RESUMO
The role(s) of one family of polypeptide growth factors in a developing organ system was examined. Renal anlagen (metanephroi) were surgically removed from 13-d-old rat embryos and grown in organ culture for up to 6 d. Over this period of time when placed in serum-free defined media, the metanephroi increased in size and morphologic complexity. Messenger RNAs for both insulin-like growth factors (IGFs), IGF I and IGF II, were present in the metanephroi. Immunoreactive IGF I and IGF II were produced by the renal anlagen and released into culture media. Levels were relatively constant during the 6 d in culture and averaged 3.5 X 10(-9) M IGF I and 8.3 X 10(-9) M IGF II in media removed from metanephroi after contact for 24 h. IGF binding protein activity was not detected in culture media. Growth and development of metanephroi in vitro was prevented by the addition of anti-IGF I or anti-IGF II antibodies to organ cultures. IGF II produced by metanephroi was active in an IGF II biological assay system and addition of anti-IGF II receptor antibodies to organ cultures prevented growth and development, consistent with the action of IGF II in metanephroi being mediated via the IGF II receptor. The data demonstrate production of both IGF I and IGF II by developing rat metanephroi in organ culture. Each of these peptides is necessary for growth and development of the renal anlage to take place in vitro. Our findings suggest that both IGF I and IGF II are produced within the developing metanephros in vivo and promote renal organogenesis.
Assuntos
Fator de Crescimento Insulin-Like II/fisiologia , Fator de Crescimento Insulin-Like I/fisiologia , Rim/embriologia , Animais , Membrana Celular/metabolismo , Técnicas de Cultura , Eletroforese em Gel de Poliacrilamida , Inositol 1,4,5-Trifosfato/biossíntese , Fator de Crescimento Insulin-Like I/biossíntese , Fator de Crescimento Insulin-Like II/biossíntese , Rim/crescimento & desenvolvimento , Rim/metabolismo , Ratos , Ratos EndogâmicosRESUMO
To address the question of insulin-like growth factor (IGF) I localization and synthesis in kidney, we used two complementary experimental approaches: immunohistochemistry of fixed paraffin-embedded rat kidney sections; and measurement of IGF I mRNA in isolated components of the rat nephron, using a highly sensitive and specific solution hybridization assay. Immunostainable IGF I was localized exclusively to principal cells of cortical and medullary collecting ducts. Administration of growth hormone to hypophysectomized rats for 8 d resulted in enhanced immunohistochemical staining of IGF I within collecting ducts, but no detectable IGF I in other portions of the nephron. The abundance of IGF I mRNA was 7-12-fold higher in isolated papillary collecting ducts than in proximal tubules or glomeruli, and was enriched 10-fold compared with whole kidney. Our data demonstrate colocalization of IGF I and IGF I mRNA in the collecting duct, consistent with focal expression of the IGF I gene at this site.
Assuntos
Fator de Crescimento Insulin-Like I/biossíntese , Túbulos Renais Coletores/metabolismo , Túbulos Renais/metabolismo , Somatomedinas/biossíntese , Animais , Autorradiografia , Peso Corporal/efeitos dos fármacos , Feminino , Substâncias de Crescimento/farmacologia , Imuno-Histoquímica , Fator de Crescimento Insulin-Like I/análise , Fator de Crescimento Insulin-Like I/genética , Rim/análise , Rim/efeitos dos fármacos , Rim/crescimento & desenvolvimento , Túbulos Renais Coletores/análise , Masculino , Hibridização de Ácido Nucleico , Tamanho do Órgão/efeitos dos fármacos , Prolactina/farmacologia , RNA Mensageiro/análise , Ratos , Ratos EndogâmicosRESUMO
The renal collecting duct is a site of insulin-like growth factor I (IGF I) synthesis. Epidermal growth factor (EGF) is also synthesized within the kidney in the thick ascending limb of Henle's loop and the distal tubule. EGF has been shown to regulate IGF I expression in nonrenal tissues. To shed light upon a role of EGF in intrarenal regulation of IGF I gene expression, plasma membranes prepared from collecting ducts isolated from rat kidney and collecting ducts themselves were incubated in the presence and absence of recombinant human EGF (hEGF). hEGF enhanced phospholipase C activity in collecting duct plasma membranes establishing the potential for EGF signal transduction at this site. Inclusion of hEGF in suspensions of collecting ducts increased production of immunoreactive IGF I in a concentration-dependent manner. Production was stimulated significantly by addition of 10(-8) or 10(-6) M hEGF to suspensions for 2 h. Levels of IGF I mRNA in collecting ducts were increased 2.8-fold after incubation with 10(-6) M hEGF in vitro. Our findings demonstrate a direct action of hEGF to enhance collecting duct IGF I gene expression in vitro. Such enhancement is likely to reflect an effect of EGF to stimulate IGF I production in the collecting duct of the intact kidney. Since EGF is produced in kidney, our findings are consistent with intrarenal paracrine regulation of IGF I gene expression by EGF.
Assuntos
Fator de Crescimento Epidérmico/farmacologia , Expressão Gênica/efeitos dos fármacos , Fator de Crescimento Insulin-Like I/genética , Túbulos Renais Coletores/metabolismo , Animais , Fosfatos de Inositol/metabolismo , Fator de Crescimento Insulin-Like I/biossíntese , Túbulos Renais Coletores/efeitos dos fármacos , Masculino , RNA Mensageiro/análise , Ratos , Ratos EndogâmicosRESUMO
Na+-independent L-arginine uptake was studied in rabbit renal brush border membrane vesicles. The finding that steady-state uptake of L-arginine decreased with increasing extravesicular osmolality and the demonstration of accelerative exchange diffusion after preincubation of vesicles with L-arginine, but not D-arginine, indicated that the uptake of L-arginine in brush border vesicles was reflective of carrier-mediated transport into an intravesicular space. Accelerative exchange diffusion of L-arginine was demonstrated in vesicles preincubated with L-lysine and L-ornithine, but not L-alanine or L-proline, suggesting the presence of a dibasic amino acid transporter in the renal brush border membrane. Partial saturation of initial rates of L-arginine transport was found with extravesicular [arginine] varied from 0.005 to 1.0 mM. L-Arginine uptake was inhibited by extravesicular dibasic amino acids unlike the Na+-independent uptake of L-alanine, L-glutamine, glycine or L-proline in the presence of extravesicular amino acids of similar structure. L-Arginine uptake was increased by the imposition of an H+ gradient (intravesicular pH less than extravesicular pH) and H+ gradient stimulated uptake was further increased by FCCP. These findings demonstrate membrane-potential-sensitive, Na+-independent transport of L-arginine in brush border membrane vesicles which differs from Na+-independent uptake of neutral and acidic amino acids. Na+-independent dibasic amino acid transport in membrane vesicles is likely reflective of Na+-independent transport of dibasic amino acids across the renal brush border membrane.
Assuntos
Arginina/metabolismo , Membrana Celular/metabolismo , Rim/metabolismo , Microvilosidades/metabolismo , Sódio/farmacologia , Aminoácidos/farmacologia , Animais , Transporte Biológico/efeitos dos fármacos , Carbonil Cianeto p-Trifluormetoxifenil Hidrazona/farmacologia , Concentração de Íons de Hidrogênio , Cinética , Microvilosidades/efeitos dos fármacos , Concentração Osmolar , CoelhosRESUMO
Dietary phosphate (Pi) restriction increases renal Pi reabsorption and induces resistance to the phosphaturic action of parathyroid hormone. Na+-gradient-stimulated Pi transport in membrane vesicles isolated from the renal brush border of experimental animals has been shown to parallel changes in renal Pi reabsorption induced by dietary Pi restriction and in vivo administration of parathyroid hormone. Dietary Pi restriction has been shown to markedly inhibit the phosphaturic response to parathyroid hormone in rats and dogs. Parathyroid hormone has been reported not to decrease the Na+-gradient-stimulated transport of Pi in brush border membrane vesicles isolated from dietary Pi restricted rats unless the rats were administered an acute Pi load prior to killing, however, thyroparathyroidectomy of rats fed a low Pi diet has been reported to increase Na+-gradient-stimulated Pi transport. Using the dietary Pi restricted dog, we demonstrated no significant decrease in renal reabsorption of Pi in response to parathyroid hormone administration. However, significant decreases in Pi transport in brush border membrane vesicles isolated from the kidneys of dietary Pi restricted dogs were observed in response to in vivo parathyroid hormone administration. These data demonstrate that the resistance to the phosphaturic action of parathyroid hormone observed in vivo does not include resistance to the inhibitory effect of parathyroid hormone on Pi transport in brush border membrane vesicles. Thus, the data suggest that parathyroid hormone continues to alter Pi transport characteristics of the brush border membrane in states of Pi depletion despite the resistance to parathyroid hormone seen in vivo.
Assuntos
Rim/metabolismo , Hormônio Paratireóideo/farmacologia , Fosfatos/deficiência , Animais , Transporte Biológico/efeitos dos fármacos , Cálcio/sangue , Creatinina/metabolismo , Cães , Microvilosidades/metabolismo , Fosfatos/metabolismo , Sódio/farmacologiaRESUMO
The uptake of glycine in rabbit renal brush border membrane vesicles was shown to consist of glycine transport into an intravesicular space. An Na+ electrochemical gradient (extravesicular greater than intravesicular) stimulated the initial rate of glycine uptake and effected a transient accumulation of intravesicular glycine above the steady-state value. This stimulation could not be induced by the imposition of a K+, Li+ or choline+ gradient and was enhanced as extravesicular Na+ was increased from 10 mM to 100 mM. Dissipation of the Na+ gradient by the ionophore gramicidin D resulted in diminished Na+-stimulated glycine uptake. Na+-stimulated uptake of glycine was electrogenic. Substrate-velocity analysis of Na+-dependent glycine uptake over the range of amino acid concentrations from 25 microM to 10 mM demonstrated a single saturable transport system with apparent Km = 996 microM and Vmax = 348 pmol glycine/mg protein per min. Inhibition observed when the Na+-dependent uptake of 25 microM glycine was inhibited by 5 mM extravesicular test amino acid segregated dibasic amino acids, which did not inhibit glycine uptake, from all other amino acid groups. The amino acids D-alanine, D-glutamic acid, and D-proline inhibited similarly to their L counterparts. Accelerative exchange of extravesicular [3H]glycine was demonstrated when brush border vesicles were preloaded with glycine, but not when they were preloaded with L-alanine, L-glutamic acid, or with L-proline. It is concluded that a single transport system exists at the level of the rabbit renal brush border membrane that functions to reabsorb glycine independently from other groups of amino acids.
Assuntos
Membrana Celular/metabolismo , Glicina/metabolismo , Rim/metabolismo , Microvilosidades/metabolismo , Sódio/farmacologia , Aminoácidos/farmacologia , Animais , Transporte Biológico/efeitos dos fármacos , Cátions Monovalentes , Cinética , Microvilosidades/efeitos dos fármacos , CoelhosRESUMO
We have previously demonstrated that the cAMP-dependent phosphorylation of two or more proteins (bands IX and V) in membrane vesicles isolated from the renal brush border from kidneys of dogs, was associated with decreased Na+-dependent Pi transport. In the present studies a specific dephosphorylation of band IX was demonstrated in brush border vesicles incubated in the absence of F- which had been used in previous studies to inhibit phosphoprotein phosphatase activity. Dephosphorylation of band IX was 80% complete after 5 min of incubation at which time inhibition of Pi transport in membrane vesicles which had been phosphorylated in the presence of cAMP could no longer be demonstrated. Dephosphorylation of band IX was no different in vesicles from kidneys originating from parathyroidectomized dogs prior to or following the administration of parathyroid hormone in vivo and normal dogs. We conclude that the cAMP-dependent phosphorylation of brush border membrane proteins may mediate a phosphaturic effect of the hormone. Parathyroid hormone-induced phosphaturia may be terminated through the action of a specific membrane phosphoprotein-phosphatase.
Assuntos
Membrana Celular/enzimologia , AMP Cíclico/farmacologia , Rim/enzimologia , Microvilosidades/enzimologia , Fosfatos/metabolismo , Proteínas Quinases/metabolismo , Animais , Transporte Biológico , Cálcio/sangue , Creatinina/metabolismo , Cães , Túbulos Renais/metabolismo , Cinética , Nefrectomia , Glândulas Paratireoides/fisiologia , Fosfatos/sangue , Fosfoproteínas Fosfatases/metabolismo , Fosforilação , TireoidectomiaRESUMO
Renal phosphate (Pi) reabsorption is increased by growth hormone (GH) and decreased by parathyroid hormone (PTH). Na+-stimulated Pi transport across the brush border membrane of the proximal tubule is the initial step in the process of Pi reabsorption. To determine whether changes in Pi reabsorption induced by GH or PTH are accompanied by changes in brush border membrane Na+-gradient-stimulated Pi transport, we examined the effect of in vivo GH and PTH administration and thyroparathyroidectomy on Pi transport by isolated brush border membrane vesicles prepared from canine kidney. In experiments in which the effect of PTH administration was examined, the same animal provided the control kidney (before PTH administration) and the experimental kidney (after PTH administration). The Na+-gradient Pi overshoot in vesicles isolated from normal, GH-treated and thyroparathyroidectomized dogs was increased after in vivo PTH administration. GH administration and thyroparathyroidectomy increased the height of the overshoot compared to normal. PTH administration decreased the apparent V value by 44% in vesicles from normal animals. The apparent V value was increased, compared to normal, by GH (34%) and thyroparathyroidectomy (57%). PTH administration decreased the apparent V in both the latter groups. GH administration to thyroparathyroidectomized dogs further increased the apparent V. Changes in the apparent V paralleled changes in Pi reabsorption in vivo induced by experimental manipulations. We conclude that changes in renal Pi reabsorption induced by GH were like those induced by PTH, accompanied by changes in the Na+-stimulated Pi transport system in the renal brush border membrane, and that the effect of PTH on vesicular Pi transport in GH-treated dogs did not differ from the effect on vesicles from normal animals.
Assuntos
Hormônio do Crescimento/fisiologia , Rim/metabolismo , Hormônio Paratireóideo/fisiologia , Fosfatos/metabolismo , Animais , Transporte Biológico , Cães , Glucose/metabolismo , Técnicas In Vitro , Cinética , Microvilosidades/metabolismo , Sódio/fisiologiaRESUMO
We previously demonstrated inhibition of Na+-dependent 32Pi transport in canine renal brush-border membranes in association with NAD+-induced ADP ribosylation of membrane protein(s) and postulated that NAD+ inhibits Pi transport across the brush-border membrane via ADP ribosylation. Recently it was shown that incubation of rat brush-border membrane with NAD+ resulted in release of Pi which was prevented by EDTA. It was proposed that NAD+-mediated inhibition of 32Pi transport might occur through this mechanism. To determine whether NAD+ inhibited 32Pi transport by a mechanism other than or in addition to release of Pi, we compared Na+-dependent 32Pi counterflow in brush-border membrane equilibrated with Pi or with Pi generated from NAD+. Release of Pi from NAD+ incubated with brush-border membrane was confirmed. The increased uptake of 32Pi which was demonstrated in brush-border membrane equilibrated with Pi was not measured when intravesicular Pi was generated from a concentration of NAD+ which effected ADP-ribosylation of brush border membranes (100 microM NAD+). In contrast, increased uptake of 32Pi was demonstrated when intravesicular Pi was generated from 1 microM NAD+ which did not effect ADP ribosylation. Mg2+-dependent ADP ribosylation of brush-border membrane incubated with NAD+ was demonstrated which persisted during the time interval of 32Pi uptake measurements. Our findings are compatible with the hypothesis that NAD+-induced ADP ribosylation of brush-border membrane protein(s) results in inhibition of Pi transport across the membrane in vivo. EDTA may act to prevent this inhibition in brush-border membrane by chelation of Mg2+ and decreased ADP ribosylation.
Assuntos
Membrana Celular/metabolismo , Rim/metabolismo , Microvilosidades/metabolismo , NAD/farmacologia , Fosfatos/metabolismo , Adenosina Difosfato Ribose/metabolismo , Animais , Transporte Biológico/efeitos dos fármacos , Cães , Magnésio/farmacologia , Proteínas de Membrana/isolamento & purificação , Proteínas de Membrana/metabolismo , Microvilosidades/ultraestrutura , Peso Molecular , NAD/metabolismo , Sódio/metabolismoRESUMO
The direct effect of insulin on the high-affinity Ca2+-Mg2+-ATPase was studied in kidney proximal tubular basolateral membranes (BLM) obtained from control and streptozocin-induced non-insulin-dependent diabetes mellitus (NIDDM) rats. Plasma glucose of the diabetic animals was only mildly elevated (217 +/- 9 vs. 138 +/- 3 mg/dl). Both high- and low-affinity calcium-dependent Ca2+-Mg2+-ATPase activities were identified in the BLM. Enzyme activity in BLM from diabetic rats was higher at all Ca2+ concentrations tested due to a higher maximum velocity of the enzyme from NIDDM rats. The high-affinity Ca2+-Mg2+-ATPase activity was inhibited by trifluoroperazine (TFP) in both membranes. No difference in calmodulin content was found in the membranes from the diabetic and control rats. Insulin (16-200 microU/ml) significantly increased the high-affinity Ca2+-Mg2+-ATPase activity (17-40%) in membranes from control animals but had no effect on the enzyme activity in the membranes from the NIDDM rats. The basal activity of the enzyme at 0.1 microM free Ca2+ was higher in the BLM from the NIDDM animals compared to controls (17.8 +/- 0.5 vs. 14.7 +/- 0.8 nM Pi X mg-1 X min-1; P less than .02). There was no effect of insulin on the Ca2+-independent ATPase activity of BLM preparations. These findings demonstrate a defect in the ability of insulin to regulate the high-affinity Ca2+-Mg2+-ATPase activity in BLM from diabetic rats. Such a defect in enzyme activity may play a role in the mechanism of impaired insulin action observed in these NIDDM rats.
Assuntos
ATPase de Ca(2+) e Mg(2+)/metabolismo , ATPases Transportadoras de Cálcio/metabolismo , Diabetes Mellitus Experimental/enzimologia , Insulina/farmacologia , Rim/enzimologia , Animais , Membrana Basal/enzimologia , ATPase de Ca(2+) e Mg(2+)/antagonistas & inibidores , Cálcio/metabolismo , ATPases Transportadoras de Cálcio/antagonistas & inibidores , Diabetes Mellitus Tipo 2/enzimologia , Cinética , Ratos , Ratos Endogâmicos , Trifluoperazina/farmacologiaRESUMO
Pretreatment of rats with insulin-like growth factor I (IGF-I) ameliorates the course of acute ischemic renal injury. Differential display PCR was used to identify genes that are expressed in kidney after induction of acute ischemic renal injury in rats pretreated with vehicle or IGF-I. One amplification product that showed enhanced expression in kidneys of rats rendered ischemic compared to kidneys of sham-operated rats was identified as osteopontin. Sequence analysis of full-length complementary DNAs revealed a single species. Renal tissue was obtained for study 12 h and 1, 2, 3, 5, 7, 14, and 28 days postinjury. Levels of whole kidney osteopontin messenger RNA (mRNA) in rats rendered ischemic 1 day previously were elevated approximately 18-fold compared to levels measured in sham-operated controls, as determined by Northern analysis. No differences were noted 12 h postinjury. Levels of osteopontin mRNA remained elevated for 14 days after ischemia, but were no longer elevated at 28 days. IGF-I pretreatment resulted in enhanced levels of osteopontin mRNA 12 h, 1 day, and 5 days postinjury. In situ hybridization demonstrated that the elevated expression of osteopontin 1 day postinjury was localized predominantly to cells in the distal tubule and medullary thick ascending limb of Henle's loop. Immunostaining showed an identical localization for elevated protein expression. Five days postinjury, osteopontin peptide and mRNA were clearly detected in regenerating proximal tubules in addition to distal tubule and medullary thick ascending limb. We propose that endogenous osteopontin serves to promote tissue regeneration and tissue remodeling within 1 day after acute ischemic injury of kidney. IGF-I enhanced expression of osteopontin at an earlier time postischemia may ameliorate the course of injury.
Assuntos
Expressão Gênica , Fator de Crescimento Insulin-Like I/farmacologia , Isquemia/complicações , Nefropatias/etiologia , Rim/irrigação sanguínea , Rim/metabolismo , Sialoglicoproteínas/genética , Animais , Sequência de Bases , Northern Blotting , Hibridização In Situ , Rim/química , Nefropatias/metabolismo , Nefropatias/terapia , Masculino , Dados de Sequência Molecular , Osteopontina , Reação em Cadeia da Polimerase , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Distribuição TecidualRESUMO
High affinity (Ca2+ + Mg2+)ATPase activity was demonstrated in proximal tubule basolateral membranes (BLM) obtained from canine kidney. The Km of the enzyme for free Ca2+ was 0.12 +/- 0.02 microM, and at 3 microM free Ca2+, the enzyme reached its maximal velocity. To evaluate hormonal regulation of this enzyme, we studied the in vitro effects of several polypeptide hormones on enzyme activity. We measured the effects of insulin and human (h) PTH-(1-34) and their inactive analogs desoctapeptide insulin, bovine (b) PTH-(3-34), and oxidized hPTH-(1-34); insulin-like growth factors (IGFs) I and II; calcitonin; and the common cellular mediator for PTH and calcitonin, cAMP. At 0.1 microM free Ca2+, insulin (25-100 microU/ml) increased (Ca2+ + Mg2+)ATPase activity in a dose-dependent manner by 35-52% (P less than 0.01) and by 8-13% (P less than 0.05 to P less than 0.01) at a 3-microM free Ca2+ concentration; hPTH-(1-34) PTH (10(-9)-10(-7) M) increased the enzyme activity at a free Ca2+ concentration of 0.1 microM by 13-25% (P less than 0.05 to P less than 0.01). IGF-I increased (Ca2+ + Mg2+)ATPase activity by 40% (P less than 0.05) at 0.1 microM free Ca2+ at high peptide concentrations (10 ng/ml). No effect was obtained at 2 ng/ml IGF-I. cAMP (10(-7)-10(-4) M) stimulated enzyme activity by 18-27% (P less than 0.05 to P less than 0.02) at 0.1 microM Ca2+ and by 8-12% (P less than 0.05 to P less than 0.01) at 3 microM Ca2+. The effects of insulin and cAMP on (Ca2+ + Mg2+)ATPase activity were additive. No effect on the enzyme activity was obtained by the inactive analogs desoctapeptide insulin, bPTH-(3-34), and oxidized hPTH-(1-34), or by calcitonin or IGF-II. The data suggest that insulin and PTH have a specific stimulatory effect on (Ca2+ + Mg2+)ATPase activity in canine kidney proximal tubular BLM. While the insulin action is independent of cAMP, a role of cAMP in the regulatory effect of PTH on this enzyme cannot be ruled out. The direct stimulatory effect of insulin and PTH on (Ca2+ + Mg2+)ATPase in canine kidney proximal tubular BLM suggests that these hormones mediate their cellular effects in part by changes in cellular calcium homeostasis.
Assuntos
ATPase de Ca(2+) e Mg(2+)/metabolismo , ATPases Transportadoras de Cálcio/metabolismo , Córtex Renal/enzimologia , Animais , Calcitonina/farmacologia , AMP Cíclico/farmacologia , Cães , Relação Dose-Resposta a Droga , Insulina/farmacologia , Fator de Crescimento Insulin-Like I/farmacologia , Fator de Crescimento Insulin-Like II/farmacologia , Cinética , Hormônio Paratireóideo/farmacologia , Relação Estrutura-AtividadeRESUMO
Since its proposal three decades ago, the evidence in favor of the somatomedin hypothesis has been compelling. It is clear that somatotrophic actions of growth hormone are mediated through generation of insulin-like peptides and interaction of these peptides with plasma membrane receptors on sensitive cells. It is possible that such actions result from effects of circulating insulin-like peptides and/or insulin-like peptides generated in proximity to their sites of action (autocrine or paracrine effects). Most or all of circulating somatomedin activity in humans can be accounted for by insulin-like growth factors I and II (IFGs I and II). These peptides have considerable structural homology with insulin but, unlike insulin, they circulate in tight, noncovalent association with specific carrier protein. Levels of circulating IGF I and IGF II are affected by growth hormone, but the former peptide is the more sensitive to growth hormone. Levels of circulating IGF I in humans are low at birth, rise progressively during childhood, and peak during midadolescence. The increase in stature that occurs normally during adolescence probably results from this increase in circulating IGF I. Following adolescence, levels of circulating IGF I fall progressively as a function of age. There is good evidence that the reduction in levels of circulating IGF I is related to decreased secretion of growth hormone that accompanies aging. Although it has been suggested that decreased function of the growth hormone-somatomedin axis may cause changes in anabolic indices that accompany the aging process, definitive proof for this hypothesis is lacking. In contrast to IGF I, circulating IGF II reaches "adult" levels early in childhood, and changes are relatively small as a function of increasing age. Counterparts of IGF I and IGF II are present in rats. Dynamics of the growth hormone-somatomedin axis are similar in rats and humans for IGF I. In contrast, levels of IGF II in rat fall precipitously following birth, suggesting a role for rat IGF II in fetal growth and development. The rat has been used as an experimental animal to define the role of the growth hormone-somatomedin axis in aging. As in human studies, no firm relationship between somatomedins and aging has been established in the rat.
Assuntos
Envelhecimento/fisiologia , Somatomedinas/fisiologia , Adulto , Animais , Feminino , Hormônio do Crescimento/metabolismo , Humanos , Fator de Crescimento Insulin-Like I/sangue , Fator de Crescimento Insulin-Like II/sangue , Masculino , Pessoa de Meia-Idade , Somatomedinas/análiseRESUMO
The formation of all organs during embryogenesis, including the kidney, is dependent on the timed and sequential expression of a number of polypeptide growth factors. Synthesis and actions of one or more members of the insulinlike growth factor (IGF), epidermal growth factor/transforming growth factor-alpha (EGF/TGF-alpha), transforming growth factor-beta (TGF-beta), platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), hepatocyte growth factor (HGF), and nerve growth factor (NGF) families have been characterized in the developing metanephric kidney. Studies originating from a number of laboratories have defined the localization of growth factor mRNAs, receptors and peptides, have delineated patterns of growth factor synthesis, have established the growth factor-dependency of embryonic kidney development, and have identified abnormalities of growth factor-expression as potentially causative of aberrancies in metanephrogenesis. The results of these investigations are summarized in this review.
Assuntos
Substâncias de Crescimento/fisiologia , Rim/embriologia , Animais , Apoptose/fisiologia , Células Cultivadas , Células Epiteliais , Epitélio/embriologia , Epitélio/fisiologia , Substâncias de Crescimento/biossíntese , Humanos , Rim/citologia , Rim/fisiologia , Neoplasias Renais/embriologia , Tumor de Wilms/embriologiaRESUMO
Strategies for the treatment of acute renal failure in humans are directed toward supportive care to permit regeneration of damaged tissue to occur. There exists a need for new therapeutic approaches that can speed recovery and reduce mortality. Induction of acute renal injury in rats results in changes in expression of a panoply of growth factor genes. The administration of several of these growth factors to animals with ischemic or toxic acute renal failure accelerates recovery of renal function and normal histology. One of these agents, IGF-I, has been used to treat humans at risk to develop acute renal failure. A greater understanding of the patterns of gene expression post-injury will lead to the generation of more effective therapeutic strategies for acute renal failure.
Assuntos
Injúria Renal Aguda/tratamento farmacológico , Injúria Renal Aguda/prevenção & controle , Substâncias de Crescimento/fisiologia , Substâncias de Crescimento/uso terapêutico , Injúria Renal Aguda/fisiopatologia , Animais , Humanos , Fator de Crescimento Insulin-Like I/fisiologia , Fator de Crescimento Insulin-Like I/uso terapêutico , Rim/efeitos dos fármacos , Rim/patologiaAssuntos
Fator de Crescimento Insulin-Like I/fisiologia , Rim/fisiologia , Acromegalia/fisiopatologia , Sequência de Aminoácidos , Animais , Diabetes Mellitus Tipo 1/fisiopatologia , Humanos , Hipertrofia , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/uso terapêutico , Rim/crescimento & desenvolvimento , Rim/patologia , Nefropatias/tratamento farmacológico , Nefropatias/fisiopatologia , Dados de Sequência Molecular , Receptor IGF Tipo 1/fisiologiaRESUMO
BACKGROUND: Transplantation therapy for human diabetes is limited by a shortage of donor organs, and transplant function diminished over time by cell death and limited potential for expansion of beta cells in pancreas or islets. Outcomes are complicated by immunosuppression. A way to overcome supply and expansion problems is to xenotransplant embryonic tissue. Previously, we have shown that beta cells originating from embryonic day (E) 28 (E28) pig pancreatic primordia transplanted into the mesentery of streptozotocin (STZ)-diabetic (type 1) Lewis rats or Zucker Diabetic Fatty (ZDF) diabetic (type 2) rats engraft and normalize glucose tolerance without the need for host immune-suppression. METHODS: In this study, we transplant E28 pig pancreatic primordia in the mesentery of STZ-diabetic rhesus macaques. RESULTS: Long-term engraftment of pig beta cells within liver, pancreas and mesenteric lymph nodes post-transplantation of E28 pig pancreatic primordia into STZ-diabetic rhesus macaques is demonstrated by electron microscopy, positive immune-histochemistry for insulin, and positive RT-PCR and in situ hybridization for porcine proinsulin mRNA. Insulin requirements were reduced in one macaque followed over 22 months post-transplantation and porcine insulin detected in plasma using sequential affinity chromatography, HPLC and mass spectrometry. Of potential importance for application of this transplantation technology to treatment of diabetes in humans and confirmatory of our previous findings in Lewis and ZDF rats, no host immunosuppression is required. CONCLUSIONS: Under selected circumstances, pancreatic primordia elicit a muted immune response relative to more differentiated tissue, such that engraftment occurs in non-immunosuppressed hosts. Our findings that pig pancreatic primordia engraft long-term in non-immunosuppressed STZ-diabetic rhesus macaques establishes the potential for their use in human diabetics.
Assuntos
Diabetes Mellitus Experimental/cirurgia , Diabetes Mellitus Tipo 1/cirurgia , Sobrevivência de Enxerto , Transplante de Pâncreas/fisiologia , Transplante Heterólogo/fisiologia , Animais , Primers do DNA , Éxons , Íntrons , Linfonodos/imunologia , Linfonodos/patologia , Macaca mulatta , Transplante de Pâncreas/patologia , Proinsulina/genética , SuínosRESUMO
Insulin is known to regulate both metabolic and transport functions in the renal proximal tubule. Insulin present in plasma and in glomerular ultrafiltrate is known to be degraded at this nephron site. This paper summarizes what is known about these processes and about the mechanisms by which the actions of insulin and the degradation of insulin are effected in the proximal tubular epithelial cell. Recent studies have characterized the binding of insulin to specific receptors present in the proximal tubular basolateral membrane. Binding of insulin to its receptor in this membrane is thought to initiate events that lead to the phosphorylation of that receptor. Such insulin-stimulated phosphorylation may mediate hormonal action. A possible role for insulin-like growth factor II in the modulation of the actions of insulin has been suggested by observations in the kidney and in nonrenal cells. These findings are integrated into a model characterizing the nature of the interaction of insulin with the renal proximal tubular cell.
Assuntos
Insulina/fisiologia , Túbulos Renais Proximais/metabolismo , Animais , Sítios de Ligação , Transporte Biológico Ativo , Permeabilidade da Membrana Celular , Células Epiteliais , Gluconeogênese , Humanos , Insulina/metabolismo , Túbulos Renais Proximais/citologia , Microvilosidades/metabolismo , Modelos Biológicos , Peptídeos/fisiologia , Ésteres de Forbol/metabolismo , Fosfatos/metabolismo , Fosforilação , Receptor de Insulina/metabolismo , Sódio/metabolismo , Somatomedinas/fisiologiaRESUMO
The transport of phosphate across the plasma membrane of the renal proximal tubular epithelial cell is thought to take place through the activities of specific transporters located in the membrane. The activities of these transporters are essential to effect the reabsorption of phosphate present in glomerular ultrafiltrate. In addition, their activities are thought to be important for the maintenance of metabolic functions in the proximal tubular cell. Studies utilizing proximal tubular brush-border and basolateral membranes isolated from mammalian kidney have provided significant insights into the mechanisms by which phosphate transport across the brush-border and basolateral membranes of the intact proximal tubular cell occurs and is modulated. In this editorial review, the results of many of these studies are summarized. Particular emphasis is placed on studies that utilized isolated membranes to determine the mechanism by which the phosphaturic action of parathyroid hormone is mediated in the renal proximal tubule. On the basis of studies conducted in isolated membranes and in more physiologically intact preparations, models are constructed to integrate the role of the brush-border and basolateral membranes in the transport of phosphate into and out of the renal proximal tubular cell.
Assuntos
Túbulos Renais Proximais/metabolismo , Fosfatos/metabolismo , Difosfato de Adenosina/metabolismo , Animais , Transporte Biológico , Fenômenos Biomecânicos , Membrana Celular/metabolismo , AMP Cíclico/fisiologia , Técnicas In Vitro , Túbulos Renais Proximais/citologia , Túbulos Renais Proximais/ultraestrutura , Microvilosidades/metabolismo , Modelos Biológicos , Hormônio Paratireóideo/fisiologia , Fosforilação , Ribose/metabolismo , Sódio/fisiologiaRESUMO
We have previously demonstrated cAMP-dependent 32P phosphorylation and dephosphorylation of a 62,000 relative molecular weight (Mr) protein in autoradiograms of sodium dodecyl sulfate polyacrylamide gels originating from canine renal brush border membranes. In the current studies 32P phosphorylation of the 62,000 Mr protein that was independent of cAMP was noted in the presence of Zn2+. Under these conditions, cAMP inhibited the 32P phosphorylation of this protein. Concentration-dependent photoaffinity labeling of a band with Mr 60,000 in autoradiograms of gels resulted from incubation of membranes with cyclic 8-azidoadenosine-3',5'-monophosphate (8-N3-[32P]cAMP) followed by exposure to light. In the presence of Zn2+ and ATP, an apparent shift in the Mr of a portion of the photoaffinity-labeled band to 62,000 was seen. The 62,000 Mr phosphoprotein in detergent-solubilized supernatants of brush border membranes was immunoprecipitated with antibodies directed against the regulatory subunit of type II cAMP-dependent protein kinase. Our observations strongly suggest that the 62,000 Mr protein is the regulatory subunit.