Histochemical distribution of digestive enzymes in intestine of goldline, Sarpa salpa L. 1758

Histochemical localization of non‐specific esterase, alkaline and acid phosphatase in the intestine of free‐living goldline (Sarpa salpa L. 1758) was investigated. Fish were caught in the vicinity of the town of Zadar (Adriatic Sea, Croatia), and samples of three parts of the intestine proper (anterior, middle and posterior) as well as the rectum were used for presentation of non‐specific esterases, alkaline phosphatase and acid phosphatase. Non‐specific esterase activity was found in the cytoplasm and brush border of enterocytes in all investigated intestinal segments and the rectum. The activity was stronger in the middle and posterior part of the intestine but weaker in the anterior segment of the intestine as well as in the rectum. Intestinal alkaline phosphatase was detected in the brush border and supranuclear cytoplasm of enterocytes of all investigated intestinal segments. Enzymatic activity gradually decreased in a posterior direction. Acid phosphatase activity was observed as a fine granular reaction product in the supranuclear region of enterocytes and was almost equal in all investigated intestinal segments as well as in the rectum. The possible role of enzymes in intracellular digestion and transport is discussed.     

Histological and ultrastructural study of the digestive tract of rice field eel, Monopterus albus

The histological characteristics of the digestive tract and the ultrastructure of mucosal cells of the stomach and intestine of rice field eel, Monopterus albus, are described to provide a basis for future studies on its digestive physiology. The digestive tract of the rice field eel is a long and coiled tube composed of four layers: mucosa, lamina propria‐submucosa, muscularis and serosa. The pharynx and oesophagus mucosa is lined with a stratified epithelium. The stomach includes the cardiac and pyloric portions and the fundus. Many gastric pits are formed by invaginations of the mucosal layer and tubular gastric glands formed by the columnar cells in the fundus. The intestine is separated from the stomach by a loop valve and divided into a proximal portion and a distal portion. The proximal intestinal epithelium consists of columnar cells with microvilli towards the lumen and goblet cells. The enterocytes are joined at the apical surface by the junctional complex, including the evident desmosomas. Numerous lysosomes and some vesicles are evident in the upper cytoplasm of the cells, and a moderate amount of endoplasmic reticulum and lysosomes are scattered in the supranuclear cytoplasm. The epithelium becomes progressively thicker and the folds containing large numbers of goblet cells are fewer and shorter in the distal portion of the intestine. At the ultrastuctural level, the columnar cells of the tubular gastric glands have numerous clear vacuoles and channels. A moderate amount of pepsinogen granules are present in the stomach. The enterocytes of the intestinal mucosa display a moderate amount of endoplasmic reticulum and lysosomes, and long and regular microvilli.     

Histology and ultrastructure of the gut epithelium of the neotenic cave salamander, Proteus anguinus (Amphibia, Caudata)

Histological, histochemical, and ultrastructural features of the gut of the European endemic cave salamander Proteus anguinus were studied. The gut is a relatively undifferentiated muscular tube lined with a simple columnar epithelium containing numerous goblet cells. The mucosa and underlying lamina propria/submucosa are elevated into a number of high longitudinal folds projecting into the lumen. The enterocytes are covered apically with uniform microvilli. Irregularity in the arrangement of microvilli was observed. Occasionally, irregular protrusions of the cytoplasm appear between groups of microvilli. Pinocytotic activity occurs at the bases of the intermicrovillous space. Mitochondria are numerous in the apical cytoplasm and basally beneath the nuclei. The supranuclear cytoplasm contains most of the cell organelles. The lateral plasma membranes of adjacent cells interdigitate and are joined by junctional complexes. The periodic acid-Schiff (PAS) reaction, indicating neutral mucosubstances, is positive only in the apical brush border of enterocytes and in goblet cells. The goblet cells also stained with Alcian blue (AB), at pH 2.5, thus revealing the presence of carboxylated glycosaminoglycans. Compact aggregations of AB- and PAS-negative cells are situated directly below the epithelium. Mitotic figures are present in individual clusters of cells. The fine structure of cells in these clusters indicated that these cells could be responsible for renewal of intestinal epithelium. Numerous endocrine-like cells could also be seen. The closely packed smooth muscle cells and amorphous extracellular material with collagen fibrils constitute a net-like structure under the basal lamina that is very closely associated with the epithelium. There are numerous acidophilic granular cells between epithelial cells, in the lamina propria/submucosa, and between cells aggregations. They seem to be associated with nematode infections and possibly constitute a humoral defense mechanism.     

Immunocytochemical and enzyme histochemical localization of kallikrein-like enzymes in colon, intestine, and stomach of rat and cat

Kallikrein was localized in goblet (or mucous) cells of rat colon and in rat and cat small intestine and stomach by two immunocytochemical techniques. A kallikrein-like enzyme was also localized by enzyme histochemistry in mast cells of colon, intestine, and stomach of the cat, where they appeared to be associated with blood vessels in the lamina propria. The mast cell enzyme, however, was not detected by immunocytochemistry using antibodies to kallikrein. Modification in the enzyme histochemical procedure (pH, fixation) yielded positive results for a kallikrein-like protease in goblet cells of the intestine and colon. The possible physiological and pathological significance of kallikrein-like enzyme in the gastrointestinal tract and elsewhere is discussed.     

Evidence for vasopressin production in the human gastrointestinal system.

In the present study we performed immunohistochemical examination of segments from the human gastrointestinal system for the presence of cells containing vasopressin (VP) and vasopressin-associated human neurophysin (VP-HNP). VP immunoreactivity was found in crypt cells of the stomach and small intestine, and in mononuclear cells within the lamina propria and submucosa. VP-HNP was demonstrated in the crypt and lamina propria regions of the small intestine, and was colocalized with vasopressin in crypt cells. This colocalization indicates local vasopressin synthesis by these cells and raises the possibility that they may perform an endocrine or exocrine function in the human gastrointestinal system.     

Structure of the digestive tract of tornaria larva inEnteropneusta (Hemichordata)

The ultrastructure of the digestive tract of tornaria larva of enteropneusts was investigated. It showed that the digestive tract consists of three parts: esophagus, stomach, and intestine. The esophagus epithelium consists of two types of multiciliated epithelial cells and solitary muscle cells. Axonal tracts and neurons were found in the ventral wall of the esophagus. The cardiac sphincter contains an anterior band of strongly ciliated cells and a posterior band of cells with long vacuolized processes which partition the sphincter lumen. The stomach consists of three cell types: (1) cells with electron-opaque cytoplasm, bearing a fringed border on their apical sides; (2, 3) sparse cells with electron-light cytoplasm and different patterns of apical microvilli. Cells of the pyloric sphincter bear numerous cilia and almost no microvilli. The intestine consists of three parts. The anterior part is formed of multiciliated cells which bear the fringed border. The middle part consists of flattened cells bearing rare cilia and vast numbers of mace-like microvilli. The posterior part of the intestine is formed of cells bearing numerous cilia and few microvilli. Muscle cells were not found in either stomach or intestine epithelium. One noticed that the structure of the digestive tract of enteropneust tornaria larva differs from that of echinoid pluteus larva.     

Histochemical and Biometric Study of the Gastrointestinal System of Hyla Orientalis (Bedriaga, 1890) (Anura,Hylidae)

This study was carried out to assess the localization of hyaluronic acid (HA) and the distribution of glycoproteins in the gastrointestinal system of adult Hyla orientalis. Histochemical analysis of the gastrointestinal system in H. orientalis showed that mucous content included glycogene and/or oxidable dioles [periodic acid/Schiff (PAS)+], neutral or acid-rich (PAS/AB pH 2.5+), sialic acid residues (KOH/PAS+) and acid sulphate [Aldehyde fuchsin (AF)+] glycoproteins. However the mucus content was not the same in stomach, small and large intestine. The mucus content of stomach included only glycogene and/or oxidable dioles and sialic acid residues. Besides these histochemical methods, the localization of HA was detected using biotinylated hyaluronic acid binding protein labeled with streptavidin-fluorescein isothiocyanate (FITC). In the extracellular matrix of the submucosa, the reaction for HA was evident. Since HA was located in submucosa beneath the epithelial layer of gastrointestinal system, it has a significant role in hydric balance, and essential to provide the gastrointestinal system integrity and functionality. According to biometric results, there were statistical differences between small and large intestine in terms of the amount of material stained positive with PAS/AB, PAS, KOH/PAS and AF/AB. Additionally, number of goblet cells in the small and large intestine was significantly different.Key words: Gastrointestinal system, goblet cell, glycoproteins, hyaluronic acid, amphibian, Hyla.     

牛蛙消化道黏膜5种酶的分布

目的研究牛蛙(Rana catesbeinana)消化道黏膜碱性磷酸酶(ALP)、酸性磷酸酶(ACP)、ATP酶、酯酶和脂酶的分布。方法在消化道的8个部位取材,采用冰冻切片技术和酶的组织化学方法。结果 ALP主要分布于十二指肠、空肠和回肠,胃中酶反应呈弱阳性。ACP主要分布于胃中,食道和肠道酶反应呈弱阳性。ATP酶在消化道各部位均有较多分布,十二指肠、空肠和回肠显著较多,胃各部位其次。酯酶和脂酶均主要分布于肠道,胃各部位其次。结论牛蛙消化道黏膜酶的分布同其它动物有相似之处,也有其自身特点。十二指肠、空肠和回肠是牛蛙的主要消化吸收部位。     

高山倭蛙消化道结构初步观察

高山倭蛙食道粘膜上皮为复层柱状上皮,且固有膜中有丰富的复泡状腺;胃贲门部粘膜上皮无杯状细胞,PAS反应显示固有膜中有深红色颗粒分布,胃体中胃腺丰富;肠分为小肠、大肠、直肠3部分.小肠和直肠上皮中杯状细胞数量多.无尾类消化道结构与海拔高度无明显相关关系.     

变色树蜥消化道5-羟色胺细胞的免疫组织化学定位

用免疫组织化学ABC法,应用5-羟色胺(5-HT)特异性抗血清,对变色树蜥(Calotes versicolor)消化道内含有5-HT的内分泌细胞进行了免疫组织化学的定位研究和形态学观察。结果显示,5-HT细胞在变色树蜥消化道的各个部位均有分布,分布密度近似呈“M”形,其中以空肠分布密度最高,胃体次之,食管最低。5-HT细胞的形态多样,呈圆形、椭圆形、锥体形、长锥体形等,其中胃、幽门和直肠以圆形和椭圆形为主,小肠则以长锥体形为主,其细长突起指向肠腔或固有膜;细胞分布于上皮基部、上皮细胞之间、腺泡上皮细胞之间或固有膜内;多数细胞以内分泌功能为主,少数细胞具有外分泌功能。比较分析表明,5-HT细胞分布型可能与动物的食性有关。     

Histological and morphological evaluations of the digestive tract and associated organs of haddock throughout post-hatching ontogeny

At hatching, the oesophagus of haddock Melanogrammus aeglefinus lacks goblet cells, the intestine is a simple undifferentiated tube, the liver is present as a rounded mass caudal to the heart, and numerous zymogen granules are present in the pancreas. The first intestinal convolution appears at day 2, at the posterior end of the digestive tract. The oesophagus displays alcian blue and PAS positive mucus secreting cells on day 12, which become numerous by day 15. By day 18, epithelial cells of the posterior intestine show evidence of protein absorption in the form of supranuclear vacuoles. The swimbladder inflates in 50% of the larvae by day 22, although inflation rate is highly variable. By day 35, or 10 mm, a pyloric caecal ridge appears which separates the presumptive stomach, which is now showing evidence of gastric gland formation, from the intestine. This marks the beginning of digestive features characteristic of the juvenile stage.     

长鬣蜥胃肠道5-羟色胺细胞的免疫组织化学定位

研究长鬣蜥消化道内含有的5一羟色胺（5-HT）内分泌细胞的位置和形态,采用ABC（avidin—biotin—peroxidase complex）免疫组织化学法,应用5-HT特异性抗血清,对其消化道内的5-HT细胞进行免疫化学定位。结果：5-HT细胞从胃部到直肠的胃肠道各段均有分布,细胞密度分布以胃部最高,十二指肠次之,回肠最低。5-HT细胞的形态呈圆形、椭圆形和锥体形等,其中胃体部和直肠部以圆形和椭圆形为主,小肠部以锥体形为主;5-HT细胞广泛分布于上皮基部、上皮细胞之间、腺泡上皮及腺泡之间,有时可见于固有膜内。结论：5-HT细胞的密度分布与其食性、食物组成和生活环境有关,5-HT细胞的形态与其内、外分泌是相适应的。     

Fine structure of the intestine development in cultured sea bream larvae

At hatching, the gut cells of Sparus aurata are quite undifferentiated; however, slight ultrastructural differences can already be distinguished between the presumptive intestinal regions. The hindgut cells are more differentiated than midgut cells and the rectal cells show rather particular ultrastructural features. During days 1 (D1) and 2 (D2) after hatching, major changes occur that lead to full differentiation of the epithelial cells. Shortly before the onset of exogenous feeding (D3), the anterior intestine enterocytes can synthesize lipoprotein particles (LP) from endogenous lipids. The posterior intestine enterocytes show morphological features indicating a role in absorption and intracellular digestion of nutrients, whereas the rectal cells do not. Transient ciliated cells occur at hatching (D0) in the presumptive intestine, except in the caudal rectum, and disappear at the start of the late endotrophic phase about 3 days after hatching (D3). At hatching, very scarce enteroendocrine and leucocyte-like cells are found at the base of the gut epithelium. Their number increases throughout development. At D3 (late endotrophic phase), LP synthesized mainly in the periblast invade the circulatory system, interstitial spaces of the subepithelial tissue and intercellular spaces of the gut epithelium. When the endo-exotrophic phase begins (D4), the enterocytes can absorb exogenous food. Acid phosphatase activity was detected in microvilli, pical vacuoles and Golgi complex in both anterior and posterior enterocytes, as well as in supranuclear vacuoles (SNV) of posterior enterocytes, but not in the apical tubulovesicular system (TVS). During the exotrophic phase, large lipid droplets (LD) are found in anterior enterocytes, and the SNV occupy a large cell volume in posterior enterocytes. LP accumulate first in extracellular spaces and then are transferred to the circulatory system. Mucous and rodlet cells appear in the intestinal epithelium during the exotrophic phase, from D15.     

Organogenesis of the digestive tract in the white seabream,Diplodus sargus. Histological and histochemical approaches

The ontogeny of the digestive tract of the white seabream, Diplodus sargus during the larval development up to day 45 post-hatching (dph) has been studied using histological and histochemical techniques. The oesophageal goblet cells appeared around 6 dph and contained neutral and acid mucosubstances (PAS/diastase-PAS and Alcian Blue pH 2.5 positive reactions). An incipient stomach can be distinguished from 2 dph but the first sign of gastric gland development was detected around 13-15 dph, increasing in number and size by 22-23 dph. Gastric glands were concentrated in the cardiac stomach region and they had a high content of protein rich in tyrosine, arginine and tryptophan. Acidophilic supranuclear inclusions related to pynocitosis of proteins, were already observed in the intestinal cells of the posterior intestine around 4-6 dph (exogenous feeding) and they were present until 25 dph. The intestinal mucous cells appeared between 15-18 dph and contained a mixture of neutral and acid mucosubstances/glycoconjugates, carboxylated ones being more abundant than the sulphated ones. The stomach and gastric glands were fully developed by the first month of life marking the beginning of digestive features characteristic of the juvenile stage. Around 4-6 dph, glycogen, proteins and neutral lipids were observed in the granular cytoplasm of hepatocytes. Strongly acidophilic zymogen granules were also present, at this time, in the basophilic cytoplasm of the exocrine pancreatic acinar cells and contained abundant proteins, especially rich in arginine, tyrosine and tryptophan.     

Development of the digestive system of Argentine hake,Merluccius hubbsi,larvae

The Argentine hake Merluccius hubbsi is an important fishery resource of the Southwestern Atlantic Ocean and it is also a potential species for cultivation. In this work, the digestive system development in field-caught hake larvae was studied using histological and histochemical approaches. The digestive tract of larvae was divided into: oropharyngeal cavity (OPC), esophagus, stomach (that develops in the preflexion stage), and intestine. The annexed digestive glands consisted of the liver and the exocrine pancreas. At the beginning of the preflexion stage, teeth were developed in the OPC. There were mucous cells in the esophagus secreting different glycoconjugates from hatching. The enterocytes in the posterior intestine exhibited supranuclear vesicles associated with protein absorption. Mucous cells were observed in the posterior intestine in the preflexion stage and, in the anterior region, ending the flexion stage. Each type of glycoconjugates has a specific role. Acidic mucins lubricate and protect from mechanical damage, sialomucines protect from bacterial infections and neutral mucins regulate the acidity of mucus secretion, protect against abrasion and participate in the formation of the chyme, indicating a pregastric digestion. The liver was present since hatching with pancreatic tissue inside and increased in size acquiring the typical structure with hepatocyte cords, sinusoids, vacuoles, and hepatic duct. The hepatocytes vacuolization increased with larval development. The pancreas became extra-hepatic, with basophilic acinar cells and acidophilic zymogen granules. Throughout the ontogeny, the increased structural and functional complexity of the digestive system reflected the transition to exogenous feeding and nutritional increasing needs.     

Identification and regional distribution of the dopamine D(1A) receptor in the gastrointestinal tract

Dopamine (DA) is regarded as an important modulator of enteric function. Recent experiments have suggested that newly cloned DA receptor subtypes are widely expressed in peripheral organs, including the gastrointestinal tract. In the present studies, the D(1A) receptor subtype was identified in rat gut regions through localization of receptor protein by means of light microscopic immunohistochemistry and Western blot analysis and receptor mRNA by RT-PCR and in situ amplification and hybridization (3SR in situ). D(1A) receptor immunoreactivity was shown to have a diverse distribution in the gastrointestinal tract, being present in the gastroesophageal junction, stomach, pylorus, small intestine, and colon. The receptor has a transmural distribution present in both epithelial and muscle layers as well as in blood vessels and lamina propria cells of different gastrointestinal regions. Western blot analysis demonstrated a single 50-kDa band for esophagus, stomach, duodenum, jejunum, and colon. The in situ hybridization signal was localized to the same sites revealed by D(1A) receptor immunoreactivity. RT-PCR revealed an appropriate sized signal in similar regions. This study is the first to identify expression of the central D(1A) receptor throughout the normal mammalian gastrointestinal tract.     

Non-specific carboxylic esterase activity in the digestive tract of the perch, Perca fluviatilis L.

The distribution of non-specific carboxylic esterases (Ec 3.1.1) in the digestive tract of perch, Perca fluviatilis L., was investigated histochemically using 1-naphthyl acetate as the substrate. Strong enzymatic activity was present in the gastric glands and surface cells of the stomach, intestinal mucosa of the pyloric caeca, upper and middle intestine, pancreas (exocrine cells) and liver. The enzymatic activity in the lower intestine and rectum was weak. The activity was not demonstrated in the oesophagus or pyloric sphincter. In the intestine, the activity was localized in the columnar cells especially in the supranuclear cytoplasm. The enzymatic activity demonstrated in the digestive tract of perch using 1 -naphthyl acetate represents combined esterolytic and lipoproteolytic activity.     

鲇肠道黏液细胞的类型、分布、发育及分泌方式研究

利用阿利新蓝-过碘酸雪夫氏试剂(AB-PAS,AB染液pH2．6),对鲇(Silurus asotus)肠道黏液细胞的类型与分布进行了研究,同时采用扫描和透射电镜,研究了黏液细胞的发育与分泌方式。结果显示:鲇肠道黏液细胞染色后呈深蓝色、淡蓝色和浅蓝色,主要含有酸性黏多糖:形态上进一步可将其分为囊状、梨状和杯状三种形态。肠道中黏液细胞的分布密度以中肠最高。其次是后肠,前肠最低;肠道黏液细胞由肠黏膜基底层细胞衍生后,不断向肠腔表层推移而发育成熟;发育过程中黏液细胞内富含大小不等的黏原颗粒,其分泌方式为顶浆分泌。