Abstract
mRNA levels encoding lactase were detected by Northern blot analysis using two different probes in developing rat intestine. Probe I and probe II corresponding to second half of prolactase gene showed a 6.8 kb mRNA transcript in 7, 14, 21 and 30 day old rat intestine. There was no change in quantity of lactase mRNA detected using probe II, but hybridization with probe I showed a progressive decrease in mRNA transcript encoding lactase with age. At day 7 and 14 of postnatal development, the lactase mRNA was quite high, but it reduced upon weaning. Thein vitro translation products of RNA detected by Western blot analysis using brush border lactase antibodies showed several isoforms of lactase antigen with molecular weight ranging from 100–220 kDa. Analysed at days 7 and 30 of postnatal development, lactase isoforms of molecular weight 130 kDa and 220 kDa were similar to those found in purified brush border membranes. The translation of RNA to 220 kDa lactase protein was high in 7 and 14 day old pups, but it was markedly reduced in 30 day old animals. These findings support the contention that translation of mRNA to lactase is impaired in weaned animals, which may also be responsible for the maturational decline in lactase activity in adult rat intestine
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Abbreviations
- LPH:
-
Lactase phlorizin hydrolase
- MOPS:
-
3-(N-morpholino) propane-sulphonic acid
References
Büller H A, Kothe M J, Goldman D A, Grubman S A, Sasak W V, Matsudaira P T, Montgomery R K and Grand R J 1990 Coordinate expression of Lactate-Phlorizin Hydrolase mRNA and enzyme levels in rat intestine during development;J. Biol. Chem. 265 6978–6983
Büller H A, Van Wassenaer A G, Raghavan S, Montgomery R K, Sybicki M A and Grand R J 1989 New insights into lactase and glycosylceramidase activities of rat lactase-phlorizin hydrolase;Am. J. Physiol. 257 G616-G623
Chomenzynski P and Sacchi N 1987 Single step method of RNA isolation by acid guanidium thiocyanate phenol chloroform extraction;Anal. Biochem. 162 156–159
Duluc I, Boukamel R, Mantei N, Semenza G, Raul F and Freund J N 1991 Sequence of the precursor of intestinal lactase-phlorizin hydrolase from fetal rat;Gene 103 275–276
Freund J N, Duluc I, Foltzer-Jourdainne C, Gosse Fand Raul F 1990 Specific expression of lactase in the jejunum and colon during postnatal development and hormone treatments in the rat;Biochem. J. 268 99–103
Goda T, Yasutake H, Takase Tand Takase S 1999 Lactase-Phlorizin hydrolase and sucrase-isomaltase genes are expressed differently along the villus-crypt axis of rat jejunum;J. Nutr. 129 1107–1113
Keller P, Zecca L, Boukamel R, Zwicker E, Gloor S and Semenza G1995 Furin, PC1/3, and/or PC6A process rabbit, but not human, Pro-lactase-phlorizin-hydrolase to the 180-kDa intermediate;J. Biol. Chem. 270 25722–25728
Kessler M, Acuto O, Storelli C, Murer M and Semenza G 1978 A modified procedure for the rapid preparation of efficiently transporting vesicle from small intestinal brush border membrane: their use in investigating some properties of D-glucose and choline transport systems;Biochim. Biophys. Acta 506 136–154
Krasinski S D, Estrada G, Yeh K Y, Yeh M, Traber P G, Verhave M, Rings E H, Buller H A, Montgomery R K and Grand R J 1994 Transcriptional regulation of intestinal hydrolase: biosynthesis during postnatal development in rats;Am. J. Physiol. 267 G584-G594
Kuokkanen M, Enattah N S, Oksanen A, Savilahti E, Orpana A and Jarvela I 2003 Transcriptional regulation of the Lactase-Phlorizin Hydrolase gene by polymorphisms associated with adult-type hypolactasia;Gut 52 647–652
Lammeli U K 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T4;Nature (London)227 680–685
Lee M, Russell R M, Montgomery R K and Krasinski S D 1997 Total intestinal Lactase and Sucrase activities are reduced in aged rats;J. Nutr. 127 1382–1387
Lloyd M, Mevissen G, Fischer M, Olsen W, Goodspeed D, Genini M, Boll W, Semenza G and Mantei N 1992 Regulation of intestinal lactase in adult hypolactasia;J. Clin. Invest. 89 524–529
Lowry O H, Rosebrough N J, Farr A L and Randall R J 1951 Protein measurement with the Folin-Phenol reagent;J. Biol. Chem. 193 265–275
Mahmood A, Mahmood S, Desehryver-KecsKimeti K and Alpers D H 1993 Characterization of proteins in rat and human intestinal surfactant particles;Arch. Biochem. Biophys. 300 280–286
Mantei N, Villa M, Enzler T, Wacker H, Boll W, James P, Hunziker W and Semenza G 1988 Complete primary structure of human and rabbit lactase-phlorizin hydrolase: implications for biosynthesis, membrane anchoring and evolution of the enzyme;EMBO J. 7 2705–2713
Nsi-Emvo E, Launay J F and Raul F 1987 Is adult type hypolactasia in intestine of mammals related to changes in the intracellular processing of lactase;Cell. Mol. Biol. 33 335–344
Nudell D M, Santiago N A, Zhu J S, Cohen M L, Majuk Z and Gray G M 1993 Intestinal lactase: maturation excess expression of mRNA over enzyme protein;Am. J. Physiol. 265 G1108-G1115
Rings E H, de Boer P A, Moorman A F, Van Beers E H, Dekker J, Montgomery R K, Grands R J and Buller H A 1992 Lactase gene expression during early development of rat small intestine;Gastroenterology 103 1154–1161
Shulman R J, Tivey D R, Sunitha I, Dudley M A and Henning S J 1992 Effect of oral insulin on lactase activity, mRNA and posttranscriptional processing in the newborn pig;J. Pediatr. Gastroenterol. Nutr. 14 166–172
Vasseur M, Frangne R, Cauzac M, Mahmood A and Alvarado F 1990 pH dependent inhibitory effects of Tris and Li ions on intestinal brush border sucrase;J. Enzyme Inhibition 4 15–26
Villa M, Menard D, Semenza G and Mantei N 1992 The expression of lactase enzymatic activity and mRNA in human fetal jejunum. Effect of organ culture and of treatment with hydroxycortisone;FEBS Lett. 301 202–206
Vrese M D, Stegelmann A, Richter B, Fenselau S, Laue C and Schrezenmeir J 2001 Probiotics-Compensation for lactase insufficiency;Am. J. Clin. Nutr. (Suppl.) 73 421S-429S
Wering H M, Huibregtse I L, Zwan S M, Bie M, Dowling L N, Boudrean F, Rings E H, Grand R J and Krasinski S D 2002 Physical interaction between GATA-5 and Hepatocyte Nuclear Factor-1 α Results in synergistic Activation of human Lactase-Phlorizin Hydrolase Promoter;J. Biol. Chem. 277 27659–27667
Yeh K Y, Yeh M, Montgomery R K, Grand R J and Holt P R 1991 Cortisone and thyroxine modulates intestinal lactase and sucrase mRNA levels and activities in the suckling rat;Biochem. Biophys. Res. Commun. 180 174–180
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Kaur, J., Kaur, K., Mahmood, A. et al. In vitro translation of RNA to lactase during postnatal development of rat intestine. J. Biosci. 30, 183–189 (2005). https://doi.org/10.1007/BF02703698
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DOI: https://doi.org/10.1007/BF02703698