Apoptosis in the digestive tract of herbivorous Rana pipiens larvae and carnivorous Ceratophrys ornata larvae: An immunohistochemical study

The lifespan of herbivorous Rana pipiens larvae is ～3 months, while that of carnivorous Ceratophrys ornata larvae is only about 2 weeks. During metamorphic climax, the larval gut shortens dramatically, especially in R. pipiens, and its luminal epithelium is replaced by adult‐type epithelium. To determine when programmed cell death occurs during the metamorphic restructuring of the gut, we prepared cross‐sections of the stomach, small intestine, and large intestine from representative larval stages and from juvenile frogs of both species. The sections were incubated with monoclonal antibody against active caspase‐3, one of the key enzymes in the apoptotic cascade. We observed apoptosis in some luminal epithelial cells in each of the three regions of the larval gastrointestinal tract of both species. However, apoptotic cells appeared earlier in larval stages of R. pipiens than C. ornata and few were seen in juvenile frogs of either species. The results demonstrate the occurrence of apoptosis in the metamorphic remodeling of the gut of both R. pipiens larvae and C. ornata larvae. J. Morphol., 2011. © 2011 Wiley Periodicals, Inc.     

Anatomy and histology of the gastrointestinal tract of the neo-tropical opossum (Didelphis marsupialis insularis,Allen 1902)

The morphology of the gastrointestinal tract (GI) is a strong indicator of the dietary habits of a species. The goal of this study was to describe the gross and microanatomy of the digestive tract of the neo-tropical opossum (Didelphis marsupialis insularis) and relate them to the animals’ food habits. GI tracts from 12 adult animals were used for this study. Results found the small intestine made up 65.9% of the GI tract, and the stomach was simple with a prominent caecum. Histologically, the oesophageal mucosa was non-keratinized and glands were found throughout the oesophagus. The large intestine showed a great number of goblet cells, the jejunum possessed well-developed villi, and Peyer's patches were absent in the ileum. The absence of keratinization of the epithelial lining of the oesophagus and stomach and a high lymphocytic infiltration throughout the small and large intestine reflected a more carnivorous diet, whereas the presence of a well-developed caecum in the large intestine indicated the ability to digest plant matter. Overall, the morphology of GI tract of D. m. insularis displayed both carnivorous and herbivorous features, allowing us to conclude that it is an omnivorous animal.     

Digestion in relation to feeding strategies exhibited by crustacean larvae

Decapod crustaceans have adopted a full range of reproductive strategies from the release of large numbers of small eggs (Penaeoidea) to the release of relatively low numbers of large advanced larvae (Nephropidae). As larval size determines trophic position in planktonic food webs, all food sources from phyto- to zooplankton are exploited, with many species changing trophic level during ontogenetic development. Comparative studies on digestive enzymes, levels of activity and changes during ontogeny, together with measurements of gastroevacuation rates and food energy values appear to reveal a general pattern. While herbivorous decapod larvae adapt to low food energy values with high enzyme activity levels, rapid food turnover and low assimilation efficiency, carnivorous larvae exhibit low levels of enzyme activity but compensate by extending retention time of high-energy food to maximise assimilation efficiency. New studies on digestive enzyme levels during development in the penaeid Litopenaeus vannamei, the caridean Lysmata debelius and the cirriped Elminius modestus, appear to agree with previous observations.     

黄喉拟水龟消化道的组织学观察

观察黄喉拟水龟消化道的组织结构.采用常规石蜡切片和HE染色方法对黄喉拟水龟的消化道进行观察.除了口咽腔以外,消化道的管壁是由粘膜层、粘膜下层、肌肉层和外膜组成;各部分的主要区别在于粘膜层,食道和大肠的是复层柱状上皮,胃和小肠的是单层柱状上皮.黄喉拟水龟的舌桔红色,不能伸缩;食管中无食管腺,扩张性强;胃呈囊状,有大量胃腺,腔面皱襞较多;小肠较长,是消化的主要场所,表面有大量的绒毛,在绒毛中可见肠腺;大肠无绒毛,也存在皱襞.     

Characterization of the ultrastructure of the gastrointestinal tract mucosa, stomach contents and liver enzyme activity of the pinfish during development

Carnivorous juveniles (<16 mm L_S) of pinfish Lagodon rhomboides apparently lacked gastric glands in the stomach while larger fish, intermediates (30-33 mm L_S) and herbivorous adults (>80 mm L_S) had numerous gastric glands. Two cell types were identified in the gastric glands of larger fish: mucous secreting cells and secretory cells which had ultrastructure features typical of digestive enzymes and acid secretion. Lipid absorption occurred throughout the caeca and intestine in all sizes of fish. Microvilli found on the rectal epithelial cells of the intermediate and adult pinfish occurred on stalks and were possibly associated with water reabsorption. Liver enzyme activities changed in small fish (26–39 mm L_S) compared to the adult fish. Alanine amino transferase (ALT) and fructose diphosphotase (FDP) activities declined while pyruvate kinase (PK) activities increased significantly. These changes were consistent with a change in diet from a carnivorous (high protein) diet in juveniles to an omnivorous (lower protein) diet determined by stomach content analyses.     

Programmed cell death and heterolysis of larval epithelial cells by macrophage-like cells in the anuran small intestine in vivo and in vitro.

The degenerative processes in the larval small intestine of Xenopus laevis tadpoles during spontaneous metamorphosis and during thyroid hormone-induced metamorphosis in vitro were examined by electron microscopy. Around the beginning of spontaneous metamorphic climax (stages 59-61), both apoptotic bodies derived from larval epithelial cells and intraepithelial macrophage-like cells suddenly increase in number. The macrophage-like cells become rounded and enlarged because of numerous vacuoles containing the apoptotic bodies. Mitotic profiles of the macrophage-like cells, however, are localized in the connective tissue where different developmental stages of macrophage-like cells are present. After stage 62, the intraepithelial macrophage-like cells decrease in number, while large macrophage-like cells which include the apoptotic bodies and retain intact cell membranes and nuclei appear in the lumen. Degenerative changes similar to those during spontaneous metamorphosis described above could be reproduced in vitro. In tissue fragments isolated from the small intestine of stage 57 tadpoles and cultured in the presence of thyroid hormone, the number of intraepithelial macrophage-like cells reaches its maximum around the 3rd day of cultivation when the larval epithelial cells most rapidly decrease in number. These results suggest that the rapid degeneration of larval epithelial cells occurs not only because of apoptosis of the epithelial cells themselves but also from heterolysis by macrophages. The macrophages probably originate in the connective tissue, actively proliferate, migrate into the larval epithelium around the beginning of metamorphic climax, and are finally extruded into the lumen.     

Histological structure of the digestive tract,liver, and pancreas of Ambassis nalua (Hamilton, 1822) with ultrastructural details of the gastric gland

The scalloped perchlet Ambassis nalua is one of the dominant fishes in the Estuarine Pranburi River, Thailand. It is suggested that this fish is in the secondary trophic level with a carnivorous nature. Studies on digestive system will help us further identify the niche of this species in the food web/food chain. The present study therefore aimed to report the detailed structure and ultrastructure of A. nalua digestive system. Fish samples (n = 30) with a total length of 5.7 ± 0.5 cm were obtained using beach seines from the Estuarine Pranburi River. Their digestive tract length and intestine coeficient were 3.6 ± 0.07 cm and 0.91, respectively. Light microscopic observation showed that the digestive wall comprised four layers, namely mucosa, submucosa, muscularis, and serosa. The prominent mucous-secreting cells were found in the mucosal oesophagus. The stomach had many gastric folds, with height and width being 649.76 ± 85.15 and 370.30 ± 68.56 μm, respectively. Gastric glands were found in the anterior stomach but not in the posterior stomach. Each gastric gland was made up of a single type of columnar cells. The gastric cells were ultrastructurally characterized by numerous mitochondria and well-developed secretory granules of varying sizes. A few small vacuoles were also identified in the apical area of the gastric cells. The intestine had two regions (anterior and posterior intestines), and pyloric caecum was absent. The density of the goblet cell was significantly higher in the posterior intestine. These results provide basic knowledge of the digestive system of A. nalua, and the low intestine coefficient and the absence of pyloric caecum suggest the carnivorous feeding habit of this species.     

Ontogeny of some endocrine cells of the digestive tract in sea bass (Dicentrarchus labrax): An immunocytochemical study

Serotonin- and ten peptide-immunoreactive (IR) cell types were identified in the digestive tract of sea bass (Dicentrarchus labrax L.) larvae of four morphofunctional phases ranging in age from hatching to 61 days. The sequence of appearance and location of endocrine cells during ontogenetic development of the larvae was determined. The differentiation of endocrine cells followed a distal-proximal gradient in the gut which paralleled the morphofunctional differentiation. Serotonin-IR cells were identified in the last portion of the digestive tract from phase I onwards and in the gastric region from phase III, before these regions were morphofunctionally differentiated; met-enkephalin-IR cells were identified from phase II onwards in both the differentiated rectum and the undifferentiated intestine; cholecystokinin (CCK)- and synthetic human gastrin-34-IR cells were located only in the intestine and first found in the undifferentiated intestine of phase II; human gastrin-17-, peptide YY (PYY)- and neuropeptide Y (NPY)-IR cells appeared in the intestine from phase II and in stomach in phase IV, when it showed gastric glands; pancreatic polypeptide (PP)- and glucagon-IR cells were observed in both intestine and stomach, but insulin- and somatostatin-IR cells only in stomach, from phase III, during which the intestine but not the stomach was differentiated. PP- and PYY-, PP- and glucagon-, and PYY- and glucagon-like immunoreactivities coexisted from their first appearance in some cells of the gut.     

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.     

Ontogeny of the digestive tract during larval development of yellowtail flounder: a light microscopic and mucous histochemical study

The histological development and mucous histochemistry of the alimentary tract in larval yellowtail flounder were studied using light microscopy. Samples were taken when the larvae were first offered food at 3 days post-hatch, then at 7, 10, 29, 36, and 46 days post-hatch, at which time they were metamorphosing. Regional partitioning of the digestive tract into the buccal cavity, pharynx, oesophagus, post-oesophageal swelling (PES), intestine, and rectum was complete by day 10. Goblet cells were present only in the buccal cavity, pharynx and intestine by day 7, but increased in number and distribution as development continued. By day 29, the posterior zone of the oesophagus had a marked increase in goblet cell density and mucosal folding. At the transition from oesophagus to PES/stomach stratified epithelium with goblet cells changed abruptly to a columnar epithelium with no goblet cells. Multicellular glands in the PES of 36-day larvae allowed it to be defined as a stomach. The distinct brush border of columnar epithelium and the presence of goblet cells characterize the intestine and rectum. All goblet cells throughout the digestive tract were strongly positive for acid mucins as was the luminal layer of the stratified epithelia lining the buccal cavity, pharynx and oesophagus. The PES/stomach epithelium stained weakly for neutral mucins. No mucin staining was associated with the gastric glandular epithelium. The brush borders of the intestine and rectum were strongly positive for combinations of neutral and acid mucins.     

Changes in lectin-binding pattern in the digestive tract of Xenopus laevis during metamorphosis. II. Small intestine.

The binding of seven lectins (concanavalin A, Con A; Dolichos biflorus agglutinin, DBA; peanut agglutinin, PNA; Ricinus communis agglutinin I, RCA-I; soybean agglutinin, SBA; Ulex europeus agglutinin, UEA-I; and wheat germ agglutinin, WGA) to the small intestine in metamorphosing Xenopus laevis was studied by the avidin-biotin-peroxidase (ABC) method. The staining pattern of the epithelium with all lectins except for UEA-I and Con A changed gradually during metamorphic climax; the main component of the epithelium, absorptive cells, gradually became positive for DBA, PNA, and SBA and the scattered goblet cells for RCA-I and WGA. On the other hand, the change of the staining pattern in the connective tissue occurred only for Con A, RCA-I, and WGA, and this change took place rapidly at the beginning of climax (stage 60). Increased staining for Con A and WGA at stage 60 was observed only in a group of connective tissue cells close to the epithelium and in the basement membrane. As metamorphosis progressed, this localization of the staining intensity became less clear. At the completion of metamorphosis (stage 66), the absorptive cells were stained with all lectins except for UEA-I, whereas the goblet cells stained only with RCA-I and WGA. These results indicate that lectin histochemistry can distinguish between larval and adult cells of both two epithelial types (absorptive and goblet cells). The technique may also identify a group of connective tissue cells, close to the epithelium, that possibly induce the metamorphic epithelial changes.     

Distribution of endocrine cells in the digestive tract of Alligator sinensis during the active and hibernating period

The digestive tract is the largest endocrine organ in the body; the distribution pattern of endocrine cells varies with different pathological and physiological states. The aim of the present study was to investigate the distributed density of 5-hydroxytryptamine (5-HT), gastrin (GAS), somatostatin (SS) and vasoactive intestinal peptide (VIP) immunoreactive (IR) cells in the digestive tract of Alligator sinensis during the active and hibernating period by immunohistochemical (IHC) method. The results indicated that 5-HT-IR cells were distributed throughout the entire digestive tract, which were most predominant in duodenum and jejunum. The density increased significantly in stomach and duodenum during hibernation. GAS-IR cells were limited in small stomach and small intestine. The density decreased significantly in small stomach during hibernation, while increased in duodenum. What's more, most of the endocrine cells in duodenum were generally spindle shaped with long cytoplasmic processes ending in the lumen during hibernation. SS-IR cells were limited in stomach and small stomach. The density increased in stomach while decreased in small stomach during hibernation, meanwhile, fewer IR cells occurred in small intestine. VIP-IR cells occurred in stomach and small stomach. The density decreased in small stomach, while increased in stomach during hibernation. These results indicated that the endocrine cells in different parts of digestive tract varied differently during hibernation, their changes were adaptive response to the hibernation.     

Cytochemical features of the digestive tract mucosa of Hemisorubim platyrhynchos (Siluriformes: Pimelodidae)

Membranous organelles, acid glycoconjugates and lipids were characterized in the digestive tract mucosa of Hemisorubim platyrhynchos by cytochemistry techniques. Two types of mucous‐secreting cells were observed in the digestive tract epithelium: goblet cells in the oesophagus and intestine and epithelial cells in the stomach. These cells had a Golgi apparatus more developed than the other cell types. The cytochemical analysis revealed that secretory granules are reactive to acid glycoconjugates, varying in reaction intensity according to the region of the digestive tract. Acid glycoconjugate reactions were also observed in oesophageal epithelial cell microridges and in enterocyte microvilli. In the digestive tract, acid glycoconjugates act to protect the epithelial surface, increasing mucous viscosity, which facilitates the passage of food, prevents the binding of parasites and facilitates their removal. Through lipid staining, a coated membrane was observed around each secretory granule of the oesophageal and intestinal goblet cells, while gastric epithelial cells granules were fully reactive. Oxynticopeptic cells of the gastric glands showed lipid droplets in the cytoplasm and also in the mitochondrial matrix, which act as an energy reserve for these cells that have a high energy demand. Enterocytes showed a well‐developed smooth endoplasmic reticulum, especially in the apical region of the cell, being related to absorption and resynthesis of lipids.     

刀鲚幼鱼消化系统的组织形态学结构

采用光镜和扫描电镜观察长江刀鲚(Coilia nasus)幼鱼消化系统组织形态学结构。结果显示,刀鲚体长,口裂大,含有犬齿状的颌齿和尖锥状的腭齿,具有5对鳃弓,鳃耙长度明显大于鳃丝且表面附着不规则绒毛状细齿;胃呈"Y"型,胃与肠连接处具有16～21个指状环形幽门盲囊;肠为直肠,较短,比肠长为0.241±0.080;肝分为两叶,胰为独立的器官。刀鲚口咽腔为复层鳞状上皮,含有腺体、大量椭圆形黏液细胞、少量杯状细胞及味蕾;胃黏膜都为典型的单层柱状上皮,含有较多由上皮凹陷形成的胃小凹和胃腺;幽门盲囊具有20～25个丰富的褶皱,占满大部分幽门盲囊腔,黏膜层具有微绒毛;中肠黏膜上皮最发达,形成的褶皱细长且连接成网状,单层柱状上皮与复层扁平上皮交替分布。观察结果表明,刀鲚消化系统具有典型肉食性鱼类特征。     

Does diet variation determine the digestive tract length of Capoeta banarescui Turan,Kottelat, Ekmekci and Imamoglu, 2006?

This study tested the spatial variations in the digestive/intestine tract length of Capoeta banarescui, with regard to their diets in different habitats. Highly varied diets observed in a previous study within the same river system posed the question whether this flexibility is reflected in the digestive tract and intestine length of the species in the Ye?il?rmak River, Turkey. Totals of 382 specimens (standard length 4.6–19.1 cm) were captured by electro‐fishing along the river in September 2012 at 11 locations spanning elevations from 34 to 992 m. The stomach, intestine and total digestive tract lengths were measured, and stomach contents analysed from 196 specimens. For statistical analyses, the stomach, intestine and total digestive tract length were expressed as percentages of total weight and standard length. The data provided evidence that the digestive tract and intestine lengths varied significantly among locations in association with the diet. Fish having dominantly carnivorous diets (e.g. chironomid larvae/invertebrates) in two locations had significantly shorter intestines and digestive tracts than those with diets dominated by benthic algae and other plants. The data indicated that C. banarescui showed broad flexibility in their feeding habits. Feeding heavily on plant materials might lead to the development of longer digestive tracts, increasing the active surface area for digestion; alternatively, there may be less invested in development of the digestive tract when feeding primarily on carnivorous diets where the respective digestive enzymes are readily available. The data suggest that phenotypic plasticity in the digestive tract length of C. banarescui is associated more with the abundant protein‐rich carnivorous food sources in the studied habitats. Whether this digestive tract plasticity has a genetic background remains to be verified in future studies.     

Development of digestive secretions in white sturgeon juveniles (Acipenser transmontanus)

Development of the digestive enzyme complement corresponded with this species' carnivorous habits and a natural dietary shift at metamorphosis from larval pelagic zooplanktivory to benthic carnivory. During the first developmental stanza when the digestive tract was differentiating and the larvae were dependent on endogenous nutritional reserves, digestive enzyme concentrations were low. Initiation of gastric secretion (pepsin and acid) was concomitant with onset of exogenous feeding. Highest amylolytic and lipolytic activities were recorded from feeding larvae prior to their metamorphosis to juveniles.     

Energetics of metamorphic climax in the southern toad (<Emphasis Type="Italic">Bufo terrestris</Emphasis>)

During metamorphic climax, anuran larvae must rely on stored energy because changes in oral and digestive morphology prevent foraging and efficient assimilation. Thus, the time required to store adequate energy for metamorphic climax may set a lower limit on age at which it can occur. Therefore, the amount and type of energy used during metamorphic climax must be determined. To quantify the energetic costs of metamorphic climax in Bufo terrestris, oxygen consumption during climax was measured. Wet mass, dry mass, and lipid mass for a group of individuals at the initiation of climax (forelimb emergence, FL) and for another group at the end of climax (complete tail resorption, TR) were also measured to determine whether lipids were used to fuel metamorphic climax. The total amount of energy used, maintenance costs, and development costs during metamorphic climax varied considerably among individuals. Variation in energy metabolism during climax was not related to differences in energy metabolism during larval development or body mass at initiation of climax. TR individuals were significantly lighter in terms of wet mass and had less body water than FL individuals. However, the two groups did not differ in dry mass or lipid mass. Therefore, lipid catabolism is not a major source of energy during metamorphic climax in B. terrestris. As a result, decreases in age at metamorphosis may not be constrained by the need to store energy in the form of lipids.     

Neurotrophin receptors and enteric neuronal development during metamorphosis in the amphibian<Emphasis Type="Italic"> Xenopus laevis</Emphasis>

During metamorphosis, the frog intestine goes through a dramatic shortening with extensive apoptosis and regeneration in the epithelial layer and connective tissue. Our aim was to study changes in the enteric nervous system represented by one inhibitory (vasoactive intestinal polypeptide; VIP) and one excitatory (substance P, neurokinin A; SP/NKA) nerve population and concomitant changes in neurotrophin receptor occurrence during this development in the gut of Xenopus laevis adults and tadpoles at different stages of metamorphosis (NF stages 57–66). Sections were incubated with antibodies against the neurotrophin Trk receptors and p75^NTR, and the neurotransmitters VIP and SP/NKA. Trk-immunoreactive nerves increased dramatically but transiently in number during early metamorphic climax. Nerves immunoreactive for p75^NTR were present throughout the gut, decreased in number in the middle intestine during climax, and increased in the large intestine during late metamorphosis. The percentage of VIP-immunoreactive nerves did not change during metamorphosis. SP/NKA-immunoreactive nerves were first apparent at NF stages 61–62 in the middle intestine and increased in the stomach and large intestine during metamorphosis. Endocrine cells expressing SP/NKA increased in number in stomach, proximal, and middle intestine during metamorphic climax. Thus, neurotrophin receptors are expressed transiently in neurons of the enteric nervous system during metamorphosis in Xenopus laevis and SP/NKA innervation is more abundant in the intestine of the postmetamorphic frog than in the tadpole.This study was supported by grants from the Swedish Research Council to S. Holmgren