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P Falardeau

Publications and source records attributed to P Falardeau.

At least 55 records · Page 3Linked to original sources
PubMed1988

Purification and characterization of the D2-dopamine receptor from bovine anterior pituitary.

The D2-dopamine receptor from bovine anterior pituitary has been purified approximately 33,000-fold to apparent homogeneity by sequential use of affinity chromatography on immobilized carboxymethyleneoximinospiperone-Sepharose, Datura stramonium lectin-agarose, and hydroxylapatite chromatography. The purification yields a single polypeptide band of Mr approximately 120,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed by labeling with radioiodinated Bolton-Hunter reagent, Coomassie Blue, or silver staining. The purified D2 receptor preparations display a specific activity of approximately 5.3 nmol of [3H]spiperone bound per mg of protein. In detergent solutions, the purified receptor has a KD for [3H]spiperone of 5-8 nM; however, after reinsertion of the purified protein into phospholipid vesicles, a KD of approximately 160 pM is obtained, similar to that found for the receptor in crude membrane preparations. Several lines of evidence document that this polypeptide contains the ligand binding site as well as the functional activity of the D2 receptor. The Mr approximately 120,000 peptide can be covalently labeled by the affinity probe, 125I-bromoacetyl-N-(p-aminophenethyl)spiperone, with the pharmacological specificity expected of a D2-dopamine receptor. Agonist and antagonist ligands compete for [3H]spiperone binding to purified receptors in phospholipid vesicles with a rank order of potency and selectivity typical of a D2-dopamine receptor. Moreover, when reinserted into phospholipid vesicles with purified brain Gi/Go, the purified D2 receptors mediate the agonist stimulation of 35S-labeled guanosine 5'-O-(thiotriphosphate) binding to brain G-proteins with a typical D2-dopaminergic order of potency. These data suggest that we have purified an intact functional D2-dopamine receptor.

Affinity Labels↗
PubMed1988

Repeated stimulation of D-1 dopamine receptors increases the circling response to bromocriptine in rats with a 6-OHDA lesion.

We have studied the effect of repeated injections of a specific D-1 agonist, (SKF 38393), on circling behaviour in response to a specific D-2 agonist (bromocriptine) and on striatal D-2 DA receptors in rats with a unilateral lesion of the nigrostriatal pathway. Two groups of rats were formed on the basis of their circling response to a first injection of SKF 38393: clear contralateral circling (SKF-positive) and no (or ipsiversive) circling (SKF-negative). A few days later, these rats were tested once for bromocriptine-induced circling. A repeated treatment of 8 injections of SKF 38393 over 4 days was started the next day. The animals were tested again for bromocriptine-induced circling at the end of the repeated treatment with the D-1 agonist. A third group of rats received only the 2 test injections of bromocriptine and a fourth group included intact control animals. During the repeated treatment, the D-1 agonist induced an increase of the circling response up to the fourth injection and a decrease thereafter in the SKF-positive group. This was followed by a pronounced increase of circling in response to the second injection of bromocriptine. This increased sensitivity appears to be due to the chronic treatment with SKF 38393 since animals that did not respond initially to the D-1 agonist (SKF-negative group) and those that received only 2 injections of bromocriptine showed no increase of their circling response. The treatments did not increase the affinity of [3H]spiperone binding for the striatal D-2 dopamine receptor.(ABSTRACT TRUNCATED AT 250 WORDS)

2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-ben↗
PubMed1988

Behavioral and biochemical effect of chronic treatment with D-1 and/or D-2 dopamine agonists in MPTP monkeys.

Monkeys developed a severe parkinsonian syndrome after intravenous administration of (MPTP). L-DOPA/carbidopa (D-1 and D-2) or bromocriptine (D-2) treatment relieved the parkinsonian symptoms, whereas SKF 38393 (D-1) was ineffective. No dyskinesia was seen in monkeys receiving bromocriptine or SKF 38393 as opposed to the L-DOPA-treated animals, in which the dyskinetic response appeared to increased with time. MPTP induced a significant increase (25%, P less than 0.01) in the number of [3H]spiperone binding sites (Bmax) in the caudate nucleus and in putamen. The Bmax of spiperone binding in the L-DOPA-treated monkeys was on average 18% lower (P less than 0.01) than that of the animals treated with MPTP alone. The Bmax for the bromocriptine-treated group was 29% (P less than 0.01) less than that in the MPTP-treated group or 11% (P less than 0.05) less than that in the L-DOPA-treated monkeys. The SKF 38393 treatment induced a 23% (P less than 0.01) decrease in the Bmax as compared to that of animals treated with MPTP alone, and no significant change compared to the L-DOPA- or bromocriptine-treated animals. These results suggest that stimulation of D-1 and D-2 dopamine receptors can differently influence the mechanisms controlling dopamine agonist-induced dyskinesia in MPTP-treated monkeys.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗
PubMed1988

Relation between brain dopamine loss and D2 dopamine receptor density in MPTP monkeys.

The relationship between dopamine (DA), dihydroxyphenylacetic acid (DOPAC) or homovanillic acid (HVA) concentrations and [3H]spiperone binding to D2 DA receptors in caudate nucleus, putamen and nucleus accumbens following DAergic lesion with MPTP in monkeys was investigated. The level of DA depletion varied from 37% to 100% while a mean elevation in [3H]spiperone binding density of 114.8 +/- 4.7% of control (P less than 0.01) was observed when DA depletion was at least 90% of control values. A logarithmic correlation (y = -7.19 In x +111.15; r = -0.54, P less than 0.01) between DA concentrations (x) and the density of [3H]spiperone binding sites (y) was observed in all brain regions. The correlation between DOPAC or HVA concentrations and [3H]spiperone binding was best represented by linear rather than logarithmic equations. These results indicate that supersensitivity of DA receptors develops after MPTP lesion in monkeys when the number of remaining nerve fibers becomes too few to compensate for the DA loss.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗
PubMed1988

Striatal D-2 dopamine agonist binding sites fluctuate during the rat estrous cycle.

Striatal D-2 dopamine (DA) antagonist and agonist binding sites were measured during the rat estrous cycle and compared to ovariectomized (OVX) rats. Dopaminergic D-2 antagonist binding sites were constant during the estrous cycle while agonist binding sites show a rapid and significant decrease of the ratio of high to low D-2 agonist binding sites from proestrus AM (PAM) to diestrus 1 (D1) and return to OVX value in diestrus 2 (DII). Thus, physiological fluctuations of hormones as occur during the estrous cycle can modulate extrahypothalamic biogenic amine activity, namely striatal DA systems which are not involved in the control of hormone secretion.

Animals↗
PubMed1988

Enzymatic oxidation of all-trans retinal to retinoic acid in rat tissues.

The 100,000 x g supernatant (cytosolic) fraction of rat tissue homogenates catalyzes the oxidation of all-trans retinal to retinoic acid. Kidney, testis, and lung were the most active of the tissues examined. The presence of enzyme activity in liver and intestine could be detected only when a substrate concentration beyond the saturation point for retinal reductase was used. Spleen, brain, and plasma had no activity. Boiled supernatants did not catalyze the reaction. The enzymatic product was chemically and physically identified as retinoic acid. The cytosol of kidney tissue also catalyzed the conversion of retinol to retinoic acid. These data indicate that kidney tissue has the highest retinal oxidase activity and suggest that it may play a major role in the oxidative metabolism of retinol in the body.

Aldehyde Oxidoreductases↗
PubMed1987

Guanine nucleotide regulation of dopamine receptor agonist affinity states in rat estradiol-induced pituitary tumors.

We have investigated dopamine (DA) receptor agonist high- and low-affinity states in female rat estradiol-induced prolactin (PRL)-secreting pituitary tumors and intact pituitary tissue. Estradiol treatment increased the anterior pituitary weight 9-fold and plasma prolactin levels 74-fold and these measures are correlated (R = 0.745, n = 73, p less than 0.001). Competition for [3H]-spiperone binding to the DA receptor by apomorphine was compared in normal and adenomatous pituitary tissue. The inhibition constants (Ki) and the proportions of the two apomorphine sites are unchanged in tumors compared to intact pituitary tissue. Guanosine 5'-[beta-gamma-imino]triphosphate (Gpp(NH)p) causes complete conversion of the high into low affinity dopaminergic agonist site in normal pituitary and in tumors. These results suggest that rats with primary estradiol-induced pituitary tumors have normal and functional DA receptors.

Animals↗
PubMed1987

Purification and quantitative analysis of urinary prostanoids in human and in rat by packed and capillary gas chromatography-negative-ion chemical-ionization mass spectrometry.

We describe a method for the quantitative analysis of prostaglandin (PG) E2 and the major urinary metabolites PGI2 and thromboxane (Tx) A2 in human and in rat by combined gas chromatography and negative-ion chemical-ionization mass spectrometry. The procedure is based on the sequential use of small columns with distinct properties combined with a thin-layer chromatography step, for the extraction and the purification of urinary prostaglandins. The compounds are then analysed as their pentafluorobenzyl ester-O-methyloxime-trimethylsilyl ether derivatives, using either packed or capillary columns. Deuterated analogues are used as internal standards. The method was established by using tritiated prostaglandins covering the extremes of polarity in order to optimize the recovery of prostanoids as well as the quality of the chromatograms and spectra. The overall recovery was 24%. Standard curves were obtained by the same procedure and found to be reproducible, with a maximal day-to-day variation of +/- 5%. The relatively simple approach required for the sequential extraction and purification of prostaglandins on small columns of distinct properties, combined with the highly specific and highly sensitive method of detection, places this procedure among the most reliable method for measuring urinary prostanoids in both humans and animals. In addition, the procedure is faster than classical approaches and necessitates smaller amounts of samples and solvents.

6-Ketoprostaglandin F1 alpha↗
PubMed1987

Regional effect of estradiol on rat caudate-putamen dopamine receptors: lateral-medial differences.

The effect of a chronic 17 beta-estradiol (10 micrograms, b.i.d.) treatment of 2 weeks to ovariectomized rats was investigated on lateral and medial caudate-putamen dopamine receptors. Dopamine D2 receptors were assayed with [3H]spiperone binding to caudate-putamen homogenates or by autoradiography of forebrain sections. Estradiol treatment leads to a significant increase in the density of lateral caudate-putamen dopamine receptors while for the medial dopamine receptors the increase is non-significant. This effect is observed when plasma levels of 17 beta-estradiol and prolactin are increased. These results indicate that the effect of estradiol on striatal dopamine receptors is heterogeneous and that those in the lateral part are more susceptible to this steroid.

Animals↗
PubMed1987

Behavioral and biochemical evidence for a different effect of repeated administration of L-dopa and bromocriptine on denervated versus non-denervated striatal dopamine receptors.

The effect of repeated administration of bromocriptine and L-3,4-dihydroxyphenylalanine (L-DOPA) was studied behaviorally and biochemically in rats with a unilateral lesion of the nigrostriatal pathway. Groups of rats injected eight times with bromocriptine or L-DOPA significantly increased their contraversive circling. Rats receiving only two injections of bromocriptine did not. Animals receiving two injections of L-DOPA showed a slight but significant increase in circling. The affinity of the binding of [3H]spiperone to the dopamine receptors was unchanged by the lesion or the treatments, while the density of the binding was significantly modified. Chronic treatment with bromocriptine induced a significant decrease in the density of D2 dopamine receptors in the intact striata, while on the lesioned side, it remained unchanged. By contrast, chronic administration of L-DOPA induced a significant increase in density of the striatal dopamine receptors in the lesioned striata in addition to that caused by denervation, while the decrease on the intact side was not significant. It seems that contrary to the intact striatum, the lesioned side had a defective down-regulation mechanism in response to chronic treatment with a dopamine agonist. The results also show that L-DOPA was more potent than bromocriptine in inducing agonist supersensitivity in a denervated striatum. This may explain why chronic treatment with bromocriptine has a lesser tendency to induce dyskinesia in patients with Parkinson's disease.

Animals↗
PubMed1987

Unilateral nodular diabetic glomerulosclerosis: recurrence of an experiment of nature.

The case of a 60-year-old diabetic patient with a fully developed diabetic glomerulosclerosis in one kidney, but only ischemic lesions in the contralateral one, which was 'protected' by a renal artery stenosis, is presented. The only other report of such a peculiar observation was made by Berkman and Rifkin on a patient who died in 1940. Because of the rather high incidence of diabetes and of renal artery stenosis, the scarcity of this 'experiment of nature' is astonishing and can be barely explained by a precise timing of the two pathological conditions. Despite rather detailed information on the clinical and paraclinical evolution of the present patient, the exact sequence of events could not be determined with certitude. Both cases bring a strong support for the role of glomerular hyperperfusion-hypertension in the pathogenesis and evolution of diabetic nephropathy and provide a theoretical basis for the importance of keeping the arterial pressure low in diabetics.

Diabetes Mellitus, Type 2↗
PubMed1987

Contrasting dopaminergic patterns in two forms of genetic hypertension.

Dopaminergic mechanisms in genetic hypertension were explored via the measurement of catecholamine (CA) turnover, tissue concentration and urinary excretion of dopamine (DA) and its metabolites. In salt-sensitive (S) Dahl rats, the tissue concentration and urinary excretion of DA and its metabolites were decreased in response to salt loading, while adrenal dopamine-beta-hydroxylase (D beta H) activity and aldosterone responsiveness to angiotensin II (A II) were increased. In contrast, spontaneously-hypertensive rats (SHR) showed elevated tissue levels and urinary excretion of DA and its metabolites, adrenal DA turnover and ganglionic DA generation following cholinergic stimulation, but D beta H activity and aldosterone responsiveness to A II were diminished. These two patterns, the hypodopaminergic state in Dahl S rats and the hyperdopaminergic state in SHR, account for two distinct mechanisms of hypertension and precede its development. We detected striking dopaminergic activity-related similarities between Dahl S rats and low-renin essential hypertension (EH) on the one hand, and SHR and non-modulator EH patients on the other.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1986

Chronic treatment with L-DOPA, but not bromocriptine induces dyskinesia in MPTP-parkinsonian monkeys. Correlation with [3H]spiperone binding.

A group of 5 monkeys developed a severe parkinsonian syndrome after intravenous administration of the toxin MPTP. One remained untreated while two animals were treated daily for 5 months with supramaximal doses of Sinemet and two with bromocriptine orally. Both drugs relieved the parkinsonian symptoms but the animals on Sinemet developed after 2 weeks prominent lingual dyskinesia which remained visible after each dose until the end of the experiment. In the two animals on bromocriptine no dyskinesia was observed. After sacrifice, the levels of dopamine and [3H]spiperone binding were studied bilaterally in the anterior and posterior caudate nucleus, anterior and posterior putamen and in the nucleus accumbens. The loss of dopamine was equivalent in the Sinemet and the bromocriptine treated animals (more than 90%) and there was a complete disappearance of the substantia nigra pars compacta. In all structures studied, the Bmax for [3H]spiperone binding was on average 10% higher in the Sinemet than in the bromocriptine-treated animals. We therefore believe that L-DOPA and bromocriptine affect denervated postsynaptic dopamine receptors differently, that bromocriptine is less likely to induce agonist supersensitivity and that this probably explains the lesser tendency to induce dyskinesia after chronic treatment.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗
PubMed1986

Effect of indomethacin treatment on prostaglandin excretion and tissue phospholipid fatty acid composition in rats bearing mammotropic pituitary tumor, MtT-F4.

The hypothesis that hormonal changes induced by the mammotropic pituitary tumor, MtT-F4, might accelerate the conversion of n-6 fatty acids to prostaglandins, resulting in a partial depletion of n-6 fatty acids was examined. In tumor-bearing rats, the administration of indomethacin induced a 50% reduction of the urinary prostaglandin levels, but exerted no significant effect on the fatty acid composition of the tissue phospholipids. It is concluded that the observed depletion of n-6 fatty acids in tumor-bearing rats is not caused by an increased production of prostaglandins.

6-Ketoprostaglandin F1 alpha↗
PubMed1985

Simultaneous determination of glucose turnover, alanine turnover, and gluconeogenesis in human using a double stable-isotope-labeled tracer infusion and gas chromatography-mass spectrometry analysis.

We have developed and validated a new method to measure simultaneously glucose turnover, alanine turnover, and gluconeogenesis in human, in steady and non-steady states, using a double stable-isotope-labeled tracer infusion and GC-MS analysis. The method is based on the concomitant infusion and dilution of D-[2,3,4,6,6-2H5]glucose and L-[1,2,3-13C3]alanine. The choice of the tracers was done on the basis of a minimal overlap between the ions of interest and those arising from natural isotopic abundances. Alanine was chosen as the gluconeogenic substrate because it is the major gluconeogenic amino acid extracted by the liver and, with lactate, constitutes the bulk of the gluconeogenic precursors. The method was validated by comparing the results obtained during simultaneous infusion of trace amounts of both stable isotope labeled compounds with the radioactive tracers (D-[3-3H]glucose and L-[1,2,3-14C3]alanine) in a normal and a diabetic subject; the radiolabeled tracers were used as the accepted reference procedure. A slight overestimation of glucose turnover (7.3 versus 6.8 in normal and 10.8 versus 9.2 mumol/kg min in diabetic subject) was noticed when the stable isotope-labeled tracers were used. For the basal turnover rate of alanine, similar values were obtained with both methods (6.2 mumol/kg min). For gluconeogenesis, higher values were observed in the basal state with the stable isotopes (0.42 versus 0.21 mumol/kg min); however, these differences disappeared in the postprandial period after the ingestion of a mixed meal. Despite those minor differences, the overall correlation with the reference method was excellent for glucose turnover (r = 0.87) and gluconeogenesis (r = 0.86).(ABSTRACT TRUNCATED AT 250 WORDS)

Alanine↗
PubMed1985

Urinary levels of 2,3-dinor-6-oxo-PGF1 alpha: a reliable index of the production of PGI2 in the spontaneously hypertensive rat.

The urinary levels of 2,3-dinor-6-oxo-PGF1 alpha (PGI2-M), a major metabolite of PGI2, are determined by the balance between the amount of PGI2 synthesized and the extent of its further metabolic oxidation. The purpose of the present study was to determine if the urinary excretion of PGI2-M can be used as a reliable index of the in vivo production of PGI2 in both normal Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR). This involved the exclusion of differences in metabolism between these two strains of rats. In order to do so, we monitored the urinary excretion of PGI2-M during paired intravenous infusions of 6-oxo-PGF1 alpha (the stable product of the spontaneous hydrolysis of PGI2) in conscious, unrestrained SHR and WKY rats aged 12-15 weeks, in doses ranging from 250 to 700 ng. In one experiment, PGI2 was infused instead of 6-oxo-PGF1 alpha. The results of these experiments indicate that SHR and WKY rats are equal with regard to the transformation of 6-oxo-PGF1 alpha and PGI2 into PGI2-M. For both groups, there is a good correlation between the amount of 6-oxo-PGF1 alpha infused and the amount of PGI2-M excreted in urine. These observations confirm the validity of using the urinary levels of 2,3-dinor-6-oxo-PGF1 alpha as an index of PGI2 production in both WKY and SHR. In addition, they support the conclusions drawn from our previous studies, namely that SHR do not produce more PGI2 than WKY rats in vivo, contrary to the situation prevailing in vitro.

6-Ketoprostaglandin F1 alpha↗
PubMed1985

Modulation of brain and pituitary dopamine receptors by estrogens and prolactin.

Ovariectomized rats were treated for 2 weeks with 17 beta-estradiol (0.0002-100 micrograms/day). [3H]spiperone striatal dopamine receptor binding was maximally increased by 30% after 0.05 micrograms/day of 17 beta-estradiol and a similar increase is observed at higher doses. By contrast, plasma prolactin concentrations of these rats are unchanged after 0.05 micrograms/day and increased after 100 micrograms/day. A chronic estradiol treatment at very low doses (0.0002-0.001 microgram/day) leads to increases in pituitary dopamine receptor binding while plasma prolactin levels are unchanged. At higher doses (1-100 micrograms/day) binding is decreased and plasma prolactin concentrations are elevated. [3H]spiperone striatal dopamine receptor binding is elevated in lactating female rats compared to intact or ovariectomized female rats. Anterior pituitary dopamine receptor concentrations fluctuate during the estrous cycle while striatal dopamine receptors are unchanged. An injection of 30 ng of estradiol, which reproduces the estradiol proestrus surge, leads as in proestrus, to a decrease of anterior pituitary dopamine receptors.

Animals↗