely conserved neural architecture, though this view is based on limited sampling.ResultsTo gain better understanding of nervous system evolution within Clitellata, we used immunohistochemistry and confocal laser scanning microscopy to describe the nervous system architecture of 12 species of the basally branching family Naididae. Although we found considerable similarity in the nervous system architecture of naidids and that of other clitellate groups, our study identified a number of features that are variable within this family, including some that are variable even among relatively closely related species. Variable features include the position of the brain, the number of ciliary sense organs, the presence of septate ventral nerve cord ganglia, the distribution of serotonergic cells in the brain and ventral ganglia, and the number of peripheral segmental nerves.ConclusionsOur analysis of patterns of serotonin immunoreactive perikarya in the central nervous system indicates that segmental units are not structurally homogeneous, and preliminary homology assessments suggest that whole sets of serotonin immunoreactive cells have been gained and lost across the Clitellata. We also found that the relative position of neuroectodermal and mesodermal segmental components is surprisingly evolutionarily labile; in turn, this revealed that scoring segmental nerves by their position relative to segmental ganglia rather than to segmental septa clarifies their homologies across Annelida. We conclude that fine taxonomic sampling in comparative studies aimed at elucidating the evolution of morphological diversity is fundamental for proper assessment of trait variability.Electronic supplementary materialThe online version of this article (doi:10.1186/s12983-015-0100-6) contains supplementary material, which is available to authorized users.

Histochemical and Immunohistochemical Analysis of the Stomach of Rhinella icterica (Anura, Bufonidae)

The stomach of Rhinella icterica was analyzed at light microscopy, employing histochemical techniques, lectin histochemistry, and immunohistochemistry for identifying enteroendocrine cells (EC). Although the stomach was composed of fundic and pyloric regions, its wall is formed by mucosa, submucosa, muscularis, and serosa. The mucosa was lined by a simple columnar mucous epithelium, supported by loose connective tissue. Several tubular, simple glands were composed of mucous neck cells, containing oxynticopeptic cells and EC cells. The mucous neck cells were rich in neutral glycoconjugates. The oxynticopeptic cells were predominant in fundic glands, exhibiting weaker alcianophilic reaction at their apical cytoplasm. Serotonin (5-HT) immunoreactive (IR) cells occurred throughout the entire stomach, preferentially located among mucous cells at upper part of the fundic glands. The muscularis mucosae, formed of smooth muscle, separated the mucosal layer from the submucosa, both of which were constituted by loose connective tissue, but without glands. Lymphoid modules occurred in the mucosa at the boundary at the stomach and the gut. In addition, the muscularis was constituted by two sublayers, the circular internal and the longitudinal external, being recovered by the connective tissue of the serosa.

Stomach antral endocrine cells in patients with irritable bowel syndrome.

To the best of our knowledge, stomach antral endocrine cells have not previously been investigated in patients with irritable bowel syndrome (IBS). Thus, in the present study, 76 patients with IBS were examined (designated as IBS-total). Diarrhoea was the predominant symptom in 26 of these patients (IBS-D), while in 21 patients, the predominant symptoms were both diarrhoea and constipation (IBS-M) and in 29 patients the predominant symptom was constipation (IBS-C). Forty-three healthy subjects were enrolled as the controls. Stomach antral biopsy samples obtained from all of the subjects were immunostained using the avidin-biotin-complex method for serotonin, gastrin, somatostatin and serotonin transporter (SERT). The immunopositive cell densities and immunoreactivity intensities were determined by computer-aided image analysis. The density of the serotonin-immunoreactive cells was significantly decreased in the IBS-M patients and increased in the IBS-C patients relative to the controls. The immunoreactivity intensity did not differ significantly between the controls and IBS-total. The density of the gastrin-immunoreactive cells was significantly greater in the IBS-D, IBS-M and IBS-C patients than in the controls. The immunoreactivity intensity of gastrin was significantly greater in the IBS-D patients than in the controls. The density of the somatostatin-immunoreactive cells cells was significantly lower in the IBS-total, IBS-D, IBS-M and IBS-C patients than in the controls. The immunoreactivity intensities of both somatostatin and SERT did not differ significantly between the controls and IBS-total. The increase in gastrin cell density and the decrease in somatostatin cell density in all IBS subtypes may cause high levels of gastric secretion, which may in turn contribute to the high incidence of dyspepsia and gastro-oesophageal reflux observed in patients with IBS.

Immunohistochemical study on localization of serotonin immunoreactive cells in the gastrointestinal tract of the European catfish (Silurus glanis, L.).

In this study, it was aimed to identify the distribution of serotonin immunoreactive cells within the gastrointestinal tract (GIT) of European catfish (Silurus glanis). For this purpose, the tissue samples were taken from the stomach (cardia, fundus and pylorus region) and intestine (anterior, middle and posterior region). They were examined by applying the avidin-biotin-immunoperoxidase method. The serotonin containing immunoreactive cells are presented in all regions of the GIT. It was determined to be localized generally in different distribution within the stomachs and intestines of S.glanis. It was found that the most intensive regions of immunoreactive cells were the cardia stomach and posterior of intestine.

Red kidney bean (Phaseolus vulgaris) lectin stimulation increases the number of enterochromaffin cells in the small intestine of suckling piglets

Abstract The quantities and distribution patterns of serotonin-immunoreactive (serotonin-IR) enterochromaffin cells (EC) were studied immunohistochemically in the small intestine of suckling piglets stimulated with red kidney bean lectin, and in nonstimulated, control animals. The co-expression patterns of serotonin with somatostatin (SOM) or corticotropin releasing-factor (CRF) were also studied. After the lectin treatment, the increased numbers of EC were noted in the duodenum of experimental animals. Lectin stimulation did not change the proportions of EC in the jejunum and ileum. In the duodenal epithelium of the lectin-stimulated piglets, the vast majority of serotonin-IR EC were distributed at the basis of crypts. After the lectin administration, the proportions of serotonin-IR/SOM-IR EC were statistically similar in all sections of the small intestine. No upregulation of CRF was found in duodenal, jejunal, and ileal EC of lectin-treated animals. The findings demonstrated that red kidney bean lectin increased the serotonin reservoir in the duodenum, and thus may be an effective stimulant of the gut maturation in suckling mammals.

향사양위탕이 시스플라틴 유발 랫트의 위장관 기능 장애에 미치는 영향

Beneficial Effect of Hyangsayangwi-tang on the Cisplatin-Induced Gastrointestinal Dysfunctions in Rats Eun-Hee Seo, Seong-Tae Kim, Na-Young Bae, Ae-Ryun Choi Dept. of Sasang Constitutional Medicine, College of Oriental Medicine, Daegu Haany Univ. Dept. of Sasang Constitutional Medicine, Div. of clinical medicine, School of Korean Medicine, Pusan National University Objectives This study aimed to observe the effect of Hyangsayangwi-tang on the cisplatin-induced gastrointestinal dysfunctions in rats. Methods Four groups(each of 8 rats per group) were used in this study. Saline and distilled water treated control rats are Intact vehicle control group. Delayed gatrointestinal mortility was induced by intraperitoneal treatment of cisplatin 2mg/kg, once a week for 5 weeks(Cisplatin control group). Hyangsayangwi-tang aqueous extracts(HY) were orally administered in a volume of 5ml/kg, once a day for 14 days from 4th ciplatin treatmernt(HY group). Ondansetron 1mg/kg was subcutaneously treated, in a volume of 1ml/kg, as same as HY(ondansetron group). We measured the body weights, intestinal charcoal transit ratio, fecal parameters, fundus MDA, GSH contents and SOD, CAT activities, TPH and MAO activities, pyloric gastrin and serotonin contents with their immunoraective cells, colonic serotonin-immunoreactive cells, the histopathology of pylorus, fundus mucosa and colon. Results and Conclusions (1) The body weight gains, the small intestinal charcoal transfer rates, the fecal parameters(numbers, weights and water contents) were increased in HY, ondansetron group. (2) The inhibit of fundus antioxidant defense systems by cisplatin were decreased in HY, ondansetron group. (3) The pyloric TPH activities were increased and the pyloric MAO activities were decreased in HY group. (4) The pyloric gastric contents and the gastrin-immunoreactive cells were increased in HY group. And the pyloric serotonin contents and the pyloric and colonic serotonin-immunoreactive cells were decreased in HY group. (5) The pyloru atrophic changes and the gastric surface erosive damage regions by cisplatin were favorably inhibited by treatment of HY. HY, a representative Soeumin prescription improve GI dysfunctions and constipation retarded by cisplatin through modulations of GI enterochromaffin cells, serotonin and gastrin-producing cells and antioxidative systems. Especially HY showed the highest favorable effects more than those of ondansetron.

Postnatal changes in the number of serotonin-immunoreactive cells in midbrain raphe nuclei of male rats

To clarify the developmental changes in serotonergic neurons in the subdivisions of the dorsal (DR) and median raphe (MR) nuclei before puberty, the extent of the nuclei and the number of serotonin (5-HT) immunoreactive (ir) cells were measured in 5-, 15-, and 30-day-old rats and 8-week-old (adult) castrated male rats. The brains were fixed and 50μm frozen sections prepared. After immunostaining for 5-HT, the number of 5-HT-ir cells in a 0.2×0.2mm frame in the dorsal, ventral and lateral subdivisions of the DR (dDR, vDR and lDR, respectively) and MR were counted. Total numbers of 5-HT-ir cells counted in the frame of three sections in each rat were expressed as the number of cells per cubic millimeter (density). The results indicated that the densities of 5-HT-ir cells in the MR were almost the same in all age groups. On the other hand, among the subdivisions of the DR, the mean density of 5-HT-ir cells in 15-day-old rats was higher than that in the 5-day-old group in the lDR only. The area of the three sections of the DR and of the MR was also measured. The area of the DR in 15-day-old rats was found to be twice that in the 5-day-old rats, and differed from the area in 30-day-old rats and adults. There were no differences among the age groups in the areas of the MR. The results indicate that the expression of 5-HT in the lDR and extent of the DR increased to adult levels from days 5 to 15 after birth. In the dDR, vDR and MR, expression of 5-HT at postnatal day5 was at adult levels already.

Comparative analysis of the serotonergic systems in the CNS of two lungfishes, Protopterus dolloi and Neoceratodus forsteri

The organization of the serotonergic system, one of the most important neurotransmitter systems in the brain, has been carefully analyzed in most vertebrate groups, and major shared characteristics have been described, although traits particular to each vertebrate class have also been found. The present study is the first that provides a comprehensive and detailed map of the serotonergic structures in the brain of two representative species of lungfishes, the African lungfish (Protopterus dolloi) and the Australian lungfish (Neoceratodus forsteri), as revealed by immunohistochemistry against serotonin (5-HT). Lungfishes are currently considered the closest living relatives of tetrapods and represent an interesting group for the study of evolutionary traits in the transition from fishes to tetrapods. Distinct groups of serotonin immunoreactive cells were observed in the preoptic area, nucleus of the periventricular organ, pretectum, optic tectum and the long column of the raphe. Fiber labeling was widely distributed in all main brain subdivisions but was more abundant in regions such as the striatum, septum, amygdaloid complex, preoptic area, suprachiasmatic nucleus, lateral hypothalamic area, prethalamus, thalamus, mesencephalic tegmentum and rhombencephalic reticular formation. Comparison of these results with those from other classes of vertebrates highlights numerous common traits shared by most groups of fishes but also reveals that the serotonergic system in lungfishes largely resembles those of amphibians and other tetrapods.

Gastrointestinal Viral Load and Enteroendocrine Cell Number Are Associated with Altered Survival in HIV-1 Infected Individuals

Human immunodeficiency virus type 1 (HIV-1) infects and destroys cells of the immune system leading to an overt immune deficiency known as HIV acquired immunodeficiency syndrome (HIV/AIDS). The gut associated lymphoid tissue is one of the major lymphoid tissues targeted by HIV-1, and is considered a reservoir for HIV-1 replication and of major importance in CD4+ T-cell depletion. In addition to immunodeficiency, HIV-1 infection also directly causes gastrointestinal (GI) dysfunction, also known as HIV enteropathy. This enteropathy can manifest itself as many pathological changes in the GI tract. The objective of this study was to determine the association of gut HIV-1 infection markers with long-term survival in a cohort of men who have sex with men (MSM) enrolled pre-HAART (Highly Active Antiretroviral Therapy). We examined survival over 15-years in a cohort of 42 HIV-infected cases: In addition to CD4+ T cell counts and HIV-1 plasma viral load, multiple gut compartment (duodenum and colon) biopsies were taken by endoscopy every 6 months during the initial 3-year period. HIV-1 was cultured from tissues and phenotyped and viral loads in the gut tissues were determined. Moreover, the tissues were subjected to an extensive assessment of enteroendocrine cell distribution and pathology. The collected data was used for survival analyses, which showed that patients with higher gut tissue viral load levels had a significantly worse survival prognosis. Moreover, lower numbers of serotonin (duodenum) and somatostatin (duodenum and colon) immunoreactive cell counts in the gut tissues of patients was associated with significant lower survival prognosis. Our study, suggested that HIV-1 pathogenesis and survival prognosis is associated with altered enteroendocrine cell numbers, which could point to a potential role for enteroendocrine function in HIV infection and pathogenesis.