Search PubMed⌕ Search

Biomedical subjects

J D Vincent

Publications and source records attributed to J D Vincent.

At least 109 records · Page 6Linked to original sources

Aldosterone secretion and adrenal angiotensin II receptors in the Brattleboro rat.

The basal secretion of aldosterone, measured in adrenal venous blood, was three- to fourfold lower in Brattleboro than in Long-Evans rats used as controls. Infusion of a low dose of angiotensin II (1 ng/min per 100 g body/wt) to Long-Evans rats caused a fourfold increase in aldosterone release but neither the low dose nor a tenfold higher dose changed the rate of release in Brattleboro rats. Only a very high dose (300 ng/min per 100 g body wt) succeeded in increasing the secretion of aldosterone in Brattleboro rats but throughout the time-course of the infusion, secretion remained about fivefold lower than in Long-Evans rats and the incremental response was reduced by 74.9%. Adrenal zona glomerulosa angiotensin II receptor sites had similar affinity and maximum binding capacity in the two groups of rats. It is suggested that the reduced corticosteroidogenic capacity of the adrenal cortex of Brattleboro rats results from an impairment of the post-receptor mechanisms involved in the biosynthesis of aldosterone.

Adrenal Cortex↗

Ifenprodil and SL 82.0715 as cerebral anti-ischemic agents. II. Evidence for N-methyl-D-aspartate receptor antagonist properties.

The effects of the anti-ischemic agents ifenprodil and its derivative SL 82.0715 ((+/-)-alpha-(4-chlorophenyl)-4-[(4-fluorophenyl) methyl]-1-piperidineethanol] have been analyzed in a number of models indicative of N-methyl-D-aspartate (NMDA) antagonistic potential in vitro and in vivo. Ifenprodil and SL 82.0715 potently and noncompetitively antagonize the stimulatory effects of NMDA on cyclic GMP production in immature rat cerebellar slices (IC50 values, 0.4 and 10 microM, respectively), as well as the NMDA-evoked [3H]acetylcholine release in adult rat striatal slices (IC50 values, 1.6 and 6.6 microM, respectively). Ifenprodil is 10 times more potent than (+/-)3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP) but less active than the reference noncompetitive NMDA channel blockers [MK 801, ((+)-5-methyl-10,11-dihydro-5H-dibenzo-[a,d]cyclohepten-5,10-imine ], phencyclidine and 1-[1-(2-thienyl)cyclohexyl]piperidine (TCP)] in these models. Ifenprodil and SL 82.0715 partially displace (maximal displacement 40-50% at 10 microM) the NMDA receptor ligand [3H]CPP from its binding site to rat brain membranes (IC50 values, 0.1 and 0.3 microM, respectively) in a noncompetitive manner; in the micromolar range the two agents also partially displace the NMDA channel ligand [3H]TCP from its binding site to rat brain membranes, and noncompetitively antagonize the L-glutamate-induced increase in [3H]TCP binding. Ifenprodil (0.01-1 microM) partially antagonizes the depolarizing effects of NMDA on the immature rat hemisected spinal cord in vitro. In mouse cultured spinal cord neurons, ifenprodil dose-dependently antagonizes the depolarizing effects of micropressure applied NMDA. Inhibition of the effects of NMDA in this model by ifenprodil and SL 82.0715 is noncompetitive. In vivo and after systemic i.p. administration, ifenprodil and SL 82.0715 antagonize the stimulatory effects of intrastriatally dialyzed NMDA on striatal dopamine release in rats (ID50 values, 0.9 and 0.3 mg/kg, respectively), and block the harmaline-evoked increase in cerebellar cyclic GMP production in mice (ID50 values, 3 and 4 mg/kg, respectively). These results indicate that ifenprodil is a noncompetitive NMDA antagonist which has a mechanism of action distinct from either the reference competitive NMDA receptor antagonists (CPP and 2-amino-5-phosphonovalerate) or the noncompetitive NMDA channel blockers (phencyclidine, TCP and MK 801). The potent NMDA antagonistic effects of the ifenprodil class of compounds are likely to be related to the demonstrated anti-ischemic potential of these compounds.

Animals↗

Influence of cell cycle phases on the electrical activity and hormone release in a transformed line of anterior pituitary cells.

Electrophysiological experiments have shown that about 50% of cultured GH3 cells (tumoral cell line, from the anterior pituitary gland) are inexcitable i.e. they do not display action potentials either spontaneously or when depolarized by a current pulse. We report here this inexcitability may be related to cellular kinetics. Thus we have studied the relationship between the various phases of the cell cycle, the electrophysiological properties of GH3/B6 cells and spontaneous or induced Prolactin and Growth Hormone (GH) release rates. Asynchronous populations of viable cells were stained with Hoechst 33 342 DNA fluorescent dye, and sorted using a flow cytometer into G1 and S phases. After selection intracellular potentials were recorded using a single glass micro-electrode; the basal or TRH stimulated rates of PRL and GH secretions were determined by RIA. Electrical properties of the cells i.e. resting potentials, input membrane resistance and excitability, reached a maximum for cells in G2+M phases. Only cells in G2+M displayed action potentials and TRH increased their secretion by 5 times for GH and by 6 times for PRL. In G1 and S phases the cells were electrically inactive and secretion rates remained at their basal levels. These findings demonstrate that the mechanism of stimulus secretion coupling is dependent upon the phases of the cell cycle.

Animals↗

Angiotensin II inactivation process in cultured mouse spinal cord cells.

The pattern of hydrolysis of [3H]angiotensin II ( [3H]AII; 20 nM) by intact cells was studied on cultured mouse spinal cord cells. Degradation products were identified by HPLC analysis after incubation for 2 h at 37 degrees C. In the absence of peptidase inhibitors, 70% of [3H]AII was degraded, and the main labeled metabolite was [3H]tyrosine (40% of total radioactivity). Minor quantities of [3H]AII1-5 and [3H]AII4-8 were formed. Results obtained in the presence of various inhibitors indicate that several enzymes were involved in the AII-hydrolyzing process. Dipeptidyl aminopeptidase III (EC 3.4.14.4) could play a critical role, as suggested by the formation of [3H]Val3-Tyr4 and [3H]-Tyr4-Ile5 in the presence of bestatin (2 X 10(-5) M). This hypothesis was confirmed by the potency of dipeptidyl amino-peptidase III inhibitors to inhibit both [3H]AII hydrolysis and formation of these 3H-labeled dipeptides. An arylamidase-like activity could also be participating in [3H]AII hydrolysis, because higher concentrations of bestatin (10(-4) M) in association with dipeptidyl aminopeptidase III inhibitors totally inhibited [3H]tyrosine formation, increased protection of [3H]AII and [3H]AII1-7 formed, and provoked a slight accumulation of [3H]AII2-8. These results suggest that the formation of [3H]AII2-8 is due to the action of a bestatin-insensitive acidic aminopeptidase and that the Pro7-Phe8 cleavage is also a step of AII hydrolysis, resulting from the action of an unidentified peptidase different from prolyl endopeptidase.(ABSTRACT TRUNCATED AT 250 WORDS)

Angiotensin II↗

Electrophysiological responses to dopamine of rat hypophysial cells in lactotroph-enriched primary cultures.

1. Cells from 14-day-old and lactating female rat pituitary glands were dissociated, separated and enriched on a continuous gradient of bovine serum albumin at unit gravity. They were maintained for at least 6 days in culture before perifusion and electrophysiological experiments were performed. 2. Immunofluorescent staining of the resulting gradient fractions (numbered F2 to F9) from both groups of animals indicated that the majority of lactotrophs were located in the light fractions (F3-F4). However, a second population of lactotrophs was observed in the heavy fractions (F7-F9) isolated from lactating females. 3. Basal secretion rates of prolactin were in the order of 2-40 ng 2 min-1 10(6) cells-1 and were inhibited by dopamine in a dose-dependent manner. 4. According to their electrophysiological properties, cells from 14-day-old females (first group) were categorized as follows: (1) inexcitable cells, which displayed a low resting potential of about -35 mV (39% of cells tested, n = 118); and (2) excitable cells, which displayed either triggered or spontaneous action potentials and resting membrane potentials higher than -50 mV (61% of cells tested, n = 185). 5. In the light fraction from lactating females (second group), the majority of the cells were excitable (70%) and showed high resting membrane potentials (-50 to -55 mV) and 15% of these cells displayed spontaneous action potentials. 6. Heavy fractions (third group) contained a high percentage of non-spontaneous but excitable cells (80% of the cells tested, n = 65). These cells were able to elicit action potentials after the cessation of hyperpolarizing current pulses ('off' potentials). 7. Action potentials were insensitive to the sodium channel blocker, tetrodotoxin (TTX; 5 x 10(-6) M) but were reversibly blocked by calcium channel blockers such as cobalt, manganese and cadmium (10 mM). 8. In excitable cells from the three groups, dopamine (10(-7) M) induced a hyperpolarizing response due to an increase of the membrane conductance. During this response, action potentials were inhibited. It was shown that this was not a direct effect of dopamine. The reversal potential of the dopamine-induced response in these cells was found to be at -100 mV. This value was shifted to more positive potentials (-50 mV) when high-potassium medium was used (56 mM). 9. In non-excitable cells (first group), dopamine (10(-7) M) induced a hyperpolarizing response due to a decrease of the membrane conductance.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

Structural plasticity in the rat supraoptic nucleus during gestation, post-partum lactation and suckling-induced pseudogestation and lactation.

In the supraoptic nucleus (SON) of parturient and lactating rats, large portions of the surface membranes of almost all oxytocinergic neurons are directly juxtaposed with no glial interposition. A significant number of the same neurones are also contacted by the same presynaptic terminal ('double' synapses). Our present observations have revealed that direct appositions between adjacent neurons in the SON increase quite rapidly during the day before parturition. 'Double' synapses also become visible during late gestation, but they appear more progressively. Earlier studies have shown that 1 month after weaning, as in virgin rats, there are again very few appositions and 'double' synapses in the nucleus. We show here that the SON can remain structurally modified, and to the same degree, beyond normal weaning time so long as lactation is prolonged by renewing suckling litters. However, if the mothers are deprived of their pups immediately after birth, neuronal appositions disappear within 2 days and 'double' synapses by 10 days. In non-pregnant primiparous rats, continuous exposure to suckling litters leads to pseudogestation and eventually lactation (in 16-22 days). Examination of the SON in such animals revealed that the oxytocinergic system is already modified by day 12 of dioestrus; during suckling-induced lactation, the anatomical changes are identical to those seen during a normal post-partum lactation. These observations indicate that neither gestation nor suckling alone are indispensable for the anatomical reorganization of the SON apparent at lactation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[Development of electrical activity of hypothalamic cells in culture].

The development of the electrical activity of hypothalamic cells was studied using intracellular and patch clamp recording technics on cultured hypothalamic neurones from 14 days mouse embryos. After 24 h of incubation, 15% of recorded cells were spontaneously active. During the first five days of culture this ratio increased exponentially to reach 95% at day 5. Between the 5th and the 9th day the spontaneous activity progressively decreased although the majority of cells remained excitable. Spontaneous activity reappeared after the 9th day and was underlain by a synaptic potential activity. During the first five days of culture, only a TTX sensitive inward current was observed in all cell tested. A calcium inward current appeared after the first week of incubation. It was recorded on 40% of cells at day 11th and in 80% of cells tested after a month of incubation. Our results show that electrical activity of cultured hypothalamic neurones develop in three stages. A first stage characterized by a spontaneous electrical activity without post synaptic potential, an intermediate stage during which a Calcium inward current appeared corresponding to synaptic contact development and a third stage during which post synaptic potential activity was observed, corresponding to synaptic contact maturation.

Animals↗

[Characterization by cytofluorometry of the different types of adenohypophyseal cells in the rat].

The different antehypophysical cell types which synthetize and release somatotroph (GH), corticothroph (ACTH), gonadotroph (LH-FSH) and lactotroph (PRL) hormones were analysed. The experiments were performed on hypophyses from five groups of animals: adult males, 14 days-old female, adult females, gestating adult females and lactating adult females. The cells were analysed by immunofluorescence using flow cytometry. For each of the hormones studied, there was a characteristic spectral distribution of cells. The evolution of cell size and granular content with respect to sex and physiological state of each group was studied by the analysis of diffused light. Small, slightly granular cells represented 50% of the cell population in males and 14 day-old females but only 8% in gestating or lactating females. The study of the cell cycle showed the presence of dividing cells in the population of large, granular cells from gestating and from lactating females. No features of cell division were observed in the population of small, slightly granular cells. This study indicates the potential value of multiparametric analysis in the separation of pure sub-populations of antehypophysial cells.

Animals↗

[Evaluation of perfusion technics of the magnocellular nucleus of the hypothalamus in vitro and in vivo].

In suckled rats, OT necessary for the occurrence of the neurosecretory bursts on OT cells, is probably released inside the magnocellular nuclei. In order to demonstrate this in vivo release and to precise the mechanism involved, perifusions were realized in vivo on lactating rats and in vitro with isolated magnocellular nuclei. In vivo, the push-pull perifusion of a single supraoptic nucleus (SON) was realized simultaneously with the recording of the electrical activity of OT cells in the contralateral nucleus. This would allow to determine the possible relationships between the amount of OT released in the SON and the electrical activity of OT cells (bursting activation during suckling or continuous activation during an hyperosmotic stimulation). Results obtained showed 1) that OT was released in vivo inside the SON, 2) that this release was specifically increased during the milk ejection reflex and 3) that this increase was only detectable inside the SON. However, this technique did not permit to determine either the mechanism of OT release or the neuron elements (perikaryon, dendrites, axon collaterals) responsible for this release. That is for why, in vitro perifusions were undertaken with isolated magnocellular nuclei. The first stage was to determine the stimulus able to induce a reproducible increase of OT release. Among the chemical stimuli (neurotransmitters, K+) only K+ at a 56 mM concentration increased OT release by SON but this increase was small and non significant. Repetitive electrical stimulations with short pulses (0.2 to 5 msec) were ineffective even if the pulse intensity was raised up to 10 mA and the frequency up to 80 Hz.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Vasopressin and oxytocin reduce plasma prolactin levels of conscious rats in basal and stress conditions. Study of the characteristics of the receptor involved.

Subcutaneous administration of arginine vasopressin (AVP) to conscious rats induced a dose-dependent increase of plasma ACTH and beta-endorphin levels and decrease of plasma prolactin (PRL) levels 30 min later. AVP similarly reduced PRL increase induced by exposure to a novel environment stress. Oxytocin (OT) was also active but 5-fold less potent than AVP. The study of several analogs with specific agonistic and antagonistic activity on the oxytocic, vasopressor and antidiuretic receptors of OT and AVP suggests that the receptor involved in this effect does not fit into this classification.

Adrenocorticotropic Hormone↗

Septal release of vasopressin in response to osmotic, hypovolemic and electrical stimulation in rats.

The central release of vasopressin was studied in anesthetized rats using push-pull perfusions and radioimmunoassay of the hormone. A basal release was observed in the lateral septum and in the lateral ventricle, whereas no vasopressin was detected in the perfusates from the caudate nucleus. Under osmotic stimulation, vasopressin release increased up to 12 and 60 times basal levels following i.p. injections of 5 ml and 10 ml/kg b.wt. of 2 M NaCl, respectively. This increase was blocked by using a calcium-free perfusion medium containing 0.1 mM EGTA. In the lateral ventricle, osmotic stimulation (5 ml/kg of 2 M NaCl i.p.) had the same effect as in the septum. In the caudate nucleus, no release was observed. Hemorrhage also increased the septal release of vasopressin in 5 out of 6 animals tested. Electrical stimulation of the pituitary stalk and of the supraoptic nucleus was used to evoke the release of vasopressin into the bloodstream. Septal release slightly decreased during pituitary stalk stimulation, whereas it did increase during stimulation of the supraoptic region. Our results show that systemic stimuli for vasopressin release evoke both a peripheral and a septal release of the hormone. The dissociation of the effects of electrical stimulation of the pituitary stalk and of the supraoptic nucleus suggests, however, that the vasopressinergic neurones responsible for septal release are distinct from those which project to the neurohypophysis.

Animals↗

Simultaneous determination of radio-immunoassayable methionine-enkephalin and radioreceptor-active opiate peptides in CSF of chronic pain suffering and non suffering patients.

Radio-immunoassayable methionine-enkephalin (ME) and radioreceptor-active opiate peptide levels (OP) were determined in CSF from patients, both with and without chronic pain, under investigation for vertebral disk disease. This study showed: that there was no direct correlation between ME and OP levels in CSF; OP levels were negatively correlated with the ME/OP ratio; migraine patients had higher levels of ME; ME concentrations were reduced in patients receiving anti-inflammatory drugs (nonsteroidal): patients with chronic pain (non migraine, no anti-inflammatory drug therapy) had lower ME levels than patients without pain. The data are discussed in relation to animal models of chronic pain.

Adolescent↗

[In situ hybridization of messenger RNA in the central nervous system].

Beside the standard approach for the study of peptide hormone gene expression that quantitate the amount of specific mRNA in the total RNA extracted from an homogenate of tissue by filter or solution hybridization, a technique called in situ hybridization has been developed for the location of specific mRNA populations in tissue section. The in situ technique can be useful in answering anatomical question but its greatest interest in neuroendocrinology lies in its ability to resolve hybrids at the levels of cells and therefore in its potential as a semiquantitative assay for mRNA levels in individual neurons.

Animals↗

Characterization of two angiotensin II binding sites in cultured mouse spinal cord neurones.

Characteristics of angiotensin II (AII) binding have been determined in cultured mouse spinal cord neurones using [125I]AII and [3H]AII. The Scatchard plot of equilibrium binding was curvilinear and could be described by postulating the existence of two different classes of independent binding sites (Kd1 = 0.43 nM, Bmax1 = 12.5 fmol/1.5 X 10(6) cells; Kd2 = 25.6 nM, Bmax2 = 220 fmol/1.5 X 10(6) cells). These values are in close agreement with the Kd values obtained from kinetic studies. The high affinity binding sites appeared to be similar to the single class of sites described in other studies. The relative inhibition potency of AII-related peptides was studied. Sar1,-Leu8-AII was the most potent in inhibiting specific AII binding. The characteristics of the two AII binding sites suggest that they correspond to two receptors as described in a previous electrophysiological approach using this model in our laboratory. Taken together, these data confirm that this model of neurones in primary culture is a unique and very attractive model of receptor studies. The classical criteria necessary for positive identification of a ligand-receptor have been satisfied: saturability, reversibility, specificity and most importantly correlation of the binding parameters and biological effects of AII.

Angiotensin II↗

Evidence for bursting pacemaker neurones in cultured spinal cord cells.

Intracellular recordings were made from dissociated mouse spinal cord cells in primary culture. One type of spinal cord neurone, with a large cell body (40-50 micron), 3-5 short neurites, and a mean resting potential of -65 mV, was found to fire rhythmic bursts of action potentials with a phase duration of approximately 1s when the membrane potential was depolarized to -55 mV. These bursts did not arise from spontaneous synaptic input, but appeared to result from endogenous ionic conductance properties of the membrane resembling those observed in molluscan bursting pacemaker neurones. Ionic conductances underlying this bursting activity were studied pharmacologically by local application of ionic conductance blockers. Pacemaker potentials depended on Na+ conductance, since tetrodotoxin and Na-free medium were the most potent agents for blocking spontaneous rhythmic activity. However, a Ca2+ conductance was involved in the depolarizing phase of membrane potential oscillations, since Ba2+ application increased oscillation amplitude. Action potentials observed during the bursts were Na+- and Ca2+-dependent. They did not differ significantly from those observed in other spinal cord neurones in culture. Application of tetraethylammonium, CoCl2, BaCl2 and 4-aminopyridine revealed at least three different potassium conductances which controlled this bursting pacemaker activity. A delayed potassium conductance controlled spike duration, a Ca-dependent potassium conductance controlled the duration of the burst and underlay the hyperpolarizing phase terminating the burst, and finally, a transient potassium conductance appeared to be involved in the control of phase duration. The demonstration that spinal cord neurones growing in monolayer culture display typical bursting pacemaker activity raises the possibility that bursting pacemaker neurones in the mammalian spinal cord may be involved in a phasic pattern generator that could control such activities as walking and the respiratory rhythm.

Action Potentials↗

The electrical properties of isolated human prolactin-secreting adenoma cells and their modification by dopamine.

Human prolactinoma cells were maintained in culture for a period of at least 8 days and were able to secrete PRL in large amounts. This secretion was inhibited by bromocriptine, an agonist of dopaminergic receptors, in a dose-dependent manner. The cells showed electrical activity (action potentials) which was blocked by inhibitors of calcium current (cobalt, manganese), whereas it was insensitive to blockers of sodium current (tetrodotoxin). At the resting potential of the cell, dopamine induced a hyperpolarizing response such that action potentials no longer occurred. This effect was due to increase of the membrane conductance and depended on the cell potential. The reversal potential of this response was at -100 mV, which suggests the involvement of potassium ions. Bromocriptine and RU 24213, which are strong dopaminergic receptor agonists, both induced responses identical to the dopamine-induced response. The D2 receptor antagonists (haloperidol, domperidone, and spiperone) blocked the dopamine-induced response in a reversible manner. The D1 antagonist of dopaminergic receptors flupentixol had no effect on the dopamine response. It is concluded that the dopamine modulation of electrical activity involving calcium current may be an early important step in the mechanism by which dopamine inhibits PRL release.

Adenoma↗