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J P Geloso

Publications and source records attributed to J P Geloso.

At least 19 recordsLinked to original sources

Perinatal maturation of rat kidney mitochondria.

In the rat kidney, NaK-ATPase activity increased between days 19 and 20 of gestation (+50%) and between 1 and 24 h after birth (+20%), requiring an increased energy supply. In order to determine whether mitochondrial changes were involved, renal mitochondrial development was investigated from day 19 of gestation to 1 day after birth. Slot-blot analyses of mitochondrial-DNA/nuclear-DNA ratio and determination of citrate synthase activity showed a doubling in the mitochondrial pool between days 19 and 20 of gestation. In isolated mitochondria, oxygen consumption remained unchanged between days 19 and 20 of gestation, and then it was enhanced between days 20 and 21 of gestation (+70%) and between 1 and 24 h after birth (+50%). We also focused on one of the respiratory-chain complexes, ATP synthase, and measured its activity and content during the perinatal period. We demonstrated increases in both activity and content of ATP synthase between days 20 and 21 of gestation and between 1 and 24 h after birth, thus suggesting that changes in ATP synthase activity are ascribed to an increase in the mitochondrial density of ATP synthase complexes. Moreover, the mitochondrial ATP/ADP ratio only increased between 1 and 24 h (+90%), indicating a critical step in the renal respiratory-chain maturation at that time. We therefore conclude that the postnatal enhancement of renal mitochondrial oxidative capacity might depend on protein synthesis de novo and on changes in the adenine nucleotide concentrations.

Adenine Nucleotides↗

Effect of maternal hyperglycemia on NaK ATPase activity in fetal rat kidney.

The effect of moderate hyperglycemia on renal ATP production and ATPase activity of rat fetus was investigated using the experimental procedure of maternal continuous infusion of glucose during the last 5 days of gestation. Glucose-infused mothers and their fetuses showed a high level of glycemia (8.8 and 5.5 mM, respectively) and a high level of insulinemia (3 times higher than in controls). No change in either ATP or ADP concentration was detectable but an increase in NaK ATPase activity occurred without any change in Mg ATPase activity. These modifications should be the result of an enhanced Na/glucose cotransport leading to an enhanced extrusion of Na at the basolateral membrane. These results indicate that immature kidney is able to increase NaK ATPase activity to maintain Na homeostasis.

Adenosine Triphosphatases↗

Milk-stimulated PGE2 production by isolated gastric cells: a possible role in the inhibition of histamine-induced acid secretion.

The effects of a milk diet on gastric acid secretion of rats fed raw bovine milk for 4 days were examined using dispersed gastric cells. Parietal cell acid secretion was estimated by the accumulation of 14C-aminopyrine (AP), an index of secretory function. Basal AP accumulation was significantly increased (60%) by the milk diet. There was a marked upward shift in the dose-response curve of histamine (HA; 10(-8) to 10(-3) M) in milk-fed rats, indicating enhanced sensitivity of parietal cell-H2 receptor to exogenous HA. In contrast, the dose-dependent inhibition of HA-induced AP accumulation by prostaglandin (PG) E2 was significantly reduced, indicating that the parietal cells of milk-fed rats were less sensitive to exogenous PGE2. The PGE2 content of bovine milk was low (less than 20 pg/ml), but the production of endogenous PGE2 by the gastric cells was dramatically increased by the milk diet and exhibited maximal control production rate in the presence of 10 microM arachidonic acid. The increased responsiveness to histamine and the decreased responsiveness to PGE2 indicated that the milk diet induced low histamine and high PGE2 availability in the vicinity of the parietal cell basolateral membrane. This regulation, which involves stimulation of PGE2 production in the gastric mucosa, may underly the inhibition of acid secretion observed in vivo in chronically milk-fed adult rats.

Aminopyrine↗

Effect of lipid diet on mitochondrial palmitoyl-l-carnitine oxidation in kidney at postnatal development.

Oxygen consumption (VO2) and beta-hydroxyacyl-CoA dehydrogenase (beta OAC) activity were measured in isolated mitochondria of developing rat kidney from late fetal to adult age. In the presence of palmitoyl-L-carnitine, VO2 consumption was higher in suckling than in adult rats while beta OAC activity rose during the postnatal period and declined after weaning. During postnatal development, the high level of mitochondrial fatty acid oxidation was linked to the high level of fatty acid supply and any change in lipid diet altered mitochondrial fatty acid oxidation. By contrast at adult age, a high fat diet did not change either mitochondrial fatty acid oxidation or beta OAC activity measured in two nephron structures (PCT and mTAL). Dietary lipids seem to play an important role in the evolution of mitochondrial fatty acid oxidation in developing rat kidney.

3-Hydroxyacyl CoA Dehydrogenases↗

Relationship between maternal milk PGE2 and gastric acid secretion in newborn rats.

We previously demonstrated that in rats gastric acid secretion declines after birth and drops steeply on day 12 of life. In the present study, we investigated the part played in this decline by prostaglandin E2 (PGE2) from maternal milk. PGE2 content was first measured in the milk of untreated dams 0, 1, 5, 10, 12, 15, and 18 days after parturition. PGE2 levels were high during the first 5 days (123.5-200.5 pg/ml), declined significantly between days 10 and 15 (56.6-85.4 pg/ml; P less than 0.05), and dropped to 18.4 pg/ml on day 18. We also found that depleting milk of PGE2 prevented drop of acid secretion in 12-day-old suckling rats. Injecting lactating dams with indomethacin significantly reduced milk PGE2 content by 65% vs. milk of untreated dams. Surprisingly, administration of sesame oil, the indomethacin vehicle to the dams, increased milk PGE2 content by 182%. In the pups of the indomethacin-treated dams, acid secretion did not drop. On the contrary, in vivo basal and histamine-induced acid output rose markedly by 40 and 50%, respectively, and in vitro the net movements of 36Cl and 22Na measured in the isolated stomach indicated that active Cl- secretion had resumed. Mucosal PGE2 did not appear to be significantly involved in early development of acid secretion because administration of indomethacin to pups from untreated dams did not significantly modify the secretion measured on day 12. Data indicate that maternal milk depletion of PGE2 prevents the drop of gastric acid secretion previously observed in 12-day-old pups and suggest that in infant rats maternal PGE2 plays a physiological part in regulating acid secretion.

Animals↗

Postnatal development of oxidative enzymes in various rat nephron segments: effect of weaning on different diets.

Activities of fumarase and 3-hydroxyacylCoA dehydrogenase (3-OHDH) were determined in homogenates of rat kidneys between day 21 of gestation and postnatal day 10 and in single isolated nephron segments at postnatal days 16, 21, and 30, and in adult segments. For 3-OHDH activity, main developmental changes were found in proximal convoluted (PCT) and straight tubules (PST) and were characterized by an overshoot of adult level from postnatal days 21 to 30 (59.7 +/- 3.0 and 37.5 +/- 3.4 at day 21 vs. adult values 27.1 +/- 1.5 and 22.7 +/- 1.5 mol.kg dry wt-1.h-1). When rats were precociously weaned on day 16 and fed a diet containing lipid to equal 13% of total caloric intake, a significant decrease in 3-OHDH activity was observed in some parts of the nephron. These changes could be prevented by maintaining early weaned animals on high-fat diet providing 70% of total calories as lipid. Results suggest that changes in fat content of diet during kidney maturation can in part regulate 3-OHDH activity in some nephron segments. Fumarase activity increased 2.6-fold in the medullary thick ascending limb between days 16 and 30; pattern of development was similar to the one reported for Na(+)-K(+)-ATPase activity in this segment. High levels of both enzymes were reached noticeably earlier during development in PCT and PST than in medullary thick ascending limb, which emphasizes metabolic heterogeneity of developing rat kidney nephron.

3-Hydroxyacyl CoA Dehydrogenases↗

Mitochondrial activity of rat kidney during ontogeny.

The development of oxidative metabolism was studied from the late fetal to adult stages in mitochondria isolated from rat kidney. We used the oxygen consumption rate, as an index of inner membrane activity and citrate synthase and fumarase activities as an index of matrix activity and cytochrome c oxidase activity as an index of the number of mitochondria. Fumarase and citrate synthase activities displayed different developmental patterns, suggesting that these Krebs cycle enzymes did not mature synchronously. In fetal mitochondria, net oxygen consumption measured in the presence of succinate or glutamate as substrate, was low; it increased during the day after birth and reached adult level between days 10 and 15. During this period, the levels of citrate synthase and cytochrome c oxidase activity did not change significantly in the isolated mitochondrial fraction. However, in fetal and adult kidney homogenates, these levels increased four-fold, suggesting a corresponding increase in the number of mitochondria. Most of these increases occurred during the 15 days after birth. These results suggest that in rat kidney, mitochondrial maturation precedes the maturation of reabsorptive ion transport and does not limit its development.

Adenosine Diphosphate↗

Inhibition by cow's milk of histamine-induced gastric acid secretion in the rat.

The effects of a 4-day milk diet on basal and histamine-induced gastric acid secretion were examined in vivo in adult rats. Rats were fed on raw cow's milk only. Their acid secretion was measured in the perfused stomach under basal conditions or after administration of varying doses of histamine. In milk-fed rats, basal secretion rose slightly compared with control secretion (1.86 +/- 0.09 vs. 1.52 +/- 0.06 microEq H+/10 min; p less than 0.01), but maximal secretion in response to noncumulative doses of histamine (range: 7.0-20.0 mg/kg) was inhibited by 57%. The degree of this inhibition was similar to that observed in control rats with an equimolar dose of H2-antagonist cimetidine. These results indicate that in the adult rat, chronic administration of cow's milk strongly inhibits histamine-stimulated acid output.

Animals↗

Development of acid secretory function in the rat stomach: sensitivity to secretagogues and corticosterone.

Gastric acid secretion was studied in anesthetized rats from day 6 of the postnatal period up to the time of weaning. Basal H+ secretion was detected from day 6 in the first group studied (2.4 +/- 0.2 muEq of H+/10 min/100 g of body weight, BW) and remained constant up to the time of weaning (day 18: 2.5 +/- 0.2 muEq of H+/10 min/100 g of body weight) except for the period between days 10 and 12, when it fell significantly (1.5 +/- 0.06 muEq H+/10 min/100 g of BW on day 12). Both histamine H2 receptor sensitivity and intracellular transduction mechanism activities were evaluated by studying the secretory responses to histamine, impromidine (an H2 receptor agonist), cimetidine (an H2 receptor antagonist), forskolin (a direct adenylate cyclase activator), and dibutyryl (db) cAMP (an analogue of cAMP, the intracellular messenger mediating the response to histamine). The effects of pentagastrin and carbachol were also determined. The secretory responses obtained on days 6, 8, and 18 were similar and represented about threefold increases over basal secretion for all the secretagogues used. After weaning on day 20, both the basal secretion and the response to secretagogues were significantly increased compared with those of unweaned animals. On day 12, the responses were always weaker than on both days 8 and 18. Injection of 1 mg/kg of corticosterone 21 acetate daily from day 8 resulted on day 12 in a basal secretion and a response to histamine equivalent to those measured in 18-day-old pups not injected.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effects of birth on energy metabolism in the rat kidney.

The oxygen-consumption rates and the activities of fumarase and beta-hydroxyacyl-CoA dehydrogenase were compared in mitochondria isolated from fetal- and neonatal-rat kidney. Whole-organ ATP, phosphocreatine and creatine contents were determined in parallel. Kidney mitochondrial respiratory rates in the presence of succinate, glutamate/malate and palmitoyl-L-carnitine increased between 21 days post coitum and 1 day post partum, together with activities of oxidative enzymes. However, this postnatal maturation of oxidative metabolism was not yet initiated in mitochondria isolated from kidney 1 h post partum. An increase in ATP and phosphocreatine was observed immediately after delivery; newborn-rat kidney ATP content then remained high, whereas phosphocreatine reserves decreased considerably between 6 h and 1 day post partum. It is concluded that the increase in high-energy phosphate compounds observed at birth is not initially related to an activation of oxidative phosphorylation, and probably involves a transient stimulation of anaerobic glycolysis, while a progressive mitochondrial maturation takes place in the rat kidney during the first day of newborn life.

3-Hydroxyacyl CoA Dehydrogenases↗

Development of sodium and chloride transport across fetal and newborn rat stomach in vitro.

1. Unidirectional and net Na+ and Cl- fluxes were determined across isolated fetal rat stomach 19-21 days post-coitum, and across the stomach of newborn rats aged 5 or 12 days. 2. On fetal day 19, absorption of both Na+ and Cl- was greater than the short-circuit current, Isc (net Na+ flux, JnetNa = 4.7 +/- 1.0 and net Cl- flux, JnetCl = 5.4 +/- 1.4 mu equiv cm-2 h-1 vs. Isc = 0.9 +/- 0.1 mu equiv cm-2 h-1). Mucosal addition of 10 microM-amiloride did not significantly alter JnetNa, JnetCl, Isc or total conductance. 3. However, on fetal day 20, neutral absorption of NaCl was no longer observed but amiloride had inhibited electrogenic absorption of Na+, and significant active secretion of Cl- was observed (JnetCl = -1.3 +/- 0.6 mu equiv cm-2 h-1). On day 21 (i.e. 24 h before birth), values for JnetNa, JnetCl, and Isc were not different from those determined on adult gastric mucosa. 4. After birth, NaCl transport continued to exhibit its prenatal characteristics on day 5 but not on day 12, when Na+ and Cl- were both absorbed; on that day, JnetNa-Isc was equal to both JnetCl and to the amiloride-insensitive component of Isc, indicating that neutral NaCl absorption had resumed. 5. These data show that in rat stomach, NaCl transport differentiates on fetal day 20, when H+ secretion is first observed, and thereafter undergoes biphasic development. 6. The significant Cl- absorption observed on post-natal day 12 was concomitant with the inhibition of net H+ secretion.

Amiloride↗

Ontogeny of rat gastric H+-K+-ATPase activity.

Gastric H+-K+-ATPase activity was examined in subcellular fractions (P1, 1,000 g; P2, 20,000 g; P3, 100,000 g) obtained from fundic mucosae of fetal, newborn, and adult rats. Depending on the tissue secretory state the distribution of this enzyme activity predominates in P3, i.e., resting adult, or in P2, i.e., stimulated adult, while total enzyme activity (P2 + P3) remains constant. Enzyme activity was detected as soon as day 19 of gestation and P2 + P3 reached 2.2 +/- 0.2 mumol Pi X mg prot-1 X h-1 at parturition. H+-K+-ATPase activity steadily decreased from 4.0 +/- 0.7 mumol Pi X mg prot-1 X h-1 on day 6 after birth to approximately 0 on day 12. This activity recovered as soon as day 13 (3.5 +/- 0.6 mumol Pi X mg prot-1 X h-1) and continued to rise slowly until adulthood. Incubation of subcellular fractions with 0.1 mM omeprazole resulted in a total loss of H+-K+-ATPase activity at all the stages of development examined. These studies of functional relationship during the development between gastric acid secretion and distribution of gastric H+-K+-ATPase activity among P2 and P3 fractions indicate that this last could be considered as an index of the secretory state of the parietal cell. This enzyme's pattern of development in P2 + P3, correlated with the one previously observed for stimulated H+ secretion in intact tissue, suggest that the H+-K+-ATPase is the limiting step in response to secretagogues during postnatal period.

Adenosine Triphosphatases↗

Role of catecholamines in the control of newborn kidney adenine nucleotide content.

During the 1st h of extrauterine life, the adenine nucleotide content of the rat kidney is modified: the ATP level increases (+30%) while ADP and AMP are lowered (-30 and -50%, respectively). This leads to a high value of energy charge in the newborn kidney (0.89 vs. 0.80 in the fetus). It was possible to obtain in utero a similar modification of ATP, ADP, AMP concentrations by injections to the fetuses of cAMP, dibutyryl cAMP, or isoprenaline. Conversely, the postnatal changes in adenine nucleotide content could be prevented by administration to the fetus, just before birth, of beta- or beta 1-adrenoreceptor antagonists. Therefore the rise of kidney energy charge following parturition appears to be under hormonal control. Glucagon had no effect on kidney adenine nucleotide content. It is strongly suggested that the catecholamines released at the time of parturition are the triggering factor of this evolution.

Adenine Nucleotides↗

[Mitochondrial oxidative phosphorylation in the rat kidney during the perinatal period].

Oxidative metabolism in the developing rat kidney has been studied on isolated mitochondria. An increase of about 50% in state 3 respiration has been observed at birth, using succinate, glutamate, or palmitoyl-L-carnitine as a substrate. The rate of respiration in the presence of 2,4-dinitrophenol was found identical to state 3 respiration in all cases. Cytochrome oxidase activity did not change between the fetal and newborn stages. The increase of mitochondrial respiration revealed here, which is not linked to a modification of the respiratory chain, could be involved in the rise of kidney ATP level and energy charge observed at birth.

Aging↗

[Development of gastric sensitivity to histamine in the rat].

Sensitivity of the developing rat stomach to histamine (HA) was examined on isolated gastric mucosae of rats of various ages from the fetal to adult periods. Spontaneous acid secretion in mu eq/h.cm2 occurred at all the ages studied, at a basal rate of 0.45 +/- 0.07 in fetuses to 0.22 +/- 0.03 (day 5), 0.11 +/- 0.04 (day 10), 0.12 +/- 0.04 (day 12), 0.22 +/- 0.08 (day 16) and 0.33 +/- 0.04 (adults). In the fetal rats as in the adults, marked responses to respectively 10(-5) and 10(-4) M HA were demonstrated. The H2-receptor antagonist cimetidine diminished HA-induced secretion by 66 and 57% in fetuses and adults respectively. Between these two stages (from days 5 to 12), basal secretion and the response to HA dropped significantly. On day 21 of gestation, as well as on the critical days 5 and 12 after parturition, db-cAMP (10(-4) M) caused maximal stimulation of acid secretion. These results indicate that the development of responsiveness to HA in the rat is biphasic. They suggest that after birth, the H2-receptor adenylate cyclase system undergoes major modifications which might lead to the complete lack of responsiveness to HA by day 12.

Aging↗

[Movement of Na+ and Cl- across the isolated fetal rat stomach].

Unidirectional and net isotopic fluxes of Na+ and Cl- were determined at steady state across isolated stomach of rat fetuses on days 19 and 21. On day 19, when parietal cells are not yet functional, net absorptions of Na+ and Cl- (respectively: 3.8 +/- 1.1 and 5.2 +/- 1.7 mu eq X h-1 X cm-2) were observed. By contrast, active secretion of Cl- (-2.1 +/- 1.8 mu eq X h-1 X cm-2) associated with decreased absorption of Na+ (45%) was noted on day 21, and both Na+ and Cl- net movements accounted for the short-circuit current, as observed on adult gastric mucosa. These results show that Na+ active absorption precedes Cl- secretion in fetal rat at the time of parietal cells differenciation.

Amiloride↗

Perinatal changes in adenine nucleotide content of developing rat kidney.

Adenine nucleotides have been measured in fetal and newborn kidneys of rat using the luciferine-luciferase method. In fetuses, between days 18 and 21 of gestation there is a drop of the relative amount of ATP in the renal nucleotide pool. Consequently, the kidneys of 21 days-old fetuses have lowered ATP/ADP ratio (3.6) and energy charge (0.80) compared with values found on day 18 (6.9 and 0.91, respectively); this relative energy deficit is heightened in progesterone induced postmaturity. One hour after delivery whether the gestational stage is 21, 22 or 23 days, there is a rise in ATP and a decrease in AMP content which restore a high energy level in kidney of the newborn and a 30% increase in the total adenine nucleotide pool.

Adenine Nucleotides↗