Ontogeny, distribution and amine/peptide content of chromogranin A- and B-immunoreactive endocrine cells in the small and large intestine of the chicken

Chromogranin A-(CgA-) and chromogranin B-(CgB-)-immunoreactive endocrine cells were investigated in the chicken intestine during embryonic and post-hatching life. CgA- and CgB-immunoreactive cells first appeared in the intestinal tract at various embryonic ages from day 10 in the cloaca to day 16 in the distal ileum and colon. To identify the CgA- and CgB-immunoreactive cells, each tissue section was double-immunostained using a panel of polyclonal antibodies raised against gut amine/peptides. Almost all the serotonin-immunoreactive cells co-localised CgA and CgB along the entire intestinal mucosa and at all ages examined. In contrast, substance P-, peptide tyrosine tyrosine-, neurotensin- and secretin-immunoreactive cells displayed heterogeneous co-localisation patterns. For example, either all or only some cells of a given endocrine type co-stored Cg; they did so variously-only in the embryo, only after hatching, or at both stages, and co-localizing cells were sometimes located within the mucosa only in the villi and not in the glands, and sometimes vice versa. All the CgA/CgB-immunoreactive cells also displayed argyrophilia.     

The ontogeny and distribution of glucagon- and pancreatic polypeptide-immunoreactive cells in the gastrointestinal tract of the chicken

Summary The times of first appearance and the distribution of APP- and glucagon-immunoreactive cells have been established in the embryonic chick gut between 11 days of incubation and hatching. These immunoreactive cell types appeared for the first time at 13 days of incubation, APP-immunoreactive cells in the duodenum and upper ileum and glucagon-immunoreactive cells in the proventriculus and duodenum. At 14 days, APP-immunoreactive cells were detected in the proventriculus and lower ileum and glucagon-immunoreactive cells in the pyloric region, upper and lower ileum. Thereafter both APP- and glucagon-immunoreactive cells increased in frequency until the numbers at hatching were approximated, APP-immunoreactive cells at 19 days and glucagon immunoreactive cells at 17 1/2 days of incubation. No APP-or glucagon-immunoreactive cells were found in the gizzard, caecum or rectum at any of the selected stages examined. When these types of endocrine cells first appeared, the surface epithelium of the gastrointestinal tract was relatively undifferentiated. A few glands were present in the proventriculus only, at this stage. Thereafter immunoreactive cells of both types were found in the glandular epithelium of the proventriculus, pyloric region and small intestine soon after morphogenesis had begun.This study reports part of work undertaken for the degree of PhD at the University of the Witwatersrand, Johannesburg     

An immunohistochemical study of the ontogeny of the neuroendocrine system in the chicken oesophagus

 The ontogenesis and distribution of serotonin-, chromogranin A-, chromogranin B-, galanin-, neurotensin-, bombesin- and neuropeptide Y-immunoreactive elements were studied in the chicken oesophagus during pre- and post-hatching life. Unlike positive nerve elements, that were present in pre- and post-hatching life, positive endocrine cells were observed only during embryonic life in the oesophageal epithelium. The first endocrine cells, immunoreactive for serotonin and chromogranins, appeared on day 12, in the cervical and thoracic portions of the oesophagus. At the same age, but only in its distal portion, a few bombesin- and neurotensin-immunoreactive cells also appeared. The number of the endocrine cells progressively increased, reaching a maximum on day 15. They then decreased, with a cranio-caudal progression, until they disappeared a few days after hatching. Almost all the serotonin-immunoreactive cells but only a subpopulation of bombesin- and neurotensin-immunoreactive cells colocalized chromogranins. About half of this subpopulation also colocalized serotonin. All these cells reacted positively with Grimelius argyrophile stain. The mucosa of the crop never contained positive endocrine cells. Positive nervous elements appeared first in the wall of the terminal oesophagus and only one or two days later in the proximal oesophagus including the crop. Nervous elements immunoreactive for galanin first appeared from days 6 to 7, for neurotensin from days 7 to 8, for neuropeptide Y from 13 to 15 and for bombesin from 15 to 18. At day 15 galanin-immunoreactive ganglionic cells and fibres occupied both the myenteric and submucous plexus and galanin-positive nerve fibres could be seen throughout the oesophageal wall from the adventitia to a thin subepithelial network. Neurotensin- and neuropeptide Y-immunopositive ganglionic cells and fibres, by contrast, invariably occupied the muscular and submucous layers. Scattered bombesin-immunoreactive ganglionic cells were observed only in the myenteric plexus. The number of positive nerve elements progressively increased until some weeks after birth. Density and intensity were always much higher for galanin and neurotensin than for neuropeptide Y and bombesin. Accepted: 5 November 1997     

Ontogeny of guanylin-immunoreactive cells in rat salivary glands

Guanylin-like peptides regulate electrolyte/water transport through the epithelia. Moreover, these peptides possess antiproliferative activity and regulate the turnover of epithelial cells. In an earlier study we localized guanylin immunoreactivity in secretory ducts of adult rodent salivary glands. In this study we investigated the appearance and distribution pattern of this peptide during the development of rat salivary glands. Guanylin immunoreactivity appeared at the beginning of cell differentiation from solid bud, on embryonic day 17 in the submandibular and sublingual glands and after day 18 in the parotid gland. Guanylin immunoreactivity appeared first in ductal and acinar anlage: its cell distribution pattern and fate differed in these two compartments. In the duct cells guanylin immunoreactivity spread after the duct system developed, whereas in acinar cells it disappeared after cell differentiation. The guanylin immunoreactivity we detected in adult salivary duct cells accords with guanylins role in regulating electrolyte and water transport through the various epithelia. It does so by activating guanylate cyclase-C receptor, increasing intracellular cGMP concentration, and phosphorylating the cystic fibrosis transmembrane conductance regulator (CFTR) protein by the cGMP-dependent protein kinase II. This signaling cascade couples to the ductal electrolyte/water secretion and modulates finally the electrolyte composition of the saliva. On the other hand, CFTR is also involved in mechanisms of cell growth, by regulating apoptosis, and promoting cell differentiation. The early diffuse guanylin immunoreactivity we observed in ducts and acinar anlage, before the secretory set is operative, suggests guanylin has a role in cell differentiation.     

Neurohormonal peptide immunoreactive cells in mucinous cystadenomas and cystadenocarcinomas of the ovary

Summary In 144 benign mucinous cystadenomas of the ovary, 33 mucinous cystadenomas of borderline malignancy and 64 mucinous cystadenocarcinomas, the incidence of tumours containing argyrophil (and probably endocrine) cells was 18%, 33%, and 53%, respectively. The results of a semiquantitative assessment of the number of argyrophil cells in each individual tumour indicates that the greatest numbers occurred in the cystadenocarcinomas. As, however, the tumour cell density was larger in the cystadenocarcinomas than in the cystadenomas, and as the argyrophil cells often had a patchy distribution in the tumour epithelium, the incidence figures are unreliable. In addition, visualization of the argyrophil cells depends on an adequate fixation which is difficult to achieve in the routine processing of large tumour specimens.Many argyrophil cells in the cystadenocarcinomas displayed immunoreactivity with antisera raised against gastro-entero-pancreatic (GEP) neurohormonal peptides. In ten such tumours immunohistochemical evidence was obtained for the presence of the following neurohormonal peptides in the tumour cells: somatostatin, glucagon, gastrin/CCK, neurotensin, and enkephalin. Four of these ten cystadenocarcinomas were multihormonal, in that three contained two cell populations storing GEP neurohormonal peptides, and one tumour even three such populations. In the benign cystadenomas, however, no immunoreactive tumour cells were found. In those of borderline type, only two harboured immunoreactive cells. In both cases the tumour cells stored gastrin/CCK.The general appearance of the epithelium in the mucinous tumours — a continuous single-cell layer of mucin-producing cells intermingled with argyrophil cells of open type — and the spectrum of neurohormonal peptides observed, indicate an origin from the foregut endoderm.This investigation was supported by grants from the Swedish Medical Research Council (Projects No. 4X-4499 and 12X-718), the Swedish Cancer Society (Project No. 805), the Cancer Research Fund of Malmö General Hospital, and the Medical Faculty at the University of Lund     

Granin proteins (chromogranin A and secretogranin II C23-3 and C26-3) in the intestine of reptiles

The occurrence, distribution and the possible cellular co-localizations of chromogranin A (CgA) and of two synthetic secretogranin II-peptides (SgIIC23-3 and SgIIC26-3) with several enteric neuropeptides and serotonin have been investigated immunohistochemically in turtles, lizards and snakes. The distribution of CgA-immunoreactivity was restricted only to the enteroendocrine cells in all the reptiles studied. SgII-immunoreactivity--absent in turtle--revealed nerve cells and fibers, besides enteroendocrine cells in lizard and snake guts. Moreover, the two antisera (C23-3 and C26-3) raised against the different regions of the SgII-molecule yielded distinct distribution patterns of immunoreactivity both in the lizard and snake organs. Small amounts of enteric serotonin cells co-stored CgA or SgIIC23-3 in lizards and snakes and only SgIIC26-3-peptide in snakes. CgA was found co-stored with somatostatin in a few enterocytes of the turtle duodenum. In the same gut segment of lizards and throughout the snake organ, neurotensin and the SgIIC23-3-peptide co-existed in a small number of endocrine cells. The pancreatic polypeptide-containing cells were devoid of immunoreactivity both for CgA and SgII. Bombesin immunopositive cells were absent throughout the intestines of the reptiles investigated. The above findings entirely support the heterogenous distribution of granins in neuroendocrine organs and tissues and also within the same neuroendocrine cell population. They further support the concept of a good conservation of granins during phylogeny.     

人胎结肠及直肠内胃泌素、血管活性肠肽、胰多肽、生长抑素免疫反应阳性细胞的个体发生

目的：探讨人胎结肠及直肠胃泌素(Gas)、血管活性肠肽(VIP)、胰多肽(PP)和生长抑素(SS)免疫反应阳性(IR)细胞的个体发生及其相互关系。方法：免疫组织化学SABC法及细胞计数。结果：结肠内Gas-、PP-、SS-IR细胞始见于第9w,VIP-IR细胞于第18w出现。直肠内未见Gas-IR细胞,PP-IR细胞始见于第9w,SS-、VIP-IR细胞于第11w出现。结肠内Gas-IR细胞随胎龄增长数量减少,于第26w未见。PP-IR细胞数量在结肠内随胎龄增长先少后多,直肠内未见明显变化。VIP-IR细胞在胎期结肠、直肠少,而SS-IR细胞较多,整个胎期VIP-、SS-IR细胞数量无明显增减。结论：VIP、PP和SS存在于人胎结肠、直肠的内分泌细胞,Gas仅在结肠内分泌细胞中有表达,提示各种内分泌激素对胎儿结肠及直肠的发育起调节作用。     

Granin proteins (chromogranin A and secretogranin II C23-3 and C26-3) in the endocrine pancreas of amphibians

The occurrence and cellular distribution of chromogranin A (CgA) and of two synthetic secretogranin II (SgII)-fragments (termed C23-3 and C26-3) has been investigated immunohistochemically in the endocrine pancreas of five amphibian species. Immunoreactivity for CgA was detected only in specimens of the genus Rana, whereas for SgII it was found in all the urodeles and anurans studied. Either CgA or the SgII-fragment displayed its own cellular distribution patterns in the endocrine pancreas of a given species. Moreover, immunoreactivity for both regions (C23-3 and C26-3) of the SgII-molecule exhibited by the same endocrine cell population have been encountered in newt and frog organs. Besides the interspecific heterogeneous distribution of CgA and of the two SgII-fragments in relation to the insular cell types, a striking heterogeneity of their immunostaining density among the endocrine cells of the same type was also revealed. The above findings entirely support the concept of a good conservation of granins during phylogeny; they do not support, however, the previously ascribed usefulness of these anionic glycoproteins as markers for all neuro-endocrine cells.     

Presence and distribution of ghrelin-immunopositive cells in the chicken gastrointestinal tract

The presence and distribution patterns of ghrelin, a gastric acylated peptide, were studied in the entire gastrointestinal tract of the chicken (Gallus domesticus) using the peroxidase-antiperoxidase immunohistochemical method, western blot analysis and a specific antibody against the C-terminal region of rat ghrelin. Ghrelin-immunopositive cells were observed in the mucosal layer of all segments examined. The largest numbers of ghrelin-positive cells were located at the base of lobuli of the proventriculus gland, along villi of the intestines and in crypts of the duodenum. Lower numbers of ghrelin-immunostained cells were located in crypts of jejunum and ileum and only few ghrelin-immunostained cells were detected at the base of crypts of the large intestine. Closed and open types of cells were observed in all segments. Western blot analysis confirmed the presence of ghrelin-like protein in the entire chicken gastrointestinal tract. The anatomical distribution patterns and the morphological characteristics of chicken ghrelin-positive cells suggest that they are endocrine cells. Furthermore, it is concluded that ghrelin shows a high degree of preservation during evolution from non-mammalian vertebrates to mammals.     

The pulmonary neuroepithelial endocrine system in the quail,Coturnix coturnix. Light- and electron-microscopical immunocytochemistry and morphology

Despite extensive knowledge of the neuroepithelial endocrine (NEE) system in the lungs of species of various vertebrate classes, data on avians are limited. The present investigation deals with the light-and electron-microscopical immunocytochemistry and morphology of pulmonary NEE cells in the quail, Coturnix coturnix. Light-microscopically, serotonin immunoreactivity was detected in numerous solitary and clustered NEE cells located in the cilio-mucous epithelium of primary and secondary bronchi in adult as well as in newly hatched quails. Only in newly hatched quails could a small number of bombesin- and somatostatin-like immunoreactive NEE cells be demonstrated. Electron-microscopical morphology revealed that NEE cells contained dense-cored vesicles of a wide range of diameters and electron densities. Nearly all of the NEE cells were seen to rest on the basement membrane of the cilio-mucous epithelium, lacking direct contact with the luminal surface. Nerve varicostities or nerve endings, of both afferent and efferent morphological appearance, were found directly apposed to the basal portion of NEE cells, invaginating between NEE cells or between NEE cells and adjacent epithelial cells. Often, synaptic specializations could be recognized between NEE cells and nerve terminals. Electron-microscopical immunocytochemistry confirmed that the intraepithelial serotonin-containing cells correspond to the cells with NEE characteristics. Moreover, two types of NEE cells could be distinguished in newly hatched quail lungs. Both types showed serotonin immunoreactivity selectively distributed over the dense-cored vesicles, but somatostatin- and bombesin-like immunoreactivities were only noted in one of the NEE cell types and were never seen colocalized. Thus, the avian NEE system too, harbors at least three different bioactive substances and has a morphology comparable to that of mammals, reptiles and amphibians. © 1994 Wiley-Liss, Inc.     

Life, death, and miracles: Th17 cells in the intestine

Th17 cells, a distinct subset of CD4(+) T-helper cells, are commonly associated with chronic inflammatory and autoimmune diseases; however, Th17 cells also possess a variety of beneficial functions as they maintain and defend mucosal barriers against pathogens and promote tissue repair. Furthermore, recent findings indicate that Th17 cells can also acquire immunosuppressive functions that protect against inflammatory and auto-immune diseases. A sentinel population of Th17 cells is localized in the intestine in the absence of pathology and, in response to infection, this population expands in number, and can also modulate its functions. This review discusses the beneficial and pathogenic roles played by Th17 cells in the intestine.     

Morphology and retinal distribution of tyrosine hydroxylase-like immunoreactive amacrine cells in the retina of developingXenopus laevis

Summary The development of neurons immunoreactive to tyrosine hydroxylase (TH-IR) in the retina ofXenopus laevis was investigated from stage 53 tadpoles to adult, by using an antibody against tyrosine hydroxylase. At all developmental stages, most of the immunoreactive somata were located in the inner nuclear layer, and a few in the ganglion cell layer. Immunoreactive processes arborised in the scleral and vitreal sublaminae of the inner plexiform layer, indicating that these cells were bistratified amacrine cells. However, occasionally a few immunoreactive processes were observed projecting to the outer plexiform layer, suggesting the presence of THIR interplexiform cells. The number of immunoreactive amacrine cells in the inner nuclear layer per retina increased from 204 at stage 53 tadpole to 735 in adult, while the number of immunoreactive amacrine cells in the ganglion cell layer did not change significantly over the same period. Retinal area increased from 1.95 mm² at stage 53 to 23.40 mm² in the adult, and correspondingly cell density in the inner nuclear layer decreased from 104/mm² to 31/mm². At all stages there was an increasing density towards the ciliary margin, but this gradient decreased with age. The soma size of immunoreactive amacrine cells increased with age, and was consistently larger in the central than in the peripheral retina. Dendritic field size was estimated to increase 13-fold, from stage 53 to adult. This study shows that tyrosine hydroxylase-like immunoreactive amacrine cells are generated continuously throughout life, that after metamorphosis the retina grows more by stretching than by cell generation at the ciliary margin, and that the increase of dendritic field size is proportional to the increase in retinal surface area.On leave from Department of Anatomy, Zhanjiang Medical College, Guangdong, People's Republic of China     

Nutrient sensing in the gut: interactions between chemosensory cells, visceral afferents and the secretion of satiation peptides

The chemical environment of the intestinal lumen and the presence of nutrients in the gut result in the secretion of regulatory peptides and in the discharge of intestinal sensory afferent fibers. Since afferent nerve terminals do not directly extend into the intestinal lumen, their activation presumably depends on intermediary steps — post-absorptive mechanisms, i.e. direct neuronal activation by absorbed substances and pre-absorptive mechanisms, i.e. neuronal activation by a secondary substance released from the mucosal epithelium.Entero-endocrine cells (EEC) are able to ‘taste’ the luminal content and to function as chemosensory transducers to provide the interface between the intestinal lumen and the afferent nerve terminals. Their secretory products - mainly peptide hormones - are released upon stimulation by nutrients into the extracellular space of the lamina propria to either act 1) locally in a paracrine fashion to activate afferent terminals or other cells or 2) in an endocrine fashion via intestinal capillaries and the lymphatic system to bind to specific receptors at more distant targets.The chemosensory mechanisms by which EEC sense the intestinal lumen remain, however, poorly understood. Recent information suggest that taste signaling mechanisms known from the oral epithelium also operate in the mucosal epithelium. Several nutrient-responsive G-protein coupled receptors (GPCRs) have been identified in EEC including the sweet-taste responsive T1R2/T1R3 heterodimer or GPR120, responsive to free fatty acids (FFAs).This review provides a brief overview on gastrointestinal chemosensory mechanisms and their functional involvement in the secretion of satiation peptides with a focus on human studies albeit most evidence at present comes from in vitro or animal studies.     

Electron microscopic investigations on the differentiation of Purkinje cells in the ontogenetic development of the chicken heart

Summary The ultrastructural development of subendocardial Purkinje cells of chicken left ventricle was investigated. In 9-day-old chick embryos the cell diameter and the organization of the cell organelles allow a distinction between Purkinje cells and ordinary myocardial cells. In 14-day-old chick embryos, Purkinje cells show large accumulations of myosin filaments with interspersed ribosomes in addition to normomeric myofibrils. In these accumulations actin filaments seem to be absent. The deficiency of actin filaments is supposedly the reason for the random distribution of the myosin filaments.Purkinje cells of early chick embryos show areas with densely packed glycogen granules. In older embryos the glycogen concentration declines and only separate glycogen granules are visible.At hatching time the first subsarcolemmal leptomeric fibrils were observed in Purkinje cells. Leptomeric complexes arising in a close spatial relationship to the accumulations of myosin filaments and ribosomes can be seen in 2–4 weekold chickens. With the increasing age of the chickens, the size of these accumulations declines. Adult hens exhibit smaller accumulations, mainly in the neighborhood of leptomeric complexes.Purkinje cells show a distinct ontogenetic development. They are not simple embryonic remnants of ordinary myocardial cells.     

Ontogeny of the endocrine cells of the intestine and rectum of sea bass (Dicentrarchus labrax L.): an ultrastructural study

Several endocrine cell types were ultrastructurally characterized during the differentiation of the intestine and rectum of sea bass (Dicentrarchus labrax L.) larvae. Only one cell type (type I) was found in the posterior region of the undifferentiated gut of 5-day-old larvae (phase I). Types V and VI were found in both the intestine and rectum, types II, III and IV in the intestine, and types VII and VIII in the rectum of 9- and 12-day-old larvae (phase II), the rectum alone showing signs of functional differentiation. In phase III larvae, in which both the intestine and rectum were differentiated, types IX, X, XI, XII, XIII, XIV and XV were found in the intestine, only types X, XI and XII being seen in the rectum. Besides these, a new cell type, XVI, was observed in the intestine of 55- and 60-day-old larvae (phase IV), in which the digestive tract was completely differentiated. The endocrine cells appearing in phases I and II showed very scarce secretory granules and the ultrastructural features of undifferentiated cells. Some endocrine cell types in the earliest developmental stages were related to some of those found later. A maturational process of the endocrine cell types paralleled the differentiation of the intestine and rectum, with an apparent increase in the number of secretory granules accompanying organelle development.     

Ontogeny of the endocrine cells of the stomach of sea bass (Dicentrarchus labrax L.): an ultrastructural study

The endocrine cells present in the developing stomach of sea bass larvae have been characterized ultrastructurally. Only one endocrine cell type (type I) was found in the presumptive stomach of 9- and 12-day-old larvae, one (type II) and five (types III, IV, V, VI and VII) in the aglandular stomach of 32-, and of 39- to 46-day-old larvae, respectively, and five (types III, VIII, IX, X and XI) in the differentiated stomach of 55- and 60-day-old larvae. A maturation process was established for some of these cells. Types I, II and III and types IV and X were thought to be different maturational stages of the same endocrine cell type.     

Immunohistochemistry and morphometry of gastrin cells in the rat pyloric antrum during starvation

Summary The gastrin cells (G cells) in the rat pyloric antrum after 7, 14, 21 and 28 days of starvation were investigated by immunohistochemistry and electron microscopy. In the peroxidase anti-peroxidase method for light microscopy, gastrin immunoreactive cells during starvation markedly decreased in number and size. Quantitative electron microscopy revealed that during starvation the number of electron-lucent granules were greatly reduced, but the number of electron-dense granules increased; the number of intermediate granules were not remarkably changed in G cells. These results may suggest that the synthesis of gastrin and granule maturation were greatly inhibited during long-term starvation.     

调节性T细胞促进球囊损伤颈动脉内皮化的作用及机制

目的 探讨调节性T细胞（Treg细胞）促进球囊损伤颈动脉内皮化的作用及机制。 方法 使用Treg细胞分选试剂盒分选大鼠脾脏Treg细胞;构建大鼠颈动脉损伤模型;模型制作成功后分为对照组（尾静脉注射相同体积的生理盐水）、血管内皮生长因子（VEGF）组（尾静脉注射VEGF, 20 μmol/kg）和Treg细胞组（尾静脉注射1×105个Treg细胞）。HE染色检测内皮化情况;ELISA和免疫组织化学法检测血清中白细胞介素（IL）-10、转化生长因子-β（TGF-β）、IL-1β和肿瘤坏死因子α（TNF-α）的含量;流式细胞术检测单核细胞、T细胞和内皮祖细胞（EPC）的比例。 结果 组织学染色结果显示,对照组未见内皮细胞层的形成,Treg细胞组可见较多内皮细胞覆盖在颈动脉内层;免疫组织化学结果显示,对照组与Treg细胞组IL-10、TGF-β、IL-1β和TNF-α蛋白表达差异均有统计学意义（t=8.252,P<0.01;t=3.254,P<0.05;t=6.237,P<0.01;t=7.529,P<0.01）。ELISA结果显示,对照组IL-10、TGF-β、IL-1β和TNF-α的含量分别为（17.38±2.595）μg/L、（4.750±1.549）μg/L、（11.65±1.908）μg/L和（1.163±0.3333）μg/L;Treg细胞组的含量分别为（58.43±6.060）μg/L、（14.17±2.250）μg/L、（1.550±0.3819）μg/L和（0.2100±0.06938）μg/L,差异有显著性（t=6.170,P<0.01;t=3.558,P<0.01;t=5.191,P<0.01;t=2.800,P<0.05）;流式细胞术的结果显示,对照组CD34+VEGFR-2+EPC比例为（0.2838±0.01975）%,Treg细胞组为（0.5667±0.05993）%,差异有显著性（t=4.483,P<0.01）;IL-10阻断组EPC比例为（0.4807±0.03067）%,相对于Treg细胞组,差异不具有统计学意义（t=1.278,P>0.05）;TGF-β阻断组EPC比例为(0.3082±0.02291)%,相对于Treg细胞组,差异有显著性（t=4.029,P<0.01）。 结论 Treg细胞通过诱导EPC动员,促进球囊损伤颈动脉的快速内皮化。     

Comparative distribution of NADPH-diaphorase activity and tyrosine hydroxylase immunoreactivity in the diencephalon and mesencephalon of the domestic chicken (Gallus domesticus)

We described the distribution of NADPH-diaphorase-containing neurons in relation to tyrosine hydroxylase immunoreactivity in the diencephalon and mesencephalon of the chicken. In the diencephalon, both markers were found in the lateral hypothalamus, dorsal hypothalamic area, hypothalamic periventricular nucleus, paraventricular nucleus and mamillary area. A close examination showed that the fine distribution of these markers differed slightly, so that they were never observed in the same neurons. In the mesencephalon, NADPH-diaphorase and tyrosine hydroxylase immunoreactivity were found in the ventral pedunculopontine area (nucleus tegmenti pedunculopontinus pars compacta, adjacent areas surrounding the quintofrontal tract and the nucleus mesencephalicus profundus ventralis), the coeruleus complex (locus coeruleus, ventral and dorsal subcoeruleus nuclei), the ventral tegmental area and the central gray. The majority of these neurons contained either diaphorase or tyrosine hydroxylase. Nevertheless, in a few cases both markers appeared to colocalize in the same neuron, typically in large perikarya of the ventral pedunculopontine area.Abbreviations AVT Ventral tegmental area of Tsai - DAB diaminobenzidine - DHA dorsal hypothalamic area - DMA anterior dorsomedial thalamic nucleus - DMP posterior dorsomedial thalamic nucleus - DSV ventral supraoptic decussation - GCt central gray - ICH intercalated hypothalamic nucleus - IH inferior hypothalamic nucleus - LHy lateral hypothalamic area - LoC locus coeruleus - ML lateral mamillary nucleus - MM medial mamillary nucleus - MPv ventral part of the deep mesencephalic nucleus - nI interstitial nucleus of Rendahl - NADPH-d NADPH-diaphorase - NGS normal     

Potential role of plasmacytoid dendritic cells for FOXP3 regulatory T cell development in human colorectal cancer and tumor draining lymph node

FOXP3⁺ regulatory T cells (Tregs) play an important role in the maintenance of tumor immunity tolerance. Compared with conventional myeloid dentritic cells (mDCs), plasmacytoid dendritic cells (pDCs) exhibit poor immunostimulatory ability, and their interaction with T cells often promotes the development of Tregs. The aim of this study was to determine FOXP3⁺ Tregs and CD123⁺pDCs infiltration in colorectal cancer and tumor draining lymph node (TDLN), and to evaluate the clinical significance and relationship between pDCs infiltration and Tregs development in the CRC tolerogenic milieu. An immunohistochemical assay was conducted to assess FOXP3⁺Tregs and CD123⁺pDCs infiltration in tumor tissue and in metastatic-free TDLN (mfTDLN) and metastatic TDLN (mTDLN). The results showed that FOXP3⁺ Tregs infiltration was more frequent in tumor tissue than in adjacent normal mucosa (P < 0.001). FOXP3⁺Tregs infiltration was associated with advanced TNM stage and lymph node metastasis (P < 0.01 and P < 0.01 for TNM stage and lymph node metastasis, respectively). Different from FOXP3⁺Tregs, CD123⁺pDCs frequencies were lower in most CRC tumor tissues, whereas the positive rate of CD123 expression in CRC was significantly higher than in adjacent normal mucosa tissue (P < 0.01). Compared to mfTDLN, mTDLN was significantly enriched in FOXP3⁺ Tregs (P < 0.01) and increased in pDC/mDC ratio (P < 0.01). The statistical analysis demonstrated a significant correlation in both Tregs and pDC/mDC ratio in mTDLN. These results suggest that there are more FOXP3⁺ Tregs with a stronger prognostic significance which might promote tumor tolerance, and that CD123⁺pDCs might contribute to Tregs development in the CRC tolerogenic milieu.