Nutrient Transporter Gene Expression Research Articles

Effect of dietary lead on intestinal nutrient transporters mRNA expression in broiler chickens.

Lead- (Pb-) induced oxidative stress is known to suppress growth performance and feed efficiency in broiler chickens. In an attempt to describe the specific underlying mechanisms of such phenomenon we carried out the current study. Ninety-six one-day-old broiler chicks were randomly assigned to 2 dietary treatment groups of 6 pen replicates, namely, (i) basal diet containing no lead supplement (control) and (ii) basal diet containing 200 mg lead acetate/kg of diet. Following 3 weeks of experimental period, jejunum samples were collected to examine the changes in gene expression of several nutrient transporters, antioxidant enzymes, and heat shock protein 70 (Hsp70) using quantitative real-time PCR. The results showed that addition of lead significantly decreased feed intake, body weight gain, and feed efficiency. Moreover, with the exception of GLUT5, the expression of all sugar, peptide, and amino acid transporters was significantly downregulated in the birds under Pb induced oxidative stress. Exposure to Pb also upregulated the antioxidant enzymes gene expression together with the downregulation of glutathione S-transferase and Hsp70. In conclusion, it appears that Pb-induced oxidative stress adversely suppresses feed efficiency and growth performance in chicken and the possible underlying mechanism for such phenomenon is downregulation of major nutrient transporter genes in small intestine.

Improvements in growth performance, bone mineral status and nutrient digestibility in pigs following the dietary inclusion of phytase are accompanied by modifications in intestinal nutrient transporter gene expression.

Phytase (PHY) improves growth performance, nutrient digestibility and bone structure in pigs; however, little is known about its effects on intestinal nutrient transporter gene expression. In the present study, a 44 d experiment was carried out using forty-eight pigs (11·76 (sem 0·75) kg) assigned to one of three dietary treatment groups to measure growth performance, coefficient of apparent ileal digestibility (CAID), coefficient of apparent total tract nutrient digestibility (CATTD) and intestinal nutrient transporter gene expression. Dietary treatments during the experimental period were as follows: (1) a high-P (HP) diet containing 3·4 g/kg available P and 7·0 g/kg Ca; (2) a low-P (LP) diet containing 1·9 g/kg available P and 5·9 g/kg Ca; (3) a PHY diet containing LP diet ingredients+1000 phytase units (FTU)/kg of PHY. The PHY diet increased the average daily gain (P< 0·05) and final body weight (P< 0·01) and decreased the feed conversion ratio (P< 0·05) compared with the LP diet. Pigs fed the PHY diet had a higher CAID of gross energy compared with those fed the HP and LP diets (P< 0·001). Pigs fed the PHY diet had increased CAID of P (P< 0·01) and CATTD of Ca and P (P< 0·001) compared with those fed the LP diet. The PHY diet increased the gene expression of the peptide transporter 1 (PEPT1/SLC15A1) (P< 0·05) in the ileum compared with the LP diet. The LP diet decreased the gene expression of the sodium-glucose-linked transporter 1 (SGLT1/SLC5A1) and GLUT2/SLC2A2 (P< 0·05) and increased the expression of membrane Ca channel (TRPV6) and calbindin compared with the HP diet (P< 0·001). In conclusion, feeding a diet supplemented with PHY improves growth performance and nutrient digestibility as well as increases the gene expression of the peptide transporter PEPT1.

Effects of maternal over- and undernutrition on intestinal morphology, enzyme activity, and gene expression of nutrient transporters in newborn and weaned pigs

ObjectivesIt has been suggested that maternal nutrition during gestation is involved in the offspring's intestinal development. The aim of this study was to evaluate the effects of maternal nutrition on digestive and absorptive function of offspring at both birth and weaning with pig as model. MethodsEighteen primiparous Landrace × Yorkshire gilts (ages 253 ± 9 d; body weight 148.6 ± 6.9 kg) were allocated to a normal nutrition (control) group as recommended by the National Research Council (NRC; 1998), an undernutrition (UN) group (75% of NRC recommendations), and an overnutrition (ON) group (150% of NRC recommendations); the nutritional treatments were introduced from day 1 of gestation to farrowing. ResultsBy comparison, the small intestine weight (+33%; P < 0.01) and the ratio of small intestine weight to length (+25%; P < 0.05) for the ON group were higher than those for the control group. Meanwhile, the maternal ON treatment had significant effect on the activity of brush border lactase in the jejunum (+125%; P < 0.05) at birth. Expression of mRNA for SGLT1, GLUT2, PEPT1, and GLP2R were significantly increased (+1.7∼9.6-fold; P < 0.05) in jejunum of newborn or weaned piglets from the ON group; whereas maternal UN significantly affected small intestine weight (–28%; P < 0.01), length, and the ratio of weight to length (–11% and –19%; P < 0.05, respectively), and villus height (–29% and –22%; P < 0.05, respectively) in jejunum and ileum. Furthermore, maternal UN enhanced the activity of brush border lactase in the jejunum of newborn piglets (P < 0.05). ConclusionsMaternal ON enhanced intestinal function via up-regulating digestive enzyme activities and gene expression of transporters in both newborn and weaning piglets. Maternal UN impaired fetal intestinal development that could be partially compensated during the neonatal period.

Expression of digestive enzymes and nutrient transporters in Eimeria acervulina-challenged layers and broilers

Avian coccidiosis is a disease caused by intestinal protozoa in the genus Eimeria. Clinical signs of coccidiosis include intestinal lesions and reduced feed efficiency and BW gain. This growth reduction may be due to changes in expression of digestive enzymes and nutrient transporters in the intestine. The objective of this study was to examine the differential expression of digestive enzymes, transporters of amino acids, peptides, sugars, and minerals, and an antimicrobial peptide in the small intestine of Eimeria acervulina-infected broilers and layers. Uninfected broilers and layers, in general, expressed these genes at comparable levels. Some differences included 3-fold and 2-fold greater expression of the peptide transporter PepT1 and the antimicrobial peptide LEAP2 (liver expressed antimicrobial peptide 2), respectively, in the jejunum of layers compared with broilers and 17-fold greater expression of LEAP2 in the duodenum of broilers compared with layers. In the duodenum of Eimeria-infected broilers and layers, there was downregulation of aminopeptidase N; sucrase-isomaltase; the neutral, cationic, and anionic amino acid transporters bo,+ AT/rBAT, Bo AT, CAT2, and EAAT3; the sugar transporter GLUT2; the zinc transporter ZnT1; and LEAP2. In the jejunum of infected layers there was downregulation of many of the same genes as in the duodenum plus downregulation of PepT1, bo,+ AT/rBAT, and the y+ L system amino acid transporters y+ LAT1 and y+ LAT2. In the ileum of infected layers there was downregulation of CAT2, y+ LAT1, the L type amino acid transporter LAT1, and the sugar transporter GLUT1, and upregulation of APN, PepT1, the sodium glucose transporter SGLT4, and LEAP2. In E. acervulina-infected broilers, there were no gene expression changes in the jejunum and ileum. These changes in intestinal digestive enzyme and nutrient transporter gene expression may result in a decrease in the efficiency of protein digestion, uptake of important amino acids and sugars, and disruption of mineral balance that may affect intestinal cell metabolism and Eimeria replication.

Delayed Post Hatch Feeding Affects Performance, Intestinal Morphology and Expression Pattern of Nutrient Transporter Genes in Egg Type Chickens

Effect of post-hatch (PH) feed deprivation (FD) for 6, 12, 24 and 36 hrs., on the performance, gut development and differential expression of nutrient transporter genes in egg type chickens (crosses of White Leghorn) was studied. Significant decrease (P=0.001) in yolk utilization (in all the FD chicks) and relative weight of gizzard, intestine, liver and pancreas was observed in 24 and 36 hrs FD chicks. The 36 hrs FD chicks recorded lower (P=0.001) body weight, lower feed intake and inferior FCR, decreased serum glucose, but higher cholesterol and uric acid level than the control (immediate fed group) or 6 hrs FD chicks. Villi height in duodenum, jejunum and ileum decreased (P=0.001) but villi width increased (P=0.001) with increase in FD period and significant changes were observed particularly in 24 or 36 hrs FD chicks. Relative expression of Cdx gene decreased with age of the bird and the feed restriction period. Expression of SGLT, FABP gene was not associated with the feed deprivation period, while that of EAAT3 gene increased in 24 or 36 hrs FD chicks. No difference was observed in bursa, spleen and thymus weight. In vivo humoral and cellular immune response was significantly better in chicks FD for 6, 12 and 24 hrs than control and 36 hrs FD chicks. Expression of immunity related genes IL-6 and TLR-2 increased as the FD period increased. It may concluded that PH feed deprival for first 12 hrs did not affect growth performance, intestinal morphology and immune response but feed withdrawal for 24 or 36 hrs adversely affect the intestinal morphology and few nutrient transporter genes expression in egg type chickens.

Maternal protein restriction during pregnancy affects gene expression and immunolocalization of intestinal nutrient transporters in rats

Intrauterine dietary restriction may cause changes in the functioning of offspring organs and systems later in life, an effect known as fetal programming. The present study evaluated mRNA abundance and immunolocalization of nutrient transporters as well as enterocytes proliferation in the proximal, median and distal segments of small intestine of rats born to protein-restricted dams. Pregnant rats were fed hypoproteic (6% protein) or control (17% protein) diets, and offspring rats were evaluated at 3 and 16 weeks of age. The presence of SGLT1 (sodium-glucose co-transporter 1), GLUT2 (glucose transporter 2), PEPT1 (peptide transporter 1) and the intestinal proliferation were evaluated by immunohistochemical techniques and the abundance of specific mRNA for SGLT1, GLUT2 and PEPT1 was assessed by the real-time PCR technique. Rats born to protein-restricted dams showed higher cell proliferation in all intestinal segments and higher gene expression of SGLT1 and PEPT1 in the duodenum. Moreover, in adult animals born to protein-restricted dams the immunoreactivity of SGLT1, GLUT2 and PEPT1 in the duodenum was more intense than in control rats. Taken together, the results indicate that changes in the small intestine observed in adulthood can be programmed during the gestation. In addition, they show that this response is caused by both up-regulation in transporter gene expression, a specific adaptation mechanism, and intestinal proliferation, an unspecific adaptation mechanism.

Maternal Administration of the Herbal Medicine Toki-Shakuyaku-San Promotes Fetal Growth and Placental Gene Expression in Normal Mice

Toki-shakuyaku-san (TSS), an herbal formula based on traditional Chinese medicine, is commonly used in obstetrics. To examine the effects of TSS on the normal mouse fetus and placenta, TSS was administered to normal pregnant mice and their placentas and fetuses were studied. First, the effects of maternal TSS treatment on implantation were investigated. Administration of TSS from gestation day 0.5 (G0.5) to G6.5 showed that litter size was not altered at embryonic day 11.5 (E11.5), but the number of resorbed fetuses was slightly decreased. Then, to investigate effects on fetal and placental growths after implantation, TSS was administered from G5.5. At E14.5, the body weight of fetuses from TSS-treated dams was significantly increased. Gene expression of insulin-like growth factor 2 (Igf2), one of the most important modulators of fetal growth, was significantly increased in the placentas and fetuses of TSS-treated dams. In addition, the expression of particular placental developmental genes and nutrient transporter genes was significantly increased in TSS-treated placentas. At E18.5, after longer-term administration of TSS, fetal and placental weights were not altered, but the expression of the placental developmental and nutrient transporter genes remained elevated compared with controls. These results suggest that maternal TSS treatment in normal mice enhances the expression of Igf2, placental developmental genes and nutrient transporter genes, resulting in increased fetal weight. No obvious changes were observed in the expression of these genes after longer-term maternal TSS treatment.

Gene expression of nutrient transporters and digestive enzymes in the yolk sac membrane and small intestine of the developing embryonic chick

Chick embryos derive nutrients from the yolk during incubation and transition to intestinal absorption of nutrients posthatch. The uptake of nutrients is mediated by a variety of membrane-bound transporter proteins. The objective of this study was to determine the expression profiles of nutrient transporters and digestive enzymes during incubation in the yolk sac membrane (YSM) and embryonic intestine of egg-laying (Leghorn) and meat-producing (Cobb) chickens derived from 22 to 30 wk (young) and 45 to 50 wk (old) breeder flocks. Transporters examined included the peptide transporter PepT1, the glutamate/aspartate (EAAT3), cationic (CAT-1) and neutral (B0AT) amino acid transporters, and the fructose (GLUT5) and glucose (SGLT1) transporters. Digestive enzymes included aminopeptidase N (APN) and sucrase-isomaltase (SI). Expression of these genes was assessed by real-time PCR using the absolute quantification method in YSM at embryonic day (E) 11, 13, 15, 17, 19, 20, and 21 and intestine at E15, 17, 19, 20, and 21. The PepT1 and APN gene expression in the YSM increased until E15 and then decreased until E21, whereas expression in the intestine increased from E15 to E21. The B0AT showed a similar pattern, with greatest expression in the YSM occurring at E17/E19. The CAT1 and GLUT5 genes showed decreased expression in the YSM and increased expression in the intestine until E17/E19 and then a decrease until E21. Expression of SGLT1 and EAAT3 showed increased gene expression over time in both the intestine and YSM. Expression of SI showed little to no gene expression in the YSM, whereas the intestine exhibited consistently high levels of gene expression. In YSM and intestine, SI expression was greater in Leghorn than Cobb, whereas CAT1 and GLUT5 expression was greater in Cobb than Leghorn. Expression of the APN, CAT1, and SI genes was greater in embryos from young flocks than old flocks in YSM and intestine. These results demonstrate that the YSM expresses many of the digestive enzymes and nutrient transporters typically associated with the intestine and that these genes show tissue- and development-specific patterns of expression.

The ontogeny of nutrient transporter and digestive enzyme gene expression in domestic pigeon (Columba livia) intestine and yolk sac membrane during pre- and posthatch development

To better understand the digestive capacity in domestic pigeons (Columba livia), this study was conducted to evaluate nutrient transporters and digestive enzymes gene expression in small intestine and yolk sac membrane (YSM) during pre- and posthatch development. We investigated the oligopeptide transporter Pept1, sodium glucose transporter SGLT1, glucose transporter GLUT2, aminopeptidase-N (APN), and sucrase-isomaltase (SI). Intestine was collected at embryo d 12, 14, and 16, day of hatch, and d 1, 3, 5, 8, and 14 posthatch. The YSM was collected at embryo d 12, 14, 16, and day of hatch. The cDNA fragments for Pept1, SGLT1, GLUT2, APN, and SI were isolated and cloned using reverse-transcription PCR. The sequences data showed that these genes were highly identical to the gene of chicken. The mRNA expression of each gene was assayed using real-time PCR. Expression of intestinal nutrient transporters increased linearly (P<0.001) with age whereas that of APN increased quadratically (P<0.001) and SI increased cubically (P=0.001). Levels of PepT1 mRNA were greatest in the duodenum (P=0.001), GLUT2 and SGLT1 mRNA were greatest in the jejunum (P=0.002) and APN were greatest in the ileum (P=0.004). The YSM expressed all the examined genes. The YSM-expressed genes decreased between embryo d 16 and day of hatch, whereas intestine-expressed genes increased. Our findings provide a comprehensive profile of gene expression patterns of nutrient transporter and digestive enzyme in the small intestine and YSM of pigeons and establish a foundation for future research on the nutrients requirements for young pigeons.

Messenger ribonucleic acid abundance of intestinal enzymes and transporters in feed-restricted and refed chickens at different ages

The effects of feed restriction and subsequent refeeding on the gene expression of intestinal enzymes and nutrient transporters at 2 ages, 7 and 35 d, were examined in different groups of broiler chickens. At each age, birds were feed restricted for 7 d (30% of ad libitum intake) followed by 3 d of refeeding ad libitum. Control groups were fed ad libitum. Total RNA of jejunal mucosa was extracted according to the Trizol protocol, and mRNA expression of sodium glucose transporter 1, glucose transporter 2, peptide transporter 1, aminopeptidase, maltase, and sucrase-isomaltase complex was obtained by reverse-transcription PCR. The expression of aminopeptidase, sodium glucose transporter 1, and peptide transporter 1 was higher in feed-restricted groups than in control groups at d 14 (181.4, 116.7, and 80.4%, respectively) and d 42 (143.5, 84.2, and 195.9%, respectively). The mRNA abundance of sucrase-isomaltase complex was higher (159.1%) only in chickens that were feed restricted from d 35 to 42. No statistically significant effect of feed restriction was observed for mRNA abundance of maltase and glucose transporter 2 at either age. After refeeding (d 17 and 45), the RNA abundance of enzymes and nutrient transporters was similar to that in the control group. Thus, this study suggests that an effect of upregulation in gene expression exists during feed restriction that disappears when feed is supplied ad libitum.

Effect of incubation temperature on nutrient transporters and small intestine morphology of broiler chickens

This study evaluated the effects of elevated incubation temperature on posthatch nutrient transporter gene expression, integrity of the intestinal epithelium, organ development, and performance in Ross 308 broiler chickens. Ross × Ross 308 fertile eggs (n = 900) were incubated at different eggshell temperatures during development. From embryonic day (ED) 1 to ED12, all eggs were incubated at 37.1°C, whereas from ED13 to ED21, the eggs were divided into 2 groups for incubation at 37.4°C (S) or 39.6°C (H). Performance characteristics were measured at day of hatch (DOH) and d 7, 14, 21, 30, and 42. Small intestine and residual yolk sacs were collected at DOH and d 2, 4, 6, and 10 and weighed individually. Intestinal samples from the duodenum, jejunum, and ileum were evaluated for mucosal morphology and relative nutrient transporter gene expression. No significant differences were found in performance or organ weights. The intestinal morphology results showed a temperature × age interaction in duodenum villus height (P = 0.02) and crypt depth (P = 0.05) and in ileum villus height-to-crypt depth ratios (P = 0.02). There was a main effect of temperature, resulting in deeper crypts (P = 0.02) in the jejunum of chicks incubated at H compared with S. In the nutrient gene expression evaluation, peptide transporter (PepT1) showed a temperature × age interaction. On DOH and d 2, 4, and 10, PepT1 expression was similar between chicks incubated at S and H. However, on d 6, chicks incubated at S had significantly higher expression of PepT1 than those incubated at H. This study presents the effects of elevated incubation temperature on small intestine morphology and relative expression of nutrient transporter mRNA in high-yield broiler chicks, which can be important for the availability of nutrients and distribution of energy.

Gene Expression of Nutrient Transporters in the Small Intestine of Chickens from Lines Divergently Selected for High or Low Juvenile Body Weight

Nutrient transporters in the small intestine are responsible for dietary nutrient assimilation; therefore, the expression of these transporters can influence overall nutrient status as well as the growth and development of the animal. This study examined correlated responses to selection in the developmental gene expression of PepT1, EAAT3, SGLT1, and GLUT5 in the small intestine of chickens from lines divergently selected for 48 generations for high (HH) or low (LL) 56-d BW and their reciprocal crosses (HL and LH). Duodenum, jejunum, and ileum were collected from male and female chicks on embryonic d 20, day of hatch with no access to feed, and d 3, 7, and 14 posthatch. Total RNA was extracted, and nutrient transporter expression was assayed by real-time PCR using the relative quantification method. In comparing male and female HH and LL chicks, there was a mating combination × age × sex interaction for PepT1 expression (P < 0.001), a main effect of sex for EAAT3 (P < 0.05) and SGLT1 (P < 0.001) expression, and an age × sex interaction for SGLT1 expression (P < 0.001). These results demonstrate a sexual dimorphism in the capacity to absorb nutrients from the intestine, which has implications for the poultry industry with regard to diet formulations for straight-run and sex-separate grow-out operations. Results from comparing male LL, LH, HL, and HH chicks indicate that selection for high or low juvenile BW may have influenced the gene expression profiles of these nutrient transporters in the small intestine, which may contribute to the overall differences in the growth and development of these lines of chickens.