Glucagon-like peptide 2 has limited efficacy to increase nutrient absorption in fetal and preterm pigs.

Exogenous glucagon-like peptide 2 (GLP-2) prevents intestinal atrophy and increases nutrient absorption in term newborn pigs receiving total parenteral nutrition (TPN). We tested the hypothesis that the immature intestine of fetuses and preterm neonates has a diminished nutrient absorption response to exogenous GLP-2. This was accomplished using catheterized fetal pigs infused for 6 days (87-91% of gestation) with GLP-2 (25 nmol.kg(-1).day(-1) iv; n = 7) or saline (n = 7), and cesarean-delivered preterm pigs (92% of gestation) that received TPN with GLP-2 (25 nmol.kg(-1).day(-1) iv; n = 8) or saline (n = 7) for 6 days after birth. Responses to GLP-2 were assessed by measuring intestinal dimensions, absorption of nutrients (glucose, leucine, lysine, proline) by intact tissues and brush border membrane vesicles, and abundance of sodium-glucose cotransporter mRNA. Infusion of GLP-2 increased circulating GLP-2 levels in fetuses, but did not increase intestinal mass or absorption of nutrients by intact tissues and brush border membrane vesicles, except for lysine. Administration of exogenous GLP-2 to preterm TPN-fed pigs similarly did not increase rates of nutrient absorption, yet nutrient absorption capacities of the entire small intestine tended to increase (+10-20%, P < 0.10) compared with TPN alone due to increased intestinal mass (+30%, P < 0.05). GLP-2 infusion did not increase sodium-glucose cotransporter-1 mRNA abundance in fetuses or postnatal preterm pigs. Hence, the efficacy of exogenous GLP-2 to improve nutrient absorption by the intestine of fetal and preterm pigs is limited compared with term pigs and more mature animals and humans.

Identification of microsomal triglyceride transfer protein in intestinal brush-border membrane.

Microsomal triglyceride transfer protein (MTP) is a heterodimeric complex consisting of a unique large 97-kDa protein and the multifunctional 58-kDa protein disulfide isomerase (PDI). It plays an essential role in the assembly of lipoproteins by shuttling lipids between phospholipid membranes. Based on cell fractionation, early studies have suggested the endoplasmic reticulum (ER) as the exclusive site of MTP. Focusing on the plasma membrane in this study, our attempts with immunoelectron microscopy and specific antibodies surprisingly revealed that labeling was not exclusively confined to the microsomes of rat absorptive cells. Immunogold labeling was also detected over the microvillus membrane of enterocytes. Western blot analysis and biochemical activity measurement confirmed MTP protein expression in brush-border membrane vesicles (BBMV) isolated from the intestinal epithelial cells of various species. Furthermore, MTP was coexpressed in microvilli membrane with PDI that is crucial to maintain the structure and activity of the MTP complex. The treatment of Caco-2 cells with nocodazole and colchicine blocked the appearance of MTP in the apical membrane. Similarly, the addition of BMS-197636, a known inhibitor of MTP transfer activity, suppressed the latter. In conclusion, the present studies suggest that MTP is present in the brush-border membrane of the enterocyte. Understanding the possible physiological role of MTP in this location may reveal additional functions.

Activities of gastric, pancreatic, and intestinal brush-border membrane enzymes during postnatal development of dogs.

OBJECTIVE: To measure activities of digestive enzymes during postnatal development in dogs. SAMPLE POPULATION: Gastrointestinal tract tissues obtained from 110 Beagles ranging from neonatal to adult dogs. PROCEDURE: Pepsin and lipase activities were measured in gastric contents, and amylase, lipase, trypsin, and chymotrypsin activities were measured in small intestinal contents and pancreatic tissue. Activities of lactase, sucrase, 4 peptidases, and enteropeptidase were assayed in samples of mucosa obtained from 3 regions of the small intestine. RESULTS: Gastric pH was low at all ages. Pepsin was not detected until day 21, and activity increased between day 63 and adulthood. Activities of amylase and lipase in contents of the small intestine and pancreatic tissue were lower during suckling than after weaning. Activities of trypsin and chymotrypsin did not vary among ages for luminal contents, whereas activities associated with pancreatic tissue decreased between birth and adulthood for trypsin but increased for chymotrypsin. Lactase and gamma-glutamyltranspeptidase activities were highest at birth, whereas the activities of sucrase and the 4 peptidases increased after birth. Enteropeptidase was detected only in the proximal region of the small intestine at all ages. CONCLUSIONS AND CLINICAL RELEVANCE: Secretions in the gastrointestinal tract proximal to the duodenum, enzymes in milk, and other digestive mechanisms compensate for low luminal activities of pancreatic enzymes during the perinatal period. Postnatal changes in digestive secretions influence nutrient availability, concentrations of signaling molecules, and activity of antimicrobial compounds that inhibit pathogens. Matching sources of nutrients to digestive abilities will improve the health of dogs during development.

Postnatal development of nutrient transport in the intestine of dogs.

OBJECTIVE: To measure nutrient absorption by the intestine during postnatal development of dogs. ANIMAL: 110 Beagles ranging from neonatal to adult dogs. PROCEDURE: Rates of absorption for sugars (glucose, galactose, and fructose), amino acids (aspartate, leucine, lysine, methionine, and proline), a dipeptide (glycyl-sarcosine), and linoleic acid by the proximal, mid, and distal regions of the small intestine were measured as functions of age and concentration (kinetics) by use of intact tissues and brush-border membrane vesicles. Absorption of octanoic acid by the proximal portion of the colon was measured in intact tissues. RESULTS: Rates of carrier-mediated transport by intact tissues decreased from birth to adulthood for aldohexoses and most amino acids but not for fructose and aspartate. Kinetics and characteristics of absorption suggest that there were changes in the densities, types, and proportions of various carriers for sugars and amino acids. Saturable absorption of linoleic acid in the small intestine and octanoic acid in the proximal portion of the colon increased after weaning. CONCLUSIONS AND CLINICAL RELEVANCE: Rates of absorption decreased between birth and adulthood for most nutrients. However, because of intestinal growth, absorption capacities of the entire small intestine remained constant for leucine and proline and increased for glucose, galactose, fructose, aspartate, and proline but were less than predicted from the increase in body weight. Although postnatal ontogeny of nutrient absorption was consistent with changes in the composition of the natural and commercial diets of growing dogs, rates of amino acid and peptide absorption were lower than expected.

L-Carnitine transport in mouse renal and intestinal brush-border and basolateral membrane vesicles.

We characterized the uptake of carnitine in brush-border membrane (BBM) and basolateral membrane (BLM) vesicles, isolated from mouse kidney and intestine. In kidney, carnitine uptake was Na(+)-dependent, showed a definite overshoot and was saturable for both membranes, but for intestine, it was Na(+)-dependent only in BLM. The uptake was temperature-dependent in BLM of both kidney and intestine. The BBM transporter in kidney had a high affinity for carnitine: apparent K(m)=18.7 microM; V(max)=7.85 pmol/mg protein/s. In kidney BLM, similar characteristics were obtained: apparent K(m)=11.5 microM and V(max)=3.76 pmol/mg protein/s. The carnitine uptake by both membranes was not affected within the physiological pH 6.5-8.5. Tetraethylammonium, verapamil, valproate and pyrilamine significantly inhibited the carnitine uptake by BBM but not by BLM. By Western blot analysis, the OCTN2 (a Na(+)-dependent high-affinity carnitine transporter) was localized in the kidney BBM, and not in BLM. Strong OCTN2 expression was observed in kidney and skeletal muscle, with no expression in intestine in accordance with our functional study. We conclude that different polarized carnitine transporters exist in kidney BBM and BLM. L-Carnitine uptake by mouse renal BBM vesicles involves a carrier-mediated system that is Na(+)-dependent and is inhibited significantly by specific drugs. The BBM transporter is likely to be OCTN2 as indicated by a strong reactivity with the anti-OCTN2 polyclonal antibody.