Overexpression of PKC-βI and -δ contributes to higher PKC activity in the proximal tubules of old Fischer 344 rats

2003 ◽  
Vol 285 (6) ◽  
pp. F1100-F1107 ◽  
Author(s):  
Mohammad Asghar ◽  
Tahir Hussain ◽  
Mustafa F. Lokhandwala
Keyword(s):  
Skf 38393 ◽  
Proximal Tubules ◽  
Fischer 344 Rats ◽  
Adult Rats ◽  
Altered Expression ◽  
Fischer 344 ◽  
Pkc Isoforms ◽  
Diuretic Response ◽  
Old Rats ◽  
Pkc Ζ

Previously, we reported that natriuretic and diuretic response to dopamine is diminished in old Fischer 344 rats, which is due to higher basal protein kinase C (PKC) activity and hyperphosphorylation of Na-K-ATPase in the proximal tubules (PTs) of old rats. The present study was conducted to determine whether higher PKC activity could be due to altered expression of some of the PKC isoforms in the superficial cortex (rich in PTs) of old rats. Fluorimetric measurement showed almost twofold increase in the PKC activities in homogenates and membranes of old (24 mo) compared with adult (6 mo) rats. Interestingly, in the basal state PKC-βI was overexpressed in the membranes, whereas PKC-δ expression was increased in the cytosol of old compared with adult rats. Treatment of the cortical slices with either SKF-38393, a D1-like agonist, or PDBu, a direct activator of PKC, caused translocation of PKC-βI from cytosol to membranes in adult but not in old rats. Both of these drugs caused translocation of PKC-δ from membranes to cytosol in adult but not in old rats. These drugs had no effect on translocation of PKC-ζ in both adult and old rats. Both PKC-βI and -δ coimmunoprecipiated with α1-subunit of Na-K-ATPase in adult and old rats. These observations suggest that both SKF-38393 and PDBu differentially regulate PKC-βI and -δ in adult but not in old rats. Also, PKC-βI and -δ seem to interact with Na-K-ATPase in these animals. The overexpression of both PKC-βI and -δ in old rats could be responsible for a higher basal PKC activity, which causes the hyperphosphorylation of Na-K-ATPase and contributes to the diminished inhibition of Na-K-ATPase activity by dopamine in old rats.

Higher basal serine phosphorylation of D1Areceptors in proximal tubules of old Fischer 344 rats

2002 ◽  
Vol 283 (2) ◽  
pp. F350-F355 ◽  
Author(s):  
Mohammad Asghar ◽  
Tahir Hussain ◽  
Mustafa F. Lokhandwala
Keyword(s):  
Receptor Agonist ◽  
Sch 23390 ◽  
Proximal Tubules ◽  
Serine Phosphorylation ◽  
Fischer 344 Rats ◽  
Adult Rats ◽  
Fischer 344 ◽  
Agonist Affinity ◽  
Old Rats ◽  
Cortical Slices

Dopamine (DA) and D1-like receptor agonists promote an increase in Na excretion by means of activation of the D1-like receptor signaling cascade and subsequent inhibition of the Na/H exchanger and Na-K-ATPase in renal proximal tubules. Recently, our laboratory reported that DA and the D1-like receptor agonist failed to inhibit Na-K-ATPase activity in old Fischer 344 rats because of uncoupling of D1A receptors from G proteins and that this abnormality led to a diminished natriuretic response to DA in old Fischer 344 rats. In this study, we have tested the hypothesis that the mechanism of this uncoupling may be an altered phosphorylation of D1A receptors in old rats. In experiments performed in renal cortical slices, both DA and SKF-38393, a D1-like receptor agonist, increased the serine phosphorylation of D1A receptors in adult (6 mo) but not old (24 mo) rats. Interestingly, the basal serine phosphorylation of D1Areceptors was higher in old than in adult rats. Competition ligand binding ([3H]SCH-23390) experiments on the D1-like receptor in adult and old rats with fenoldopam, a D1-like receptor agonist, revealed the presence of two affinity states of the receptors. There was a rightward shift in the agonist displacement of the ligand in old compared with adult rats, as reflected in the IC50 values (adult vs. old, 7.46 × 10−9 ± 2.26 vs. 7.93 × 10−7± 1.33 M). Also, there was a reduction in agonist affinity in the low-affinity receptors in old compared with adult rats (IC50, adult vs. old, 5.67 × 10−5 ± 1.33 vs. 12.60 × 10−5± 6.50 M). Moreover, the abundance of D1A receptor proteins was ∼47% lower in the membranes of old compared with adult rats. We speculate that higher basal serine phosphorylation of D1A receptors may have rendered the D1Areceptor uncoupled from G protein, leading to a reduced agonist affinity state and thus diminished natriuretic response to DA in old rats.

Alterations in dopamine DA1 receptor and G proteins in renal proximal tubules of old rats

1997 ◽  
Vol 273 (1) ◽  
pp. F53-F59 ◽  
Author(s):  
V. Kansra ◽  
T. Hussain ◽  
M. F. Lokhandwala
Keyword(s):  
Atpase Activity ◽  
G Proteins ◽  
Sch 23390 ◽  
Skf 38393 ◽  
Proximal Tubules ◽  
Adult Rats ◽  
Protein Activation ◽  
Proximal Tubular ◽  
Old Rats ◽  
Renal Proximal Tubular

The present study examines the effect of dopamine DA1-receptor agonists on the renal proximal tubular Na(+)-K(+)-adenosinetriphosphatase (Na(+)-K(+)-ATPase) activity and quantitates DA1 receptors and the coupled G proteins in Fischer 344 model of adult (6 mo old) and old (23 mo old) rats. Dopamine and the preferential DA1-receptor agonist, SKF-38393, produced a concentration-dependent inhibition of Na(+)-K(+)-ATPase activity in proximal tubules from adult rats, whereas the enzyme activity was unaffected by these agonists in the old rats. The binding of DA1-receptor antagonist [3H]Sch-23390 in the proximal tubular basolateral membranes showed a marked decrease (approximately 47%) in the receptor numbers in old compared with adult rats, whereas dissociation constant (Kd) values in old compared with adult rats were not significantly different. Dopamine and SKF-38393 stimulated 35S-labeled guanosine 5'-O-(3-thiotriphosphate) binding in adult rats, but there was no significant effect on the binding in the old rats. Quantification of G2 alpha and Gq/11 alpha using Western analysis revealed a significant increase in quantities of both the G proteins in old rats. The data suggest that a reduction in DA1 receptor number and subsequently reduced G protein activation may be the causative factors for the impairment in DA1 receptor-mediated inhibition of Na(+)-K(+)-ATPase activity in the proximal tubules of old rats.

Sensitivity of lungs of aging Fischer 344 rats to ozone: assessment by bronchoalveolar lavage

1996 ◽  
Vol 271 (4) ◽  
pp. L555-L565 ◽  
Author(s):  
R. Vincent ◽  
D. Vu ◽  
G. Hatch ◽  
R. Poon ◽  
K. Dreher ◽  
...  
Keyword(s):  
Bronchoalveolar Lavage ◽  
Biological Effects ◽  
Age Groups ◽  
Lavage Fluid ◽  
Lung Lavage ◽  
Ozone Exposure ◽  
Fischer 344 Rats ◽  
Adult Rats ◽  
Fischer 344 ◽  
Old Rats

Biological effects indicators in bronchoalveolar lavage fluid were studied in Fischer 344 rats of different ages after exposure to 0.4-0.8 ppm ozone for periods of 2-6 h on a single day or on 4 consecutive days. The magnitude of alveolar protein transudation induced by ozone was not different between age groups, but the interindividual variability of protein changes was higher in senescent (24-mo-old) rats. By comparison to juvenile (2-mo-old) and adult (9-mo-old) rats, senescent animals had higher increases of interleukin-6 (up to 10-fold higher) and N-acetyl-beta-D-glucosaminidase (NAGA; 2-fold higher) in lung lavage after ozone. Ascorbic acid was lower in lungs of senescent rats (one-half of juvenile values), and acute ozone exposure brought a further decrease in lung ascorbate. Whereas alveolar protein transudation was attenuated after ozone exposure on 4 days, persistent elevation of NAGA in senescent rats suggested only partial adaptation. Injection of endotoxin did not modify the patterns of effects. Incorporation of 18O-ozone into macrophages and surfactant was not different between age groups, indicating that the magnified biological responses in senescent rats were not dominated by differences in internal dose of ozone. The results indicate that senescent rats respond differently than juvenile and adult rats to lung injury.

scholarly journals Hyperphosphorylation of Na-Pump Contributes to Defective Renal Dopamine Response in Old Rats

2001 ◽  
Vol 12 (2) ◽  
pp. 226-232
Author(s):  
MOHAMMAD ASGHAR ◽  
VIKRAM KANSRA ◽  
TAHIR HUSSAIN ◽  
MUSTAFA F. LOKHANDWALA
Keyword(s):  
Atpase Activity ◽  
Phorbol Ester ◽  
Receptor Agonist ◽  
Receptor Activation ◽  
Skf 38393 ◽  
Proximal Tubules ◽  
Adult Rats ◽  
Basal Activity ◽  
Na Pump ◽  
Old Rats

Abstract. Dopamine D1-like receptor activation causes phosphorylation and inhibition of Na,K-ATPase (Na-pump) activity in the proximal tubules, which is associated with an increase in sodium excretion. It has been shown that dopamine and SKF 38393, a D1-like receptor agonist, caused inhibition of Na,K-ATPase activity in the proximal tubules of adult (6 mo) but not of old (24 mo) Fischer 344 rats. The present study demonstrated that SKF 38393 and PDBu, a phorbol ester and protein kinase C (PKC) activator, increased phosphorylation of the α1-subunit of Na,K-ATPase in adult but not in old rats. In adult rats, SKF 38393-mediated phosphorylation was antagonized by SCH 23390, a D1-like receptor antagonist. Similarly, Na,K-ATPase activity was inhibited by SKF 38393 and PDBu in adult but not in old rats. The basal activity of Na,K-ATPase was decreased and the basal phosphorylation state of the enzyme was increased in old compared with adult rats. Basal activity of PKC was higher in old compared with adult rats, and SKF 38393 and PDBu stimulated PKC activity in adult but not in old rats. The conclusion is that the failure of D1-like receptor agonist and phorbol ester to stimulate PKC and inhibit Na,K-ATPase activity in old rats is due, at least in part, to the higher basal PKC activity and Na,K-ATPase phosphorylation in old compared with adult rats.

Age-dependent activation of PKC isoforms by angiotensin II in the proximal nephron

2001 ◽  
Vol 281 (3) ◽  
pp. R861-R867 ◽  
Author(s):  
Dianne M. Boesch ◽  
Jeffrey L. Garvin
Keyword(s):  
Specific Binding ◽  
Age Groups ◽  
Proximal Tubules ◽  
Ang Ii ◽  
Fluid Absorption ◽  
Adult Rats ◽  
Young Rats ◽  
Age Related ◽  
Pkc Isoforms ◽  
Pkc Β

ANG II increases fluid absorption in proximal tubules from young rats more than those from adult rats. ANG II increases fluid absorption in the proximal nephron, in part, via activation of protein kinase C (PKC). However, it is unclear how age-related changes in ANG II-induced stimulation of the PKC cascade differ as an animal matures. We hypothesized that the response of the proximal nephron to ANG II decreases as rats mature due to a reduction in the amount and activation of PKC rather than a decrease in the number or affinity of ANG II receptors. Because PKC translocates from the cytosol to the membrane when activated, we first measured PKC activity in the soluble and particulate fractions of proximal tubule homogenates exposed to vehicle or 10−10 M ANG II from young (26 ± 1 days old) and adult rats (54 ± 1 days old). ANG II increased PKC activity to the same extent in homogenates from young rats (from 0.119 ± 0.017 to 0.146 ± 0.015 U/mg protein) ( P < 0.01) and adult rats (from 0.123 ± 0.020 to 0.156 ± 0.023 U/mg protein) ( P < 0.01). Total PKC activity did not differ between groups (0.166 ± 0.018 vs. 0.181 ± 0.023). We next investigated whether activation of the α-, β-, and γ-PKC isoforms differed by Western blot. In homogenates from young rats, ANG II significantly increased activated PKC-α from 40.2 ± 6.5 to 60.2 ± 9.5 arbitrary units (AU) ( P < 0.01) but had no effect in adult rats (46.1 ± 5.1 vs. 48.5 ± 8.2 AU). Similarly, ANG II increased activated PKC-γ in proximal tubules from young rats from 47.9 ± 13.2 to 65.6 ± 16.7 AU ( P < 0.01) but caused no change in adult rats. Activated PKC-β, however, increased significantly in homogenates from both age groups. Specifically, activated PKC-β increased from 8.6 ± 1.4 to 12.2 ± 2.1 AU ( P < 0.01) in homogenates from nine young rats and from 19.0 ± 5.5 to 25.1 ± 7.1 AU ( P < 0.01) in homogenates from 12 adult rats. ANG II did not alter the amount of soluble PKC-α, -β, and -γ significantly. The total amount of PKC-α and -γ did not differ between homogenates from young and adult rats, whereas the total amount of PKC-β was 59.7 ± 10.7 and 144.9 ± 41.8 AU taken from young and adult rats, respectively ( P < 0.05). Maximum specific binding and affinity of ANG II receptors were not significantly different between young and adult rats. We concluded that the primary PKC isoform activated by ANG II changes during maturation.

scholarly journals Aging and arteriosclerosis. I. Development of myointimal hyperplasia after endothelial injury.

1986 ◽  
Vol 164 (4) ◽  
pp. 1171-1178 ◽  
Author(s):  
R J Hariri ◽  
D R Alonso ◽  
D P Hajjar ◽  
D Coletti ◽  
M E Weksler
Keyword(s):  
Vascular Injury ◽  
Experimental Models ◽  
Endothelial Injury ◽  
Vascular Lesions ◽  
Fischer 344 Rats ◽  
Cell Nuclei ◽  
Fischer 344 ◽  
Endothelial Denudation ◽  
Old Rats ◽  
Intimal Thickness

Old Fischer 344 rats are more susceptible to vascular lesions after arterial endothelial injury than are young animals. Thus, 20-26-mo-old Fischer 344 rats developed greater and more persistent intimal proliferative lesions than did 2-5-mo-old rats after aortic endothelial denudation. 3 d after deendothelialization, intimal thickness was increased two-fold in both old and young animals. However, 14 d after endothelial injury, intimal thickness had increased nearly five times in old animals, but had regressed to normal in young animals. Intimal thickness of young aortic grafts transplanted into young recipients did not differ significantly from adjacent host aorta or autotransplanted aortic segments 6 wk after surgery. In contrast, intimal thickness of old grafts transplanted into young recipients was eight times greater than adjacent young host aorta 6 wk after surgery. The density of cell nuclei in the intima of old grafts was also much greater than that in young grafts. Thus, in two experimental models of vascular injury, old rats have consistently had greater myointimal hyperplasia than young rats. The increased proliferative response of aortic smooth muscle cells after vascular injury of old animals may contribute to the increased prevalence of vascular disease with age.

NAD(P)H oxidase-generated superoxide anion accounts for reduced control of myocardial O2 consumption by NO in old Fischer 344 rats

2003 ◽  
Vol 285 (3) ◽  
pp. H1015-H1022 ◽  
Author(s):  
Alexandra Adler ◽  
Eric Messina ◽  
Ben Sherman ◽  
Zipping Wang ◽  
Harer Huang ◽  
...  
Keyword(s):  
Oxidant Stress ◽  
Heart Weight ◽  
Fischer 344 Rats ◽  
No Donor ◽  
Fischer 344 ◽  
Old Rats ◽  
Fischer Rats ◽  
Enos Protein

We investigated the role of nitric oxide (NO) in the control of myocardial O2 consumption in Fischer 344 rats. In Fischer rats at 4, 14, and 23 mo of age, we examined cardiac function using echocardiography, the regulation of cardiac O2 consumption in vitro, endothelial NO synthase (eNOS) protein levels, and potential mechanisms that regulate superoxide. Aging was associated with a reduced ejection fraction [from 75 ± 2%at4moto66 ± 3% ( P < 0.05) at 23 mo] and an increased cardiac diastolic volume [from 0.60 ± 0.04 to 1.00 ± 0.10 ml ( P < 0.01)] and heart weight (from 0.70 ± 0.02 to 0.90 ± 0.02 g). The NO-mediated control of cardiac O2 consumption by bradykinin or enalaprilat was not different between 4 mo (36 ± 2 or 34 ± 3%) and 14 mo (29 ± 1 or 25 ± 3%) but markedly ( P < 0.05) reduced in 23-mo-old Fischer rats (15 ± 3 or 7 ± 2%). The response to the NO donor S-nitroso- N-acetyl penicillamine was not different across groups (35%, 35%, and 44%). Interestingly, the eNOS protein level was not different at 4, 14, and 23 mo. The addition of tempol (1 mmol/l) to the tissue bath eliminated the depression in the control of cardiac O2 consumption by bradykinin (25 ± 3%) or enalaprilat (28 ± 3%) in 23-mo-old Fischer rats. We next examined the levels of enzymes involved in the production and breakdown of superoxide. The expression of Mn SOD, Cu/Zn SOD, extracellular SOD, and p67phox, however, did not differ between 4- and 23-mo-old rats. Importantly, there was a marked increase in gp91phox, and apocynin restored the defect in NO-dependent control of cardiac O2 consumption at 23 mo to that seen in 4-mo-old rats, identifying the role of NADPH oxidase. Thus increased biological activity of superoxide and not decreases in the enzyme that produces NO are responsible for the altered control of cardiac O2 consumption by NO in 23-mo-old Fischer rats. Increased oxidant stress in aging, by decreasing NO bioavailability, may contribute not only to changes in myocardial function but also to altered regulation of vascular tone and the progression of cardiac or vascular disease.

Age-related immunosenescence in Fischer 344 rats: influence of exercise training

1992 ◽  
Vol 73 (5) ◽  
pp. 1932-1938 ◽  
Author(s):  
I. Nasrullah ◽  
R. S. Mazzeo
Keyword(s):  
Endurance Training ◽  
Interleukin 2 ◽  
Cytolytic Activity ◽  
Treadmill Running ◽  
Target Cells ◽  
Fischer 344 Rats ◽  
Middle Aged ◽  
Fischer 344 ◽  
Age Related ◽  
Old Rats

The present investigation examined the extent to which 15 wk of endurance training could influence immune function in young, middle-aged, and older animals. Forty-eight male Fischer 344 rats were divided into trained and untrained groups. Training consisted of treadmill running at 75% maximal running capacity for 1 h/day, 5 days/wk, for 15 wk. Animals were killed at 8, 17, and 27 mo, at which time splenocytes were isolated. The capacity for lymphocyte proliferation in response to mitogen (concanavalin A, ConA), interleukin-2 (IL-2) production, and cytolytic activity against YAC-1 target cells was determined. ConA-induced proliferation declined significantly with age. Training suppressed the proliferative response in the young (-41%) and middle-aged animals (-27%) compared with the age-matched controls; however, training improved this response (+58%) in the older group. IL-2 production followed a pattern similar to that for mitogen-induced proliferation, such that production declined with age and was reduced with training in young and middle-aged animals but was significantly more improved in the older animals than in age-matched controls. The ability to lyse target cells, measured as percent cytotoxicity, declined steadily with advancing age at all effector-to-target cell ratios tested: 52, 14, and -16% for 8-, 17-, and 27-mo-old rats, respectively. It was concluded that the capacity for ConA-induced splenocyte proliferation, IL-2 production, and cytolytic activity declines significantly with advancing age. Furthermore, 15 wk of endurance training suppressed proliferation and IL-2 production in young animals but improved these responses in older animals. Training had no effect on cytolytic activity.

Effect of age on induction of hepatic phosphoenolpyruvate carboxykinase by fasting

1994 ◽  
Vol 267 (2) ◽  
pp. G195-G200 ◽  
Author(s):  
H. Van Remmen ◽  
W. F. Ward
Keyword(s):  
G Protein ◽  
Time Course ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Age Groups ◽  
Fischer 344 Rats ◽  
Fischer 344 ◽  
Fasting And Refeeding ◽  
Old Rats ◽  
Rate Limiting ◽  
Effect Of Age

This study examines the effect of age on the induction of the rate-limiting enzyme in gluconeogenesis, phosphoenolpyruvate carboxykinase (PEPCK), in response to fasting and refeeding in male Fischer 344 rats aged 3-18 mo. The rats were fasted for 30 h to increase the activity of PEPCK and subsequently were refed for 24 h to lower activity toward basal levels. PEPCK activity increased 2.2-fold in the 3-mo-old rats and 2.3-fold in the 18-mo-old rats during the 30-h fast. Therefore PEPCK induction during the 30-h fast was not altered with age. Similarly, refeeding resulted in a significant decrease in PEPCK activity at all ages. After the 24-h refeeding period, the rats were fasted a second time, and the time course of induction from the basal refed level was measured. In the young rats (6 mo), the activity of PEPCK increased rapidly from 18.12 +/- 1.61 to 42.66 +/- 5.94 U/g protein (P < 0.01) within 8 h of fasting. However, in the 18-mo-old rats, the initiation of the induction of PEPCK activity was delayed, and, after 12 h, PEPCK activity had increased from 17.34 +/- 1.34 to only 32.50 +/- 3.21 U/g protein (P < 0.01). Furthermore, the rate of induction appears to be decreased in the older animals. The activity after 24 h of fasting was equivalent in all four age groups (ranging from 44.72 +/- 5.38 at 3 mo to 40.18 +/- 5.42 U/g protein at 18 mo).(ABSTRACT TRUNCATED AT 250 WORDS)

An age-related difference in hyperoxia lethality: role of lung antioxidant defense mechanisms

1995 ◽  
Vol 268 (4) ◽  
pp. L539-L545 ◽  
Author(s):  
A. T. Canada ◽  
L. A. Herman ◽  
S. L. Young
Keyword(s):  
Defense Mechanisms ◽  
Antioxidant Defense ◽  
Fischer 344 Rats ◽  
Fischer 344 ◽  
Related Difference ◽  
Age Related ◽  
65 H ◽  
Old Rats ◽  
Gpx Activity

The role of animal age in the lethal response to > 98% oxygen has been extensively studied, with the observation that neonatal rats were resistant while mature animals were sensitive. Antioxidant enzymes increased during the oxygen exposure in neonatal but not in mature rats, suggesting they were important in the age-related toxicity difference. Because no studies had compared the response of mature and old rats to hyperoxia, we exposed Fischer 344 rats, aged 2 and 27 mo, to > 98% oxygen. Unexpectedly, the old rats lived significantly longer than young, 114 and 65 h, respectively. No histopathological differences were found to explain the results. Of the antioxidants, only glutathione peroxidase (GPx) activity was higher in the lungs of nonexposed old rats. Superoxide dismutase (SOD) was higher in the young, results opposite those expected if SOD was important in the lethality difference. No antioxidant induction occurred in the old oxygen-exposed rats. These results suggest that although there may be a role for GPx, mechanisms in addition to antioxidant protection and inflammation are likely responsible for the age-related difference in hyperoxia lethality.

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