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PubMed1991

Angiotensin receptor subtypes in rat, rabbit and monkey tissues: relative distribution and species dependency.

The displacement of [125I]Sar1, Ile8 angiotensin II binding by the receptor subtype selective angiotensin II antagonists, DuP-753 and WL-19 (PD121981) was used to define the relative proportion of angiotensin subtype AT1 and subtype AT2 receptors, respectively in various tissues (aorta, heart, adrenal cortex, kidney cortex and brain) of the rat, rabbit and monkey. The relative abundance of these receptor subtypes varied greatly not only among different tissues of the same species but also within the same tissue of different species. The relative affinity of the DuP-753 and WL-19 for the angiotensin receptor subtypes did not vary markedly suggesting that the two angiotensin receptor subtypes in these tissues and species are similar.

Adrenal Cortex↗
PubMed1992

Debrisoquine 4-monooxygenase and bufuralol 1'-monooxygenase activities in bovine and rabbit tissues.

The tissue distributions of debrisoquine 4-monooxygenase and bufuralol 1'-monooxygenase activities in microsomes from bovine and rabbit tissues were analysed. Debrisoquine 4-monooxygenase and bufuralol 1'-monooxygenase activities were found in liver, and at low levels in cerebral cortex, kidney cortex, lung, small intestine and spleen. Other tissues, such as kidney medulla, adrenocortex, adrenomedulla, blood vessels, thyroid gland, heart, ovary, uterus and testis, showed low levels of bufuralol 1'-monooxygenase activity but not detectable debrisoquine 4-monooxygenase activity. The bufuralol/debrisoquine monooxygenase activity ratios were higher in kidney and lung, and lower in cerebral cortex and spleen than in liver. Both monooxygenase activities in several bovine tissues including liver were inhibited strongly by phenylisocyanide (0.1 mM) and quinidine (0.5 mM), moderately by metyrapone (1 mM), and not at all by KCN (1 mM). NaN3 (5 mM) and sodium cholate (0.5% w/v) inhibited debrisoquine 4-monooxygenase activity strongly and moderately, but bufuralol 1'-monooxygenase activity moderately and strongly, respectively. No effect of a hydroxyl radical scavenger or of superoxide dismutase on either monooxygenase activity was observed. It was concluded from these results, as well as the NADPH dependency of the reactions, that the two monooxygenase reactions observed in these tissues were catalysed by cytochrome P450s.

Animals↗
PubMed2026

Longitudinal multiorgan transcriptomic atlas of salt-induced hypertension.

High dietary salt intake elevates blood pressure and drives multiorgan damage. However, the molecular programs underlying progressive organ injury remain poorly defined. Here, we present a longitudinal multiorgan transcriptomic atlas of salt-induced hypertensive injury. We profiled kidney cortex, kidney medulla, heart, and liver across 4 stages, spanning early hypertension to advanced pathology in Dahl salt-sensitive rats. We identified dynamic and tissue-specific molecular trajectories, including a shared early proliferative response that converges on proinflammatory and fibrotic remodeling. Notably, we uncovered compartment-specific renal responses, showing that the cortex and medulla, despite their proximity, follow distinct molecular trajectories during disease progression. We further identified 79 stage- and tissue-specific transcription factors that drive gene expression dynamics in salt-induced hypertensive injury. Integration with human genome-wide association studies revealed conserved pathways in endocrine signaling, ion transport, lipid metabolism, and detoxification, establishing cross-species relevance and highlighting mechanistic targets of clinical importance. Compound-transcriptome analysis revealed stage- and organ-specific therapeutic opportunities, prioritizing kinase and epigenetic modulators as candidates to rebalance maladaptive gene programs. Overall, this study provides a resource for understanding molecular mechanisms from early salt-induced hypertension to tissue-specific injury and underscores the need for precision interventions.

Animals↗
PubMed1990

Expression of mRNA coding for kidney and red cell water channels in Xenopus oocytes.

The existence and identity of protein water transporters in biological membranes has been uncertain. Osmotic water permeability (Pf) was measured in defolliculated Xenopus oocytes microinjected with water or mRNA from kidney cortex, kidney papilla, reticulocyte, brain, and muscle. Pf was measured by quantitative image analysis from the time course of oocyte swelling in response to an osmotic gradient. When assayed at 10 degrees C, Pf in water-injected oocytes increased from (3.6 +/- 0.9) x 10(-4) cm/s (S.D., n = 16) to 74 x 10(-4) cm/s with addition of amphotericin B, showing absence of unstirred layers. At 48-72 h after injection of 50 ng of unfractionated mRNA, Pf (in cm/s x 10(-4] was: 4.0 +/- 1.5 (rabbit brain, n = 15), 4.2 +/- 1.8 (rabbit muscle, n = 10), 18.4 +/- 6.3 (rabbit reticulocyte, n = 20), 16.1 +/- 5.6 (rat renal papilla, n = 24), 12.9 +/- 6.3 (rat renal cortex, n = 20), 14.4 +/- 6.1 (rabbit renal papilla, n = 15), and 11.8 +/- 3.4 (rabbit renal cortex, n = 8). In oocytes injected with mRNA from rat renal papilla, Pf was inhibited reversibly by 0.3 mM HgCl2 (4.1 +/- 1.6, n = 10); expressed water channels from kidney and red cell had activation energies of less than 4 kcal/mol. These results show functional oocyte expression of water channels from red cell, kidney proximal tubule (cortex), and the vasopressin-sensitive kidney collecting tubule (papilla), indicating that water channels are proteins, and providing an approach for the expression cloning of water channels.

Animals↗
PubMed2025

Longitudinal Multi-Organ Transcriptomic Atlas of Salt-Induced Hypertension.

BACKGROUND: Salt-sensitive hypertension is a prevalent and clinically significant subtype of hypertension, where increased dietary salt intake elevates blood pressure and causes injury to multiple organ systems. Despite extensive research, dynamic molecular changes and conserved versus organ-specific transcriptional programs in hypertensive multi-organ damage remain poorly understood. Defining complex molecular pathways both in a temporal sequence and in an organ-specific manner is essential for developing targeted, precision therapies to mitigate hypertensive disease burden. METHODS: We generated a longitudinal multi-organ transcriptomic atlas of salt-sensitive hypertension using RNA sequencing of kidney cortex, kidney medulla, heart, and liver from Dahl salt-sensitive rats across four disease stages. A comprehensive bioinformatic analysis mapped dynamic transcriptional programs, evaluated 50 biological pathways, and defined upstream regulators. Histological and biochemical assays complemented transcriptomic analysis, while integration with human genome-wide association studies (GWAS) and compound-transcriptome analysis provided translational insights and identified candidate therapeutics. RESULTS: Salt-induced hypertension elicited both shared and tissue-specific transcriptional programs that evolved with disease progression. The kidney medulla showed robust early immune activation with metabolic suppression, while the cortex exhibited transient metabolic activation before declining and initiating immune activation. The liver and heart showed time-dependent metabolic and inflammatory remodeling. Cross-organ comparisons revealed a shared early proliferative response that converged on proinflammatory and fibrotic signatures. Upstream regulator analysis identified 79 time- and tissue-specific transcription factors associated with gene expression dynamics. GWAS integration analysis revealed endocrine signaling, ion transport, lipid metabolism, and detoxification as conserved pathways across species, underscoring the translational relevance of the model and study. Predictive compound-transcriptome analyses identified kinase inhibitors targeting phosphoinositide 3-kinase, mechanistic target of rapamycin and cyclin-dependent kinases as top candidates to counteract maladaptive transcriptional programs. CONCLUSIONS: This study defines temporal and tissue-specific transcriptomic remodeling in salt-sensitive hypertension and highlights the need for precision interventions to prevent progressive organ damage.

Journal Article↗
PubMed1970

A radiochemical assay for glutamine synthetase, and activity of the enzyme in rat tissues.

1. A radiochemical assay for glutamine synthetase has been developed in which an ATP-regenerating system is incorporated to prevent accumulation of inhibitory amounts of ADP. It is particularly suitable for assay of the enzyme in crude tissue extracts containing high adenosine triphosphatase activity. 2. A survey of the distribution of the enzyme in tissues from normal male rats showed that activity is present in liver, brain cortex, kidney cortex, spleen, testis and retina. 3. The K(m) of the enzyme for l-glutamate is approx. 1.5x10(-2)m.

Adenine Nucleotides↗
PubMed1991

Molecular cloning and expression of a cDNA encoding the rabbit ileal villus cell basolateral membrane Na+/H+ exchanger.

A cDNA clone encoding a rabbit ileal villus cell Na+/H+ exchanger was isolated and its complete nucleotide sequence was determined. The cDNA is 4 kb long and contains 322 bp of 5'-untranslated region, 2451 bp of open reading frame and 1163 bp of 3'-untranslated area, with 70%, 91% and 40% identity to the human sequence, respectively. Amino acid sequence deduced from the longest open reading frame indicated a protein of 816 residues (predicted Mr 90,716) which exhibits 95% amino acid identity to the human Na+/H+ exchanger. The two putative glycosylation sites in the human Na+/H+ exchanger are conserved in this protein, suggesting that it is a glycoprotein. Stable transfection of the cDNA into an Na+/H+ exchanger deficient fibroblast cell line, established Na+/H+ exchange. The Na+/H+ exchanger was stimulated by serum and a phorbol ester but not by 8-Br-cAMP. In Northern blot analysis, the cDNA hybridized to a 4.8 kb message in rabbit ileal villus cells, kidney cortex, kidney medulla, adrenal gland, brain and descending colon and to a 5.2 kb message in cultured human colonic cancer cell lines, HT29-18 and Caco-2. In immunoblotting, a polyclonal antibody raised against a fusion protein of beta-galactosidase and the C-terminal 158 amino acids of the human Na+/H+ exchanger identified a rabbit ileal basolateral membrane protein of 94 kd and only weakly interacted with the ileal brush border membrane. In immunocytochemical studies using ileal villus and crypt epithelial cells, the same antibody identified basolateral and not brush border epitopes. Restriction analysis of genomic DNA with a 462 bp PstI-AccI fragment of the rabbit Na+/H+ exchanger strongly suggests the existence of closely related Na+/H+ exchanger genes. The near identity of the basolateral Na+/H+ exchanger and the human Na+/H+ exchanger plus the ubiquitous expression of this message suggests that the ileal basolateral Na+/H+ exchanger is the 'housekeeping' Na+/H+ exchanger.

Amino Acid Sequence↗
PubMed1994

Comparisons between the inorganic content of healthy and hypertensive rat tissues by inductively coupled plasma-mass spectrometry.

The inorganic contents of bone, brain, erythrocyte, heart, kidney cortex, kidney medulla, liver, lung, muscle and plasma from spontaneously hypertensive rats were compared with those of the same tissues from healthy Sprague-Dawley rats. A general inductively coupled plasma-mass spectrometry method developed for multi-element determinations of most of the elements present in biological tissues was used. Variations were found not only for major elements, as expected, but also for many trace elements in several tissues.

Animals↗
PubMed1987

Cadmium, zinc and copper relationships in kidney and liver of humans exposed to environmental cadmium.

Cadmium, copper and zinc have been determined in kidney cortex, kidney medulla and liver from 51 cadmium-polluted and 122 nonpolluted autopsied subjects. Liver cadmium and zinc were significantly higher in the cadmium-polluted subjects than in the nonpolluted subjects. Kidney cadmium, copper and zinc were lower in the cadmium-polluted subjects because of renal damage. Significant correlations of the three metals were found in the kidney. In the liver the correlation coefficients were significant only between cadmium and zinc.

Adolescent↗
PubMed1976

Effects of daily oral subtoxic doses of a wet lead-based paint on cattle.

Cattle were dosed daily with a wet lead-based paint at a rate of about 6 mg of lead per kg per day for 60 weeks. The cattle did not exhibit overt symptoms of lead toxicosis. Blood and fecal samples collected during the study, and tissue samples obtained at slaughter of the animals were analyzed for lead by atomic absorption spectrophotometer equipped with a graphite furnace. Levels of lead in whole blood generally did not exceed 0.5 ppm. The residues of lead in feces reached as high as 1000 ppm. Highest residues in tissues were in calcified bone, the kidney cortex, kidney medulla, and then in liver. Residues of lead in muscle, brain, spleen, and bone marrow were similar to residues of lead in the control animal. Calves born to 2 test animals had elevated levels of lead in bone, kidney and liver.

Animals↗
PubMed1997

Identification of a cDNA/protein leading to an increased Pi-uptake in Xenopus laevis oocytes.

In a previous report we documented an increased Na(+)-dependent transport of inorganic phosphate (P(i)) in Xenopus laevis oocytes injected with mRNA isolated from rabbit duodenum (Yagci et al., Pfluegers Arch. 422:211-216, 1992; ref 24). In the present study we have used expression cloning in oocytes to search for the cDNA/mRNA involved in this effect. The identified cDNA (provisionally named PiUS; for P(i)-uptake stimulator) lead to a 3-4-fold stimulation of Na(+)-dependent P(i)-uptake (10ng cRNA injected, 3-5 days of expression). Na(+)-independent uptake of P(i) was also affected but transport of sulphate and L-arginine (in the presence or absence of sodium) remained unchanged. The apparent K(m)-values for the induced Na(+)-dependent uptake were 0.26 +/- 0.04 mM for P(i) and 14.8 +/- 3.0 mM for Na+. The 1796 bp cDNA codes for a protein of 425 amino acids. Hydropathy analysis suggests a lack of transmembrane segments. In vitro translation resulted in a protein of 60 kDa and provided no evidence of glycosylation. In Northern blots a mRNA of approximately 2 kb was recognized in various tissues including different intestinal segments, kidney cortex, kidney medulla, liver and heart. Homology searches showed no similarity to proteins involved in membrane transport and its control. In conclusion, we have cloned from a rabbit small intestinal cDNA library a novel cDNA encoding a protein stimulating P(i)-uptake into Xenopus laevis oocytes, but which is not a P(i)-transporter itself.

Amino Acid Sequence↗
PubMed1982

Studies on the range of tissue protein synthesis in pigs: the effect of thyroid hormones.

The effects of thyroid hormones on the range of tissue protein synthesis in growing pigs using the constant infusion technique with [14C]lysine were studied. 2. During a 6 h infusion, samples were taken from blood and, at the end of the infusion, from liver, pancreas, stomach, small and large intestines, kidney cortex, kidney medulla, muscle and skin. 3. Lower relative specific radioactivities of free leucine and lysine in several tissues were observed in the hormone-treated group than in the untreated one. 4. The range of protein synthesis rate and the daily amount of protein synthesized in tissues was higher in all tissues after application of thyroid hormones. 5. Assuming that the organs analysed represented 70% of the total trichloroacetic acid-precipitable protein of the pig, the estimated range of daily protein synthesis was 251-490 and 312-880 g in untreated and hormone-treated pigs respectively.

Animals↗
PubMed1984

Comparison of four different carboxylterminal tracers in a radioimmunoassay specific to the 68-84 region of human parathyroid hormone.

Two synthetic carboxylterminal fragments, [tyr52]hPTH(52-84) and [tyr63]hPTH(63-84), and purified bPTH(1-84) were iodinated with 125Iodine to be compared as tracers in a late carboxylterminal radioimmunoassay. Tracer 125I-bPTH(41-84) was generated in vitro by incubating 125I-bPTH(1-84) with plasma membranes of rat kidney cortex. Region specificity was achieved by saturating the unwanted middle component of our multivalent antiserum with a molar excess of hPTH(44-68). A charcoal-dextran separation was worked out for each tracer. The titer of the antiserum giving approximately equal to 30% specific binding of each tracer was used in all experiments. Displacement of each tracer with increasing molar concentration of hPTH(1-84), hPTH(53-84), hPTH(41-84) and of hPTH (64-84) was studied. hPTH(41-84) was also generated by incubating hPTH(1-84) with rat cortex kidney membranes and was calibrated against a commercial preparation of bPTH(37-84). A progressive increase in the titer of the antiserum was seen as the molecular weight of the tracers decreased from a titer of 1/20,000 with 125I-bPTH(1-84) to a titer of 1/50,000 with the two synthetic tracers. Similarly the so-called damage seen during the charcoal-dextran separation in absence of antibody was reduced from 16.0 +/- 6.2% (mean +/- SD) with 125I-bPTH(1-84) to 1.3 +/- .2 with the two synthetic tracers. 50% displacement of the 125I-bPTH(1-84) tracer was achieved at 13.2 +/- .8 fmol/tube for hPTH(1-84) and at 6.3 +/- 1.0 fmol/tube for hPTH(41-84), reflecting the greater reactivity of fragments in that system. With the two synthetic tracers, a concentration of 5.0 +/- .4 fmol/tube of hPTH(1-84) or of 3.5 +/- 1.2 fmol/tube of hPTH(41-84) was necessary to achieve the same goal. With 125I-bPTH(41-84) results were between the two extremes. These results indicated that an increase in antiserum titer, a decrease in assay damage, an improvement in assay sensitivity and in comparative molar reactivity of the various circulating forms of hPTH can be achieved by using synthetic carboxylterminal fragments as tracers in region specific radioimmunoassays of hPTH.

Animals↗
PubMedDate not supplied

Fine tuning of rabbit equilibrative nucleoside transporter activity by an alternatively spliced variant.

The full-length cDNA encoding an equilibrative nucleoside transporter (rbENT2) and its novel C-terminal variant, rbENT2A, were isolated from rabbit trachea. Rabbit ENT2 protein consists of 456 amino acid residues; rbENT2A is shorter by 41 residues. Both rbENT2 and rbENT2A transcripts are found in rabbit tissues including intestine, kidney cortex, kidney, and trachea, at varying levels of expression. When transfected in a heterologous expression system-Madin Darby canine kidney (MDCK) epithelial cell line-both rbENT2 and rbENT2A were expressed. rbENT2 had a molecular mass of 49 kDa; rbENT2A had a molecular mass of 44 kDa. Clones of both transporters yielded functional proteins that were capable of mediating uridine uptake and efflux without the needing to be coupled to a secondary ion (e.g. Na(+)). Remarkably, rbENT2A displayed a higher affinity (K(m) = 41 microM) and a lower capacity (V(max) = 0.6 nmol/mg protein/5 min) towards substrates than rbENT2 (K(m) = 272.8 microM, V(max) = 1.26 nmol/mg protein/5 min). Pharmacological profiles showed that nitro-benzyl-mercapto-purine-ribose (NBMPR) potently inhibited (3)H-uridine uptake mediated by rbENT2A, but not uptake mediated by rbENT2. The constitutive splicing, broad expression, markedly different kinetics, and distinct pharmacological characteristics of rbENT2A appear to act in conjunction with the wild type, rbENT2, to fine-tune basolateral nucleoside transport function in rabbit trachea.

Alternative Splicing↗
PubMed1986

Tissue distribution of subcutaneously administered aluminum chloride in weanling rabbits.

The purpose of our investigation was to determine blood and tissue levels of aluminum (Al) in normal young rabbits. Furthermore, we wished to determine tissue distribution and accumulation of Al as related to its blood concentration in Al-dosed rabbits. The levels of Al accumulated were determined in different tissues of growing rabbits after continuous subcutaneous administration of Al chloride (3.78 mg/d) for 28 d. No signs of toxicity were apparent from comparisons of hematocrit or weight gain between control and Al-dosed rabbits. The largest concentration of the Al was observed in bone, which was also found to have the highest levels in the control rabbit tissues. Following bone, the experimental animals showed the greatest increase of Al levels in kidney cortex, kidney medulla, liver, testes, skeletal muscle, heart, brain white matter, and brain hippocampus, in that order. No significant difference was found in brain grey matter between control and experimental animals. As the brain tissue of the Al-treated animals had the lowest Al level of the tissues measured, it appears that there is a partial blood-brain barrier to entry of Al.

Aluminum↗
PubMed1991

In vivo measurement of proton diffusion in the presence of coherent motion.

Measurement of the self-diffusion coefficient D of water in tissue has been performed traditionally using the technique proposed by Stejskal and Tanner. A variant of that technique is shown here, employing flow-compensated gradients that significantly reduce the sensitivity to small coherent motions that are common in body imaging. An interleaved sequence with four values of diffusion-sensitizing gradient (b) minimizes registration errors. Eddy currents and other systematic errors are reduced, permitting the measurement of standards in an imaging context within 5% of nonimaging values in the literature. The flow-compensated sequence permits the measure of D for tissues in the abdominal cavity of the rat. We present in vivo measurements of D for the following rat tissues; liver, kidney (cortex), kidney (medulla) muscle, brain, fat.

Acetone↗
PubMed1980

Effects of synthetic parathyroid hormone on vascular beds of dogs.

Blood flow rates and vascular resistance in dogs were determined in 14 tissues with 15 +/- 2 microns radioactive microspheres (141Ce, 51Cr, 85Sr). Intravenous injections of synthetic bovine parathyroid hormone containing the NH2-terminal 1--34 amino acids [bPTH-(1--34)] at the dose of 10 USP units/kg body weight produced hypotension and increases in blood flow rate and decreases in resistance in kidney cortex, kidney medulla, heart, liver, stomach and pancreas at 1/2 and 2 min after injection. 4 U bPTH-(1--34)/kg body weight produced less vasodilation. Saline injection had no effect on the tissues studied.

Animals↗
PubMed2006

Increased blood pressure in adult offspring of families with Balkan endemic nephropathy: a prospective study.

BACKGROUND: Previous studies have linked smaller kidney dimensions to increased blood pressure. However, patients with Balkan Endemic Nephropathy (BEN), whose kidneys shrink during the course of the disease, do not manifest increased blood pressure. The authors evaluated the relationship between kidney cortex width, kidney length, and blood pressure in the offspring of BEN patients and controls. METHODS: 102 offspring of BEN patients and 99 control offspring of non-BEN hospital patients in the Vratza District, Bulgaria, were enrolled in a prospective study and examined twice (2003/04 and 2004/05). Kidney dimensions were determined using ultrasound, blood pressure was measured, and medical information was collected. The parental disease of BEN was categorized into three groups: mother, father, or both parents. Repeated measurements were analyzed with mixed regression models. RESULTS: In all participants, a decrease in minimal kidney cortex width of 1 mm was related to an increase in systolic blood pressure of 1.4 mm Hg (p = 0.005). There was no association between kidney length and blood pressure. A maternal history of BEN was associated with an increase in systolic blood pressure of 6.7 mm Hg (p = 0.03); paternal BEN, +3.2 mm Hg (p = 0.35); or both parents affected, +9.9 mm Hg (p = 0.002). There was a similar relation of kidney cortex width and parental history of BEN with pulse pressure; however, no association with diastolic blood pressure was found. CONCLUSION: In BEN and control offspring, a smaller kidney cortex width predisposed to higher blood pressure. Unexpectedly, a maternal history of BEN was associated with average increased systolic blood pressure in offspring.

Adult↗