Molecular evolution of a novel marsupial S100 protein (S100A19) which is expressed at specific stages of mammary gland and gut development.

S100 proteins are calcium-binding proteins involved in controlling diverse intracellular and extracellular processes such as cell growth, differentiation, and antimicrobial function. We recently identified a S100-like cDNA from the tammar wallaby (Macropus eugenii) stomach. Phylogentic analysis shows wallaby S100A19 forms a new clade with other marsupial and monotreme S100A19, while this group shows similarity to eutherian S100A7 and S100A15 genes. This is also supported by amino acid and domain comparisons. We show S100A19 is developmentally-regulated in the tammar wallaby gut by demonstrating the gene is expressed in the forestomach of young animals at a time when the diet consists of only milk, but is absent in older animals when the diet is supplemented with herbage. During this transition the forestomach phenotype changes from a gastric stomach into a fermentation sac and intestinal flora changes with diet. We also show that S100A19 is expressed in the mammary gland of the tammar wallaby only during specific stages of lactation; the gene is up-regulated during pregnancy and involution and not expressed during the milk production phase of lactation. Comparison of the tammar wallaby S100A19 protein sequence with S100 protein sequences from eutherian, monotreme and other marsupial species suggest the marsupial S100A19 has two functional EF hand domains, and an extended His tail. An evolutionary analysis of S100 family proteins was carried out to gain a better understanding of the relationship between the S100 family member functions. We propose that S100A19 gene/protein is the ancestor of the eutherian S100A7 gene/protein, which has subsequently modified its original function in eutherians. This modified function may have arisen due to differentiation of evolutionary pressures placed on gut and mammary gland developmental during mammal evolution. The highly regulated differential expression patterns of S100A19 in the tammar wallaby suggests that S100A19 may play a role in gut development, which differs between metatherians and eutherians, and/or include a potential antibacterial role in order to establish the correct flora and protect against spiral bacteria in the immature forestomach. In the mammary gland it may protect the tissue from infection at times of vulnerability during the lactation cycle.

Melanocortinergic activation by melanotan II inhibits feeding and increases uncoupling protein 1 messenger ribonucleic acid in the developing rat.

The hypothalamic neurocircuitry that regulates energy homeostasis in adult rats is not fully developed until the third postnatal week. In particular, fibers from the hypothalamic arcuate nucleus, including both neuropeptide Y (NPY) and alpha-MSH fibers, do not begin to innervate downstream hypothalamic targets until the second postnatal week. However, alpha-MSH fibers from the brainstem and melanocortin receptors are present in the hypothalamus at birth. The present study investigated the melanocortin system in the early postnatal period by examining effects of the melanocortin receptor agonist melanotan II (MTII) on body weight, energy expenditure, and hypothalamic NPY expression. Rat pups were injected ip with MTII (3 mg/kg body weight) or saline on postnatal day (P) 5 to P6, P10-P11, or P15-P16 at 1700 and 0900 h and then killed at 1300 h. Stomach weight and brown adipose tissue uncoupling protein 1 mRNA were determined. In addition, we assessed central c-Fos activation 90 min after MTII administration and hypothalamic NPY mRNA after twice daily MTII administration from P5-P10 or P10-P15. MTII induced hypothalamic c-Fos activation as well as attenuating body weight gain in rat pups. Stomach weight was significantly decreased and uncoupling protein 1 mRNA was increased at all ages, indicating decreased food intake and increased energy expenditure, respectively. However, MTII had no effect on NPY mRNA levels in any hypothalamic region. These findings demonstrate that MTII can inhibit food intake and stimulate energy expenditure before the full development of hypothalamic feeding neurocircuitry. These effects do not appear to be mediated by changes in NPY expression.

Influence of diet type on the inclusion of plant origin active substances on morphological and histochemical characteristics of the stomach and jejunum walls in chicken.

Three hundred and thirty-six 1-day-old male Hubbard HI-Ye broiler hybrids, kept in battery cages, were fed with diets based on maize (groups I and II) or wheat and barley (groups III and IV) and supplemented with or without plant extract (XT* 100 mg/kg) containing 5% carvacrol, 3% cinnamaldehyde and 2% of capsicum oleoresin. The morphological and histochemical examinations were carried out on days 21 and 42 of bird's age. The middle part of glandular part of the stomach and 30 mm long segment from the central part of the small intestine (jejunum) were taken out and then prepared for morphometrical and histochemical assays. Mobilization of mucocytes in superficial epithelium of the glandular stomach and increased secretion of neutral mucopolysaccharides and small amounts of sialomucins with or without local cell disruption with releasing of large amounts of mucus were observed in both 'grain' groups of 21-day-old birds fed with extract. In some animals, particularly those fed mixtures with plant extract, the folds of the proventriculum mucosa were fused into large, unshaped structures. In groups fed with plant extract the mucus secretion intensity and accumulation inside cells of the gastrointestinal mucosa were slightly higher. Morphological changes on gastrointestinal mucosa observed in young chickens fed XT were reduced in older animals. The results of this study showed that the increased releasing of large amounts of mucus and the creation of a thick layer of mucus on glandular stomach and wall of jejunum in chickens fed diets with plant extract could suggest villi-related protective properties of the use of the carvacrol, cinnamaldehyde and capsaicin mixture. This can explain the reduced possibility of adhesion to epithelium and number of Escherichia coli, Clostridium perfringes and fungi in the intestinal content of bird fed with XT supplemented diet. In morphometrical parameters of depth of jejunum crypt and height of villi, the influence of kind of grain and extract supplementation was observed in 21-day-old chickens only. The significant interaction between higher jejunum wall villi layer was observed only in chickens fed on maize diet supplemented with plant extract.

Systemic and luminal influences on the perinatal development of the gut.

At birth, the mammalian gastrointestinal tract (GIT) must be able to support a shift from mainly parenteral nutrition in the fetus (via the placenta) to enteral nutrition in the neonate. In the perinatal period the GIT therefore undergoes enhanced growth as well as morphological and functional differentiation, and this maturational programme is influenced by a complex interplay of local, systemic and luminal factors. This review shows how systemic and luminal factors may influence GIT development in the perinatal period of the pig and sheep, two long-gestation species. Adrenocortical hormones play a pivotal role in the prepartum maturation of the GIT in addition to their better known effects on the development of many other tissues and body systems. More particularly, in the fetal pig and sheep, the prenatal development of gastric acid and gastrin secretion, and of GIT hydrolase activities (chymosin, pepsin, amylase, lactase, aminopeptidases) is influenced by cortisol. Additionally, glucocorticoids exert effects throughout the GIT by influencing morphological, cytological, and functional differentiation. Since the GIT epithelial cells comprise a renewing cell population there are also changes in cell kinetics. In addition to systemic factors, the presence of growth factors, hormones and nutrients from swallowed amniotic fluid (fetus) and colostrum (neonate) may influence GIT development. In utero, fetal fluid ingestion has been shown to modulate tissue growth, macromolecule and immunoglobulin transport, enterocyte differentiation, cell turnover and activity of brush-border hydrolases. These effects may be mediated via regulatory peptides (e.g. insulin-like growth factor I, gastrin-releasing peptides, insulin, epidermal growth factor, gastrin). A physiological role of luminally derived growth factors is supported by a number of unique structural and functional adaptations of the GIT in the fetus and neonate (low luminal proteolysis, intestinal macromolecule transport). Thus, in the pig and sheep, both systemic and luminal factors appear to play critical roles in GIT development in the perinatal period.

Development of the newborn GI tract and its relation to colostrum/milk intake: a review.

During the immediate postnatal period, the gastrointestinal (GI) tract undergoes profound growth, morphological changes and functional maturation. The oesophagus shows an accelerated cell proliferation in the epithelium and an increased production and accumulation of mucus in the glands. The stomach shows a rapid tissue growth and a marked increase in acid secretion capacity. The intestine shows increased tissue growth and marked epithelial modifications; the latter include the loss of the ability by the epithelial cells of the small intestine to absorb macromolecules, and the loss of the ability by the epithelial cells of the large intestine to synthesize digestive enzymes and to absorb amino acids and glucose. These changes are apparently related to the onset of colostrum ingestion, because starved or water-fed newborns showed little changes in the GI tract. A number of hormones and growth-promoting peptides, such as insulin, cortisol, epidermal growth factor (EGF) and insulin-like growth factor I (IGF-I), have been found at high concentrations in the maternal colostrum. There is evidence that colostrum-bone EGF and IGF-I play a role in postnatal GI development in newborns. The role of other colostrum-borne hormones and growth-promoting peptides remains to be assessed. Further studies are also required to demonstrate if colostrum-borne EGF and IGF-I can be used therapeutically to those newborns with immature or diseased GI tract, such as in cases of premature birth or prenatal growth retardation or cases requiring total parenteral nutrition.

Effects of Yersinia enterocolitica infection on growth of the body and internal organs in newborn colostrum-deprived piglets.

Yersinia enterocolitica enteritis in newborn, colostrum-deprived piglets fed a human milk formula caused a reduced milk intake and decreased gain in weight but not length of the body. In infected piglets, the weight of the liver was less than in controls but other abdominal organs were unaffected. The weight of the caecum and small intestinal muscle were greater, but the length and weight of the small intestine and colon were not different. In infected piglets, the RNA concentration of the small intestinal mucosa was elevated whereas the concentrations of DNA and protein were unchanged, and total lactase activity was reduced. After antibiotic therapy, the liver weight was greater and the body weight increased at the same rate as the controls but was still lower at 14 days. The body weight of the control piglets increased linearly over the 14 days after birth. The stomach, pancreas and spleen grew more rapidly, but the liver more slowly, than the body as a whole but the kidneys had a minimum relative weight at 5 days. The total lactase activity and protein and RNA concentrations of the small intestinal mucosa decreased with age, the RNA to a greater extent than the protein.

Effects of oral IGF-I or IGF-II on digestive organ growth in newborn piglets.

To study whether colostrum-borne growth factors are responsible for the rapid GI tissue growth in naturally suckled newborn animals, newborn unsuckled piglets were bottle-fed for 24 h with infant milk formula with or without addition of 2 micrograms/ml of recombinant human insulin-like growth factor-I (IGF-I) or insulin-like growth factor-II (IGF-II), a level which approximated that of porcine colostrum. The animals were then sacrificed for measurements of their digestive organ weights and contents of protein, RNA and DNA in the organs. The treatment with IGF-I or IGF-II failed to show any significant effect on the weight of the esophagus, stomach, small intestine, large intestine, mandibular glands, kidneys and the spleen, and had no effects on the contents of protein, RNA and DNA in the small intestinal mucosa, the liver and the spleen. However, piglets fed with infant formula containing IGF-I (n = 7) or IGF-II (n = 7) had a heavier pancreas (p < 0.05) compared to formula-fed controls (n = 7). The DNA content in the stomach and the pancreas were greater in animals treated with IGF-I or IGF-II than in controls. Using a cell labelling technique it was shown that both IGF-I and IGF-II stimulated cell proliferation in the small intestinal crypts. The results indicate that the substantial GI tissue growth reported in newborn animals is unlikely due to colostrum-borne IGF-I or IGF-II alone. On the other hand the study does suggest that oral IGF-I and IGF-II are capable of stimulating cell proliferation in the GI tract.

Postprandial lipid response following a high fat meal in rats adapted to dietary fiber.

Rats were adapted to diets containing 5 g/100 g cellulose (CL), 5 g/100 g oat bran fiber (OB) or 5 g/100 g psyllium husk (Psy) for 4 wk. Following a 12-h fast, animals were either killed at 0 h (baseline) or fed 4.5 g of a test meal that provided 50% energy from fat, then killed at 1, 4 or 6 h postprandially. Fasting plasma and HDL cholesterol concentrations were lower in Psy-fed animals than in rats fed either CL or OB. Plasma triglycerides increased significantly from baseline (0 h) in all groups but did not differ among diet treatments. Increases in triglyceride content of the treatments. Increases in triglyceride content of the chylomicron/VLDL fraction occurred in the CL- and OB-fed groups and in the HDL fraction of the Psy-fed group during the postprandial period. In unfed animals the hepatic and intestinal levels of apolipoprotein A-IV mRNA were higher in the CL-fed group than in the groups fed OB and Psy. Apolipoprotein B mRNA was higher in the intestine of the OB-fed group than in the groups fed CL and Psy and had a significant gradient along the small intestine, increasing in the distal third. The results suggest that chronic consumption of fiber is less likely to modify the acute plams triglyceride response to a fat-containing test meal than if a fiber supplement is incorporated into the meal.

Stimulation by colostrum or mature milk of gastrointestinal tissue development in newborn pigs.

Porcine colostrum and milk were orally administered to newborn pigs to evaluate their gastrointestinal growth-promoting activity. Five piglets per treatment group were gavage-fed 5% lactose (L), defatted colostrum (C) or defatted mature milk (d 16 of lactation) (M) at 3-h intervals over a 24-h period. Colostrum and milk were administered on equivalent dry matter basis and all piglets received 1 mCi of [3H]thymidine per kg BW at the onset of feeding. Small intestines of C- and M-fed pigs were 1.6-and 1.3-fold, respectively, the weight of small intestines of those fed L (P less than .01). Total DNA content of intact small intestines were not different among groups; however, cpm [3H]thymidine/mg intestinal DNA of C and M piglets exceeded (P less than .05) that for L piglets. DNA content and cpm [3H]thymidine of intestinal mucosa did not differ among groups. Total protein in the intestines and intestinal mucosa of C pigs exceeded (P less than .01) that for L and M pigs. Total RNA in the small intestine and intestinal mucosa were similar for C and M groups but less (P less than .01) for L piglets. Stomach and pancreas weights among all pigs were similar, although C and M pigs exceeded L pigs in stomach (P less than .01) and pancreas (P less than .01) RNA content. In contrast, no differences in stomach DNA, protein and cpm [3H]thymidine or in pancreatic DNA, protein and cpm [3H]thymidine were detected. SDS-polyacrylamide gel electrophoresis was used to identify qualitative and quantitative differences in the protein compositions of porcine colostrum and mature milk.(ABSTRACT TRUNCATED AT 250 WORDS)

Tissue-specific sex differences in galanin-like immunoreactivity and galanin mRNA during development in the rat.

Galanin-like immunoreactivity (Gal-LI), as determined by radioimmunoassay, was detectable in the brain and gastrointestinal tract by day 15 of gestation. Concentrations of Gal-LI increased after birth in the hypothalamus but decreased in the stomach and duodenum. A sex difference in Gal-LI concentrations appeared during puberty in the median eminence, neurointermediate lobe, and the anterior pituitary (AP), where females had higher Gal-LI concentrations compared to males. This difference was most pronounced in the AP; adult females had up to 4-fold greater Gal-LI concentrations and 5-fold more abundant rGal-specific mRNA compared to males.

Comparison of the gastrointestinal growth- promoting effects of rat colostrum and mature milk in newborn rats in vivo.

The purpose of this study was to compare the effects of rat colostrum and mature milk on newborn rat gastrointestinal growth under conditions that controlled the possible confounding effects of energy intake and mode of feeding. Newborn Sprague-Dawley rat pups were tubefed equicaloric amounts of rat colostrum or mature rat milk for 40 h before they were killed. Compared to littermates that were killed immediately after birth, both groups of fed rats demonstrated increases in the weights of stomach and intestine, but there was no organ weight difference detected between colostrum-fed and mature milk-fed rat pups. However, both the concentration of DNA and the rate of synthesis of DNA in the intestines were greater in rats fed colostrum than in those at birth or those mature milk. Although the pancreas exhibited no detectable increase in weight by 41 h, the DNA concentration and total DNA content increased and RNA/DNA ratio decreased in both fed groups, also without apparent difference between rats fed colostrum and those fed mature milk. The rate of 3H-thymidine incorporation into DNA in pancreas, however, was greater in colostrum-fed pups than in mature milk-fed pups. These differences at 40 h age in intestinal and pancreatic cell replication activity, but not organ weights, can be ascribed to feeding colostrum.