牛蛙胃肠胰系统内分泌细胞的免疫组织化学鉴定与定位

应用过氧化物酶标记的链霉卵白素(S-P)免疫组织化学方法对牛蛙(Rana catesbeiana)胃肠胰系统5种内分泌细胞进行了鉴定与定位.在消化道中检测到了5-羟色胺(5-HT)、生长抑素(SS)、胃泌素(Gas)和胰高血糖素(Glu)细胞.5-HT细胞主要分布于胃幽门部和空肠,食道中偶见.SS细胞主要分布于胃,幽门部较密集,小肠各段少量,直肠和食道偶见.Gas细胞主要分布于小肠各段,胃和直肠中偶见,食道中未检测到.Glu细胞主要分布于胃和直肠,小肠各段偶见,食道中未检测到.在胰腺中检测出了5-HT、SS、Gas、Glu和胰多肽(PP)细胞.SS、Glu和PP细胞数量较多,成簇分布于胰岛中,5-HT和Gas细胞少量,散在分布于胰腺腺泡之间.胃腺部和胰腺内分泌细胞多呈圆形、椭圆形或形态不规则,有的可见明显胞突伸向邻近细胞,胃肠道上皮中的内分泌细胞多呈梭形、楔形或锥形,有的可见明显胞突伸向消化腔.与其它两栖动物相比,牛蛙胃肠胰系统内分泌细胞的存在与分布有一些共性,也存在着种间差异.     

Localization in the gastrointestinal tract of immunoreactive prosomatostatin

Antisera against 5 different regions of the entire prosomatostatin molecule were used for immunohistochemical mapping of prosomatostatin-containing structures in the pig gastrointestinal tract, and for radioimmunological and chromatographical analysis of the products of prosomatostatin in extracts of ileal mucosa. The latter showed that the antisera were capable of identifying components containing N-terminal as well as C-terminal parts of prosomatostatin. Endocrine cells were identified with all antisera in most parts of the gastrointestinal tract, and varicose nerve fibres were observed in all parts of the small intestine but not in the stomach and the colon. The colon contained very few immunoreactive structures. Immunoreactive nerve cell bodies were found in the submucous plexus of the small intestine. All immunoreactive endocrine cells in the stomach and the duodenum and all immunoreactive nerves were stained by all 5 antisera whereas the small intestinal endocrine cells did not stain for the most N-terminal region of prosomatostatin. The results suggest that all gastrointestinal somatostatin is derived from the same precursor molecule, which, however, in the small intestinal endocrine cells is processed differently from that of the other tissues.     

A comparative study of neuropeptides in the intestine of two stomachless teleosts (Poecilia reticulata,Leuciscus idus melanotus) under conditions of feeding and starvation

Summary Endocrine cells containing bombesin-, enkephalin-, gastrin/CCK-, 5-HT-, and substance P-like material were demonstrated in the alimentary tract of Poecilia reticulata and Leuciscus idus melanotus. Endocrine cells with neuropeptide-Y-like immunoreactivity were found only in P. reticulata, those with VIP-like immunoreactivity only in L. idus melanotus. Gut nerves showing bombesin-, G/CCK-5-HT-, neurotensin-, substance P-and VIP-like immunoreactivity were observed in both species investigated, enkephalin- and neuropeptide Y-like immunoreactivity in P. reticulata alone. The distribution and amount of endocrine cells and nerves along the gut as visualized with the appropriate antisera varied in both teleosts. Histologically, the intestinal tract of these stomachless fish can be divided into three regions. A large number of endocrine cells with VIP-like immunoreactivity was noted in the rectum of L. idus melanotus. Endocrine cells containing bombesin-, enkepha-lin- and substance P-like material were found only in intestinal parts I and II in L. idus melanotus. Neuropeptide Y-like immunoreactivity was absent from intestinal part I of P. reticulata. The influence of starvation on the immunoreactivity of nerves and enteroendocrine cells in the teleost intestine was examined. After a starvation period of more than 6 weeks, no alterations were observed either in the appearance or amount of nerve and endocrine cell immunoreactivity.     

The development and functioning of embryonic pancreatic islet cells in the anterior chamber of rats

In the present study the morphogenesis and differentiation processes in embryonic pancreatic gland implants into Wistar line rat anterior eye chamber have been investigated. The conditions therein were found to be favourable for the endocrine tissue functioning; a number of morphogenetic changes resulting in the formation of islet structure acting as a morphophysiological unit were noted as well. Endocrine cells possess some selective properties as compared to the exocrine tissue. Alloxan diabetic animals demonstrated the most optimum conditions for the endocrine cells development and functioning.     

Expression Patterns of the Drosophila Neuropeptide CCHamide-2 and Its Receptor May Suggest Hormonal Signaling from the Gut to the Brain

The insect neuropeptides CCHamide-1 and -2 are recently discovered peptides that probably occur in all arthropods. Here, we used immunocytochemistry, in situ hybridization, and quantitative PCR (qPCR), to localize the two peptides in the fruitfly Drosophila melanogaster. We found that CCHamide-1 and -2 were localized in endocrine cells of the midgut of larvae and adult flies. These endocrine cells had the appearance of sensory cells, projecting processes close to or into the gut lumen. In addition, CCHamide-2 was also localized in about forty neurons in the brain hemispheres and ventral nerve cord of larvae. Using qPCR we found high expression of the CCHamide-2 gene in the larval gut and very low expression of its receptor gene, while in the larval brain we found low expression of CCHamide-2 and very high expression of its receptor. These expression patterns suggest the following model: Endocrine CCHamide-2 cells in the gut sense the quality of food components in the gut lumen and transmit this information to the brain by releasing CCHamide-2 into the circulation; subsequently, after binding to its brain receptors, CCHamides-2 induces an altered feeding behavior in the animal and possibly other homeostatic adaptations.     

Endocrine cells of stomach epithelium in the ontogenesis of albino rats

By means of the light and electron microscopy methods structural bases of differentiation and contents of endocrine cells of the stomach epithelium have been studied during the white rat pre- and postnatal development. The development of the endocrine epithelial cells occurs in parallel with the exocrinic ones. Endocrine EC-, G-, D-, P-cells are the first to be revealed in composition of the embryonal stomach epithelium, the others--during the early postembryonic period. The differentiation of the endocrine cells is characterized with asynchronism, is accompanied with a gradual development of organells, increasing amount of secretory granules and appearance of polarity in their distribution. By the 30th day of the postnatal development, the amount of the endocrine cells in the stomach epithelium increases by 5 times in comparison to those in the 1-day-old animals, with simultaneous reaching of the high level in specific differentiation.     

An histological and immunocytochemical study of the neuroendocrine cells in the intestine of Podarcis hispanica Steindachner, 1870 (Lacertidae)

Summary Numerous endocrine cells can be observed in the gut of the lizard Podarcis hispanica after application of the Grimelius silver nitrate technique. The argyrophilic endocrine cells are usually tall and thin in the small intestine but short, basal, and round in the large intestine. Eleven types of immunoreactive endocrine cells have been identified by immunocytochemical methods. Numerous serotonin-, caerulein/gastrin/cholecystokinin octapeptide-and peptide tyrosine-tyrosine-immunoreactive cells; a moderate number of pancreatic polypeptide-, neurotensin-, somatostatin-, glucagon-like peptide-1-and glucagon-immunoreactive cells, and few cholecystokinin N-terminal-and bombesin-immunoreactive cells were found in the epithelium of the small intestine. Coexistence of glucagon with GLP-1 or PP/PYY has been observed in some cells. In the large intestine a small number of serotonin-, peptide tyrosine-tyrosine-, pancreatic polypeptide-, neurotensin-, somatostatin-and glucagon-like peptide-1-immunoreactive cells were detected. Vasoactive intestinal peptide immunoreactivity was found in nerve fibers of the muscular layer. Substance P-immunoreactive nerve fibers were detected in lamina propria, submucosa and muscular layer. Chromogranin A-immunoreactive cells were observed throughout the intestine, although in lower numbers than argyrophilic cells.     

Quantitative characteristics of the endocrine cells of the duodenal glands of Carnivora

In the duodenal glands of the Carnivora investigated endocrine elements have been revealed, a part of them is presented as serotonin-producing EC-cells. Endocrine cells are situated in terminal parts and in glandular ducts, among them elements of open and close types are distinguished. Distribution of these cells in the glandular lobules is subjected to the distal gradient regularity, specific for the gastrointestinal tract mucosal membrane. Amount of endocrinocytes in the glands is much less than in the gut crypts. There is no correlation between distribution of the endocrine cells in the glands and in the crypts. The results of unifactor analysis of variance demonstrate a slight effect of the taxonomic position of the species on the number of endocrine cells in the duodenal glands. The proper endocrine apparatus of the duodenal glands is supposed to produce a local regulatory influence on the secretory activity of exogenic glandulocytes, as well as ensure humoral connections of the duodenal glands with other parts of the gastrointestinal tract.     

三种鲤科鱼类肠道内分泌细胞的初探

使用Grimelius嗜银染色法对草鱼,鲤,翘嘴红鳇3种不同食性的鲤科鱼的肠道内分泌细胞进行了研究,在3种鱼的整个肠道上均发现有内分泌细胞的分布。在前肠前段中,内分泌细胞分布最多,愈向后分布愈少,在肠褶各处均有内分泌细胞分布,以基部分布最密,内分泌细胞几乎都为开放型,位于上皮细胞和杯状细胞之间,将胞突伸向肠腔,有极少数内分泌细胞兼有开放型和封闭型细胞的特点,在它们顶端胞突伸向腔的同时,基部也伸出突起将分泌物送入邻近细胞或细胞间隙中,肠上皮中还发现一种与内分泌细胞具有同样嗜银特性的圆形颗粒。     

Morphology of the endocrine portion of the teleost pancreas]

Endocrine component of the crucian (Carassius carassius), carp (Cyprinus carpio), tench (Tinca tinca) and silurus (Silurus glanis) pancreas is structurally organized in the form of pancreatic islets. Gorbusha (Oncorhynchus gorbuscha) has, besides the islets, some Brockman's bodies. Endocrine component of the pancreas of Teleostei possesses A-, B-, D and acinar-islet cells "B". All types of cells are shoot-shaped and all have contacts with the capillaries. Extrusion of the hormones from the endocrine cells is carried out via emiocytosis, and in gorbusha at the time of migration--by microapocrine means. Secretory granules were observed to get into the capillaries and make hormonal storage necessary for fish migration. It was demonstrated that endocrine component of the pancreas in Teleostei is highly rich in innervation, neuronal fibers containing small granular vesicles.     

Transformation of the intestinal epithelium of the larval anadromous sea lamprey Petromyzon marinus L. during metamorphosis

The events in the transformation of the intestine of the larval lamprey into the adult intestine were followed through the seven (1–7) stages of metamorphosis in anadromous Petromyzon marinus L. Light and electron-microscope observations demonstrated that the processes of degeneration, differentiation, and proliferation are involved in the transformation. In the anterior intestine, degeneration of cells and the extrusion of others into the lumen results in the disappearance of secretory (zymogen) cells and the decline in numbers of endocrine and ciliated cells. Larval absorptive cells, with a prominent brush border, are believed to dedifferentiate into unspecialized columnar cells with few microvilli. Degeneration and removal of cells occurs by both autophagy and heterography and cells extruded into the lumen in the anterior intestine are phagocytosed by epithelial cells of the posterior intestine. The loss of epithelial cells during transformation results in the folding and degradation of parts of the basal lamina and in an extensive widening of the lateral intercellular spaces in all parts of the intestine. As metamorphosis is a nontrophic period of the lamprey life cycle, the possible morphological effects of starvation on the intestinal epithelium are discussed. The development of longitudinal folds is a consequence of the events of metamorphic transformation of the intestinal mucosa. Although an interaction between the epithelium and the underlying tissues is believed to be importent, the actual mechanism of fold development is unknown. The intestinal epithelium of adult lampreys develops from surviving cells of the larval (primary) epithelium. Unlike the situation in amphibians, there does not appear to be a group (nest) of undifferentiated larval cells which differentiate into the adult (secondary) epithelium. Instead, in lampreys, columnar cells that persist through the degradative processes seem to be the source of absorptive and ciliated cells and probably are responsible for mucous and secretory cells. Preliminary observations indicate that the intestinal epithelium of feeding adults is specialized into an anterior region which liberates a secretion, absorbs lipid, and possesses the machinery for ion transport. A posterior region absorbs lipid, secretes mucus, and likely is involved in some protein absorption.     

Microtopographic relation between the lymphatic capillaries and glands of the small intestine

Microtopographic interrelationships of the lymphatic capillaries and glands of the small intestine have been investigated taking into account new data on the diffuse endocrine system, which includes endocrine cells of the gastro-intestinal tract. An immediate contact of the lymphatic capillaries with the intestinal glands, resembling microtopographic interrelationships of the lymphatic capillaries in the endocrine glands, has been revealed in total translucent and histological preparations. A suggestion is made that the microtopographic interrelationships of the lymphatic capillaries and the intestinal glands can be regarded as a morphologic basis for absorption of hormones produced by the endocrine cells of the intestinal glands.     

Immunocytochemical detection of orexin A in endocrine cells of the developing mouse gut.

Orexins are novel neuropeptides that were originally localized in neurons of the hypothalamus and neuronal fibers of the brain. Recently orexin A and its receptor have also been reported in neurons and endocrine cells of the gastrointestinal tract. Because no studies have been done at the embryonic period, we studied the appearance and distribution of orexin A during the development of mouse gastrointestinal tract using immunocytochemical methods. Immunoreactivity to orexin A was detected in neuroendocrine cells of the pyloric region of the stomach at gestational Day 14 and 1 day after in the small intestine. The numbers of immunoreactive cells progressively increased through development until the adult pattern was reached. Staining of reverse-face sections demonstrated that orexin A and serotonin co-localized in some endocrine cells of the mouse stomach and small intestine. These findings suggest that orexin A may be relevant in the growth and maturation of the gastrointestinal tract during intrauterine life.     

Distribution of peptide-containing neurons and endocrine cells in the rabbit gastrointestinal tract,with particular reference to the mucosa

Summary The distribution patterns of peptide-containing neurons and endocrine cells were mapped in sections of oesophagus, stomach, small intestine and large intestine of the rabbit, by use of standard immunohistochemical techniques. Whole mounts of separated layers of ileum were similarly examined. Antibodies raised against vasoactive intestinal peptide (VIP), substance P (SP), somatostatin (SOM), neuropeptide Y (NPY), enkephalins (ENK) and gastrin-releasing peptide (GRP) were used, and for each of these antisera distinct populations of immunoreactive (IR) nerve fibres were observed. Endocrine cells were labelled by the SP, SOM or NPY antisera in some regions.VIP-IR nerve fibres were common in each layer throughout the gastrointestinal tract. With the exception of the oesophagus, GRP-IR nerve fibres also occurred in each layer of the gastrointestinal tract; they formed a particularly rich network in the mucosa of the stomach and small intestine. Fewer nerve fibres containing NPY-IR or SOM-IR were seen in all areas. SOM-IR nerve fibres were very scarce in the circular and longitudinal muscle layers of each area and were absent from the gastric mucosa. The SP-IR innervation of the external musculature and ganglionated plexuses in most regions was rather extensive, whereas the mucosa was only very sparsely innervated. ENK-IR nerve fibres were extremely rare or absent from the mucosa of all areas, although immunoreactive nerve fibres were found in other layers.These studies illustrate the differences in distribution patterns of peptide-containing nerve fibres and endocrine cells along the gastrointestinal tract of the rabbit and also show that there are some marked differences in these patterns, in comparison with other mammalian species.     

Secretion of soluble leptin receptors by exocrine and endocrine cells of the gastric mucosa

Leptin is a hormone secreted by the gastric mucosa into the lumen of the stomach. It is present in its intact form in the intestine where it regulates nutrient absorption and intestinal mucosa integrity. We have identified the binding protein that protects leptin from the harsh conditions of the gastric juice. Immunoprecipitations and Western blot analyses demonstrated that leptin is present in the gastric mucosa and the gastric juice, bound to a protein corresponding to the extracellular domain of the leptin receptor. In the absence of this soluble receptor, leptin is rapidly degraded. Immunocytochemistry on rat gastric mucosa identified the cells and intracellular compartments involved in secretion of this complex. Leptin receptor extracellular domain and leptin are present along the rough endoplasmic reticulum-Golgi-granules secretory pathways and form a complex in the secretory granules of Chief and specific endocrine cells. The long-form membrane leptin receptor OB-Rb, the protease activator furin, and proprotein convertase 7 were found in Chief cell granules but not in those of endocrine cells. The shedding of the receptor occurs in the immature granules. It is concluded that in the immature secretory granules of Chief cells, furin activates proprotein convertase 7 that, in turn, cleaves the extracellular portion of membrane-bound leptin receptors. Leptin bound to its soluble receptor forms a complex that is resistant to the gastric juice. Endocrine cells, on the other hand, generate a soluble leptin receptor by mechanisms different from those of the exocrine cells.     

Neuropeptides in the gastrointestinal canal of Necturus maculosus

Summary The presence and distribution of regulatory peptides in nerves and endocrine cells of the stomach, intestine and rectum of a urodele amphibian, the mudpuppy, Necturus maculosus, was studied immunohistochemically in sections or whole-mount preparations of the gut wall. The effect of the occurring peptides on gut motility was studied in isolated strip preparations of circular and longitudinal smooth muscle from different parts of the gut.Bombesin-, neurotensin-, substance P- and VIP-like immunoreactivity was present in abundant nerve fibres in the myenteric plexus of both stomach, intestine and rectum. Single fibres or bundles were present in the circular muscle layer and in a well-developed deep muscular plexus in the intestine and rectum. Immunoreactive nerve cells were found in the myenteric plexus of the stomach, intestine (neurotensin only) and rectum. Gastrin/CCK-like immunoreactivity was observed only in a few fibres in stomach and rectum.Endocrine cells containing bombesin-, met-enkephalin-, gastrin/CCK-, neurotensin-, somatostatin- or substance P- like immunoreactivity were present in the mucosa.The effect of bombesin was an inhibition of the rhythmic activity in circular muscle preparations and in longitudinal muscle from the rectum, while longitudinal muscle from the stomach usually responded with a weak increase in tonus. Neurotensin, like bombesin, was inhibitory on the spontaneous rhythmic activity of circular muscle throughout the gut, while the effect on longitudinal muscle was an increase in tonus. Met-enkephalin and substance P increased the tonus of all types of preparations, and often, in addition, initiated a rhythmic activity superimposed on this maintained tonus. VIP had a general inhibitory effect on the preparations, decreasing tonus and/or abolishing rhythmic activity.It is concluded that bombesin-, neurotensin-, substance P- and VIP-like peptides are present in nerves throughout the urodele gut and may have physiological functions in regulating the motility of the gut. The gastrin/CCK-like peptide present in nerves of the stomach and rectum may affect the function of these parts of the gut. The regulatory peptides present in endocrine cells may, perhaps with the exception of the somatostatin-like peptide, affect the motility humorally.     

VIP-, substance P-, gastrin/CCK-, bombesin-, somatostatin- and glucagon-like immunoreactivities in the gut of the rainbow trout,Salmo gairdneri

Summary The presence of peptides in the gastrointestinal tract of the rainbow trout, Salmo gairdneri, was investigated immunocytochemically. VIP-like immunoreactivity was demonstrated in nerves in all layers of the stomach and the intestine, whereas substance P-like immunoreactivity was localized to endocrine cells, predominantly in the mucosa of the stomach, and to nerves mainly concentrated in the myenteric plexus throughout the gut. Endocrine cells reactive to gastrin/CCK antiserum were demonstrated in the intestinal mucosa, while no immunoreactivity was found in the stomach. Bombesin-immunoreactive and somatostatin-immunoreactive cells were localized in the stomach mucosa, and cells reactive to glucagon antiserum in the intestinal mucosa. Radioimmunoassay of stomach mucosa and muscle confirmed the presence of VIP-like and substance P-like immunoreactivity in these tissues, while gastrin/CCK-like immunoreactivity was low and bombesin-like immuno-reactivity was insignificant. In conclusion, molecules resembling the mammalian brain-gut peptides may be involved in the neuronal and hormonal control of gut function in fish.     

An immunohistochemical study on the distribution of endocrine cells in the gastrointestinal tract of the musk shrew, Suncus murinus

The endocrine cells in the gastrointestinal tract of the musk shrew were studied immunohistochemically. Eleven kinds of endocrine cells, immunoreactive for serotonin, somatostatin, gastrin, cholecistokinin, gastric inhibitory polypeptide, motilin, secretin, neurotensin, pancreatic glucagon, enteroglucagon and bovine pancreatic polypeptide, were revealed. In the stomach, serotonin-, somatostatin-, gastrin-, pancreatic glucagon- and enteroglucagon-immunoreactive cells were detected. The first three types of cells predominated and were more abundant in the pyloric glands than in the other stomach regions. In the small intestine, all types of endocrine cells were found, each having different distributions and relative frequencies. In the large intestine, 10 types of endocrine cells except cholecystokinin-immunoreactive cells were detected. Serotonin- and bovine pancreatic polypeptide-immunoreactive cells were more numerous in the large intestine than in the small intestine.     

The retinoblastoma protein, RB, is required for gastrointestinal endocrine cells to exit the cell cycle, but not for hormone expression

Important functions of the RB family proteins include inhibition of cell cycle progression and regulation of terminal differentiation. We have examined the role of RB and the related protein, p107, in regulating cell cycle activity and differentiation of gastrointestinal endocrine cells, a relatively quiescent cell population, by conditionally disrupting the RB gene in neurogenin3 (Ngn3)-expressing cells in both p107^+/+ and p107^−/− mice. Endocrine cells in the small intestine, colon, pancreas, and stomach were present in normal numbers in RB and RB-p107 mutants except for an increase in serotonin cells and decrease in ghrelin cells in the antral stomach. Deletion of RB resulted in a dramatic increase in proliferating serotonin cells in the antral stomach and intestine, whereas other enteroendocrine cell types exhibited much lower cell cycle activity or remained quiescent. The related p107 protein appears dispensable for enteroendocrine differentiation and does not functionally compensate for the loss of RB. Our results suggest that RB is required for enteroendocrine cells, particularly serotonin cells, to undergo cell cycle arrest as they terminally differentiate. RB has relatively subtle effects on enteroendocrine cell differentiation and is not required for the expression of the normal repertoire of hormones in the gastrointestinal tract.