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Y G Huang

Publications and source records attributed to Y G Huang.

At least 19 recordsLinked to original sources
PubMed2000

Renin expression in COX-2-knockout mice on normal or low-salt diets.

Experiments were performed in mice to investigate whether cyclooxygenase-2 (COX-2) in epithelial cells near the tubulovascular contact point (macula densa and TAL cells) may regulate renin gene expression in juxtaglomerular granular cells. Renin activity, afferent arteriolar granularity, and renin mRNA were determined in wild-type mice and in COX-2-knockout mice on control and low-NaCl diets. Renin activity in microdissected glomeruli assessed as angiotensin I formation in the presence of excess substrate and afferent arteriolar granularity determined by direct visualization and immunostaining were significantly reduced in COX-2 -/- compared with wild-type animals. Similarly, renal cortical mRNA levels were lower in COX-2 -/- than in wild-type mice. Maintaining mice on a low-salt diet for 14 days induced an increase in renin mRNA, afferent arteriolar granularity, and renin activity in wild-type mice. In contrast, renin mRNA and renin granularity did not significantly increase in low-salt-treated COX-2 -/- mice, whereas the increase in juxtaglomerular renin enzyme activity was markedly attenuated, but not fully blocked. In additional experiments we found that COX-2 mRNA was increased in angiotensin type 1A receptor-knockout mice compared with wild-type mice. We conclude that COX-2 in the tubulovascular contact region is a critical determinant of renin synthesis in granular cells under resting conditions and that it participates in the stimulation of renin expression caused by a low-NaCl intake.

Animals↗
PubMed1999

Localization of PEPT1 and PEPT2 proton-coupled oligopeptide transporter mRNA and protein in rat kidney.

To determine the renal localization of oligopeptide transporters, Northern blot analyses were performed and polyclonal antisera were generated against PEPT1 and PEPT2, the two cloned rat H+/peptide transporters. Under high-stringency conditions, a 3.0-kb mRNA transcript of rat PEPT1 was expressed primarily in superficial cortex, whereas a 3.5-kb mRNA transcript of PEPT2 was expressed primarily in deep cortex/outer stripe of outer medulla. PEPT1 antisera detected a specific band on immunoblots of renal and intestinal brush-border membrane vesicles (BBMV) with an apparent mobility of approximately 90 kDa. PEPT2 antisera detected a specific broad band of approximately 85 kDa in renal but not in intestinal BBMV. PEPT1 immunolocalization experiments showed detection of a brush border antigen in S1 segments of the proximal tubule and in the brush border of villi from all segments of the small intestine. In contrast, PEPT2 immunolocalization was primarily confined to the brush border of S3 segments of the proximal tubule. All other nephron segments in rat were negative for PEPT1 and PEPT2 staining. Overall, our results conclusively demonstrate that although PEPT1 is expressed in early regions of the proximal tubule (pars convoluta), PEPT2 is specific for the latter regions of proximal tubule (pars recta).

Animals↗
PubMed1999

Tubuloglomerular feedback in ACE-deficient mice.

In these experiments, we used a strain of angiotensin converting enzyme (ACE) germline null mutant mice, generated by J. H. Krege and co-workers (J. H. Krege, S. W. M. John, L. L. Langenbach, J. B. Hodgin, J. R. Hagaman, E. S. Bachman, J. C. Jennette, D. A. O'Brien, and O. Smithies. Nature 375: 146-148, 1995), to examine the effect of chronic ACE deficiency on the magnitude of tubuloglomerular feedback (TGF) responses. The genotype was determined by PCR on DNA extracted from the tail and was verified after each experiment by assessment of the blood pressure response to an injection of ANG I. To assess TGF responsiveness, we determined the change in stop-flow pressure (PSF) caused by increasing NaCl concentration at the macula densa by using micropuncture techniques. When loop of Henle flow rate was increased from 0 to 40 nl/min, PSF fell from a mean of 42.3 +/- 1.95 to 33.6 +/- 2.09 mmHg (n = 6, P = 0.005) in wild-type mice (+/+), fell from 40.6 +/- 2.35 to 38.6 +/- 1.93 mmHg in heterozygous (+/-) mice (n = 7, P = 0.014), and did not change in homozygous ACE (-/-) mice [36.7 +/- 2.02 mmHg vs. 36.4 +/- 2.01 mmHg; n = 4, P = not significant (NS)]. During an infusion of ANG II at a dose that did not significantly elevate blood pressure (70 ng. kg-1. min-1), TGF response magnitude (PSF 0 - PSF 40) increased from 6.5 +/- 1.4 to 9.8 +/- 1.19 mmHg in +/+ (P = 0.006), from 1.14 +/- 0.42 to 4.6 +/- 1.3 mmHg in +/- (P = 0.016), and from 0.42 +/- 0.25 to 4.02 +/- 1.06 in -/- mice (P = 0.05). Absence of TGF responses in ACE null mutant mice and restoration of near-normal responses during an acute infusion of ANG II supports previous conclusions that ANG II is an essential component in the signal transmission pathway that links the macula densa with the glomerular vascular pole.

Angiotensin II↗
PubMed1999

Differential regulation of COX-2 expression in the kidney by lipopolysaccharide: role of CD14.

Induction of the inducible cyclooxygenase isoform COX-2 is likely to be an important mechanism for increased prostaglandin production in renal inflammation. We examined the effect of lipopolysaccharide (LPS) on regional renal COX-2 expression in the rat. In the inner medulla, LPS injection (4 mg/kg ip) induced a twofold and 2.5-fold increase in the levels of COX-2 mRNA and COX-2 protein, respectively. In contrast, COX-2 expression in the renal cortex was not significantly altered. COX-2 promoter transgenic mice were created using the 2.7-kb flanking region of the rat COX-2 gene. In these animals, LPS injection induced reporter gene expression predominately in the inner medulla. The LPS receptor CD14, usually regarded as a monocyte/macrophage-specific marker, was found to be abundantly expressed in the inner medulla and in dissected inner medullary collecting duct (IMCD) cells, suggesting that it may mediate medullary COX-2 induction. CD14 was present only at low levels in cortex and cortical segments, including glomeruli. In cultured cells, it was abundant in mouse IMCD (mIMCD-K2) cells and renal medullary interstitial cells, but largely undetectable in mesangial cells and M1 cells, a cell line derived from mouse cortical collecting ducts. In the mIMCD-K2 cell line, LPS significantly induced COX-2 mRNA expression, with concomitant induction of CD14. LPS-stimulated COX-2 expression was reduced by the addition of an anti-CD14 monoclonal antibody to the culture medium. These results demonstrate that LPS selectively stimulates COX-2 expression in the renal inner medulla through a CD14-dependent mechanism.

Animals↗
PubMed1998

Long-term results of percutaneous balloon mitral valvuloplasty for mitral stenosis: a follow-up study to 11 years in 202 patients.

We studied the first 202 patients with rheumatic mitral stenosis (MS) who underwent percutaneous balloon mitral valvuloplasty (PBMV) with the Inoue balloon catheter for a follow-up (FU) period of 5-11 years. Pre- and post-PBMV and at FU, the mean left atrial pressure was 21.3+/-7.4, 10.2+/-5.6, and 11.2+/-4.1 mm Hg; mean diastolic mitral gradient was 18.4+/-7.3, 2.9+/-3.2, and 5.1+/-4.3 mm Hg; and mitral valve area was 1.0+/-0.3, 2.1+/-0.6, and 1.7+/-0.5 cm2. Functional status improved from New York Heart Association (NYHA) class IV in 3, class III in 119, and class II in 80 pre-PBMV to class I in 163, class II in 37, and class III in 2 post-PBMV, and was class I in 146, class II in 39, and class III in 17 patients at FU. In the 17 patients with NYHA class III at FU, mitral restenosis was the culprit; 4 underwent repeat PBMV, 12 had mitral valve replacement for severe mitral calcification and subvalvular fusion, and 1 refused further intervention. Thus PBMV using the Inoue balloon catheter is an effective method of relieving MS with excellent long-term results in patients without severe mitral calcification and subvalvular fusion.

Adolescent↗
PubMed1998

Tubular localization and tissue distribution of peptide transporters in rat kidney.

PURPOSE: To define the tubular localization and tissue distribution of PEPT1 (low-affinity, high-capacity transporter) and PEPT2 (high-affinity, low-capacity transporter) in rat kidney. METHODS: mRNA expression of PEPT1 and PEPT2 was assessed with reverse transcription-polymerase chain reaction (RT-PCR) methods using cDNA prepared from microdissected nephron segments in rat. Tissue localization of rat renal PEPT1 and PEPT2 mRNA was further assessed by in situ hybridization with radiolabeled probes. RESULTS: RT-PCR analysis of microdissected segments from rat nephron showed that both PEPT1 and PEPT2 are confined to a proximal tubule. While PEPT1 is specific for early regions of the proximal tubule (pars convoluta), PEPT2 is overwhelmingly but not exclusively expressed in latter regions of the proximal tubule (pars recta). All other segments along the nephron were negative for PEPT1 or PEPT2 mRNA transcripts. These finding were supported by in situ hybridization results in which PEPT1 was selectively expressed in kidney cortex and PEPT2 in the outer stripe of outer medulla. CONCLUSIONS: Contrary to current opinion, the data suggest that peptides are handles in a sequential manner in proximal regions of the nephron, first by the low-affinity, high-capacity transport system and second by the high-affinity, low-capacity transport system.

Animals↗
PubMed1998

Tubuloglomerular feedback: new concepts and developments.

Luminal [NaCl] at the macula densa (MD) has two established effects: regulation of glomerular arteriolar resistance through tubuloglomerular feedback (TGF) and control of renin secretion. TGF acts as a minute-to-minute stabilizer of distal salt delivery, thereby minimizing the impact of random perturbations in filtration and absorption forces on NaCl excretion. During long-lasting perturbations of MD [NaCl], control of renin secretion becomes the dominant function of the MD. The potentially maladaptive effect of TGF under chronic conditions is prevented by TGF adaptations permitting adjustments in glomerular filtration rate to occur. TGF adaptation is mechanistically coupled to the endpoint targeted by chronic deviations in MD [NaCl], the rate of local and systemic angiotensin II generation. Studies of TGF in transgenic mice are expected to provide further insights into the mechanisms mediating between luminal [NaCl] and afferent arterioles. TGF responses are virtually abolished in mice in which either the AT1A gene or the angiotensin converting enzyme gene is rendered nonfunctional by homologous recombination. In contrast, TGF responses are unaltered in nitric oxide synthase I knockout mice. Thus, an intact renin-angiotensin system appears to be critical for the TGF signaling pathway.

Animals↗
PubMed1998

Inhibition of adenosine-1 receptor-mediated preglomerular vasoconstriction in AT1A receptor-deficient mice.

The effect of the adenosine type 1 receptor agonist N6-cyclohexyladenosine (CHA) on glomerular vascular reactivity was studied in male angiotensin II type 1A (AT1A) receptor knockout mice (9). Vascular reactivity was assessed as the response of stop-flow pressure (PSF) to infusion of CHA into loops of Henle using micropuncture techniques. In AT1A +/+ mice at ambient arterial blood pressure (96.7 +/- 2.8 mmHg), the presence of CHA (10 (-5) M) in the perfusate increased PSF responses from 6.8 +/- 0.6 to 14.3 +/- 0.9 mmHg when the loop of Henle of the index nephron was perfused and from 0.7 +/- 0.3 to 12.3 +/- 1.0 mmHg when the loop of an adjacent nephron was perfused. At reduced arterial blood pressure (82.8 +/- 1. 3 mmHg), index nephron perfusion with CHA increased PSF responses from 4.5 +/- 0.3 to 9.4 +/- 0.4 mmHg. In AT1A -/- mice with a mean arterial blood pressure of 80 +/- 1.9 mmHg, CHA increased PSF responses only from 0.1 +/- 0.3 to 3.6 +/- 0.54 mmHg during index nephron perfusion and from 0.25 +/- 0.2 to 2.7 +/- 0.55 mmHg during adjacent nephron perfusion, significantly less than in wild-type animals (P < 0.001). Responses to CHA were intermediate in AT1A +/- mice. Thus AT1A receptor knockout mice show a markedly reduced constrictor response to CHA both in the presence and absence of simultaneous activation of the tubuloglomerular feedback system. These data support the notion of a functional interaction between adenosine and angiotensin II in the regulation of afferent arteriolar tone.

Adenosine↗
PubMed1997

Expression of PTHrP, PTH/PTHrP receptor, and Ca(2+)-sensing receptor mRNAs along the rat nephron.

To provide a frame of reference for studies of renal divalent cation and phosphate metabolism, we assessed the cellular localization of kidney calcium receptor (RaKCaR), parathyroid hormone-related protein (PTHrP), and parathyroid hormone/ parathyroid hormone-related protein (PTH/PTHrP) receptor mRNA. The studies used using reverse transcription-polymerase chain reaction (RT-PCR) applied to cDNA prepared from dissected rat nephron segments and from primary cultures of mouse juxtaglomerular granular cells. With species-specific primers, PCR products of expected size were obtained for RaKCaR (967 bp), PTHrP (420 bp), and PTH/PTHrP receptor (817 bp), with product identity being confirmed by restriction digestion. RaKCaR mRNA was found in medullary and cortical thick ascending limbs (MTAL and CTAL, respectively), the macula densa-containing segment, distal convoluted tubules (DCT), and, to a lesser extent, in cortical collecting ducts (CCD). It was not found in glomeruli, proximal convoluted and straight tubules (PCT and PST, respectively), outer and inner medullary collecting ducts (OMCD and IMCD, respectively), or in juxtaglomerular granular cell isolates. PTHrP mRNA was predominantly expressed in glomeruli and at lower levels in PCT and the macula densacontaining segment but was not detectable in CTAL, MTAL, DCT, and CD segments. Presence of PTH/PTHrP receptor mRNA was demonstrated in glomeruli, PCT, PST, CTAL, MTAL, and DCT but not in CD segments. These results suggest that the function of TAL and DCT cells, in addition to being affected by PTH, may be directly altered by extracellular divalent cations through RaKCaR and that PTHrP may act in the glomerulus and proximal tubule as an autocrine or paracrine regulator of hemodynamics and phosphate transport.

Animals↗
PubMed1997

Absence of tubuloglomerular feedback responses in AT1A receptor-deficient mice.

Experiments were performed in a recently generated strain of mice with an angiotensin II AT1A-receptor null mutation (M. Ito, M. I. Oliverio, P. J. Mannon, C. F. Best, N. Maeda, O. Smithies, and T. M. coffman. Proc. Natl. Acad. Sci. USA 92: 3521-3525, 1995) to examine the effects of chronic AT1A receptor deficiency on tubuloglomerular feeback (TGF) responses. All animals were genotyped by polymerase chain reaction using primers designed to amplify sequences from the deleted AT1A gene and from the neomycin resistance gene. Normal mice (AT1A +/+) and mice heterozygous (AT1A +/-) and homozygous (AT1A -/-) for the gene disruption were anesthetized, and stop-flow pressures (PSF) were determined during changes in loop perfusion rate with previously established micropuncture methods. In five AT1A +/+ mice (26 tubules) mean PSF at zero loop flow was 37.2 +/- 1.5 mmHg, falling to 28.2 +/- 1.9 mmHg at a flow of 45 nl/min (P < 0.0001). Flow rate causing the half-maximum response (V1/2) was 8.7 +/- 0.4 nl/min. In four AT1A +/- animals (19 tubules) mean PSF at zero flow was 39.9 +/- 2.4 mmHg, falling to 34.8 +/- 2.7 mmHg at 45 nl/min (mean V1/2 8.6 +/- 1.04 nl/min). In five AT1A -/- mice (24 tubules) PSF was not significantly affected by loop flow with PSF averaging 33.9 +/- 1.7 mmHg at zero flow and 33.2 +/- 1.6 mmHg at 45 nl/min (not significant). Mean arterial blood pressures in the anesthetized and laparotomized mice were 91.8 +/- 2.2, 97.1 +/- 3, and 80.7 +/- 3.2 mmHg in the AT1A +/+, AT1A +/-, and AT1A -/- animals, respectively. Blood pressure responses to exogenous angiotensin II were greatly blunted in the AT1A -/- mice. We conclude that AT1A receptor-mediated effects of angiotensin II are in essential component of TGF responsiveness under chronic conditions. Our studies show the feasibility of using complex micropuncture methods in mice, an approach that widens the potential of genetically altered mouse strains as experimental models.

Animals↗
PubMed1996

A study on carcinogenesis of endogenous nitrite and nitrosamine, and prevention of cancer.

Cancer patients show a salivary NO2- density that is twice that of healthy individuals. This indicates that endogenous NO2- is correlated with the cancer. By applying a culture medium modified by vitamin B1 or KCIO3, we have found that many bacteria in the human body (especially the bowels) can nitrify NH4+ to NO2-. NO2- can be combined with a second amine to form nitrosamine which is a carcinogen. We fed mice using G+ coccus, which can lead to a precancerous lesion and mammary carcinoma, and proved that nitrification in vivo is carcinogenic. We have tested a method to prevent cancer in 7392 workers. Patients with a salivary density of NO2- over 10 ppm for 3 consecutive days and with light symptoms were our targets for preventive treatment using antibiotics and nitrosamine destroyers. The result shows that the average cancer incidence rate in the control group is 58.4% higher than in the experimental group over 9 years. The difference between the two groups is statistically significant (x2 = 8.5, p < 0.01). This is reliable evidence that preventive treatment is effective when based on the idea that endogenous NO2- and nitrosamine are the carcinogens.

Adult↗
PubMed1996

Percutaneous balloon valvuloplasty for pulmonic stenosis in adolescents and adults.

BACKGROUND: Percutaneous balloon valvuloplasty has been the accepted first-line treatment for congenital pulmonic stenosis in children. Its efficacy in adolescents and adults is less well defined. METHODS: Between December 1985 and July 1995 we performed percutaneous pulmonic valvuloplasty with a single Inoue balloon catheter in 53 adolescent or adult patients 13 to 55 years of age (mean [+/- SD], 26 +/- 11). Follow-up studies were performed 0.2 to 9.8 years after the procedure (mean, 6.9 +/- 3.1) by Doppler echocardiography (in all the patients) and by cardiac catheterization and angiography (in nine patients). RESULTS: After balloon valvuloplasty, the systolic pressure gradient across the pulmonic valve decreased from 91 +/- 46 mm Hg to 38 +/- 32 mm Hg (P < 0.001), and the diameter of the pulmonic-valve orifice increased from 8.9 +/- 3.6 mm to 17.4 +/- 4.6 mm (P < 0.001). In the nine patients catheterized at follow-up, the systolic gradient decreased from 107 +/- 48 mm Hg before valvuloplasty to 50 +/- 29 mm Hg after valvuloplasty and to 30 +/- 16 mm Hg at follow-up (P < 0.001 for the comparison of the gradient before and after valvuloplasty; P < 0.001 for the comparison before valvuloplasty and at follow-up; and P < 0.05 for the comparison after valvuloplasty and at follow-up). In the same nine patients, the diameter of the pulmonic valve, as measured by right ventricular angiography, increased from 8.3 +/- 1.4 mm before valvuloplasty to 17.2 +/- 2.0 mm after valvuloplasty (P < 0.001) and to 18.4 +/- 1.4 mm at follow-up (P = 0.08). Incompetence of the pulmonic valve was noted in 7 of the 53 patients (13 percent) after balloon valvuloplasty, but it had disappeared at follow-up in all of them. CONCLUSIONS: Patients with congenital pulmonic stenosis who present in late adolescence or adult life can be treated with percutaneous balloon valvuloplasty with excellent short-term and long-term results that are similar to those in young children.

Adolescent↗
PubMed1996

Effect of transmembrane Ca2+ gradient on the coupling of beta-adrenergic receptors and adenylyl cyclase.

In order to investigate the effect of transmembrane Ca2+ gradient on Gs mediated coupling of beta-AR and adenylyl cyclase, beta-AR from duck erythrocytes and Gs and adenylyl cyclase from bovine brain cortices were co-reconstituted into asolectin liposomes with different transmembrane Ca2+ gradient. These proteoliposomes were proven to be impermeable to water-soluble substances. The results obtained indicate that a physiological transmembrane Ca2+ gradient (1000-fold) is essential for higher stimulation of adenylyl cyclase by hormone-activated beta-AR via coupling to Gs and can be further enhanced by the decrease of such Ca2+ gradient within certain range (100 fold) following Ca2+ influx into cells during signal transduction. Fluorescence polarization of DPH revealed that transmembrane Ca2+ gradient modulates adenylyl cyclase and its stimulation by hormones through mediating a change in lipid fluidity. Correspondent conformational changes of beta-AR were also detected from the fluorescence spectra and quenching of Acrylodan-labelled beta-AR in those proteoliposomes. It is suggested that a proper transmembrane Ca2+ gradient is essential for the optimal fluidity of the phospholipid bilayer in the proteoliposomes, which favors the formation of a suitable conformation of the reconstituted beta-AR and thus promotes the stimulation of adenylyl cyclase activities by hormone-activated beta-AR via Gs.

Adenylyl Cyclases↗
PubMed1996

A comparative study of early postoperative hypoxemia in infants, children, and adults undergoing elective plastic surgery.

To determine the influence of age on postoperative hypoxemia, we studied postoperative hypoxemia in 1152 patients, from infants to adults, ASA physical status I, undergoing elective plastic surgery. Subjects were divided into four groups on the basis of age: Group 1, infants aged 1 yr or less (n = 108); Group 2, children aged 1-3 yr (n = 240); Group 3, children aged 3-14 yr (n = 482); and Group 4, adults aged 14-58 yr (n = 322). Arterial oxygen saturation (Spo2) levels were recorded while patients were breathing room air in the postanesthesia recovery room shortly after arrival (0 min), and 5, 10, 15, 20, 30, 40, 50, 60, 120, and 180 min thereafter. Younger patients showed lower Spo2 levels and a higher incidence of hypoxemia during the early postoperative period. The incidences of hypoxemia (Spo2 = 86%-90%) and severe hypoxemia (Spo2 < or = 85%) in the recovery room were 30.6% and 16.7%, respectively, in Group 1, 20.0% and 10.0% in Group 2, 14.1% and 3.3% in Group 3, and 7.8% and 0.6% in Group 4. Hypoxemia occurred most commonly within 1 h after anesthesia, particularly during the first 40 min in infants and during the first 15 min in older children and adults. A significant correlation was found by linear regression analysis between low Spo2 levels on admission to the recovery room and children's age. Thereafter, Spo2 levels and the incidence of hypoxemia during the early postoperative period were related only to infants' recovery scores.

Adolescent↗
PubMed1996

Observation of early postoperative hypoxaemia in children undergoing elective plastic surgery.

The incidence, severity and duration of early postoperative hypoxaemia were determined in 420 healthy infants and children undergoing elective plastic surgery. Mean value of preoperative SpO2 was 98.3 +/- 0.87%. However, it decreased significantly to 92.7 +/- 4.01% immediately following anaesthesia in the postanaesthesia recovery room (PAR) when children breathed room air. Then SpO2 gradually increased to 93.8 +/- 4.25% at 10 min, 95.4 +/- 3.14% at 30 min, 96.6 +/- 2.25% at 60 min, 96.9 +/- 1.69% at 120 min, and 97.4 +/- 1.55% at 180 min, respectively. The incidence of hypoxaemia and severe hypoxaemia, which were 14.8% and 6.2% respectively on arrival in the PAR, decreased significantly to 6.6% and 1.9%, 30 min later. The hypoxaemia occurred most commonly within 1 h, particularly the first 30 min following anaesthesia. The hypoxaemia was more common and more severe in children undergoing cleft palate repair than other kinds of plastic surgery. A significant correlation was found between low SpO2 levels on admission to the PAR and children's age, fentanyl doses, and the infants' recovery score.

Adolescent↗
PubMed1996

Localization of bumetanide- and thiazide-sensitive Na-K-Cl cotransporters along the rat nephron.

The present study was undertaken to investigate the mRNA localization of the two major kidney-specific Na-K-Cl transport proteins, the bumetanide-sensitive cotransporter (NKCC2 in rabbit and BSC1 in rat) and the thiazide-sensitive cotransporter (TSC). NKCC2 from rabbit and mouse has been shown to exist in three isoforms (designated A, B, and F) that differ only in a 96-bp region. The divergent region of each of the three NKCC2 isoforms was cloned from rat kidney by a polymerase chain reaction (PCR)-based strategy, and isoform-specific primers were chosen. RNA and cDNA were prepared from renal cortex and medulla and from microdissected nephron segments. Using reverse transcription (RT)-PCR, the B isoform was detected only in cortex and the F isoform only in medulla, whereas the A from was found in both. In dissected nephron segments, the B form was found exclusively in cortical thick ascending limb (CTAL) and macula densa-containing segment (MDCS), the F form only in medullary thick ascending limb (MTAL) and outer medullary collecting duct, and the A form in CTAL, MDCS, and MTAL. An additional isoform including both A and F exons was identified by direct sequencing of a 592-bp product from medulla. The AF product was found only in the medulla and was localized exclusively in MTAL. TSC mRNA was detected exclusively in the distal convoluted tubule. Differential nephron localization of NKCC2 isoforms suggests that Na-K-Cl cotransporters may differ in their transport characteristics to explain regulation of salt transport along the nephron.

Animals↗
PubMed1996

Regulation of endothelin production and secretion in cultured collecting duct cells by endogenous transforming growth factor-beta.

Confluent cultures of two renal collecting duct cell lines (M-1 and mIMCD-K2 cells derived from cortical and inner medullary collecting ducts, respectively) express endothelin1 (ET1), transforming growth factor-beta (TGF beta; both TGF beta 1 and TGF beta 2), and both types of the TGF beta receptor. Experiments were performed to test whether endogenous TGF beta may be a paracrine modulator of ET1 expression in these cells. Treatment of M-1 and mIMCD-K2 cells with TGF beta 2 antisense oligodeoxynucleotides (ODN) significantly reduced ET1 messenger RNA (mRNA) and ET secretion (as well as TGF beta 2 mRNA) in a concentration-dependent manner, whereas control ODN were without significant effects. To produce ET inhibition, antisense ODN had to be present in the basolateral medium, whereas its sole presence in the apical medium was without effect. In addition, a pan-specific TGF beta antibody caused a significant reduction of ET1 mRNA expression and ET1 secretion. M-1 cells were found to express high levels of the mRNA for plasminogen activator of both tissue and urokinase types. Addition of the nonspecific serine protease inhibitor aprotinin (50 micrograms/ml) to the medium for 24 h significantly reduced the secretion of ET1. These results suggest that secretion of endogenous TGF beta, at least in part activated by the plasminogen/plasmin system, participates in the regulation of ET1 synthesis and secretion by collecting duct cell lines.

Animals↗