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Appearance of somatostatin and vasoactive intestinal peptide along the developing chicken gut.

The appearance of somatostatin (SOM)-immunoreactive (IR) and vasoactive intestinal peptide (VIP)-IR neurons in different regions of the embryonic chicken gut was studied by immunostaining wholemounts. The patterns of expression of these peptides in myenteric neurons showed a number of similarities. Both peptides first appeared in the region of the proventriculus-gizzard: SOM at embryonic day (E)4, VIP at E5.5. At later times both peptides were found in positions both rostral and caudal to the gizzard. Both peptides appeared independently in cells at a second site, the cecum of the hindgut: SOM was observed at E6.5 and VIP at E7.5. VIP-IR and SOM-IR cells appear throughout the cecum, then in the rectum, and finally in the ileum. Differences in the patterns of expression were also found. SOM- and VIP-IR neurons appeared at different times along the length of the gut. VIP-IR cells populated the entire gut by E11.5, whereas SOM-IR cells were not present throughout the gut until E13.5. SOM-IR cells appeared in the terminal part of the ganglion of Remak at E4.0. At E6 these SOM-IR cells sent fibers into the wall of the hindgut and later into the midgut. No VIP-IR cells were found in the ganglion of Remak. These findings suggest that neural crest-derived cells first express SOM- and VIP-IR in particular regions of the gut, namely, the proventriculus-gizzard and the cecum. Certain conditions must exist at these sites which favor the expression of these neuropeptides by neural crest-derived cells. The observation of SOM- and VIP-IR cells in the cecum at a stage of development before cells are seen in the ileum supports the concept that sacral neural crest cells contribute precursors for enteric neurons of the avian hindgut.

Animals↗

Comparative and functional morphology of the stomach in the adult and newborn pigmy hippopotamus (Choeropsis liberiensis).

Examination of the topographical anatomy of the stomach complex and intestinal tract of an adult male and a newborn female pigmy hippopotamus (Choeropsis liberiensis) shows that the stomach consists of four chambers, the first three constituting the proventriculus and the fourth the glandular stomach. The proventriculus is made up of a visceral and parietal blind sac opening into a connecting chamber, which in turn opens into the glandular compartment. The walls of the proventricular chambers are covered with villi and non-glandular mucous membrane. The significant difference between the animals is that whereas the connecting chamber of the adult stomach lies transversely deep in the cranial part of the abdomen and connects with the glandular chamber on the right side, that of the newborn lies almost vertically and connects ventrally with the glandular compartment situated on the floor of the abdomen. A groove which in the adult runs more or less horizontally from the cardia through the visceral sac and connecting chambers is aligned almost vertically in the newborn. In the adult the connecting chamber is the largest compartment, but in the neonate the visceral blind sac and the glandular compartment are proportionally larger. Functional aspects of these anatomical differences are discussed in relation to suckling behavior.(ABSTRACT TRUNCATED AT 250 WORDS)

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Role for cGATA-5 in transcriptional regulation of the embryonic chicken pepsinogen gene by epithelial-mesenchymal interactions in the developing chicken stomach.

A gene encoding embryonic chicken pepsinogen (ECPg), a zymogen of the digestive enzyme pepsin, is expressed specifically in epithelial cells of glands of embryonic stage proventriculus (glandular stomach) under the influence of mesenchyme. We found four GATA and one Sox binding motifs in 1.1 kb of the 5' flanking region of the ECPg gene which are essential to the organ-specific expression of the gene. The expression of cGATA-5 and cSox2 in the proventriculus from day 6 to day 12 of incubation was therefore analyzed. cGATA-5 was more strongly expressed in glandular epithelial cells than in luminal epithelial cells, while cSox2 gene expression was weaker in glandular epithelial cells. Using heterologous recombination explants we also discovered that the expression of cGATA-5 and cSox2 in epithelial cells was affected by mesenchyme when the latter induced ECPg gene expression in epithelial cells. Introduction of expression constructs into epithelial cells by electroporation demonstrated that cGATA-5 upregulated transcription of a reporter luciferase gene via a cis element in the 5' flanking region of the ECPg gene. The gel mobility shift assay revealed that the cGATA-5 protein specifically binds to the GATA binding sites. cSox2 downregulated the activity of luciferase but it was not through the Sox binding motif. These results suggest that cGATA-5 positively regulates transcription of the ECPg gene and is involved in spatial regulation of the pepsinogen gene during development.

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Lead exposure in Laysan albatross adults and chicks in Hawaii: prevalence, risk factors, and biochemical effects.

Prevalence of lead exposure and elevated tissue lead was determined in Laysan albatross (Diomedea immutabilis) in Hawaii. The relationship between lead exposure and proximity to buildings, between elevated blood lead and droopwing status, and elevated liver lead and presence of lead-containing paint chips in the proventriculus in albatross chicks was also examined. Finally, the effects of lead on the enzyme delta-aminolevulinic acid dehydratase (ALAD) was determined. There was a significant association between lead exposure or elevated tissue lead and proximity to buildings in albatross chicks and presence of lead paint chips in the proventriculus and elevated liver lead in carcasses. Although there was a significant association between elevated blood lead and droopwing chicks, there were notable exceptions. Prevalence of elevated tissue lead in albatross chicks was highest on Sand Island Midway and much less so on Kauai and virtually nonexistent in other areas. Prevalence of lead exposure decreased as numbers of buildings to which chicks were exposed on a given island decreased. Laysan albatross adults had minimal to no lead exposure. There was a significant negative correlation between blood lead concentration and ALAD activity in chicks. Based on ALAD activity, 0.03-0.05 microg/ml was the no effect range for blood lead in albatross chicks.

Aging↗

Appearance and some neurochemical features of nitrergic neurons in the developing quail digestive tract.

Using immunocytochemistry, NADPH-diaphorase (NADPHd) histochemistry and electron microscopy, the appearance of nitrergic enteric neurons in different digestive tract regions of the embryonic, neonatal and adult quail was studied in whole mounts and sections. NADPHd was first expressed by embryonic day 4-5 in two distinct locations, namely the mesenchyme of the gizzard primordium and at the caeco-colonic junction. At embryonic day 6, nitrergic neurons had already begun to form a myenteric nerve network in the wall of the proventriculus, gizzard and proximal part of the large intestine and by embryonic day 9, a myenteric network was visualized along the entire digestive tract of the quail. At the level of the stomach, this network was confined to the area covered by the intermediate muscles. By embryonic day 12-13, the NADPHd-positive myenteric neurons in the wall of the distal parts of the blind-ending paired caeca also became organized into ganglia. From this developmental stage on, a submucous nitrergic nerve network, sandwiched between the lamina muscularis mucosae and the luminal side of the outer muscle layer, became prominent in the proventriculus and intestinal walls. In the adult quail, only a minority of the NADPHd-positive neurons stained for vasoactive intestinal polypeptide (VIP) along the intestine. VIP-immunoreactive (IR) cell bodies were frequent in the myenteric plexus but not in the submucous plexus, whereas there were considerable numbers of NADPHd-positive neurons in both these plexuses. Nitrergic fibres were also observed in the outer muscle layer, but were almost absent from the lamina muscularis mucosa and lamina propria, in contrast to the dense VIP-ergic innervation encircling the bases of the intestinal crypts.

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Postnatal development of intra-epithelial leukocytes in the chicken digestive tract: phenotypical characterization in situ.

In the present study, we characterized intra-epithelial leukocytes in the digestive tract of chickens during postnatal development. Their phenotype was characterized by monoclonal antibodies in cryostat sections and the numbers of the different cell-types were counted in the epithelium of the esophagus, proventriculus, duodenum, jejunum, cecum, and colon. All intra-epithelial leukocytes bore the leukocyte-common antigen CD45; 35% were T lymphocytes, and 50% bore a B-cell marker. However, no immunoglobulin-bearing cells were detected in the epithelium. Monocytes and macrophages were found only in the epithelium of the esophagus. A remaining population of non-B, non-T, non-monocyte cells (15%) was present in all parts of the digestive tract. The number of intra-epithelial leukocytes was greatest in the duodenum and jejunum, and decreased in the proximal part of the cecum and in the colon. Intra-epithelial leukocytes were only sporadically detected in the proventriculus. The total number of intra-epithelial leukocytes increased until 8 weeks after hatching and then decreased at 18 months. In the esophagus, the total number of intra-epithelial leukocytes changed little during aging. We found that the intra-epithelial leukocytes of chickens and rodents are distinct in that chicken intra-epithelial leukocytes comprise a cell population that bears a B-cell antigen but that lacks surface immunoglobulins.

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Bombesin-like immunoreactivity in the avian gut and its localisation to a distinct cell type.

The distribution of a bombesin-like immunoreactive peptide in the avian gastro-intestinal tract was analysed by combined radioimmunoassay and immunocytochemistry. Radioimmunoassay of tissue extracts showed that the largest quantities of bombesin-like immunoreactivity were present in the proventriculus (64.5 +/- 6.0 pmol/g) with smaller but still considerable amounts in the gizzard (40.0 +/- 6.0 pmol/g). Immunocytochemically the extractable bombesin-like immunoreactivity was localised in numerous endocrine cells. These, in the proventriculus, were found mainly in the deeper layers of the mucosa. Further study of these cells by the semi-thin/thin technique revealed the presence of characteristic secretory granules. The functional name BN is proposed for this cell type.

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The prevalence and retention of lead pellets in Japanese quail.

Thirty-six Japanese quail (18 control birds, 18 lead-dosed birds) were used. The 18 quail were dosed with #4 lead weight that were orally inserted into the proventriculus. Delta-aminolevulunic acid dehydrase (ALAD) activity in erythrocytes in the dosed quail decreased 90% (p < 0.01) after one week as compared with the undosed quail. This inhibition of ALAD activity in erythrocytes indicates lead exposure. Radiographics were obtained at 0, 1, 4, 9, 22, and 32 days. The lead pellets remained in the gizzard and became smaller in 4 days. At day 22, after the lead treatment, in 8 quail of the treated quail 12, the lead pellets disappeared. At day 32, all lead pellets disappeared. These findings indicate that the ingested lead pellets are absorbed gradually in the intestine. The lead concentrations in the blood, liver, kidney, and femur of the lead-dosed quail were significantly higher than in the undosed quail until the 6th week. At week 2, the lead concentration of the proventriculus, gizzard, gizzard contents, duodenum, small intestine, and cecum in the dosed quail was significantly higher. Lead concentration of feces was significantly higher at weeks 2 and 4 (p < 0.01). Throughout this study, no lead pellets were found in the feces.

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Noncoordinate expression of Drosophila glue genes: Sgs-4 is expressed at many stages and in two different tissues.

The glue genes of Drosophila melanogaster comprise a family of genes expressed at high levels in the salivary glands of late third instar larvae in response to the insect hormone ecdysone. We present evidence that, in contrast to the other glue genes, Sgs-4 is turned on throughout Drosophila development and is not expressed exclusively in the larval salivary glands. Larvae transformed with an Sgs-4/Adh (alcohol dehydrogenase) hybrid gene exhibit Sgs-4-directed Adh expression in the larval proventriculus as well as in the salivary glands as early as the first instar. Sgs-4-specific RNA can be detected at very low levels during all stages of development. During late third instar, levels of Sgs-4 RNA in the salivary glands increase several-thousand-fold, thereby accounting for the large amounts of Sgs-4 protein present in the glue produced by the salivary glands. This pattern of expression is unique to the Sgs-4 gene. While expression of several of the other glue genes can be detected in embryos and early larvae, they appear to be expressed neither throughout development nor in the larval proventriculus. Appearance of the glue gene RNAs in mid third instar salivary glands is noncoordinate, even for the chromosomally clustered genes Sgs-3, Sgs-7, and Sgs-8.

Alcohol Dehydrogenase↗

The action of bombesin on gastric secretion of the chicken.

The natural tetradecapeptide bombesin at a threshold dose of 3--5 ng/kg/min i.v. stimulates gastric secretion in the chicken with either an acute or a chronic proventriculus fistula; this effect is blocked by atropine. The occurrence of tachyphylaxis in the acute, but not in the chronic preparation, and the inhibition of the response by proventriculus acidification, in the presence of an intact sensitivity to caerulein -- a directly stimulating agent -- support the hypothesis of an indirect mechanism of action of bombesin, namely the release of endogenous gastrin, as well as of the existence of gastrin also in the chicken.

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Presynaptic inhibitory effects of catecholamines on cholinergic transmission in the smooth muscle of the chick stomach.

In the vagus nerve--smooth muscle preparation isolated from the chick proventriculus, adrenaline, clonidine (10(-8) - 2.5 x 10(-7) M), noradrenaline (10(-7) - 2.5 x 10(-6) M) and dopamine (10(-5) - 10(-4) M) inhibited the contraction induced by low frequency (0.5 Hz) stimulation of the vagus nerve, but they did not inhibit the contraction elicited by acetylcholine (5 x 10(-8) - 5 x 10(-7) M). The concentration producing 50% inhibition was 10(-7) M for adrenaline and clonidine, 10(-6) M for noradrenaline, and 5 x 10(-5) M for dopamine. Isoproterenol (5 x 10(-8) - 5 x 10(-7) M) inhibited the responses induced by both stimulation of the vagus nerve and acetylcholine. The inhibitory effects of the catecholamine and clonidine were blocked by phentolamine (2.7 x 10(-6) M) but not by 5-(3-tert-Butylamino-2-hydroxy)-propoxy-3, 4-dihydrocarbostyril hydrochloride (OPC 1085) which blocked the effect of isoproterenol. It is suggested that presynaptic alpha-receptors are present in the myenteric plexus of the chick proventriculus, and that the catecholamines and clonidine exert their inhibitory effects on cholinergic transmission via these receptors.

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Graded inhibition of cell disintegration of juvenile hormone.

The proventriculus of Galleria mellonella larvae consists of 260-300 polyploid cells. The cells grow throughout larval development and at each ecdysis produce a new cuticular lining of the proventriculus. Each cell secretes a large sclerotized plaque covered with thorns. At the end of the last larval instar, the cells disintegrate unless the larvae is treated with juvenile hormone or implanted with active corpora allata. Depending on the time of treatment and the hormone dose, these insects preserve their proventricular cells to graded degrees. In some cases, the cells continue to grow or at least preserve their full function, in other instances they secrete but small plaques with fewer thorns or a smooth cuticle without any thorns. Very late treatments preserve healthy cells incapable of secreting the cuticle.

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Distribution of neurotensin-immunoreactive neurons in the digestive tract of the chicken.

The origin of the neurotensin-containing nerve fibers in the digestive tract of the chicken has been investigated with the use of colchicine and immunohistochemistry. Neurotensin-immunoreactive nerve fibers were found in the smooth muscle layers from the esophagus to the duodenum. Their density of distribution was very high in the esophagus and crop (maximum mean value: 1315/mm2 of sectional area in the lamina muscularis mucosae of the crop) and decreased progressively to the duodenum. Neurotensin-immunoreactive neuronal cell bodies were observed after colchicine treatment in the submucosal plexuses of the esophagus and crop and in the myenteric plexuses of the esophagus, crop, proventriculus and gizzard, and they extended varicose fibers. The number of neurotensin-immunoreactive cell bodies was high in the myenteric plexus of the gizzard (28.3 +/- 2.7/ganglion) but low in the plexuses of the esophagus, crop and proventriculus. Seven days after cutting the glossopharyngeal nerve and vagus nerve unilaterally, the number and extent of neurotensin-immunostained structures in the smooth muscle layers from the esophagus to the gizzard did not show any significant difference between operated and unoperated sides. These results indicate that in the chicken the great majority of neurotensin-immunoreactive enteric fibers originate in the intramural plexuses of the upper digestive wall and are mainly distributed to smooth muscle cells.

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Hypergastrinaemia evoked by omeprazole stimulates growth of gastric mucosa but not of pancreas or intestines in hamster, guinea pig and chicken.

Treatment of chickens, hamsters and guinea-pigs with the long-acting anti-secretagogue omeprazole resulted in elevated levels of serum gastrin. The chickens received 400 mumol/kg by i.m. injection once daily, the hamsters and guinea-pigs received the same dose by the oral route once daily. In all 3 species omeprazole raised the intragastric pH to 4, measured 12-14 h after the administration of the drug. After 10 weeks of treatment, trophic changes were observed in the stomach of hamster and guinea pig and in the proventriculus of chicken. The trophic changes were manifested in a greatly increased stomach weight and gastric mucosal mass. There were no trophic effects outside the stomach (or proventriculus). The results are in agreement with previous observations in the rat and support the view that long-lasting sustained hypergastrinaemia causes trophic changes in the stomach but not in the pancreas or in the intestines.

Animals↗

Distribution of immunoreactive vasotocin and mesotocin in the chicken gastrointestinal tract.

Immunoreactive arginine vasotocin (AVT) and mesotocin (MT) were measured in heart, breast muscle, adrenals, testes, and different parts of the gastrointestinal tract in adult male chickens. Neither of the peptides were detected in liver, testis, heart and breast muscle. The amounts of AVT and MT in the adrenals were 167 +/- 25 and 669 +/- 198 pg/gland, respectively. Considerable amounts of immunoreactive peptides were found in the gastrointestinal tract with the highest concentration in the proventriculus (4.18 +/- 0.31 ng AVT and 16.58 +/- 0.86 ng MT per organ). Dose-response curves of duodenal and proventriculus extracts were parallel with synthetic AVT and MT standards.

Animals↗

Can proventricular mesenchyme promote differentiation of endocrine cells in gizzard endoderm?

Previous findings prompted the suggestion that avian proventricular mesenchyme might induce differentiation of endocrine cells with gastrin-releasing peptide (GRP)-like immunoreactivity in endoderm from an organ which, at hatching, is deficient in such cells. Therefore gizzard endoderm and proventricular mesenchyme from chick embryos of 5 days' incubation were combined and grown as chorio-allantoic grafts. Controls comprised re-associated endoderm and mesenchyme of the gizzard and of the proventriculus. In the experimental grafts, as in proventricular controls, immunocytochemistry revealed not only GRP cells, but also the other endocrine cell types characteristic of proventriculus. All these cell types were either absent or very rare in gizzard controls.

Animals↗

Cooperation of JAK/STAT and Notch signaling in the Drosophila foregut.

Temporal and spatial regulation of morphogenesis is pivotal to the formation of organs from simple epithelial tubes. In a genetic screen for novel genes controlling cell movement during posterior foregut development, we have identified and molecularly characterized two alleles of the domeless gene which encodes the Drosophila Janus kinase (JAK)/STAT receptor. We demonstrate that mutants for domeless or any other known component of the canonical JAK/STAT signaling pathway display a failure of coordinated cell movement during the development of the proventriculus, a multiply folded organ which is formed by stereotyped cell rearrangements in the posterior foregut. Whereas the JAK/STAT receptor is expressed in all proventricular precursor cells, expression of upd encoding its ligand and of STAT92E, the signal transducer of the pathway, is locally restricted to cells that invaginate during proventriculus development. We demonstrate by analyzing gene expression mediated by a model Notch response element and by studying the expression of the Notch target gene short stop, which encodes a cytoskeletal crosslinker protein, that JAK/STAT signaling is required for the activation of Notch-dependent gene expression in the foregut. Our results provide strong evidence that JAK/STAT and Notch signaling cooperate in the regulation of target genes that control epithelial morphogenesis in the foregut.

Animals↗

Cleavage of trypanosome surface glycoproteins by alkaline trypsin-like enzyme(s) in the midgut of Glossina morsitans.

EP and GPEET procyclin, the major surface glycoproteins of procyclic forms of Trypanosoma brucei, are truncated by proteases in the midgut of the tsetse fly Glossina morsitans morsitans. We show that soluble extracts from the midguts of teneral flies contain trypsin-like enzymes that cleave the N-terminal domains from living culture-derived parasites. The same extract shows little activity against a variant surface glycoprotein on living bloodstream form T. brucei (MITat 1.2) and none against glutamic acid/alanine-rich protein, a major surface glycoprotein of Trypanosoma congolense insect forms although both these proteins contain potential trypsin cleavage sites. Gel filtration of tsetse midgut extract revealed three peaks of tryptic activity against procyclins. Trypsin alone would be sufficient to account for the cleavage of GPEET at a single arginine residue in the fly. In contrast, the processing of EP at multiple sites would require additional enzymes that might only be induced or activated during feeding or infection. Unexpectedly, the pH optima for both the procyclin cleavage reaction and digestion of the trypsin-specific synthetic substrate Chromozym-TRY were extremely alkaline (pH 10). Direct measurements were made of the pH within different compartments of the tsetse digestive tract. We conclude that the gut pH of teneral flies, from the proventriculus to the hindgut, is alkaline, in contradiction to previous measurements indicating that it was mildly acidic. When tsetse flies were analysed 48 h after their first bloodmeal, a pH gradient from the proventriculus (pH 10.6+/-0.6) to the posterior midgut (pH 7.9+/-0.4) was observed.

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