RESUMO
We describe the clinical course of a female adolescent who was followed because of isolated microhematuria and hypocomplementemia before admission to hospital with a sudden onset of acute renal failure. At presentation, she exhibited complement consumption through the complement alternative pathway (AP) while other serologic tests were negative. Renal biopsy revealed dense deposit disease (DDD) with a crescentic pattern. Intravenous methylprednisolone, followed by plasma exchange (PE), and intravenous cyclophosphamide pulses were started shortly after admission. C3NeF and anti-factor H antibody tests were negative. Serum factor H and I levels were normal as well as factor H activity. Screening for mutation in the factor H gene revealed the H402 allele variant. Clinical remission, defined as normalization in renal function and in the activity levels of the complement AP, was noted at one month post-presentation and throughout the follow-up. A repeat renal biopsy showed the disappearance of crescent formation, whereas electron microscopy revealed no regression in dense transformation of the lamina densa. In summary, our patient was successfully treated with immunosuppressant and PE. The absence of known factors associated with DDD suggests that, in this particular case, other regulatory mechanisms of complement AP might have been involved in the disease process.
Assuntos
Injúria Renal Aguda/terapia , Ciclofosfamida/uso terapêutico , Glomerulonefrite Membranoproliferativa/terapia , Imunossupressores/uso terapêutico , Metilprednisolona/uso terapêutico , Troca Plasmática , Injúria Renal Aguda/genética , Injúria Renal Aguda/imunologia , Injúria Renal Aguda/patologia , Adolescente , Biópsia , Terapia Combinada , Ativação do Complemento , Fator H do Complemento/genética , Ciclofosfamida/administração & dosagem , Análise Mutacional de DNA , Quimioterapia Combinada , Feminino , Glomerulonefrite Membranoproliferativa/genética , Glomerulonefrite Membranoproliferativa/imunologia , Glomerulonefrite Membranoproliferativa/patologia , Humanos , Imunossupressores/administração & dosagem , Metilprednisolona/administração & dosagem , Mutação , Pulsoterapia , Resultado do TratamentoRESUMO
The role of angiotensin subtype-1 (AT1) and -2 (AT2) receptors in mediating the effects of angiotensin II (ANG II) on several K+ transporters was studied in rat distal colon using an Ussing chamber. Angiotensin II induced K+ secretion at two different doses. Secretion occurred at 10-(8) and 10-(4) M, as a result of an increase in serosal-to-mucosal flux (Js-m). The ANG II-induced stimulation of Js-m at a low dose (10-(8) M) was abolished by PD123319 while losartan did not alter the low-dose ANG II-dependent increase in Js-m. In contrast, the increase in Js-m induced by a high-dose of ANG II (10-(4) M) was blocked by losartan, whereas PD123319 partially reduced the stimulatory effect. In the presence of both blockers, high-dose ANG II induced an inhibition of basal Js-m. Low-dose ANG II activated the barium-sensitive K+ channels, whereas the Na+, K+, 2Cl- cotransporter and the Na+, K+ -ATPase pump were unchanged. At the high dose, ANG II activated the barium-sensitive K+ channels and the Na+, K+, 2Cl- cotransporter and inhibited the Na+, K+ -ATPase pump. These data indicate that ANG II stimulates serosal-to-mucosal K+ flux in the rat distal colon at high and low doses via different receptors and K+ transporters.
Assuntos
Angiotensina II/farmacologia , Colo/efeitos dos fármacos , Colo/metabolismo , Potássio/metabolismo , Receptores de Angiotensina/metabolismo , Antagonistas de Receptores de Angiotensina , Animais , Transporte Biológico Ativo , Proteínas de Transporte/metabolismo , Relação Dose-Resposta a Droga , Imidazóis/farmacologia , Técnicas In Vitro , Losartan/farmacologia , Masculino , Canais de Potássio/efeitos dos fármacos , Canais de Potássio/metabolismo , Piridinas/farmacologia , Ratos , Ratos Sprague-Dawley , Receptor Tipo 1 de Angiotensina , Receptor Tipo 2 de Angiotensina , Simportadores de Cloreto de Sódio-Potássio , ATPase Trocadora de Sódio-Potássio/metabolismoRESUMO
Pyelonephritis is one of the most common febrile diseases in children. If not treated appropriately, it causes irreversible renal damage and accounts for a large proportion of end stage renal failures. Renal scarring can occur in the absence of inflammatory cells, indicating that bacteria may have a direct signalling effect on renal cells. Intracellular calcium ([Ca2+]i) oscillations can protect cells from the cytotoxic effects of prolonged increases in intracellular calcium. However, no pathophysiologically relevant protein that induces such oscillations has been identified. Here we show that infection by uropathogenic Escherichia coli induces a constant, low-frequency oscillatory [Ca2+]i response in target primary rat renal epithelial cells induced by the secreted RTX (repeats-in-toxin) toxin alpha-haemolysin. The response depends on calcium influx through L-type calcium channels as well as from internal stores gated by inositol triphosphate. Internal calcium oscillations induced by alpha-haemolysin in a renal epithelial cell line stimulated production of cytokines interleukin (IL)-6 and IL-8. Our findings indicate a novel role for alpha-haemolysin in pyelonephritis: as an inducer of an oscillating second messenger response in target cells, which fine-tunes gene expression during the inflammatory response.
Assuntos
Proteínas de Bactérias/fisiologia , Cálcio/metabolismo , Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli , Escherichia coli/patogenicidade , Proteínas Hemolisinas/fisiologia , Rim/microbiologia , Pielonefrite/microbiologia , Animais , Bloqueadores dos Canais de Cálcio/farmacologia , Canais de Cálcio Tipo L/metabolismo , Linhagem Celular , Células Epiteliais/microbiologia , Infecções por Escherichia coli/imunologia , Infecções por Escherichia coli/metabolismo , Estrenos/farmacologia , Feminino , Humanos , Interleucina-6/biossíntese , Interleucina-8/biossíntese , Rim/citologia , Nifedipino/farmacologia , Pirrolidinonas/farmacologia , Ratos , Ratos Sprague-DawleyRESUMO
We recently demonstrated that the ratio between colonic K+ absorptive and K+ secretive pathways was higher in infant than in adult rats. To test the hypothesis that hormones selectively affect these pathways during ontogeny we examined the effect of adrenergic agonists on cellular K+ uptake in distal colon from infant (10-day-old) and adult (50-day-old) rats. Here we describe that adrenaline (10(-5) M) increased total and ouabain-insensitive 86Rb uptake in both age groups, but it did not affect ouabain-sensitive 86Rb uptake. This stimulation was more pronounced in adult than in infant rats. The effect of adrenaline was mediated via beta-adrenergic receptors. Incubation in vitro with beta-agonist, isoproterenol, stimulated SCH-28080-sensitive, i.e. H+, K(+)-ATPase-dependent, 86Rb uptake in adult but not in infant rats. The threshold dose of beta-agonist was at 10(-7) M, and the maximal activation was observed at 10(-5) M. In vivo inhibition of beta-adrenergic system with propranolol caused a significant decrease in H+, K(+)-ATPase-dependent 86Rb uptake in infant but not in adult colon. In conclusion, this study suggests that the higher colonic K+ absorption in infant rats may be as a result of a selective beta-adrenergic up-regulation leading to stimulation of the apical H+, K(+)-ATPase.
Assuntos
Agonistas Adrenérgicos beta/farmacologia , Colo/efeitos dos fármacos , Isoproterenol/farmacologia , Potássio/metabolismo , Animais , Colo/crescimento & desenvolvimento , Colo/metabolismo , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Epinefrina/farmacologia , ATPase Trocadora de Hidrogênio-Potássio/metabolismo , Imidazóis/farmacologia , Ouabaína/farmacologia , Propranolol/farmacologia , Inibidores da Bomba de Prótons , Ratos , Ratos Sprague-Dawley , Rubídio/metabolismoRESUMO
Vasopressin plays an essential role for the regulation of water balance by activating the collecting duct-specific water channel, aquaporin-2 (AQP2). Here we present evidence that vasopressin may also act as a long-term, transcriptional regulator of AQP2. The studies were performed on LLC-PK1 cells, which normally express V2 receptor (V2R) and which were transfected with a fragment of the human AQP2 promoter. Activation of the adenylate cyclase-coupled V2R in LLC-PK1 cells induced phosphorylation of adenosine 3',5'-cyclic monophosphate (cAMP) responsive element binding protein (CREB) and expression of c-Fos. Binding of these factors to the CRE and AP1 site did, in combination, lead to AQP2 promoter activation. These results establish the role of vasopressin as a regulator of transcription and are the first example of how a message from a highly specific receptor is, via a dual effect of the cAMP signal on CREB and immediate early gene expression, transduced to the transcription of a final target protein with known biological effects.
Assuntos
Adenilil Ciclases/metabolismo , Aquaporinas , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Canais Iônicos/genética , Regiões Promotoras Genéticas , Receptores de Vasopressinas/fisiologia , Fator de Transcrição AP-1/metabolismo , Vasopressinas/farmacologia , Animais , Aquaporina 2 , Aquaporina 6 , Sítios de Ligação , Linhagem Celular , Cloranfenicol O-Acetiltransferase/biossíntese , AMP Cíclico/metabolismo , Desamino Arginina Vasopressina/farmacologia , Regulação da Expressão Gênica , Humanos , Canais Iônicos/biossíntese , Cinética , Modelos Biológicos , Dibutirato de 12,13-Forbol/farmacologia , Fosforilação , Regiões Promotoras Genéticas/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-fos/biossíntese , Proteínas Proto-Oncogênicas c-jun/biossíntese , Proteínas Recombinantes de Fusão/biossíntese , Transdução de Sinais , TransfecçãoRESUMO
The development of gastric H,K-ATPase from fetal to adult life was studied in the rat. The alpha and beta H,K-ATPase mRNA abundance, the protein abundance, and the enzyme activity increased postnatally. The sharpest increase in mRNA and enzyme activity was observed in the weaning period. Several intestinal enzymes are known to be stimulated by glucocorticoids at the time of weaning. To study the role of glucocorticoids in the maturation of gastric H,K-ATPase, we treated 10-d-old rats with a single injection of betamethasone. Twenty-four hours after betamethasone injection, the enzyme activity was significantly higher than in the control animals (2.6-fold, p < 0.05). The abundance of catalytic alpha H,K-ATPase protein was also increased (2.5-fold, p < 0.01). The time-dependent effect of betamethasone on alpha H,K-ATPase mRNA was determined from 6 to 24 h after treatment. Glucocorticoids did not significantly alter the mRNA abundance within 18 h. Twenty-four hours after injection, the gastric H,K-ATPase mRNA was significantly increased compared with controls (2.8- and 2.2-fold increase for alpha and beta subunits, respectively, P < 0.01 for both). In conclusion this study indicates that glucocorticoids may regulate the long-term maturation of gastric H,K-ATPase by indirectly stimulating enzyme synthesis.
Assuntos
Betametasona/administração & dosagem , ATPase Trocadora de Hidrogênio-Potássio/metabolismo , Estômago/enzimologia , Animais , Feminino , Glucocorticoides/administração & dosagem , ATPase Trocadora de Hidrogênio-Potássio/genética , Gravidez , Ratos , Ratos Sprague-DawleyRESUMO
Infants need to retain more K+ than adults to avoid growth retardation. Since the K+ requirements are different in infants (I) and in adults (A), the mechanisms regulating K+ homeostasis should also be different. The colon plays an important role for the regulation of K+ homeostasis. Colonic K+ transport is bidirectional. In this study we have examined the development of colonic K+ transport with special reference to the contribution of different K(+)-transporting pathways. The net colonic K+ uptake, as determined by in vivo perfusion studies and by 86Rb uptake, was significantly higher in I than in A rats. In both I and A colon, approximately 80% of total 86Rb uptake was dependent on vanadate-sensitive P-type adenosinetriphosphatases (ATPases), but the contribution of these different ATPases changes during development. The activity of colonic Na(+)-K(+)-ATPase, measured as ouabain-sensitive Na(+)-dependent ATP hydrolysis and as 86Rb uptake, was lower in I than in A rats. In contrast, the activity of K(+)-ATPases located in apical membrane and measured as ouabain insensitive and SCH-28080 sensitive, as ouabain-sensitive Na(+)-independent ATP hydrolysis, and as 86Rb uptake was significantly higher in I than in A rats. The ratio between apically located K(+)-ATPases and basolateral Na(+)-K(+)-ATPase activities was almost 3.2-fold higher in I than in A colon. We identified with Northern blot the expression of the colonic H(+)-K(+)-ATPase and the Na(+)-K(+)-ATPase alpha-subunits. The alpha-mRNA expression of both ATPases was significantly higher in I than in A rats. The pH and K+ sensitivity of the ouabain-insensitive, SCH-28080-sensitive K(+)-ATPase was the same in I and A colons. In conclusion, the relative activity of apical K+ absorbing ATPases is higher in the I than in the A colon, which should aid infants in retaining K+.
Assuntos
Envelhecimento/metabolismo , Animais Recém-Nascidos/metabolismo , Colo/metabolismo , Potássio/metabolismo , Adenosina Trifosfatases/metabolismo , Animais , Animais Recém-Nascidos/crescimento & desenvolvimento , Transporte Biológico , Proteínas de Transporte de Cátions , ATPase Trocadora de Hidrogênio-Potássio/metabolismo , Microvilosidades/metabolismo , Ratos , Ratos Sprague-Dawley , Rubídio/farmacocinética , ATPase Trocadora de Sódio-Potássio/metabolismoRESUMO
Previous studies of hormonal regulation of renal Na(+)-K(+)-ATPase have indicated that the activity of the sodium pump is regulated by phosphorylation-dephosphorylation reactions. Here we report that okadaic acid (OA) and calyculin A (CL-A), inhibitors of protein phosphatase (PP)-1 and PP-2A, inhibited Na(+)-K(+)-ATPase activity in cells from the rat thick ascending limb (TAL) of loop of Henle in a dose-dependent manner. CL-A was 10-fold more potent than OA. On the basis of the inhibitory constant values of CL-A and OA for PP-1 and PP-2A, it is concluded that the tubular effect is mainly due to inhibition of PP-1. In situ hybridization studies with oligonucleotide probes revealed very strong PP-1 alpha and PP-1 gamma 1 mRNA labeling in the outer stripe of the outer medulla, strong labeling in the inner stripe of the outer medulla, and weak labeling in the inner medulla. Very weak labeling was demonstrated in the outer cortex. PP-1 beta mRNA labeling was very strong in the inner stripe of the outer medulla, whereas the outer stripe had weaker labeling, and the inner medulla had weak labeling. PP-1 alpha, PP-1 beta, and PP-1 gamma 1 mRNA were also demonstrated in the transitional epithelium of the ureter. The abundance of the PP-1 alpha and PP-1 gamma isoforms as measured by immunoblotting was very high in tissue from the outer medulla, which also has a high abundance of the endogenous dopamine-regulated PP-1 inhibitor, DARPP-32.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Medula Renal/enzimologia , Rim/enzimologia , Fosfoproteínas Fosfatases/fisiologia , ATPase Trocadora de Sódio-Potássio/metabolismo , Animais , Sequência de Bases , Immunoblotting , Hibridização In Situ , Masculino , Dados de Sequência Molecular , Sondas de Oligonucleotídeos/genética , Fosfoproteínas Fosfatases/antagonistas & inibidores , Proteína Fosfatase 1 , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
Several epithelial ion transporters are developmentally regulated in the preweaning period, at the time when the circulating levels of glucocorticoid and mineralocorticoid hormones increase. The specific role of glucocorticoids and mineralocorticoids in the maturation of epithelial ion transport is still disputed. In this study, we investigated the effect of corticosteroids on the mRNA expression of ion transporters in the infant rat colon, a glucocorticoid- and mineralocorticoid-sensitive organ. The expression of the Na,K-ATPase, the H,K-ATPase and the amiloride-sensitive Na+ channel mRNA was investigated in control rats from fetal to adult life. We found that the mRNA of the three transporters is temporarily up-regulated in the preweaning period. Rats were then injected with a single dose of betamethasone or aldosterone at 10 d of age. The main effect was the glucocorticoid stimulation of the Na,K-ATPase mRNA within 6 h (4-fold). Glucocorticoids did not alter H,K-ATPase nor Na+ channel mRNA within 6 h. Aldosterone moderately (1.7-fold) stimulated Na+ channel within 6 h, but dit not alter Na,K-ATPase nor H,K-ATPase mRNA. Twenty-four hours after injection, both glucocorticoids and mineralocorticoids had less pronounced and distinct effects. In tissue with lower aldosterone receptor abundance (renal cortex) or with no aldosterone receptor (stomach), glucocorticoids induce a similarly rapid increases in Na,K-ATPase mRNA (4-fold within 6 h), whereas aldosterone had no effect within 6 h. However, glucocorticoids did not stimulate Na,K-ATPase mRNA in the brain, a tissue rich in glucocorticoid receptors.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Colo/efeitos dos fármacos , Glucocorticoides/metabolismo , ATPase Trocadora de Hidrogênio-Potássio/metabolismo , Mineralocorticoides/metabolismo , RNA Mensageiro/biossíntese , ATPase Trocadora de Sódio-Potássio/metabolismo , Amilorida/farmacologia , Animais , Colo/metabolismo , Estudos de Avaliação como Assunto , Transporte de Íons/fisiologia , Masculino , Ratos , Ratos Sprague-Dawley , Canais de Sódio/efeitos dos fármacosRESUMO
Glucocorticoids modulate the maturation of Na(+)-K(+)-ATPase mRNA in a tissue- and age-dependent manner. In this study, we report the effect of glucocorticoids on Na(+)-K(+)-ATPase gene transcription in the infant rat kidney. Ten-day-old rats were treated with one intraperitoneal injection of betamethasone. In glucocorticoid-treated rats, there was a significant increase in renal cortical alpha 1- and beta 1-mRNAs (3.08 +/- 0.34- and 4.06 +/- 0.10-fold). Pretreatment with cycloheximide, an inhibitor of protein synthesis, did not abolish the increase in alpha 1- and beta 1-mRNA after glucocorticoids. The alpha 1- and beta 1-gene transcription rates were significantly increased in nuclei isolated from kidneys of glucocorticoid-treated rat (2.16 +/- 0.05- and 3.12 +/- 0.50-fold). Interaction between nuclear proteins and Na(+)-K(+)-ATPase alpha 1-promoter was studied by gel retardation assay. Nuclear protein from glucocorticoid-treated rats retarded a fragment of alpha 1-promoter that includes a half-consensus glucocorticoid response element (GRE) at position -750 bp but did not retard a fragment including a half-consensus GRE at position -481. Retardation of alpha 1-promoter was inhibited by incubation with molar excess of GRE or with a monoclonal antibody against glucocorticoid receptor. We conclude that in the infant kidney, glucocorticoids directly stimulate the transcription of alpha 1- and beta 1-Na(+)-K(+)-ATPase subunits. It is likely that the binding of glucocorticoid receptor to alpha 1-Na(+)-K(+)-ATPase promoter requires the presence of an auxiliary factor.
Assuntos
Glucocorticoides/farmacologia , Rim/fisiologia , ATPase Trocadora de Sódio-Potássio/genética , Transcrição Gênica/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Betametasona/farmacologia , Northern Blotting , Cicloeximida/farmacologia , Rim/enzimologia , Masculino , Proteínas Nucleares/fisiologia , Regiões Promotoras Genéticas , Ratos , Ratos Sprague-DawleyRESUMO
The coupling between the Na+/glucose cotransporter and Na(+)-K(+)-ATPase (NKA) described for epithelial cells (1) prompted us to study in rats with streptozocin-induced diabetes the effect of increased tubular glucose load on tubular Na+ reabsorption, NKA-dependent O2 consumption (QO2), and NKA activity. Filtered glucose is mainly reabsorbed in the proximal tubuli via the phlorizin-sensitive Na+/glucose cotransporter. In this study, the diabetic rats had a significantly higher renal blood flow (RBF), glomerular filtration rate (GFR), and Na+ reabsorption than the control rats. Total renal QO2 as well as QO2 in cortical tissue, which consists mainly of proximal tubular cells, was significantly higher in diabetic than in control rats. The increase in tissue QO2 was entirely caused by increased NKA-dependent QO2. NKA activity, measured as rate of ATP hydrolysis, was increased in cortical tubular but not glomerular tissue from diabetic rats. Phlorizin treatment abolished the increase in NKA activity, Na+ reabsorption, and QO2, as well as the increase in RBF and GFR in diabetic rats. We conclude that diabetes is associated with increased renal O2 metabolism secondary to the increase in coupled Na+ reabsorption via the Na+/glucose cotransporter and NKA. The increased oxygen consumption might contribute to the hyperperfusion and hyperfiltration in the diabetic kidney.
Assuntos
Diabetes Mellitus Experimental/metabolismo , Córtex Renal/metabolismo , Túbulos Renais Proximais/metabolismo , Consumo de Oxigênio , ATPase Trocadora de Sódio-Potássio/metabolismo , Sódio/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Diabetes Mellitus Experimental/fisiopatologia , Taxa de Filtração Glomerular , Rim/fisiopatologia , Cinética , Masculino , Proteínas de Transporte de Monossacarídeos/metabolismo , Ouabaína/farmacologia , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Fluxo Sanguíneo Regional , Circulação Renal , ATPase Trocadora de Sódio-Potássio/biossínteseRESUMO
This review focuses on the ontogeny of factors involved in the transcriptional regulation of Na+,K(+)-ATPase expression. The Na+,K(+)-ATPase enzyme is of vital importance for cell function. It is likely that the limited availability of Na+,K(+)-ATPase in infant tissue is the major limiting factor for adaptation to extra-uterine life in several organs. The factors regulating Na+,K(+)-ATPase gene transcription in infancy are discussed. Special emphasis is given to the role of circulating hormones such as glucocorticoids and thyroxine.
Assuntos
Regulação Enzimológica da Expressão Gênica , Rim/enzimologia , ATPase Trocadora de Sódio-Potássio/genética , Animais , Diferenciação Celular , Glucocorticoides/fisiologia , Humanos , Tiroxina/fisiologia , Transcrição GênicaRESUMO
We examined the effect of a high-salt (HS) diet on the regulation of renal cortical Na(+)-K(+)-adenosinetriphosphatase (Na(+)-K(+)-ATPase) in young Dahl salt-sensitive (DS) and salt-resistant (DR) rats. The activity of Na(+)-K(+)-ATPase, determined in permeabilized proximal tubule segments, was similar in DS and DR rats on normal salt (NS) diet. HS diet resulted in a twofold increase in proximal tubule Na(+)-K(+)-ATPase activity in DS rats but not in DR rats. The mRNA abundance, which was also similar in DS and DR rats on NS diet, increased after 2 days on HS diet in both innervated and denervated kidneys from DS rats but had no effect in DR rats. The activity of Na(+)-K(+)-ATPase and the content of alpha 1- and beta-protein in cortical homogenate were similar in DS and DR rats on both NS and HS diets. Treatment with benserazide, an inhibitor of dopa decarboxylase, upregulated proximal tubule Na(+)-K(+)-ATPase activity and increased Na(+)-K(+)-ATPase mRNA in DR rats on HS diet. Taken together, these data indicate that there is a primary defect in the dynamic hormonal regulation of Na(+)-K(+)-ATPase activity in intact tubular cells, which might stimulate Na(+)-K(+)-ATPase transcription.
Assuntos
Córtex Renal/enzimologia , RNA Mensageiro/metabolismo , Sódio na Dieta/administração & dosagem , ATPase Trocadora de Sódio-Potássio/genética , Animais , Western Blotting , Denervação , Rim/inervação , Túbulos Renais Proximais/enzimologia , Masculino , Ratos , Ratos Endogâmicos , ATPase Trocadora de Sódio-Potássio/metabolismoRESUMO
The postnatal maturation of Na+,K(+)-ATPase alpha- and beta-subunit genes can be accelerated in the rat kidney by the administration of glucocorticoid hormones (GC). In heart, Na+,K(+)-ATPase alpha-isoform and beta-subunit genes exhibit a complex pattern of expression during development. This study examines the role of GC in the regulation of Na+,K(+)-ATPase mRNA abundance in rat heart during infancy. In 10-d-old rats given injections with a single intraperitoneal dose of betamethasone or diluent, the Na+,K(+)-ATPase activity was 2-fold higher in treated than in control rats after 24 h. GC differentially regulated the mRNA for Na+,K(+)-ATPase subunits. A significant increase in Na+,K(+)-ATPase mRNA occurred with a dose of 2.5 micrograms betamethasone/100 g body weight. The following experiments were performed with a saturating dose of 60 micrograms betamethasone/100 g body weight. The alpha 1 mRNA was moderately but significantly increased (1.5-fold) 6 h after treatment. The mRNA for the alpha 2 subunit increased 2.2-fold after betamethasone treatment. The mRNA for beta 1 was numerically increased after 20 min (1.3-fold); it was 1.5-fold higher (p < 0.05) after 1 h and was 3-fold higher after 6 h (p < 0.01). Betamethasone treatment did not significantly change the abundance of the mRNA for the alpha 3 subunit. The expression of actin mRNA was not altered after GC. These data indicate that GC hormones may act as a "molecular switch" in the developmental expression of the mRNA for the Na+,K(+)-ATPase alpha-isoforms and contribute in stimulating the maturation of rat heart during the preweaning period.
Assuntos
Betametasona/farmacologia , RNA Mensageiro/metabolismo , ATPase Trocadora de Sódio-Potássio/metabolismo , Animais , Animais Lactentes , Regulação da Expressão Gênica/efeitos dos fármacos , Coração/efeitos dos fármacos , Coração/crescimento & desenvolvimento , Isoenzimas/genética , Isoenzimas/metabolismo , Masculino , Miocárdio/enzimologia , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , ATPase Trocadora de Sódio-Potássio/genética , Regulação para Cima/efeitos dos fármacosRESUMO
We have previously reported that in the infant rat renal cortex, a saturating dose of glucocorticoid hormones (GC) rapidly increases the abundance of Na+,K(+)-ATPase mRNA. We now show that this effect is dose dependent. In the renal cortex of 10-d-old rats, an increase in renal Na+,K(+)-ATPase mRNA occurs with 2.5 micrograms betamethasone/100 g body weight. In subsequent experiments, performed 6 h after a saturating dose (60 micrograms/100 g body weight), we show that the effect is age dependent. The most marked effects on renal cortical alpha-mRNA were found at 10 d of age (5.3- +/- 0.9-fold). A significant increase was also found in 20-d-old rats (1.6- +/- 0.2-fold), but no effect was found in fetal and 5-d-old rats. Studies were also performed on the lung, where the most marked effect was noted in the perinatal period (2.0- +/- 0.1-fold 2 d before birth and 1.76 +/- 0.2 at 5 d of age), but no effect on alpha-mRNA was found at 10 and 20 d. In one protocol, the effect of betamethasone on renal Na+K(+)-ATPase mRNAs abundance was determined in adult adrenalectomized rats. In these rats, betamethasone induced a significant 1.6- +/- 0.2-fold and 1.8- +/- 0.3-fold increase in renal Na+,K(+)-ATPase mRNA. This effect, however, was significantly smaller than the increase induced in intact 10-d-old rats. GC induction of Na+,K(+)-ATPase mRNA is age and tissue dependent and is dependent on factors other than GC-receptor availability. The GC-sensitive period appears to coincide with the physiologic need for organ maturation.
Assuntos
Betametasona/farmacologia , RNA Mensageiro/metabolismo , ATPase Trocadora de Sódio-Potássio/metabolismo , Fatores Etários , Animais , Animais Recém-Nascidos , Feminino , Feto/efeitos dos fármacos , Feto/enzimologia , Regulação da Expressão Gênica/efeitos dos fármacos , Rim/efeitos dos fármacos , Rim/enzimologia , Rim/crescimento & desenvolvimento , Pulmão/efeitos dos fármacos , Pulmão/enzimologia , Pulmão/crescimento & desenvolvimento , Gravidez , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , ATPase Trocadora de Sódio-Potássio/genética , Regulação para Cima/efeitos dos fármacosRESUMO
The mechanisms by which adreno-corticoid hormones regulate Na+,K(+)-ATPase in adult kidney were studied in adrenalectomized (Adx) rats. Five days after adrenalectomy, Na+,K(+)-ATPase activity was significantly reduced in the renal cortex homogenate (C = 13.0 +/- 0.8 vs. Adx = 7.1 +/- 0.7 mumol Pi mg-1 protein h-1) and in renal microsomes (C = 30.3 +/- 1.9 vs Adx = 14.6 +/- 1.3 mumol Pi mg-1 protein h-1). Glucocorticoid replacement treatment of adrenalectomized rats with betamethasone (20 micrograms kg-1 body wt twice daily for 5 days) effectively counteracted the observed reduction in Na+,K(+)-ATPase activity. In cortical homogenate the protein level of alpha 1 and beta 1 subunits measured in immunoblots was not significantly different in Adx and control rats, indicating that 5 days after adrenalectomy the alpha 1 and beta 1 subunits were present in renal cortical cells to almost normal extent but could not be assembled into a transmembrane functional unit. In support of this conclusion we found that the protein level of both the alpha 1 and beta 1 subunits was significantly lower (P less than 0.001 for both subunits) in microsomes from Adx than in control rats. The mRNA abundance for alpha 1 and beta 1 subunits were not lower in Adx as compared to control rats 1 and 5 days after surgery. However, if Adx rats were given a single dose of betamethasone (600 micrograms kg-1 body wt), a significant 2-fold increase in both alpha 1 and beta 1 mRNAs was observed (P less than 0.05 for both subunits).(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Córtex Renal/enzimologia , Processamento de Proteína Pós-Traducional , RNA Mensageiro/metabolismo , ATPase Trocadora de Sódio-Potássio/metabolismo , Adrenalectomia , Animais , Betametasona/farmacologia , Immunoblotting , Injeções Intraperitoneais , Córtex Renal/metabolismo , Masculino , Microssomos/enzimologia , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Ratos , Ratos Endogâmicos , Regulação para CimaRESUMO
Dopamine was shown to affect Na+,K(+)-ATPase activity in basolateral membranes of the rat kidney via a pertussis toxin dependent mechanism. In order to examine if some form of pertussis toxin sensitive G-protein is present exclusively in the basolateral membrane of the rat renal cortex we examined the G-protein composition of both apical and basolateral membrane vesicles. Western blots showed an essentially uniform distribution of G alpha total, G alpha S and G beta over the two membranes. Go could not be detected with western blot technique in the vesicle preparations. By contrast, the distribution of ADP-ribosylation with the bacterial toxins pertussis toxin and cholera toxin depended on the amount of detergent in the assay and perhaps other factors, and thus could not be used to evaluate the relative amounts of G-protein subunits. Thus, in contrast to the situation in cultured renal cells, unequal distribution of receptor and G-protein substrates is apparently not paralleled by an unequal distribution of the detected forms of G-proteins under physiological conditions.
Assuntos
Proteínas de Ligação ao GTP/metabolismo , Túbulos Renais/metabolismo , Adenosina Difosfato Ribose/metabolismo , Animais , Toxina da Cólera/farmacologia , Proteínas de Ligação ao GTP/química , Técnicas In Vitro , Córtex Renal/efeitos dos fármacos , Córtex Renal/metabolismo , Túbulos Renais/efeitos dos fármacos , Masculino , Toxina Pertussis , Conformação Proteica , Ratos , Ratos Endogâmicos , Receptores Dopaminérgicos/metabolismo , ATPase Trocadora de Sódio-Potássio/metabolismo , Distribuição Tecidual , Fatores de Virulência de Bordetella/farmacologiaRESUMO
The administration of glucocorticoid hormone (GC) accelerates the postnatal maturation of renal Na(+)-K(+)-ATPase activity. This study examines the role of GC for the regulation of the Na(+)-K(+)-ATPase mRNA abundance in renal cortex during development. In 12- to 14-day-old rats an upsurge in serum GC concentration was accompanied by an increase in Na(+)-K(+)-ATPase activity and by an apparent increase in mRNA abundance. In 10-day-old rats injected with a single intraperitoneal dose of betamethasone (T) or diluent (C) the abundances of alpha 1- and beta-mRNAs were 1.8- to 2-fold higher in T than in C rats after 20 min. The mRNA abundance of both subunits was threefold higher after 1 h (P less than 0.01), and it was six- to sevenfold higher after 6 h (P less than 0.01). In any given sample there was a coordinate change in alpha 1- and beta-mRNAs relative to C rats. GC did not appear to induce the expression of any alternative catalytic subunit. The alpha 2-mRNA was not detectable in any experimental protocol. Furthermore, the ouabain inhibition of the Na(+)-K(+)-ATPase, partially purified from the renal cortex, was the same before and after GC. In adult rats injected with betamethasone neither the alpha 1- nor the beta-mRNA abundance was different at any time after injection from those in adult C rats. The rapid onset of the GC effect on mRNA abundance in infant rats suggests that the hormone directly activates the gene for Na(+)-K(+)-ATPase alpha 1-subunit, as well as beta-subunit in the developing kidney, and that GC thereby plays an important role for the postnatal maturation of the kidney.
Assuntos
Betametasona/farmacologia , Rim/metabolismo , RNA Mensageiro/metabolismo , ATPase Trocadora de Sódio-Potássio/genética , Animais , Animais Recém-Nascidos , Northern Blotting , Indução Enzimática , Córtex Renal/metabolismo , Masculino , Ouabaína/farmacologia , Ratos , Ratos Endogâmicos , ATPase Trocadora de Sódio-Potássio/antagonistas & inibidoresRESUMO
Rats unilaterally nephrectomized in infancy (Nx5) or in adulthood (Nx55) and fed a normal (21%) protein diet were studied at 2, 3 and 6 months after surgery with regard to glomerular filtration rate (GFR) and the development of both albuminuria and focal glomerulosclerosis (FGS) in the remnant kidney. Nx rats were compared with sham-operated animals (S) of corresponding age. The incidence of FGS never exceeded 1.4% in S rats. In Nx5 rats the incidence of FGS was not increased at 2 and 3 months after surgery whereas it was significantly higher (range 10%-27%) than in S rats 6 months after surgery. Urinary albumin excretion was significantly increased in Nx5 rats 2 months after surgery and was even higher 6 months after surgery. In Nx55 rats the incidence of FGS was the same as in Nx5 rats 2, 3 and 6 months after surgery, but urinary albumin excretion was significantly lower than in Nx5 rats 6 months after surgery. The GFR expressed per unit of body weight decreased in both Nx5 and S5 rats from 2 to 6 months, but the decrease was more pronounced in Nx5 rats than in S5 rats. The GFR factored by kidney weight was significantly lower in Nx5 than in any of the other groups at the 6-month follow-up study. We conclude that, as in adults, when unilateral nephrectomy is performed in infancy, FGS will develop in the remnant kidney. More pronounced albuminuria and reduction of GFR may indicate that the glomerular lesion is more severe in rats nephrectomized in infancy than in adulthood.
Assuntos
Glomerulosclerose Segmentar e Focal/patologia , Nefrectomia , Envelhecimento , Animais , Glomerulosclerose Segmentar e Focal/fisiopatologia , Rim/patologia , Testes de Função Renal , Masculino , Ratos , Ratos EndogâmicosRESUMO
In rats unilaterally nephrectomized (NX) in infancy, the compensatory growth of the remnant kidney is due first to hypertrophy and hyperplasia, but after 2 wk only to hyperplasia. We studied proximal tubular adaptation (reabsorption, Na-K-ATPase activity, length, and basolateral membrane area) 2 and 8 wk after NX. The rats were NX at 5 days of age. The size of remnant kidney obtained from uninephrectomized rats was 125% at 2 wk and 179% at 8 wk relative to the appropriate time-related controls. Single-nephron glomerular filtration rate in the uninephrectomized group was doubled relative to controls, whereas fractional reabsorption by the proximal tubule was unchanged. Na-K-ATPase activity per millimeter of proximal convoluted tubule (PCT) was significantly increased in the uninephrectomized group relative to controls at 2 but not 8 wk. The area of the basolateral cell membrane per millimeter PCT was unchanged at both 2 and 8 wk, which suggests that the density of enzyme units inserted in the membrane was increased at 2 but not at 8 wk. PCT length was increased at 8 but not at 2 wk. There was a close correlation between total PCT Na-K-ATPase activity and filtered sodium (r = 0.999) and between total PCT Na-K-ATPase activity and PCT sodium reabsorption (r = 0.998). We conclude that the proximal tubule can adapt to an increased filtered load by increasing the density of transporting units or by increasing the tubular length. The latter stage may be attained only in young growing animals.