PubMed Health⌕ Search

Biomedical subjects

S Lemaire

Publications and source records attributed to S Lemaire.

At least 73 records · Page 4Linked to original sources
PubMed1992

Characterization of [3H]desmethylimipramine binding in bovine adrenal medulla: interactions with sigma- and (or) phencyclidine-receptor ligands.

High-affinity binding sites (apparent KD 2.87 nM) for [3H]desmethylimipramine ([3H]DMI), have been demonstrated and characterized in membrane preparations of bovine adrenal medulla. The binding of [3H]DMI improved upon pretreatment of the membrane with KCl and was saturable, sodium dependent, and potently inhibited by nisoxetine and imipramine. [3H]DMI binding was also inhibited by various phencyclidine (PCP)- and (or) sigma-receptor ligands, with the following order of potency: haloperidol > rimcazole > (-)-butaclamol > dextromethorphan > MK-801 > (+)-3-(3-hydroxyphenyl)-N-(1-propyl)piperidine ((+)-3-PPP) > PCP > N-(2-thienyl)cyclohexyl-3,4-piperidine (TCP) > (+)-SKF-10047 > (-)-SKF-10047. The inhibition produced by sigma ligands was not attributed to stimulation of either sigma 1- or sigma 2-receptors, owing to inactivity of the selective sigma-receptor ligands (+)-pentazocine and 1,3-di(2-tolyl)guanidine (DTG). The inhibition of [3H]DMI binding by sigma- and PCP-receptor ligands was not attributed to PCP1- or PCP2-receptor stimulation, owing to the decreased potency (100-fold) of these ligands in [3H]DMI assays compared with the affinity for brain PCP1 sites, and the ineffectiveness of the PCP2-ligand N-(1-(2-benzo(b)thiophenyl)cyclohexyl)piperidine (BTCP). Scatchard analysis of the inhibition by the sigma-ligands haloperidol and (+)-3-PPP, as well as the PCP1 receptor ligand MK-801, demonstrated noncompetitive interaction with the site bound by [3H]DMI. These studies indicate that bovine adrenomedullary membranes possess a specific receptor for the noradrenaline uptake inhibitor [3H]DMI, which is sensitive to allosteric modulation produced by PCP and sigma-ligands.

Adrenal Medulla↗
PubMed1992

Central non-opioid physiological and pathophysiological effects of dynorphin A and related peptides.

Dynorphin A (Dyn A) and related opioid peptides derived from prodynorphin possess a high affinity for kappa opioid receptors, but they also bind to other opioid receptors (mu and delta) as well as to some non-opioid receptor sites. Although the physiological role of these peptides is not well established, recent experimental data pinpoint their particular involvement in physiological and pathophysiological conditions that relate to algesia, spinal cord injury and epilepsy. In this paper, we review data which support the concept that the non-opioid behavioral effects of Dyn A and related endogenous peptides which are observed under these conditions are physiologically and pathophysiologically relevant.

Brain↗
PubMed1991

Interaction of 1,3-di(2-[5-3H]tolyl) guanidine with sigma 2 binding sites in rat heart membrane preparations.

Membrane preparations of rat hearts displayed specific binding activity for the prototypic sigma (sigma) receptor ligand, 1,3-di(2-[5-3H]tolyl) guanidine [( 3H]DTG), but not for the phencyclidine (PCP) receptor ligand, [3H]MK-801. Scatchard plot analysis of [3H]DTG binding revealed the presence of one high affinity saturable binding site with a KD of 8.7 nM and a Bmax of 100 pmol/g protein. The drug specificity profile of the receptor correlated with that of the sigma receptor with the following order of potency: DTG greater than haloperidol greater than (-)-pentazocine greater than (-)-butaclamol greater than (+)-butaclamol greater than (-) SKF-10047 greater than (+)pentazocine greater than PCP greater than TCP greater than MK-801 greater than (+)SKF-10047. [3H]DTG binding was sensitive to the Ca2+ channel blocker, verapamil (Ki 202 nM) but not to the K+ channel blocker, 4-aminopyridine. The reverse stereoselectivity of [3H]DTG binding for (-)-SKF-10047 and (-)-pentazocine (Ki of 1289 and 140 nM as compared with 17,582 and 2190 nM for (+)-SKF-10047 and (+)-pentazocine, respectively) indicated that the heart contains sigma receptors with characteristics of the sigma 2 subtype.

Animals↗
PubMed1991

Dynorphin A-(1-13)-Tyr14-Leu15-Phe16-Asn17-Gly18-Pro19 : a potent and selective kappa opioid peptide.

Dynorphin A-(1-13)-Tyr14-Leu15-Phe16-Asn17-Gly18-Pro19+ ++ (dynorphin Ia: a peptide derived from the structure of adrenal dynorphin I) was synthesized by the solid-phase procedure. The product was purified and compared with dynorphin A-(1-13) and [D-Pro10]dynorphin A-(1-11) for its ability to inhibit the electrically evoked contractions of the guinea pig ileum (GPI) and mouse vas deferens (MVD) and to compete with the binding of [3H]ethylketocyclazocine (kappa ligand), [3H][D-Ala2,MePhe4,Glyol5]enkephalin (mu ligand) and [3H][D-Ser2,Thr6]Leu-enkephalin (delta ligand) to membrane preparations of the guinea pig cerebellum or rat brain. Additionally, the antinociceptive effects of the synthetic peptide were assessed in rat paw-pressure and tail-flick tests. In the GPI, dynorphin Ia possessed a relative potency (IC50 0.5 nM) that was comparable to that of [D-Pro10]dynorphin A-(1-11) (IC50 0.5 nM) or dynorphin A-(1-13) (IC50 0.7 nM). In the delta specific MVD assay, dynorphin Ia displayed a reduced potency (IC50 235 nM) as compared with that of dynorphin A-(1-13) (IC50 20 nM) or [D-Pro10]dynorphin A-(1-11) (IC50 46 nM). The affinity of dynorphin Ia for the kappa site in the guinea pig cerebellum (Ki 0.25 nM) was comparable to those of dynorphin A-(1-13) (Ki 0.11 nM) and [D-Pro10]dynorphin A-(1-11) (Ki 0.10 nM). However, the peptide possessed reduced affinities for the mu (Ki 6.7 nM) and delta (Ki 71 nM) opioid receptors as compared with [D-Pro10]dynorphin A-(1-11) (Ki 1.7 and 1.5 nM) an dynorphin A-(1-13) (Ki 0.5 and 4.4 nM, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1991

Circadian regulation of the biosynthesis of cardiac Met-enkephalin and precursors in normotensive and spontaneously hypertensive rats.

Preproenkephalin A mRNA (ppEnk mRNA) and immunoreactive Met-enkephalin (ir-Met-Enk) were measured in the heart of 4, 8 and 16 week-old normotensive Wistar Kyoto (WKY) and spontaneously hypertensive (SHR) rats. WKY rats displayed a small decrease in their cardiac concentration of free (1.3 to 1.0 pmol/g) and cryptic (enzyme processed: 5.3 to 3.7 pmol/g) ir-Met-Enk with aging while the abundance of ppEnk mRNA increased by 3.2 fold between 4 and 16 week-old animals. Similar decreases in free (1.5 to 1.0 pmol/g) and cryptic (5.6 to 4.2 pmol/g) ir-Met-Enk levels were observed in SHR with aging but the rise in the level of ppEnk mRNA was much more pronounced reaching at 16 week-old levels of 7.3 times higher than at 4 week-old and 4.3 times higher than in age-matched WKY. The lack of correlation between the concentration of free and cryptic ir-Met-Enk and the abundance of ppEnk mRNA led us to measure the level of peptides in the heart of 16 week-old animals sacrificed at 4 hr intervals over a 24 hr period. SHR rats displayed circadian variations in their heart content of free and cryptic ir-Met-Enk and increased levels (1.6 fold) of cryptic peptide as compared with WKY at the beginnings of light (6 hr) and dark (18 and 22 hr) periods, suggesting the occurrence of cyclic and transitory upregulation of cDNA transcription and/or derepression of mRNA translation.

Animals↗
PubMed1991

Neurobehavioral evidence for kappa agonist activity of the morphinan derivative 14-beta-methyl 8-oxacyclorphan [BC (3016)].

The purpose of the present study was to determine if the in vivo neurobehavioral effects of the morphinan 14-beta-methyl 8-oxacyclorphan, [BC (3016)], would reflect the kappa agonist activity found in our previous in vitro studies. The effects of intracisternal administration of various doses (10-80 micrograms) of BC (3016) on body temperature, muscle rigidity, nociception of thermal, chemical and mechanical stimuli as well as its ability to induce catalepsy were examined. The effects of intrathecal administration of the same doses of the compound on reactivity of animals to a thermal stimulus were also assessed. Finally, the ability of BC (3016) to antagonize well known neurobehavioral effects of morphine was investigated. Results indicate that the analgesic properties of BC (3016) resemble those of typical kappa agonists: Intracisternal administration of the drug failed to affect nociception to an aversive thermal stimulus but markedly reduced the reactivity of animals subjected to noxious chemical or mechanical stimuli. On the other hand, intrathecal administration of BC (3016) significantly attenuated nociception of animals to a thermal stimulus. The in vivo neurobehavioral effects of BC (3016) appear to be kappa selective since the drug did not decrease body temperature, increase muscular tone or induce catalepsy, three effects generally attributed to mu agonists. Furthermore, BC (3016) antagonized the immobility, trunk rigidity, catalepsy and analgesia induced by morphine. In summary, the present results reveal that BC (3016) displays a profile of neurobehavioral effects similar to that of well known kappa agonists.

Animals↗
PubMed1991

Role of the sigma receptor in the inhibition of [3H]-noradrenaline uptake in brain synaptosomes and adrenal chromaffin cells.

1. Rat brain synaptosomes and cultured bovine adrenal chromaffin cells were used to monitor the inhibitory effects of phencyclidine (PCP) and sigma (sigma)-receptor ligands on the uptake of [3H]-noradrenaline ([3H]-NA). 2. A Na(+)-dependent high affinity uptake was observed in synaptosomes (30 degrees C) and chromaffin cells (37 degrees C) with Km of 0.22 and 0.56 microM and Vmax of 2.5 pmol min-1 mg-1 protein and 0.7 pmol min-1 per 10(6) cells, respectively. 3. PCP and haloperidol inhibited the high affinity uptake with IC50 of 0.17 and 0.42 microM, respectively in synaptosomes and 0.24 and 0.47 microM, respectively in adrenal chromaffin cells. 4. A close correlation (r = 0.96) was established between the ability of various PCP and sigma-receptor ligands to inhibit [3H]-NA uptake in both systems: PCP greater than TCP greater than haloperidol greater than 3-(+)-PPP greater than MK-801 greater than or equal to (-)-butaclamol greater than (+)-SKF-10047 greater than DTG. Spiperone and opioid receptor ligands were ineffective at 20 microM. 5. These results indicate that the central and peripheral inhibitory effects of PCP and sigma-receptor ligands on [3H]-NA uptake involves a receptor (sigma 1-like) which is distinct from that (PCP2) recognized for the inhibition of [3H]-dopamine uptake by PCP.

Adrenal Glands↗
PubMed1991

Expression of the preproenkephalin A gene in the heart of cardiomyopathic hamsters.

The expression of the preproenkephalin A gene (Enk gene), which codes for the precursor of enkephalins, was investigated in the heart of hamsters with a hereditary cardiomyopathy at four different stages of the disease: the prenecrotic stage (30 days), the necrotic stage (60 days), the hypertrophic stage (120 days), and the final stage (200 days). In control atria and ventricles, the relative abundance of the Enk mRNA, as assessed by Northern blot analysis, did not change upon ageing. In the ventricles of cardiomyopathic hamsters, however, it increased about two- to three-fold at the necrotic stage, but was unaltered at the other time points studied; whereas in the atria, it progressively decreased to reach about half that of control hamsters at the final stage. Enkephalin levels, as measured by radioimmunoassay, decreased at 60 days in both the atria and ventricles of control hamsters, and also in the atria of cardiomyopathic hamsters, and remained stable thereafter, corresponding to one-third to one-half of those at 30 days. However, in the ventricles of cardiomyopathic hamsters, the peptide level decreased only slightly, the consequence being that at 60, 120, and 200 days, it was about two- to three-fold that of control hamsters. The lack of correlation between peptide levels and the relative abundance of the Enk mRNA suggests that translational and (or) post-translational mechanisms are important in the control of the expression of the Enk gene in the heart of hamsters.

Aging↗
PubMed1990

Chromaffin cell scinderin, a novel calcium-dependent actin filament-severing protein.

Scinderin, a novel Ca2+-activated actin filament-severing protein, has been purified to homogeneity from bovine adrenal medulla using a combination of several chromatographic procedures. The protein has an apparent mol. wt of 79,600 +/- 450 daltons, three isoforms (pIs 6.0, 6.1 and 6.2) and two Ca2+ binding sites (Kd 5.85 x 10(-7) M, Bmax 0.81 mol Ca2+/mol protein and Kd 2.85 x 10(-6) M, Bmax 1.87 mol Ca2+/mol protein). Scinderin interacts with F-actin in the presence of Ca2+ and produces a decrease in the viscosity of actin gels as a result of F-actin filament severing as demonstrated by electron microscopy. Scinderin is a structurally different protein from chromaffin cell gelsolin, another actin filament-severing protein described. Scinderin and gelsolin have different mol. wts, isoelectric points, amino acid composition and yield different peptide maps after limited proteolytic digestion by either Staphylococcus V8 protease or chymotrypsin. Moreover, scinderin antibodies do not cross-react with gelsolin and gelsolin antibodies fail to recognize scinderin. Immunofluorescence with anti-scinderin demonstrated that this protein is mainly localized in the subplasmalemma region of the chromaffin cell. Immunoblotting tests with the same antibodies indicated that scinderin is also expressed in brain and anterior as well as posterior pituitary. Presence of scinderin and gelsolin, two Ca2+-dependent actin filament-severing proteins in the same tissue, suggests the possibility of synergistic functions by the two proteins in the control of cellular actin filament networks. Alternatively, the actin filament-severing activity of the two proteins might be under the control of different transduction and modulating influences.

Actins↗
PubMed1990

Alterations of heart dynorphin-A in the development of spontaneously hypertensive rats.

In order to investigate the pathophysiological role of heart dynorphin-A (Dyn-A) in genetic hypertension, immunoreactive (ir)-Dyn-A was measured in heart extracts of spontaneously hypertensive rats (SHR) and compared with that of age matched Wistar (WR) and Wistar Kyoto (WKY) rats. Heart ir-Dyn-A contents in 8 week-old WK (84 fmol/g tissue) were not significantly different from those of age matched WKY (109 fmol/g tissue). In control WKY, the levels of ir-Dyn-A did not significantly vary with the age (from 109 to 117 fmol/g) except in 16 week-old animals which displayed a significant increase (238 fmol/g tissue) compared to younger animals. In SHR, the heart content of ir-Dyn-A displayed a 6.5 fold increase at 8 weeks compared to age matched WKY. Older SHR showed a return of their heart ir-Dyn-A content to control (12 week-old) or below control values (16 week-old; 121 compared with 238 fmol/g tissue in WKY). Heart ir-Dyn-A in WKY and SHR eluted as a single peak on mu-Bondapak HPLC, corresponding with the retention time of synthetic Dyn-A. A local function for cardiac ir-Dyn-A is suggested by the presence in heart membrane preparations of a high affinity binding site for the kappa selective opioid ligand, [3H]U-69593, with KD of 6.4 (WKY) and 8.5 (SHR) nM and Bmax of 3.7 (WKY) and 3.6 (SHR) pmol/g protein. The alterations in the levels of cardiac ir-Dyn-A during the development of hypertension in SHR were analyzed in regard with the reported effects of Dyn-related peptides on heart natriuretic and sympathetic functions.

Animals↗
PubMed1990

Identification, characterization, and photoaffinity labeling of the dihydropyridine receptor associated with the L-type calcium channel from bovine adrenal medulla.

The dihydropyridine receptor associated with the L-type Ca2+ channel in adrenal medulla membranes has been identified and characterized. [3H]PN200-110 binds in a stereoselective, saturable manner to a single class of high affinity sites in adrenal medulla membranes, with a Kd of 0.1 nM and a Bmax of 141 fmol/mg of protein. Dihydropyridines inhibited [3H]PN200-110 binding with the rank order (+)-PN200-110 greater than nifedipine greater than nimodipine greater than usoldipine greater than or equal to nitrendipine greater than BayK8644 greater than (-)-PN200-110. [3H] PN200-110 binding was sensitive to divalent cations, as examined by the effects of Ca2+, Mg2+, and the chelators ethylene glycol bis-(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid and EDTA. [3H]PN200-110 binding was modulated by various classes of L-type Ca2+ channel effectors. Benzothiazepines modulated binding of [3H]PN200-110 in a negative or positive manner that was temperature dependent, whereas phenylalkylamines weakly inhibited [3H]PN200-110 binding. Bepridil stimulated [3H] PN200-110 binding, whereas phencyclidine was without effect. The photoaffinity probe [3H]azidopine labeled a single polypeptide that migrated with an apparent molecular weight of 185,000-190,000 in sodium dodecyl sulfate gel electrophoresis. The dihydropyridine receptor was found to bind specifically to wheat germ agglutinin columns. These results demonstrate the presence of a Ca2+ channel blocker complex in adrenal medulla. The drug receptor sites reside on a glycoprotein complex in which a polypeptide analogous to the alpha 1-subunit of the L-type Ca2+ channel from skeletal muscle has been identified.

Adrenal Medulla↗
PubMed1989

Presence of sigma and phencyclidine (PCP)-like receptors in membrane preparations of bovine adrenal medulla.

Receptor binding studies with [3H]-(+)SKF-10047 were carried out to characterize the putative sigma (sigma) and phencyclidine (PCP) receptors in membrane preparations of bovine adrenal medulla. Specific binding of the radiolabelled compound was observed after incubation with the membrane preparation at 37 degrees, the equilibrium being reached at 20 min and the maximal binding being observed with 0.6 mg/ml protein. Saturation binding studies were performed at equilibrium (30 min at 37 degrees with 0.5 mg/ml of membrane protein) in the presence of haloperidol (1 microM) or 1-[1-(2-thienyl) cyclohexyl] piperidine (TCP; 0.2 microM) to block sigma or PCP receptors, respectively. The binding of [3H]-(+)SKF-10047 was characterized by two distinct components. A high affinity binding site (haloperidol sensitive) had an apparent KD of 8.3 nM and a Bmax of 67 pmol/g protein. A lower affinity binding site (TCP sensitive) had an apparent KD of 32.7 nM and a Bmax of 83 pmol/g protein. The drug specificity of the high affinity binding site resembled that of the putative sigma receptor, being potently inhibited by haloperidol and pentazocine. The binding pharmacology of the low affinity site resembled that of the phencyclidine receptor, being potently displaced by TCP and PCP. The binding of [3H]-(+)SKF-10047 to both receptors showed marked stereoselectivity for the dextrorotatory (+) isomer of SKF-10047 and was insensitive to the receptor specific opioid ligands DAGO (mu), DSLET (delta) and U-69593 (kappa). These data indicate that bovine adrenal medulla contains sigma and PCP-like receptors.

Adrenal Medulla↗
PubMed1988

Increased content of immunoreactive Leu-enkephalin and alteration of delta-opioid receptor in hearts of spontaneously hypertensive rats.

The levels of immunoreactive Leu-enkephalin (ir-Leu-Enk) were measured in acid extract of hearts and lungs of adult normotensive Wistar Kyoto (WKY) and spontaneously hypertensive (SHR) rats. Heart contents in ir-Leu-Enk were significantly higher in SHR rats (2.67 pmol/g) than in WKY rats (1.5 pmol/g). However, no significant change was observed in lung ir-Leu-Enk levels of these same animals (0.7-0.91 pmol/g). Reverse-phase high-performance liquid chromatography (RP-HPLC) of heart extracts indicated that the major form of ir-Leu-Enk in SHR rats coelutes with synthetic Leu-Enk and is increased by an approximate factor of 4.2-fold as compared with its eluting counterpart in the WKY chromatogram. The delta-selective opioid ligand, [3H][D-Pen2, D-Pen5]-Enk [( 3H]DPDPE) bound to two distinct receptor sites in membrane preparations of hearts from WKY rats with Kd of 1.33 and 22.8 nM and Bmax of 3.9 and 14.6 pmol/g protein, respectively. The binding with the heart membrane preparation of SHR rats displayed only the high affinity delta-binding site with a Kd of 1.53 nM and a Bmax of 6.5 pmol/g protein. These data indicate that the hypertensive state in SHR rats is manifested through a rise in the cardiac level of Leu-Enk and a selective down-regulation of its low-affinity delta-opioid receptor.

Animals↗
PubMed1988

Sensitive method of detection, quantitation and purification of peptides using pre-column derivatization with phenyl isothiocyanate.

A sensitive method for the detection, quantitation and purification of peptides is described. The method is based on pre-column derivatization of peptides with phenyl isothiocyanate to form phenylthiocarbamoyl derivatives (PTC peptides). The derivatized peptides are analysed by reversed-phase high-performance liquid chromatography on a Zorbax ODS column (5 micron) and detected at 269 nm with a sensitivity limit of 1-5 pmol. The technique was utilized for the separation of a mixture of closely related synthetic peptides. The eluted PTC peptides were collected with an average recovery yield of 75% as determined by amino acid analysis. This method of separation of PTC peptides was also combined with the determination of the complete structure of recovered PTC-dynorphin A-(1-13) using the solid-phase sequenator (Sequemat). The advantages of the derivatization method are the rapidity and completeness of the reaction, the stability of the product, the sensitivity and specificity of the detection of derivatized peptides and the compatibility of the technique with subsequent analytical procedures. A particular application of this method was exemplified by the dosage of enkephalins secreted from perfused bovine adrenal glands.

Adrenal Medulla↗