Skip to main content Accessibility help

The in ovo administration of l-trans pyrrolidine-2,4-dicarboxylic acid regulates small intestinal growth in chicks

  • X.-G. Li (a1), W.-G. Sui (a1), H.-C. Yan (a1), Q.-Y. Jiang (a1) and X.-Q. Wang (a1)...


Glutamate, which is one of the most important contributors to oxidative metabolism in the intestinal mucosa, is mainly transported by the excitatory amino acids transporters (EAATs) that are expressed in enterocytes. The objective of this study was to evaluate the effects of in ovo administration of l-trans pyrrolidine-2,4-dicarboxylic acid (l-trans-PDC), a potent competitive inhibitor of glutamate uptake by EAATs, on the growth of the small intestine in chicks. Two series of experiments were conducted with hatching eggs; 100 μl of various l-trans-PDC solutions (0, 0.075 or 0.225 mg/egg for the Control group, low-dose l-trans pyrrolidine 2,4-dicarboxylic acid group (L-PDC) or high-dose l-trans pyrrolidine 2,4-dicarboxylic acid group (H-PDC), respectively) was injected into the albumen sac of these hatching eggs before incubation. Hatchlings were sacrificed by cervical dislocation to determine the embryonic development in Experiment I, whereas the birds in Experiment II were raised or sampled at hatching, days 7 and 14 (D7 and D14) for further study. Gene expression in the small intestines was determined by real-time RT-PCR; and serum concentration of free amino acids was determined by an amino acid analyzer. The results showed that the hatchability was decreased by in ovo administration of l-trans-PDC. The small intestinal weights of the H-PDC group were decreased (P<0.05) at hatching and increased (P<0.05) on D7 and D14 compared with those in the Control group. In addition, the gene expression of EAAT2 in the completed or segmental small intestines was not changed (P>0.05); EAAT3 gene expression in the duodenum (P<0.05), jejunum (P=0.084) and ileum (P=0.060) on D14 was lower in the H-PDC group than in the Control group. Furthermore, the serum concentrations of free proline, threonine and phenylalanine but not glutamate or aspartate were increased (P<0.06) in H-PDC group. In conclusion, this paper is the first to report that in ovo administration of l-trans-PDC induces small intestinal growth retardation during the embryonic period and catch-up growth after hatching.


Corresponding author



Hide All
Ahmed, I 2009. Dietary total aromatic amino acid requirement and tyrosine replacement value for phenylalanine in Indian major carp: Cirrhinus mrigala (Hamilton) fingerlings. Journal of Applied Ichthyology 25, 719727.
Barri, A, Honaker, CF, Sottosanti, JR, Hulet, RM and McElroy, AP 2011. Effect of incubation temperature on nutrient transporters and small intestine morphology of broiler chickens. Poultry Science 90, 118125.
Bertolo, RF, Brunton, JA, Pencharz, PB and Ball, RO 2003. Arginine, ornithine, and proline interconversion is dependent on small intestinal metabolism in neonatal pigs. American Journal of Physiology-Endocrinology and Metabolism 284, 915922.
Bol, VV, Delattre, A-I, Reusens, B, Raes, M and Remacle, C 2009. Forced catch-up growth after fetal protein restriction alters the adipose tissue gene expression program leading to obesity in adult mice. American Journal of Physiology − Regulatory, Integrative and Comparative Physiology 297, 291299.
Bröer, S 2008. Amino acid transport across mammalian intestinal and renal epithelia. Physiological Reviews 88, 249286.
Duarte, M, Gionbelli, M, Paulino, P, Serão, N, Martins, T, Tótaro, P, Neves, C, Valadares Filho, S, Dodson, M and Zhu, M 2013. Effects of maternal nutrition on development of gastrointestinal tract of bovine fetus at different stages of gestation. Livestock Science 153, 6065.
Dunlop, J and Butera, JA 2006. Ligands targeting the excitatory amino acid transporters (EAATs). Current Topics in Medicinal Chemistry 6, 18971906.
Ewtushik, A, Bertolo, R and Ball, R 2000. Intestinal development of early-weaned piglets receiving diets supplemented with selected amino acids or polyamines. Canadian Journal of Animal Science 80, 653662.
Had-Aissouni, L 2011. Toward a new role for plasma membrane sodium-dependent glutamate transporters of astrocytes: maintenance of antioxidant defenses beyond extracellular glutamate clearance. Amino Acids 42, 181197.
Hundal, HS and Taylor, PM 2009. Amino acid transceptors: gate keepers of nutrient exchange and regulators of nutrient signaling. American Journal of Physiology-Endocrinology and Metabolism 296, 603613.
Janz, DM and Bellward, GD 1996. In Ovo 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin exposure in three avian species: 1. Effects on thyroid hormones and growth during the perinatal period. Toxicology and Applied Pharmacology 139, 281291.
Kidd, M 2000. Nutritional considerations concerning threonine in broilers. World’s Poultry Science Journal 56, 139151.
Li, XG, Chen, XL and Wang, XQ 2013. Changes in relative organ weights and intestinal transporter gene expression in embryos from White Plymouth Rock and WENS Yellow Feather Chickens. Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology 164, 368375.
Li, XG, Yan, HC, Zeng, PL, Zhang, DX and Wang, XQ 2011. Comparison of difference and ontogenetic expression of EAATs mRNA in the small intestine of broiler chick embryo. China Agriculture Science 44, 44744480. (in Chinese).
Lievens, J, Bernal, F, Forni, C, Mahy, N and Goff, K 2000. Characterization of striatal lesions produced by glutamate uptake alteration: cell death, reactive gliosis, and changes in GLT1 and GADD45 mRNA expression. Glia 29, 222232.
Mehri, M 2012. Development of artificial neural network models based on experimental data of response surface methodology to establish the nutritional requirements of digestible lysine, methionine, and threonine in broiler chicks. Poultry Science 91, 32803285.
Montiel, T, Camacho, A, Estrada-Sanchez, A and Massieu, L 2005. Differential effects of the substrate inhibitor L-trans-pyrrolidine-2,4-dicarboxylate (PDC) and the non-substrate inhibitor DL-threo-β-benzyloxyaspartate (DL-TBOA) of glutamate transporters on neuronal damage and extracellular amino acid levels in rat brain in vivo. Neuroscience 133, 667678.
Rasoamanana, R, Darcel, N, Fromentin, G and Tome, D 2012. Nutrient sensing and signalling by the gut. The Proceedings of the Nutrition Society 71, 446455.
Ren, W, Zou, L, Ruan, Z, Li, N, Wang, Y, Peng, Y, Liu, G, Yin, Y, Li, T and Hou, Y 2013. Dietary L-proline supplementation confers immunostimulatory effects on inactivated Pasteurella multocida vaccine immunized mice. Amino Acids 45, 17.
San Gabriel, A and Uneyama, H 2013. Amino acid sensing in the gastrointestinal tract. Amino Acids 45, 451461.
Sheaffer, KL and Kaestner, KH 2012. Transcriptional networks in liver and intestinal development. Cold Spring Harbor Perspectives in Biology 4, 120.
Speier, JS, Yadgary, L, Uni, Z and Wong, E 2012. Gene expression of nutrient transporters and digestive enzymes in the yolk sac membrane and small intestine of the developing embryonic chick. Poultry Science 91, 19411949.
Vandesompele, J, De Preter, K, Pattyn, F, Poppe, B, Van Roy, N, De Paepe, A and Speleman, F 2002. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biology 3, research0034, 1-research0034.12.
Velasco, I, Tapia, R and Massieu, L 1996. Inhibition of glutamate uptake induces progressive accumulation of extracellular glutamate and neuronal damage in rat cortical cultures. Journal of Neuroscience Research 44, 551561.
Waagepetersen, HS, Shimamoto, K and Schousboe, A 2001. Comparison of effects of DL-threo-β-benzyloxyaspartate (DL-TBOA) and L-trans-pyrrolidine-2, 4-dicarboxylate (t-2, 4-PDC) on uptake and release of [3H] D-aspartate in astrocytes and glutamatergic neurons. Neurochemical Research 26, 661666.
Wang, W, Zeng, X, Mao, X, Wu, G and Qiao, S 2010. Optimal dietary true ileal digestible threonine for supporting the mucosal barrier in small intestine of weanling pigs. The Journal of Nutrition 140, 981986.
Wang, XQ, Chen, X, Tan, HZ, Zhang, DX, Zhang, HJ, Wei, S and Yan, HC 2013a. Nutrient density and slaughter age have differential effects on carcase performance, muscle and meat quality in fast and slow growing broiler genotypes. British Poultry Science 54, 5061.
Wang, XQ, Jiang, W, Tan, HZ, Zhang, DX, Zhang, HJ, Wei, S and Yan, HC 2013b. Effects of breed and dietary nutrient density on the growth performance, blood metabolite, and genes expression of target of rapamycin (TOR) signalling pathway of female broiler chickens. Journal of Animal Physiology and Animal Nutrition 97, 797806.
Welbourne, TC and Matthews, JC 1999. Glutamate transport and renal function. American Journal of Physiology 277, 501505.
Wijtten, P, Langhout, D and Verstegen, M 2012. Small intestine development in chicks after hatch and in pigs around the time of weaning and its relation with nutrition: a review. Acta Agriculturae Scandinavica, Section A-Animal Science 62, 112.
Wu, G 1998. Intestinal mucosal amino acid catabolism. The Journal of Nutrition 128, 12491252.
Wu, G 2009. Amino acids: metabolism, functions, and nutrition. Amino Acids 37, 117.
Wu, G, Bazer, FW, Burghardt, RC, Johnson, GA, Kim, SW, Knabe, DA, Li, P, Li, X, McKnight, JR and Satterfield, MC 2011. Proline and hydroxyproline metabolism: implications for animal and human nutrition. Amino Acids 40, 10531063.
Zeng, PL, Li, XG, Wang, XQ, Zhang, DX, Shu, G and Luo, QB 2011. The relationship between gene expression of cationic and neutral amino acid transporters in the small intestine of chick embryos and chick breed, development, sex, and egg amino acid concentration. Poultry Science 90, 25482556.
Zhang, J, Yin, Y, Shu, XG, Li, T, Li, F, Tan, B, Wu, Z and Wu, G 2013. Oral administration of MSG increases expression of glutamate receptors and transporters in the gastrointestinal tract of young piglets. Amino Acids 45, 11691177.



Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed