Abstract
Before weaning, arginine biosynthesis from citrulline most likely takes place in the small intestine rather than in the kidney. We studied the expression of ornithine cycle enzymes in the rat small intestine during perinatal development. The spatiotemporal patterns of expression of ornithine aminotransferase, carbamoylphosphate synthetase, ornithine transcarbamoylase, argininosuccinate synthetase, argininosuccinate lyase, and arginase mRNAs were studied by Northern blot analysis and in situ hybridization. In addition, the expression of carbamoylphosphate synthetase and argininosuccinate synthetase protein was studied by immunohistochemistry. Before birth, the developmentally more mature proximal loops of the intestine expressed the mRNAs at higher concentrations than the more distal loops. After birth, this difference was no longer obvious. The mRNAs of argininosuccinate synthetase and argininosuccinate lyase, the enzymes that metabolize citrulline to arginine, were detectable only in the upper part of the villi, whereas the other mRNAs were concentrated in the crypts. The distribution of argininosuccinate synthetase protein corresponded with that of the mRNA, whereas carbamoylphosphate synthetase protein was present in all enterocytes of the crypts and villi. Hepatic arginase mRNA could not be detected in the enterocytes. The spatial distribution of the respective mRNAs and proteins along the villus axis of the suckling small intestine indicates that the basal enterocytes synthesize citrulline, whereas the enterocytes in the upper half of the villus synthesize arginine.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- ARG:
-
arginase
- ASL:
-
argininosuccinate lyase
- ASS:
-
argininosuccinate synthetase
- CPS:
-
carbamoylphosphate synthetase
- OAT:
-
ornithine aminotransferase
- OTC:
-
ornithine transcarbamoylase
- ED:
-
embryonal day
- ND:
-
neonatal day
References
Visek WJ 1986 Arginine needs, physiological state and usual diets. A reevaluation. J Nutr 116: 36–46
Featherston WR, Rogers QR, Freedland RA 1973 Relative importance of kidney and liver in synthesis of arginine by the rat. Am J Physiol 224: 127–129
Dhanakoti SN, Brosnan JT, Brosnan ME, Herzberg GR 1992 Net renal arginine flux in rats is not affected by dietary arginine or dietary protein intake. J Nutr 122: 1127–1134
Hartman WJ, Prior RL 1992 Dietary arginine deficiency alters flux of glutamine and urea cycle intermediates across the portal-drained viscera and liver of rats. J Nutr 122: 1472–1482
Castillo L, Ajami AM, Branch S, Chapman TE, Yu Y-M, Burke JF, Young VR 1994 Plasma arginine kinetics in adult man: response to an arginine-free diet. Metabolism 43: 114–122
Dhanakoti SN, Brosnan JT, Herzberg GR, Brosnan ME 1990 Renal arginine synthesis: studies in vitro and in vivo. Am J Physiol 259:E437–E442
Windmüller HG 1982 Glutamine utilization by the small intestine. Adv Enzymol 53: 201–237
Hoogenraad N, Totino N, Elmer H, Wraight C, Alewood P, Jones RB 1985 Inhibition of intestinal citrulline synthesis causes severe growth retardation in rats. Am J Physiol 249:G792–G799
Milner JA, Wakeling AE, Visek WJ 1974 Effect of arginine deficiency on growth and intermediary metabolism in rats. J Nutr 104: 1681–1689
Hurwitz R, Kretchmer N 1986 Development of arginine-synthesizing enzymes in mouse intestine. Am J Physiol 251:G103–G110
Morris SM, Sweeney WE, Kepka DM, O'Brien WE, Avner ED 1991 Localization of arginine biosynthetic enzymes in renal proximal tubules and abundance of mRNA during development. Pediatr Res 29: 151–154
Davis TA, Fiorotto ML, Reeds PJ 1993 Amino acid compositions of body and milk protein change during the suckling period in rats. J Nutr 123: 947–956
Shenoy V, Roig JC, Kubilis P, Neu J 1996 Characterization of glutaminase in the developing rat small intestine. J Nutr 126: 1121S–1130S
Riby JE, Hurwitz RE, Kretchmer N 1990 Development of ornithine metabolism in the mouse intestine. Pediatr Res 28: 261–265
Moorman AFM, de Boer PAJ, Vermeulen JLM, Lamers WH 1993 Practical aspects of radio-isotopic in situ hybridization on RNA. Histochem J 25: 251–260
de Groot CJ, Zonneveld D, de Laaf RTM, Dingemanse MA, Mooren PG, Moorman AFM, Lamers WH, Charles R 1986 Developmental and hormonal regulation of carbamoylphosphate synthetase gene expression in rat liver: evidence for control mechanisms at different levels in the perinatal period. Biochim Biophys Acta 866: 61–67
Takiguchi M, Miura S, Mori M, Tatibana M, Nagata S, Kaziro Y 1984 Molecular cloning and nucleotide sequence of cDNA for rat ornithine carbamoyltransferase precursor. Proc Natl Acad Sci USA 81: 7412–7416
Morris MS 1988 Nucleotide sequence of the cDNA encoding the rat argininosuccinate synthetase, Nucleic Acids R. es 16: 9352
Amaya Y, Matsubasa T, Takiguchi M, Kobayashi K, Saheki T, Kawamoto S, Mori M 1988 Amino acid sequence of rat argininosuccinate lyase deduced from cDNA. J Biochem 103: 177–181
Kawamoto S, Amaya Y, Oda T, Kuzumi T, Saheki T, Kimura S, Mori M 1986 Cloning and expression in Escherichia coli of cDNA for arginase of rat liver. Biochem Biophys Res Commun 136: 955–961
Kawamoto S, Amaya Y, Murakami K, Tokunaga F, Iwanaga S, Kobayashi K, Saheki T, Kimura S, Mori M 1987 Complete nucleotide sequence of cDNA and deduced amino acid sequence of rat liver arginase. J Biol Chem 262: 6280–6283
Mueckler MM, Pitot HC 1985 Sequence of the precursor to rat ornithine aminotransferase deduced from a cDNA clone. J Biol Chem 260: 12993–12997
Chomszynski P, Sacchi N 1987 Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159
Charles R, de Graaf A, Moorman AFM 1980 Radioimmunochemical determination of carbamoylphosphate synthetase (ammonia) content of adult rat liver. Biochim Biophys Acta 629: 36–49
Schmidlin A, Kalbacher H, Wiesinger H 1997 Presence of argininosuccinate synthetase in glial cells as revealed by peptide-specific antisera. Biol Chem 378: 47–50
Nakamura H, Saheki T, Nakagawa S 1990 Differential cellular localization of enzymes of L-arginine metabolism in the rat brain. Brain Res 530: 108–112
Sternberger LA, Hardy PH, Cululis JJ, Meyer HG 1970 The unlabelled antibody enzyme method of immunohistochemistry. J Histochem Cytochem 20: 315–333
Adlung J, Lorentz K, Grazikowske H 1971 Ueber eine neue, einfache radiochemische Bestimmung von Arginase. Z Klin Chem Klin Biochem 9: 411–414
Wu G, Knabe DA 1995 Arginine synthesis in enterocytes of neonatal pigs. Am J Physiol 269: 621–629
Ryall JC, Quantz MA, Shore GC 1986 Rat liver and intestinal mucosa differ in the developmental pattern and hormonal regulation of carbamoylphosphate synthetase I and ornithine carbamoyl transferase gene expression. Eur J Biochem 156: 453–458
Yamada E, Wakabayashi Y 1991 Development of pyrroline-5-carboxylate synthase and N-acetylglutamate synthase and their changes in lactation and aging. Arch Biochem Biophys 291: 15–23
Dubois N, Cavard C, Chasse JF, Kamoun P, Briand P 1988 Compared expression levels of ornithine transcarbamylase and carbamoylphosphate synthetase in liver and small intestine of normal and mutant mice. Biochim Biophys Acta 950: 321–328
Wu G, Knabe DA, Flynn NE 1994 Synthesis of citrulline from glutamine in pig enterocytes. Biochem J 299: 115–121
Nagy LE, Kretchmer N 1988 Utilization of glutamine in the developing rat jejunum. J Neurochem 118: 189–193
Schiller CM, Southern JT, Walden R 1981 Glutamine and glutamate utilization in the hamster small intestine. J Appl Biochem 3: 147–156
Remesar X, Arola L, Palou A, Alemany M 1985 Activities of amino acid metabolizing enzymes in the stomach and small intestine of developing rats. Reprod Nutr Dev 25: 861–866
Wang T, Lawler AM, Steel G, Sipila I, Milam AH, Valle D 1995 Mice lacking ornithine-d-aminotransferase have paradoxical neonatal hypoornithinaemia and retinal degeneration. Nat Genet 11: 185–190
Herzfeld A, Raper SM 1976 Enzymes of ornithine metabolism in adult and developing rat intestine. Biochim Biophys Acta 428: 600–610
Luk GD, Marton LJ, Baylin SB 1980 Ornithine decarboxylase is important in intestinal mucosal maturation and recovery from injury in rats. Science 210: 195–198
Blachier F, M'Rabet-Touil H, Posho L, Darcy-Vrillon B, Duée P-H 1993 Intestinal arginine metabolism during development. Evidence for de novo synthesis of L-arginine in newborn pig enterocytes. Eur J Biochem 216: 109–117
Herzfeld A, Raper SM 1976 The heterogeneity of arginases in rat tissues. Biochem J 153: 469–478
M'Rabet-Touil H, Lerminiaux H, Duée PH, Blachier F 1996 Evidence for increased anionic arginase activity in pig enterocytes during development. Biochem Mol Biol Int 38: 197–204
Glass RD, Knox WE 1973 Arginase isozymes of rat mammary gland, liver, and other tissues. J Biol Chem 248: 5785–5789
Jenkinson CP, Grigor MR 1994 Rat mammary arginase: Isolation and characterization. Biochem Med Metabol Biol 51: 156–165
Gotoh T, Araki M, Mori M 1997 Chromosomal localization of the human arginase II gene and tissue distribution of its mRNA. Biochem Biophys Res Commun 233: 487–491
Vockley JG, Jenkinson CP, Shukla H, Kern RM, Grody WW, Cederbaum SD 1996 Cloning and characterization of the human type II arginase gene. Genomics 38: 118–123
Hecker M, Sessa WC, Harris HJ, Änggård EE, Vane JR 1990 The metabolism of L-arginine and its significance for the biosynthesis of endothelium-derived relaxing factor: cultured endothelial cells recycle L-citrulline to L-arginine. Proc Natl Acad Sci USA 87: 8612–8616
Hattori Y, Campbell EB, Gross SS 1994 Argininosuccinate synthetase mRNA and activity are induced by immunostimulants in vascular smooth muscle. J Biol Chem 269: 9405–9408
Wu G, Brosnan JT 1992 Macrophages can convert citrulline into arginine. Biochem J 281: 45–48
Nagy LE, Pittler A, Kretchmer N 1988 Development of glutaminase along the villus-crypt axis in the jejunum of rat. J Pediatr Gastr Nutr 7: 907–913
D'Harlingue AE, Kwong LK, Morrill JS, Sunshine P, Tsuboi KK 1986 Growth and differentiative maturation of the rat enterocyte. J Pediatr Gastr Nutr 5: 956–963
Meijer AJ, Lamers WH, Chamuleau RAFM 1990 Nitrogen metabolism and ornithine cycle function. Physiol Rev 70: 701–748
Häussinger D, Lamers WH, Moorman AFM 1992 Hepatocyte heterogeneity in the metabolism of amino acids and ammonia. Enzyme 46: 72–93
Jungermann K, Katz N 1989 Functional specialization of different hepatocyte populations. Physiol Rev 69: 708–764
Brandtzaeg P, Nilssen DE, Rognum TO, Thrane PS 1991 Ontogeny of the mucosal immune system and IgA deficiency. Gastroenterol Clin North Am 20: 397–471
Barbul A 1990 Arginine and immune function. Nutrition 6: 53–58
Cattell V, Jansen A 1995 Inducible nitric oxide synthase in inflammation. Histochem J 27: 777–784
M'Rabet-Touil H, Blachier F, Morel M, Darcy-Vrillon B, Duée PH 1993 Characterization and ontogenesis of nitric oxide synthase activity in pig enterocytes. FEBS Lett 331: 243–247
Acknowledgements
The authors thank C. Gravemeijer and C. Hersbach for photographical assistance. Dr. David Valle (Howard Hughes Medical Institute, Baltimore, MD), Dr. M. Mori (Kuwamoto University School of Medicine, Kuwamoto, Japan), and Dr. S. M. Morris (Howard Hughes Medical Institute, Baylor College, Houston, TX) are acknowledged for making cDNAs available for this study. Dr H. Wiesinger (University of Tübingen, Germany) and Dr. T. Saheki (Kagoshima University, Japan) are gratefully acknowledged for their gift of antibodies.
Author information
Authors and Affiliations
Additional information
Supported in part by the Dutch Organisation for Scientific Research (NWO), Grants 902-23-098 (W.d.J.) and 900-523-155 (M.A.D.).
Rights and permissions
About this article
Cite this article
de Jonge, W., Dingemanse, M., de Boer, P. et al. Arginine-Metabolizing Enzymes in the Developing Rat Small Intestine. Pediatr Res 43, 442–451 (1998). https://doi.org/10.1203/00006450-199804000-00002
Received:
Accepted:
Issue date:
DOI: https://doi.org/10.1203/00006450-199804000-00002
This article is cited by
-
A transgenic approach to study argininosuccinate synthetase gene expression
Journal of Biomedical Science (2014)
-
Blimp1 regulates the transition of neonatal to adult intestinal epithelium
Nature Communications (2011)
-
The human neonatal small intestine has the potential for arginine synthesis; developmental changes in the expression of arginine-synthesizing and -catabolizing enzymes
BMC Developmental Biology (2008)
