Gastric and pyloric sphincter muscle function and the developmental-dependent regulation of gastric content emptying in the rat.

Feeding intolerance is a common issue in the care of preterm neonates. The condition manifests as delayed emptying of gastric contents and represents a therapeutic challenge, since the factors accounting for its manifestations are unknown. The main goal of this study was to comparatively investigate the age-related function of rat gastric and pyloric smooth muscle and their putative regulators. We hypothesized that a reduced gastric muscle contraction potential early in life contributes to the delayed gastric emptying of the newborn. Newborn and adult rat gastric (fundus) and pyloric sphincter tissues were comparatively studied in vitro. Shortening of the tissue-specific dissociated smooth muscle cell was evaluated, and expression of the key regulatory proteins Rho-associated kinase 2 and myosin light chain kinase was determined. Gastric and pyloric smooth muscle cell shortening was significantly greater in the adult than the respective newborn counterpart. Expression of myosin light chain kinase and Rho-associated kinase 2 was developmentally regulated and increased with age. Pyloric sphincter muscle expresses a higher neuronal nitric oxide synthase and phosphorylated vasodilator-stimulated phosphoprotein content in newborn than adult tissue. Compared with later in life, the newborn rat gastropyloric muscle has a Ca(2+)-related reduced potential for contraction and the pyloric sphincter relaxation-dependent modulators are overexpressed. To the extent that these rodent data can be extrapolated to humans, the delayed gastric emptying in the newborn reflects reduced stomach muscle contraction potential, as opposed to increased pyloric sphincter tone.

High lever dietary copper promote ghrelin gene expression in the fundic gland of growing pigs.

This experiment was conducted to examine the effect of dietary copper supplementation on ghrelin mRNA expression level in the fundic gland of growing pigs. A total of 45 crossbred pigs were randomly assigned to three groups of 15 pigs, five replicates of three animals comprised each group. Pigs were allocated to diets that contained 5 mg/kg copper (as the control group), 125 mg/kg copper sulfate, or 125 mg/kg copper methionine. At the end of the experiment, five pigs were selected at random from each group, slaughtered, and collected the fundic gland for determination of ghrelin mRNA expression level. The results showed that average daily gain, average daily feed intake, absolute weight, serum growth hormone (GH) concentration, and ghrelin mRNA level were higher in pigs fed the diets with 125 mg/kg copper methionine and 125 mg/kg copper sulfate (P < 0.05), than in pigs fed a diet with 5 mg/kg copper. These data suggest that high dietary copper (125 mg/kg) appears to increase feed intake and promote weight gain by enhancing the secretion of GH and ghrelin mRNA level in growing pigs.

Self-renewal of the human gastric epithelium: new insights from expression profiling using laser microdissection.

The gastric mucosa is subject to continual bidirectional renewal by differentiation from stem and transit amplifying cells. It was the aim of this study to characterize the self-renewal of the human gastric mucosa and its two major types of glands in the fundus and antrum, respectively. Three characteristic regions (pit, proliferative, and lower neck regions) were isolated from fundic and antral units by the use of laser microdissection, and expression profiles concerning 15 marker genes were generated by RT-PCR analysis. The surface mucous cells (SMCs) of fundic and antral units differed in their expression of at least four secretory genes, i.e., gastric lipase, TFF3, FCGBP, and lysozyme. The maturation of mucous neck cells was shown to occur stepwise, first towards a mucous phenotype followed by a serous differentiation step. Also, a stepwise maturation of both the antral SMCs and antral gland cells was observed. Additionally, the presence of gastric lipase was also demonstrated for the first time in antral gland cells. In conclusion, the different expression profiles of SMCs of the fundic and antral units could be the basis for the different self-renewal rates of fundic and antral SMCs and could influence the spatial organization of the bacterial microbiota within the various parts of the gastric mucosa.

Regeneration of gastric mucosa during ulcer healing follows pathways that correspond to the ontogenetic course of rat fundic glands.

Gastric ulcers in humans are notoriously chronic and recurring lesions. Although the average individual who undergoes no treatments requires many years for healing, most studies on the healing process of the experimentally induced ulcers have mainly focused on the early stages. Natural history of the ulcer healing has not been completely revealed. We have undertaken long-term investigation up to the 150th day after the cryo-injury to shed light on the natural history of the ulcer healing process compared with developmental changes of postnatal fundic glands. By the 30th day, restitutive gastric glands were mostly seen to cover the ulcer lesions, where well-developed gland-type mucous cells, showing Griffonia simplicifolia agglutinin (GSA)-II labeling, appeared to occupy the basal portion. Most of the bromodeoxyuridine-labeled cells were superimposed on the GSA-II-positive cell zone, forming the proliferative zone. By the 150th day, the restitutive glands were complete, with all epithelial components and topology of the normal fundic glands. The process of the ulcer healing was quite compatible with the developmental changes of the postnatal fundic glands. These results imply that the regeneration of gastric epithelium during the ulcer healing follows pathways linked to the ontogenetic course of the fundic gland.

The expression of gastric H+-K+-ATPase mRNA and protein in developing rat fundic gland.

The proton pump H+-K+-ATPase is the final common pathway mediating the production and secretion of hydrochloric acid by gastric parietal cells. The present studies were undertaken to examine whether the expression of gastric H+-K+-ATPase mRNA and protein changes are associated with the development of H+-K+-ATPase activity in the rat fundic gland. H+-K+-ATPase activity was examined in rat fundic gland at different stages from gestational day 18.5 to postnatal 8 weeks. The expression of H+-K+-ATPase mRNA was detected by in situ hybridization using a digoxigenin-labelled RNA probe with a tyramide signal amplification system. The expression of H+-K+-ATPase protein was evaluated by immunoblotting and immunohistochemistry using antibodies against H+-K+-ATPase alpha- and beta-subunits. We found that H+-K+-ATPase enzyme activity was detectable from the onset of gland formation (day 19.5 of gestation) and increased with age in the developing rat fundic gland. Expression of mRNA and protein was also discernible at the same time, and a progressive increase in expressions was observed as rats developed. Our results suggested that in developing rat fundic gland, the expression of both mRNA and protein of H+-K+-ATPase increased with age in a manner that parallels the development of H+-K+-ATPase enzyme activity.

Expression of N-acetylglucosamine residues in developing rat fundic gland cells.

The development of rat fundic gland was studied by immunohistochemistry using a recently developed monoclonal antibody, HIK 1083, at both light and electron microscope levels. Antibody HIK 1083 recognized oligosaccharides with a non-reducing terminal alpha-linked N-acetylglucosamine (GlcNAc) residue. In the developing rat fundic gland, cells expressing alpha-GlcNAc residues were discernible from day 19.5 of gestation and continued to exist till adult. The distribution of the alpha-GlcNAc expressing cells was consistent with that described previously for cells reacting to Griffonia simplicifolia lectin (GSA-II) in all developmental stages. These cells were located at the bottom of the fundic gland when they first appeared. With the elongation and maturation of the gland, these cells moved upwards and were finally restricted in the neck region of the gland. Combining previous reports and the present electron microscopical observations, HIK 1083-positive cells in the adult rat fundic gland are mucous neck cells. The interaction between antibody HIK 1083 and GSA-II lectin was investigated. GSA-II prevented the subsequent binding of HIK 1083, while HIK 1083 did not prevent GSA-II binding to mucous neck cells. Our results suggested that alpha-GlcNAc residues exist in rat fundic gland from day 19.5 of gestation and continue to exist till adult. Cells expressing alpha-GlcNAc residues appeared as typical mucous neck cells from postnatal four weeks.

Developmental changes in gastric fundus smooth muscle contractility and involvement of extracellular calcium in fetal and adult guinea pigs.

Delayed gastric emptying is a common problem in preterm infants. The factors underlying this gastroparesis remain unsettled but may involve immaturity of smooth muscle contraction. The present study was designed to test this hypothesis. Muscle strips from the gastric fundus of fetal and adult guinea pigs were studied in vitro for their contractile response to receptor activation (acetylcholine and bethanechol) and membrane depolarization (potassium chloride). The dose-response curves were analyzed for differences in active force development (kg/cm2). The role of extracellular calcium (Ca2+) in the contractile responses was determined by contracting the tissues in a zero-Ca2+ physiologic saline solution and in the presence of nifedipine, a voltage-dependent Ca2+ channel blocker. The results demonstrate the following: 1) tissues from adult animals developed significantly more active force when tested with acetylcholine, bethanechol, and potassium chloride; 2) tissues from the fetal animals were relatively unresponsive to contraction with potassium chloride compared with the adult; and 3) both nifedipine and incubation in a zero-Ca2+ physiologic saline solution had a significantly greater inhibitory effect on the contractions of adult than fetal muscle strips. Our data indicate that smooth muscle in the gastric fundus develops increasing force with maturation. The increased contractility in the adult fundus appears to be due to an increased involvement of extracellular calcium influx, in part through voltage-dependent Ca2+ channels.

Bromodeoxyuridine-immunohistochemistry on cellular differentiation and migration in the fundic gland of Xenopus laevis during development.

Cellular differentiation and migration in the fundic glands of adult and larval Xenopus laevis have been examined using bromodeoxyuridine-immunohistochemistry. In the adult fundic gland, cumulative labeling with bromodeoxyuridine revealed a proliferative cell zone between the surface mucous cells and mucous neck cells, in what is referred to as the neck portion of the gland. The labeling-index of mucous neck cells had rapidly increased by week-5. The labeling-index of oxynticopeptic cells showed a more delayed increase until week-7, coincident with the decrease in the labeling of mucous neck cells. In the immature fundic glands of larvae, the labeled proliferating cells were randomly distributed throughout the developing gastric mucosa. During metamorphosis, the labeling-index of immature epithelial cells was highest at stage 63. Following administration of bromodeoxyuridine at this stage, there was no significant loss of labeled epithelial cells during the metamorphosing period. Furthermore, there was no significant difference in the labeling-indices among the epithelial cells, such as surface mucous cells/generative cells, mucous neck cells, and oxynticopeptic cells, 7 days after administration. Cellular differentiation and migration pathways of epithelial cells in the fundic gland of adult X. laevis and its larvae are discussed.

H(+)-K(+)-ATPase and carbonic anhydrase II gene expression in the developing rat fundus.

H(+)-K(+)-ATPase and carbonic anhydrase II (CA II) are two enzymes that are involved in the production and secretion of the hydrogen ion by the gastric parietal cell and maintenance of intracellular pH therein. The present studies were undertaken to examine whether H(+)-K(+)-ATPase and CA II expression change in the rat fundus in association with the development of acid secretory capacity. Changes in enzyme mRNA content in the gastric fundus of developing rat pups 1-6 wk of age were evaluated using dot blots and ribonuclease protection assays. In additional studies the localization of H(+)-K(+)-ATPase and carbonic anhydrase II mRNA was examined by in situ hybridization in Formalin-fixed gastric tissues from rats 1, 3, 6, and 8 wk of age. We observed that H(+)-K(+)-ATPase mRNA content increased with age in the developing rat fundus while CA II mRNA exhibited a reciprocal decrease. These changes in enzyme mRNA were accompanied by concomitant changes in the regional distribution of the cells expressing the genes for the two enzymes. Although the changes in H(+)-K(+)-ATPase mRNA paralleled the development of acid secretory capacity, CA II mRNA levels might be regulated by the requirement for maintenance of intracellular pH during periods of cellular proliferation and by exposure of the gastric surface epithelium to the highly acidic luminal environment of the stomach.

Inhibitory effect of prolonged administration of long-acting somatostatin on gastrin-stimulated fundic epithelial cell growth in the rat.

The effects of a 3-week administration of long-acting somatostatin were explored (a) in young rats under both normal and long-acting gastrin stimulation and (b) in adult rats with transposition of the antrum onto the colon and, therefore, chronically stimulated with endogenous gastrin. Histomorphometric parameters of the fundic mucosa were estimated at the end of the treatment. In young rats, somatostatin alone (390 micrograms/kg/day) only lowered parietal and peptic cell densities per cubic millimeter compared to controls. That it exerted an antitropic effect under physiological conditions remains questionable. However, in cases of chronic hypergastrinemia, the same dose of somatostatin obviously antagonized the growth-promoting effect of exogenous or endogenous gastrin.