Biomedical subjects

V Sorribas

Publications and source records attributed to V Sorribas.

At least 19 recordsLinked to original sources

PubMed1999

Role of thyroid hormone in regulation of renal phosphate transport in young and aged rats.

In the present study, we have examined the cellular mechanisms mediating the regulation of renal proximal tubular sodium-coupled inorganic phosphate (Na/Pi) transport by thyroid hormone (T3) in young and aged rats. Young hypothyroid rats showed a marked decrease in Na/Pi cotransport activity, which was associated with parallel decreases in type II Na/Pi cotransporter (NaPi-2) protein and messenger RNA (mRNA) abundance. In contrast, administration of long-term physiological and supraphysiological doses of T3 resulted in significant increases in Na/Pi cotransport activity, protein, and mRNA levels. Nuclear run-on experiments indicated that thyroid hormone regulates NaPi-2 mRNA levels by a transcriptional mechanism. In aged rats, although there were no changes in T3 serum levels (when compared with young animals), there were significant decreases in serum Pi concentration, renal Na/Pi cotransport activity, and NaPi-2 protein and mRNA abundance. These effects were mediated, at least in part, by a reduction in the transcriptional rate of the NaPi-2 gene, probably caused by, among other factors, a smaller response to the stimulatory action of T3. Compared with young rats, the old rats exhibited less sensitivity of the Na/Pi cotransporter to thyroid hormone, with-decreased effects in both hypothyroid (inhibitory) and hyperthyroid (stimulatory) animals.

Absorption↗

PubMed1998

The two mature transcripts of the chick calcitonin gene are expressed within the central nervous system during embryogenesis.

Calcitonin mRNA and calcitonin gene-related peptide (CGRP) mRNA both are generated from the calcitonin gene because of tissue-specific alternative splicing of the primary transcript. It is currently established that, of the two mature transcripts, calcitonin mRNA is far the predominant transcript produced in thyroid C-cells whereas only CGRP mRNA is produced in the nervous system. However, here we provide evidence that the two splicing forms of the chick calcitonin primary transcript are found within the developing central nervous system, although displaying specific patterns of expression. While CGRP mRNA is first expressed in motor neurons at rather advanced stages of embryogenesis, calcitonin mRNA is expressed in the floor plate and dorsal rhombencephalon from earliest stages.

Animals↗

PubMed1996

Study of the action of intramuscularly administered erythromycin on the L-threonine transport and the digestive enzymatic activity in rabbit jejunum.

Erythromycin has been shown to inhibit the intestinal transport of L-threonine and D-galactose in strips of mucosal jejunum when it was directly added to the incubation medium. Nevertheless, the effect of erythromycin administered therapeutically by intramuscular injection on both the intestinal absorption of nutrients and the intestinal digestive activity, remains unknown. The results obtained show that, firstly, the intestinal absorption of L-threonine is inhibited in animals treated with erythromycin. The kinetic study shows that the effect seems to be mainly due to an alteration of the affinity apparent constant (Kt) of the Na(+)-dependent system of transport located in the mucosal border. However, the Na(+)-dependent L-threonine transport in BBMV was not altered by the treatment with erythromycin. The (Na(+)-K+) ATPase activity in BLMV from treated jejunum was 40% of the activity in control BLMV. Secondly, the treatment with erythromycin did not modify the digestive enzymatic activity of sucrase and aminopeptidase N.

Aminopeptidases↗

PubMed1996

Cellular mechanisms of the age-related decrease in renal phosphate reabsorption.

The aging process in humans and in the rat is associated with an impairment in renal tubular reabsorption of Pi and renal tubular adaptation to a low Pi diet. The purposes of the present study were to determine whether changes in the abundance of type II Na-Pi contransporter (NaPi-2) protein and/or mRNA play a role in the age-related decrease in Na-Pi cotransport activity, and to further determine the cellular mechanisms of impaired adaptation to a low Pi diet. In studies performed in 3- to 4-month-old young adult rats and 32-to 16-month-old aged rats we found that there was an age-related twofold decrease in proximal tubular apical brush border membrane (BBM) Na-Pi cotransport activity, which was associated with similar decreases in BBM NaPi-2 protein abundance and renal cortical NaPi-2 mRNA level. Immunohisto-chemistry showed lower NaPi-2 protein expression in the BBM of proximal tubules of superficial, midcortical, and juxtamedullary nephrons. We also found that in response to chronic (7 days) and/or acute (4 hr) feeding of a low Pi diet there were similar adaptive increases in BBM Na-Pi cotransport activity and BBM NaPi-2 protein abundance in both young and aged rats. However, BBM Na-Pi cotransport activity and BBM NaPi-2 protein abundance were still significantly lower in aged rats, in spite of a significantly lower serum Pi concentration in aged rats. The results indicate that impaired expression of the type II renal Na-Pi cotransporter protein at the level of the apical BBM plays an important role in the age-related impairment in renal tubular reabsorption of Pi and renal tubular adaptation to a low Pi diet.

Adaptation, Physiological↗

PubMed1995

Regulation of opossum kidney (OK) cell Na/Pi cotransport by Pi deprivation involves mRNA stability.

Renal proximal tubular Na-dependent phosphate transport (Na/Pi cotransport) has been studied extensively in the opossum kidney (OK) cell line. Recently, we cloned a complementary deoxyribonucleic acid (cDNA) (NaPi-4) from OK cells encoding an apical NaPi cotransport system. OK cells exposed to a low-Pi medium, as compared to high-Pi media, responded with an increase in Na/Pi cotransport, which was followed by an increase in NaPi-4 messenger ribonucleic acid (mRNA) abundance; maximal stimulation of Na/Pi cotransport was reached in 2 h, with no further increase for up to 16 h. NAPi-4 mRNA abundance was unaltered for 2 h, then increased to a maximum after 6-16 h in cells treated with low Pi medium. NaPi-4 mRNA decay rate was lowered by low-Pi media when compared to high-Pi media, with no increase in the NaPi-4 mRNA transcription rate. These data suggest that the upregulation of Na/Pi cotransport in OK cells by low-Pi media involves two regulatory mechanisms: an immediate (early) increase (after 2 h) in the expression of Na/Pi cotransport, independent of mRNA synthesis or stability, and a delayed (late) effect (after 4-6 h), resulting in an increase in NaPi-4 mRNA abundance, due to an increased stability.

Animals↗

PubMed1995

Thyroid hormone stimulation of Na/Pi-cotransport in opossum kidney cells.

Thyroid hormone (T3), a known stimulator of renal proximal tubular brush border membrane Na-dependent phosphate (Pi) uptake (Na/Pi-cotransport), stimulated Na-dependent Pi transport in opossum kidney (OK) cells. Na/Pi-cotransport was stimulated in a time- and dose-dependent manner with maximal effects (57%) at 24 h and 10(-10) M T3. This stimulation was related to an increase in the apparent capacity (Vmax) of Na/Pi-cotransport. Treatment with T3 had no effect on Na-independent transport of Pi or of L-arginine. The stimulation of Na/Pi-cotransport was paralleled by an increase in the messenger ribonucleic acid (mRNA) encoding the OK cell apical Na/Pi-cotransporter (termed NaPi-4); the mRNA levels related to the activity of Na-independent L-arginine transport (rBAT) were unaffected by T3. Actinomycin D (10(-7) M) completely prevented the stimulatory effect of T3 on OK cell Na/Pi-cotransport and on NaPi-4 mRNA content. In conclusion, T3 stimulates apical Na/Pi-cotransport in OK cells most likely by enhancing its transcription.

Animals↗

PubMed1995

Effect of motilin on the L-leucine transport in rabbit jejunum.

Motilin is a gastrointestinal peptide that stimulates the gastrointestinal motility in several species. The aim of the present work has been to determine the effect of motilin on the L-leucine absorption in rabbit jejunum. The results show that motilin inhibits the L-leucine Na(+)-dependent system of transport located in the mucosal border mainly by diminishing the apparent Vmax. Motilin did not directly affect the Na(+)-dependent system of transport, but it seems to act across the protein kinase C (PKC). These results suggest that motilin may act as a regulatory hormone of the intestinal absorption of nutrients.

Animals↗

PubMed1995

Cloning of a rabbit renal Na-Pi cotransporter, which is regulated by dietary phosphate.

Previously, we isolated a cDNA (NaPi-1) related to a rabbit renal proximal tubular Na-Pi cotransporter (A. Werner, M.L. Moore, N. Mantei, J. Biber, G. Semenza, and H. Murer. Proc. Natl. Acad. Sci. USA 88:9608-9612, 1991.). In this study, we isolated an additional (rabbit renal) cDNA (NaPi-6), which induces Na-dependent Pi uptake in Xenopus laevis oocytes. Substrate specificity and kinetic properties corresponded to those known for rabbit renal brush-border membrane (BBM) Na-Pi cotransport. NaPi-6 was cloned by homology using NaPi-2 cDNA, a rat renal BBM Na-Pi cotransporter (S. Magagnin, A. Werner, D. Markovich, V. Sorribas, G. Stange, J. Biber, and H. Murer. Proc. Natl. Acad. Sci. USA 90: 5979-5983, 1993). NaPi-6 encodes a protein of 642 amino acids, exhibiting at least eight transmembrane domains. NaPi-6 mRNA and protein in kidneys of rabbits fed a low-Pi diet (LPD; 0.11% Pi) for 1 wk were increased by 1.5- and 4-fold, respectively, compared with those of rabbits fed a high-Pi diet (HPD; 1.20% Pi). This effect was correlated with an increase in Na-Pi cotransport of BBM vesicles isolated from animals adapted to LPD (2.5-fold with respect to HPD). In contrast, NaPi-1 mRNA and protein were not altered in response to LPD. Thus rabbit proximal tubular BBMs contain two different Na-Pi cotransport systems: NaPi-1 (type I) and NaPi-6 (type II). Only the type II transport system seems to be under regulatory control in response to low-Pi dietary intake.

Amino Acid Sequence↗

PubMed1994

Cloning of a Na/Pi cotransporter from opossum kidney cells.

Opossum kidney (OK) cells have been extensively used to study cellular mechanisms of renal proximal tubular Na/P(i) cotransport. We have cloned a cDNA (NaPi-4) most likely encoding an apical Na/P(i) cotransporter from OK cells. The cloning strategy was based on homology to the recently cloned human renal (NaPi-3) Na/P(i) cotransporter (Magagnin, S., Werner, A., Markovich, D., Sorribas, V., Stange, G., Biber, J., and Murer, H. (1993) Proc. Natl. Acad. Sci. U. S. A. 90, 5979-5983). Kinetic characterization (P(i) interaction, sodium interaction, and pH dependence) of NaPi-4-induced Na/P(i) uptake showed high similarity to apical Pi transport in OK cell monolayers. The NaPi-4 cDNA is 2548 base pairs long and encodes a protein of 70.5 kDa, containing at least 8 predicted transmembrane domains. Northern blot analysis with OK cell mRNA shows a NaPi-4-related signal (2.5 kilobases) in cells grown on impermeant and permeant supports. Hybrid depletion with NaPi-4 antisense oligonucleotides abolished the mRNA-induced Na/P(i) cotransport in oocytes. Similarly, NaPi-4 antisense oligonucleotides inhibited (up to 70%) Na/P(i) cotransport in OK cell monolayers. We presume that NaPi-4 is closely related to the OK cell apical Na/P(i) cotransporter.

Amino Acid Sequence↗

PubMed1994

Expression of rat renal sulfate transport systems in Xenopus laevis oocytes. Functional characterization and molecular identification.

Renal proximal tubular sulfate reabsorption is mediated by brush border membrane Na+/sulfate-cotransport and basolateral Na(+)-independent sulfate transport. Injection of rat kidney cortex mRNA into Xenopus laevis oocytes induced Na(+)-dependent as well as Na(+)-independent sulfate transport. The inhibition pattern of Na(+)-dependent uptake coincided with that known for the brush border membrane; the inhibition pattern of Na(+)-independent uptake suggested that this activity could be related to the basolateral cell surface. By Northern blot hybridization of size-fractionated mRNA, we provide evidence that the Na(+)-dependent uptake is induced by an mRNA species related to a recently cloned cDNA encoding rat renal cortex Na+/SO4 cotransport (NaSi-1; Markovich, D., Forgo, J., Stange, G., Biber, J., and Murer, H. (1993) Proc. Natl. Acad. Sci. U.S.A. 90, 8073-8077); the Na(+)-independent sulfate transport activity seems to be related to an mRNA species encoding a rat liver Na(+)-independent sulfate transporter (Bissig, M., Hagenbuch, B., Stieger, B., Koller, T., and Meier, P. J. (1994) J. Biol. Chem. 269, 3017-3021). Hybrid depletion experiments using antisense oligonucleotides provided further evidence for the association of the expressed transport activities to NaSi-1 and sat-1, respectively.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

PubMed1994

Expression of rat ileal Na(+)-sulphate cotransport in Xenopus laevis oocytes: functional characterization.

Small-intestinal sulphate absorption is a Na(+)-dependent process having its highest rate in the ileum; it involves brush-border membrane Na(+)-sulphate cotransport. Injection of rat ileal mRNA into Xenopus laevis oocytes induced Na(+)-dependent sulphate uptake in a dose-dependent manner, with no apparent effect on Na(+)-independent sulphate uptake. For mRNA-induced transport, the apparent Km value for sulphate interaction was 0.6 +/- 0.2 mM and that for sodium interaction was 25 +/- 2 mM (Hill coefficient: 2.3 +/- 0.3). mRNA-induced transport, was inhibited by thiosulphate, but not by phosphate or 4,4,'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS). Using a rat renal Na(+)-sulphate cotransporter cDNA as a probe [NaSi-1; Markovich et al. (1993) Proc Natl Acad Sci USA 90:8073-8077], the highest hybridization signals (2.3 kb and 2.9 kb) were obtained in size fractions showing the highest expression of Na(+)-dependent sulphate transport in oocytes. Hybrid depletion experiments using antisense oligonucleotides (from the NaSi-1 cDNA sequence), provided further evidence that rat small-intestinal (ileal) Na(+)-sulphate cotransport is closely related to rat proximal-tubular brush-border membrane Na(+)-sulphate cotransport.

Animals↗

PubMed1994

Cellular mechanisms of acute and chronic adaptation of rat renal P(i) transporter to alterations in dietary P(i).

Recently, the cDNA for a Na-P(i) cotransport system of rat kidney cortex (NaPi-2) has been identified by expression cloning. Using polyclonal antibodies raised against this renal Na-P(i) cotransport system, and using the polymerase chain reaction after reverse transcription of mRNA in microdissected nephron segments, we recently demonstrated that NaPi-2-related mRNA and protein is expressed in the brush-border membranes (BBM) of the proximal tubules of rat kidney. The purpose of the present study was to study the cellular mechanisms involved in adaptation of rat renal Na-P(i) cotransporter to acute and chronic alterations in dietary P(i). Compared with rats fed chronically (7 days) a high-P(i) diet (1.2%), in rats fed chronically a low-P(i) (0.1%) diet the 3.4-fold increase in BBM Na-P(i) cotransport rate (chronic upregulation) was associated with a 2.2-fold increase in renal cortical NaPi-2 mRNA and a 4.9-fold increase in BBM NaPi-2 protein abundances. In contrast, compared with rats fed chronically (7 day) a high-P(i) diet, in rats fed acutely (2 h) a low-P(i) diet the 1.5-fold increase in Na-P(i) cotransport rate (acute upregulation) was associated with a 1.8-fold increase in NaPi-2 protein but no change in NaPi-2 mRNA abundance. Similarly, compared with rats fed chronically a low-P(i) diet, in rats fed acutely (2 h) a high-P(i) diet the 1.9-fold decrease in Na-P(i) cotransport rate (acute downregulation) was associated with a 3.8-fold decrease in NaPi-2 protein but no change in NaPi-2 mRNA abundance.(ABSTRACT TRUNCATED AT 250 WORDS)

Acclimatization↗

PubMed1993

Expression of Na/Pi cotransport from opossum kidney cells in Xenopus laevis oocytes.

Xenopus laevis oocytes have been used for the expression of Na/Pi-cotransport activity by injections of poly(A)+ RNA (mRNA) isolated from an established renal cell line (OK cells). 3-5 days after mRNA injection, Na-dependent phosphate (Pi) uptake by oocytes was increased in a dose-dependent manner; there was no increase in Na-independent Pi uptake. Sucrose density-gradient fractionation indicated that the mRNA species encoding this activity is 2.4-2.8 kb in length. In Northern blots, using a cDNA probe related to human kidney-cortex Na/Pi-cotransport activity (NaPi-3), hybridization with a mRNA-species of 2.4-2.6 kb was obtained. Kinetic characterization ([Pi], [Na]) showed that expressed transport activity has properties similar to apical Na/Pi cotransport in OK cells.

Animals↗

PubMed1993

Action of robenidine on the intestinal transport and digestion of nutrients in rabbit.

Robenidine is an anticoccidial guanidine used as an additive in rabbit fodder. Because its action is restricted to the small intestine, the present work addresses the question whether robenidine affects the growth of the animals, sugar and amino acid intestinal transport and membrane-bound intestinal digestion. For this purpose we have determined the intestinal transport of the substrates, and the enzymatic activity of neutral aminopeptidase and sucrase. We have found that robenidine diminishes the tissue accumulation of L-leucine and D-galactose at long incubation times, and increases the transepithelial mucosal to serosal flux of both substrates. These results suggest that robenidine may stimulate the enterocyte basolateral membrane flux of sugars and neutral amino acids. These results have been corroborated by means of isolated brush border and basolateral membrane vesicles. Apart from these effects, robenidine has also been shown to increase the enzymatic activity of neutral aminopeptidase and sucrase and thus resulting in a better digestion of nutrients.

Aminopeptidases↗

PubMed1993

Expression cloning of human and rat renal cortex Na/Pi cotransport.

We have isolated two cDNA clones, NaPi-2 and NaPi-3, by screening rat kidney cortex and human kidney cortex cDNA libraries, respectively, for expression of sodium-dependent phosphate transport in Xenopus laevis oocytes. Substrate specificity and a detailed kinetic analysis (Na, Pi, H+ concentrations) suggested that expressed uptake activities relate to proximal tubular brush border membrane Na/Pi cotransport. NaPi-2 cDNA contains 2464 bp encoding a protein of 637 aa; NaPi-3 cDNA contains 2573 bp encoding a protein of 639 aa. NaPi-2- and NaPi-3-deduced protein sequences show high homology to each other but are different from the protein sequence deduced from the previously cloned NaPi-1 cDNA (from rabbit proximal tubules). Hydropathy profile predictions suggest at least eight membrane-spanning regions in NaPi-2/3-related proteins. In vitro translation results in proteins of the expected size and suggests glycosylation. Northern blot analysis shows corresponding mRNA species (approximately 2.7 kb) in kidney cortex of various species but no hybridization with RNAs isolated from a variety of other tissues (including intestinal segments); a hybridization signal (approximately 4.8 kb) was observed only in the lung (human). We conclude that we have structurally identified two closely related proteins most likely involved in human and rat renal brush border Na/Pi cotransport.

Amino Acid Sequence↗

PubMed1993

Nicarbazin stimulates intestinal transport of L-leucine in rabbit.

Nicarbazin is an anticoccidial drug used mainly in birds, but also in rabbits. Besides the anticoccidial activity, nicarbazin has shown several other effects such as inhibition of growth and feed efficiency in poultries, and stimulation of sugar intestinal absorption in rabbits. The present work has been performed in order to check whether nicarbazin also affects L-leucine intestinal absorption. The results obtained show that nicarbazin decreases L-leucine accumulation in the jejunal tissue, and increases mucosal to serosal transepithelial fluxes of this amino acid in a dose-dependent way, without modifying its diffusion across the intestinal epithelium. The drug stimulates the amino acid uptake in brush-border and basolateral membrane vesicles, thus suggesting that nicarbazin increases the absorption of L-leucine mediated by carriers.

Animals↗

PubMed1992

Effects of nicarbazin on sugar intestinal absorption in rabbits.

Nicarbazin is an anticoccidial drug, used mainly in birds, which can also be used in rabbits. It has been shown to produce several effects, such as inhibition of growth and feed efficiency in poultry. The aim of the present work was to determine whether nicarbazin alters intestinal absorption of sugar. Results obtained show that nicarbazin decreases D-galactose accumulation in the jejunal tissue and increases mucosal to serosal transepithelial fluxes of this sugar, in both cases in a dose-dependent way. Furthermore, nicarbazin seems not to modify the sugar diffusion across the intestinal epithelium. The drug also stimulates the sugar uptake in brush border and basolateral membrane vesicles. The results suggest that in rabbits nicarbazin increases sugar intestinal absorption mediated by carriers.

Animals↗

PubMed1992

Effect of erythromycin on L-threonine transport in rabbit jejunum in vitro.

Several antibiotics characterized by different molecular structures are known to affect some intestinal activities. Some of them have been described as inhibitors of the intestinal sugar and amino acid transport with different mechanisms. Erythromycin (EM) is a macrolide antibiotic acting as a motilin agonist and thus stimulating the gastrointestinal motor activity. Since several substances which increase the motor activity of the gastrointestinal tract may produce effects on the intestinal absorption of nutrients, the present study has been carried out to determine whether erythromycin affects the L-threonine intestinal absorption. The results obtained indicate that erythromycin diminishes the L-threonine intestinal transport, probably at the mucosal border level. Two groups of experiments carried out, with Na(+)-deprived medium and ouabain-enriched medium, might indicate that erythromycin action could be due to either a direct or an indirect action on the Na(+)-dependent L-threonine transport located in the brush border.

Animals↗