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G Thibault

Publications and source records attributed to G Thibault.

At least 163 records · Page 9Linked to original sources
PubMed1987

The propeptide Asn1-Tyr126 is the storage form of rat atrial natriuretic factor.

Granules from rat atria were isolated by differential centrifugation and by a 53% (v/v) Percoll gradient after tissue homogenization in 0.25 M-sucrose/50 mM-Na2EDTA. About 40% of the immunoreactive ANF (atrial natriuretic factor) sedimented with the atrial granules during differential centrifugations. On the Percoll gradient, two distinct bands were observed. Cell debris, mitochondria, lysosomes, myofilaments and microsomes were mostly contained in the lightest-density (rho) (1.03-1.07 g/ml) fraction, as demonstrated by electron microscopy and by enzymic markers such as lactate dehydrogenase, monoamine oxidase, cytochrome c reductase, beta-glucuronidase and acid phosphatase. Atrial granules were mostly contained in the denser (rho 1.11-1.15 g/ml) band and were only slightly contaminated by lysosomes, as shown by beta-glucuronidase activity. Analysis of the ANF content in these isolated granules by h.p.l.c., amino acid composition and sequencing demonstrated that it was only the pro-ANF [ANF-(Asn1-Tyr126)-peptide]. The precursor was present in all granules, as demonstrated by immunocytochemistry. Since hormonal propeptides usually undergo intracellular processing, and the matured peptides are subsequently stored in the secretory granules, these results indicate that the processing pathway of ANF may be different from that of other hormonal peptides.

Amino Acid Sequence↗
PubMed1987

A murine monoclonal antibody against rat atrial natriuretic factor (ANF) which cross-reacts with mouse ANF.

A monoclonal antibody (MAb), 2H2, against rat synthetic atrial natriuretic factor (ANF) (Arg101-Tyr126) recognizes native ANF related peptides. The lack of reactivity of 2H2 with amino-terminal truncated ANF peptides implicates the two amino terminal arginine residues of ANF in the 2H2 epitope. Similarly, poor immunoreactivity of human ANF indicates the participation of isoleucine 110. Arginines 101 and 102 and isoleucine 110 may thus participate in a conformational epitope recognized by 2H2 or alternatively, substitution for, or elimination of these residues may alter the conformation of the 2H2 epitope. The MAb shows little cross-reactivity with extracts of rabbit atria but recognizes ANF related peptides in mouse and hamster atrial extracts. 2H2 also identifies immunoreactive ANF in histological sections of rat, mouse and hamster atria.

Animals↗
PubMed1987

Immunoreactive atrial natriuretic factor is present in both atria and ventricles.

A comparative study has been made of the amount and form of immunoreactive atrial natriuretic factor (ANF) present in atria versus ventricles in situ and in cultures of atrial and ventricular cardiocytes in rats of various ages: foetus (-2 days of age), newborn (3 days of age), 10 days, 35 days and 70 days old. It was first established that ANF is present in the circulation at all ages investigated, the highest levels being found in the foetus. The atrial content of ANF increased gradually with time and was always in the microgram range. In ventricles, where it was in the nanogram range, total ANF increased with age. More ANF was found in the right than in the left atrium at all ages except in the foetus where the reverse was true. In ventricles, ANF was distributed equally between right ventricle, left ventricle and septum in the foetus, the newborn and in the adult. At 10 days and 35 days, ANF was unequally distributed: (right ventricle greater than septum greater than left ventricle). In cultured atrial cardiocytes, ANF was more abundant than in cultured ventricular cardiocytes at all ages and the amount decreased with the age of the donor animals. Atrial cardiocytes secreted more ANF than did ventricular cardiocytes at all ages investigated, the amount secreted generally decreasing with the age of the donor animals. The secretory activity of atrial cardiocytes per hour was also higher than that of their ventricular counterparts; over this time period, both types of cells secreted more ANF in the presence of serum.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗
PubMed1987

The whole heart is an endocrine gland.

The relative contribution of atria and ventricles in the release of immunoreactive (IR-) atrial natriuretic factor (ANF) and in the eventual circulating levels of the peptide was evaluated in several situations. In situ, not only atrial but also ventricular cardiocytes contain IR-ANF although the amount present in ventricles is very low, particularly in the adult rat. In cardiomyopathic hamsters with heart failure, the amount of IR-ANF decreases constantly in the atria and increases in parallel in the ventricles so that the ratio of IR-ANF which is 114:1 in control animals becomes 3.9:1 in hamsters with severe heart failure. With the Langendorff preparation, the whole heart secretes much more IR-ANF than the isolated ventricles. From the latter, IR-ANF may originate from subendocardial cells of the conduction system and, more likely, from all ventricular cardiocytes where the peptide may be secreted by two pathways: one constitutive, the other regulated. These results are in agreement with clinical studies where circulating IR-ANF was measured in the same patients following catheterization of coronary sinus and great cardiac vein (draining the left ventricle). In this situation, it was found that while the levels of IR-ANF in the great cardiac vein are higher than in the general circulation, they are much lower than in the coronary sinus. All these results tend to indicate that IR-ANF is secreted by ventricular cardiocytes in relatively low amounts even in periods of intense stimulation.

Animals↗
PubMed1987

Comparative effect of ANF and various diuretics on isolated nephron segments.

The effect of the synthetic form of ANF 0.1 to 10 microgram/ml (peptide 101-126), a diuretic and natriuretic peptide isolated from rat heart atria, on the metabolism of dog and rat kidney tubules was studied in vitro and compared to that of furosemide (0.1 to 1 mM), hydrochlorothiazide (0.5 mM) or amiloride (0.1 mM). In order to pinpoint eventual site(s) of ANF action along the nephron, proximal tubules, thick ascending limbs and papillary collecting ducts were isolated from dog kidneys as well as proximal tubules from rat kidneys. The substrate uptake (O2, lactate, glutamine, glucose) and production of metabolites (glutamate, ammonium, alanine, glucose) by these nephron segments were measured in absence or presence of the diuretic agents or the vehicle for ANF (acetate 1 mM). The total ATP turnover and the contribution of identified metabolic pathways for this turnover was calculated. It was expected that a molecule with diuretic properties reducing the permeability of cell membranes to NaCl would secondarily reduce the Na-K-ATPase activity, and therefore the oxygen and substrate utilization by affected cells. It was shown: that each nephron segment used presented the expected specific metabolic characteristics; that furosemide markedly inhibits the oxidative metabolism of thick ascending limbs; that acetate (the vehicle used for ANF) displaces the oxidation of glutamine and lactate in nephron segments with aerobic metabolism; that ANF had no effect on the metabolism of the studied segments despite the presence of specific c'GMP-generating receptors in the distal nephron. It is concluded that ANF must exert its natriuretic effect by a mechanism different from that of classical diuretics.

Adenosine Triphosphate↗
PubMed1987

The complete amino acid sequence of rat submaxillary gland tonin does contain the aspartic acid at the active site: confirmation by protein sequence analysis.

The revised amino acid sequence of rat submaxillary gland tonin, a serine protease, does contain the active site Asp residue. The active site of this kallikrein-related enzyme is thus made up of the same catalytic triad (Asp, Ser, and His) found in all known serine proteases. The important Asp residue has now been localized in a 16 amino acid peptide previously reported as missing in the tonin sequence. The complete amino acid sequence thus contains 235 residues corresponding to a molecular weight of 25,658, more in agreement with previously reported molecular weights. Moreover, the revised structure led (a) to the assignment of Arg, Asn, and Val residues instead of His, Asp, and Gly at positions 63, 165, and 169, respectively; (b) to the assignment of residues occupying an overlapping sequence at positions 165-171, and finally (c) to the localization of two N-glycosylation sites at positions 82 and 165. These results further document the close relationship of tonin to the ever expanding kallikrein family.

Amino Acid Sequence↗
PubMed1987

Renin dependency of the effect of chronically administered atrial natriuretic factor in two-kidney, one clip rats.

Conscious two-kidney, one clip rats with 150 mm Hg or higher systolic blood pressure were infused with saralasin for 60 minutes. Those with a blood pressure decline of 30 mm Hg or more were classified as saralasin-sensitive; those with a decrease of 10 mm Hg or less were considered saralasin-resistant. The animals were then housed in metabolic cages. Groups of sham-operated normotensive, saralasin-sensitive or saralasin-resistant two-kidney, one clip (2K1C) rats were infused with atrial natriuretic factor (Arg 101-Tyr 126), 100 ng/hr per rat, for 6 days. Corresponding control groups were sham-infused. Blood pressure was initially higher in the saralasin-sensitive groups (176 +/- 6 and 181 +/- 1 mm Hg, respectively) than in the saralasin-resistant groups (160 +/- 4 and 169 +/- 4 mm Hg, respectively). Atrial natriuretic factor infusion produced a gradual decline in blood pressure to 128 +/- 5 mm Hg, but only in saralasin-sensitive 2K1C animals. Urinary volume, initially higher in saralasin-sensitive hypertensive than in normotensive rats, was depressed during atrial natriuretic factor infusion. Urinary sodium excretion and water intake showed the same tendency, but the changes were not significant. No such modifications were observed in saralasin-resistant or sham-operated rats infused with atrial natriuretic factor. Body weight, which was higher in normotensive animals, was unchanged during atrial natriuretic factor infusion. Saralasin-sensitive, noninfused 2K1C rats were the only group with higher plasma renin activity than sham-operated, normotensive controls. Plasma aldosterone was higher in the former than in the other five groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Atrial natriuretic factor during development and reversal of one-kidney, one clip hypertension.

Atrial natriuretic factor (ANF) was studied in rat plasma and atria 1, 2, 4, 6, and 8 weeks after constriction of the left renal artery and removal of the contralateral kidney. Plasma ANF was elevated at all periods of investigation. A positive correlation was observed between plasma ANF and blood pressure (r = 0.56, p less than 0.001). The total atrial ANF content (microgram/atrium) in one-kidney, one clip (1K1C) rats was lower during Weeks 1 and 2, but only in the left atrium. Lower ANF concentrations (microgram/mg protein) were also evident in the left atrium at Weeks 1, 2, and 8, and in the right atrium at Week 8. A negative correlation between ANF in plasma and in the left atrium was discerned (r = 0.43, p less than 0.01). Blood pressure (184 +/- 4 vs 114 +/- 4 mm Hg), body weight, and plasma ANF were also examined in 1K1C rats and their normotensive controls before and after unclipping. Blood pressure was normalized 6 hours after unclipping. Plasma ANF declined in 1K1C rats within 6 hours after clip removal, but it was still higher than in the controls. Plasma ANF was similar in both groups on Days 9 and 13 after unclipping. There were no differences in atrial ANF between hypertensive and normotensive animals 13 days after unclipping. The high levels of plasma ANF observed in 1K1C rats probably are secondary to increased intra-atrial pressure caused by the dual mechanism of expanded plasma volume and high blood pressure.

Animals↗
PubMed1987

Role of right and left atria in natriuresis and atrial natriuretic factor release during blood volume changes in the conscious rat.

This study investigated whether excision of either the right or left atrial appendage of rats alters their natriuretic response and the release of atrial natriuretic factor during acute blood volume expansion or reduction. These animals were subjected to a thoracotomy and either had their right or left atrial appendages removed or underwent a right or left atrial sham appendectomy for comparative, control purposes. Intrajugular vein, intracarotid artery, and intravesical catheters were installed 3-4 weeks later under sodium pentobarbital anesthesia. Then, when the rats were conscious, blood volume was expanded using blood from donor rats once every 15 minutes in 3 increments of 10% of the calculated total blood volume at a rate of 5 ml/kg/min. Blood and urine samples were collected before volume expansion and at the end of each 15-minute period, with the withdrawn blood being replaced. A maximal fourfold increase in urinary volume, urinary sodium excretion, and plasma atrial natriuretic factor was observed in all but the right-atrial-appendectomized animals. Plasma atrial natriuretic factor, urinary volume, and urinary sodium excretion were correlated in all 4 groups. No significant changes in blood pressure or hematocrit were noted. Plasma vasopressin, measured at the end of volume expansion, was significantly lower in animals subjected to left atrial appendectomy. High-performance liquid chromatography of plasma from the control groups indicated that most of the released ANF during blood volume expansion corresponded to a high molecular weight peptide. Additional rats, processed as above, were subjected to 10% blood volume decrements.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Localization of 125I-atrial natriuretic factor (ANF)-binding sites in rat renal medulla. A light and electron microscope autoradiographic study.

Using light and electron microscope autoradiography in vivo, the localization of 125I-(Arg 101-Tyr 126) atrial natriuretic factor (ANF)-binding sites was studied in the renal medulla of rats. At the light microscopic level, the autoradiographic reaction was mainly distributed in patches in the outer medulla, and followed the tubular architecture in the innermost part of the inner medulla. At the electron microscopic level, binding sites were mainly found in the outer medullary descending vasa recta and inner medullary collecting ducts. These results suggest that, in rats, the renal medulla may participate in the natriuresis and diuresis produced by ANF through vascular and tubular effects; the former by changing medullary blood flow at the level of descending vasa recta and the latter by acting on electrolyte and water transport at the level of collecting ducts.

Animals↗
PubMed1987

Cardiac and plasma atrial natriuretic factor in experimental congestive heart failure.

The cardiac and plasma levels of immunoreactive (IR-) atrial natriuretic factor (ANF) in cardiomyopathic hamsters with moderate and severe congestive heart failure were measured, compared with those of controls, and correlated by HPLC analysis of IR-ANF in atria, ventricles, and plasma and with the ultrastructure of atrial and ventricular cells. Congestive heart failure in the hamster produced a significant increase in plasma IR-ANF, a significant decrease in atrial IR-ANF, and a marked increase in ventricular IR-ANF. The HPLC pattern of IR-ANF was of the high mol wt type in atria and ventricles of control and cardiomyopathic animals. High mol wt forms of ANF appeared in the plasma of animals with severe congestive heart failure, but not in controls. Severe congestive heart failure produced a tremendous increase in the size of the Golgi complex, with a decrease in the number and size of secretory granules in atrial cardiocytes. Ventricular cardiocytes also showed a less marked increase in the size of the Golgi complex. Secretory-like granules indistinguishable from lysosomes were present in about 1% of ventricular cardiocytes of control hamsters; in hamsters with severe congestive heart failure, secretory granules, identical to those of atrial cardiocytes, were present in greater number in about 20% of ventricular cardiocytes. Immunocytochemistry (immunogold technique) revealed that secretory granules containing IR-ANF are not present in control ventricular cardiocytes but are localized in relatively large number in about 20% of ventricular cardiocytes in hamsters with severe congestive heart failure. These results suggest that the increased IR-ANF levels (including the high mol wt forms) in animals with congestive heart failure may come from hypersecretion of both atrial and ventricular cardiocytes.

Animals↗
PubMed1987

Body fluids and plasma atrial peptide after its chronic infusion in hypertensive rats.

To determine the role of body fluid volume in the chronic hypotensive effect of atrial natriuretic factor (ANF), spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats were infused with the peptide (Arg 101-Tyr 126) at a rate of 100 ng/h/rat for 5 days. Blood pressure (BP) was decreased from 176 +/- 4 to 133 +/- 3 mmHg in the SHR group 4 days after ANF infusion was initiated, whereas no changes were observed in ANF-infused WKY animals. Starting 5 days after the infusion began, body fluid measurements revealed no differences in plasma, blood and extracellular fluid volumes or in interstitial spaces. BP and plasma ANF concentrations were determined in another set of experiments before, during and after chronic ANF infusion. BP declined from 169 +/- 3 to 133 +/- 5 mmHg in SHR 5 days after the infusion commenced, but returned to basal values by day 10 or 11. Plasma ANF was significantly higher in SHR than in WKY rats throughout the observation period. However, there were no discernible changes in this parameter in ANF-infused SHR compared to non-infused SHR. A 3-fold rise in plasma ANF was noted in infused WKY rats at day 3 only. It is concluded that the chronic hypotensive effect of ANF in hypertensive animals is not related to changes in either body fluid volume or distribution. Moreover, the finding that chronic ANF infusion reduces BP in SHR without altering its plasma levels suggests a rapid ANF turnover.

Animals↗
PubMed1987

[ANF receptors at the glomerular level in reno-vascular hypertension in rats].

The effect that hypertension may have on plasma, atrial ANF levels and on isolate rat renal glomeruli ANF receptors was investigated. Different models of renal hypertension were studied: 1-K, 1-C (BP = 177 +/- 7 mmHg), 2-K, 1-C (BP = 158 +/- 3 mmHg) and their respective controls (BP = 100 +/- 2 mmHg). At this stage of hypertension (two weeks) plasma ANF was 56 +/- 9 and 25 +/- 2 pg/ml in 2-K, 1-C and controls respectively and 124 +/- 22 vs 35 +/- 4 pg/ml in 1-K, 1-C and controls. Atrial ANF content was lower in hypertensive animals. A marked up-regulation of the glomerular ANF receptor density was observed in 2-K, 1-C animals. In the right kidney we found 840 fmol/mg protein and 540 fmol/mg protein, and in the left 1,070 fmol/mg protein against 608 fmol/mg protein in 2-K, 1-C and control animals respectively. No change was observed in glomerular ANF receptor density in 1-K, 1-C animals. We have then demonstrated that glomerular ANF receptor density is higher in 2-K, 1-C hypertensive than in normotensive animals. It could be possible, however, that the receptor density may change during the evolution of high blood pressure in the models of experimental hypertension currently studied.

Animals↗
PubMed1987

Atrial natriuretic factor (ANF) in experimental and human hypertension.

The plasma levels of immunoreactive (IR)-ANF have been evaluated by radioimmunoassay in several models of experimental hypertension and in human hypertension. In all models of experimental hypertension so far studied, the plasma levels of IR-ANF are consistently increased. This is accompanied by a decrease, at certain time intervals, of the IR-ANF levels in the left atrium. In human essential hypertension, the plasma levels of IR-ANF are not increased except in the severe form (diastolic blood pressure above 110 mmHg). In renovascular hypertension, the peripheral levels of IR-ANF are not different from the normal levels but are increased above normal in aortic blood.

Animals↗