PubMed Health⌕ Search

Biomedical subjects

J M Israel

Publications and source records attributed to J M Israel.

At least 37 records · Page 2Linked to original sources
PubMed1990

Contribution of arachidonate metabolites to basal and thyrotropin releasing-hormone-stimulated release of prolactin from purified lactotrophs in primary culture.

Among the different biochemical pathways which have been suggested to play a role in the control of prolactin (Prl) release from anterior pituitaries, arachidonate and its metabolites have been proposed to be involved in the process of Prl release. In this study we investigated the contribution of arachidonate metabolites to both basal and TRH-stimulated Prl release from perifused lactotrophs in culture (derived from pituitary glands of lactating female rats), which exhibit a high sustained release of Prl in absence of inhibitory input. Inhibition of the general oxidative metabolism of arachidonate by 10(-5) M ETYA or of the arachidonate lipoxygenase metabolism by 10(-5) M NDGA decreased basal Prl release to 45 +/- 10% (n = 3) and 36 +/- 4% (n = 6) of the control release, respectively. Indomethacin, an inhibitor of the cyclooxygenase pathway, was without effect. Of the lipoxygenase metabolites tested at 10(-6) M only 15-HPETE and 15-HETE induced Prl release. 15-HETE elicited prolactin release in a concentration dependent manner with a maximal effect at 10(-6) M (10.72 +/- 3 ng/ml vs control 5.1 +/- 0.8 ng/ml, n = 3). The quantity of Prl release induced by TRH was markedly decreased in the presence of NDGA. However, the fraction of Prl release elicited by TRH, calculated as a percentage of the amount of Prl released prior to TRH application, was similar under control conditions, and in the presence of NDGA. In contrast, inhibition of the protein kinases A and G by H8 (10(-5) M) failed to alter basal Prl release but inhibited the effect of TRH by 58 +/- 6% (n = 3). These data suggest that in absence of inhibitory inputs the high sustained release of Prl observed in cultures of lactotrophs derived from lactating female rats depends on the availability of lipoxygenase metabolites, and that the blockade of lipoxygenase reduces the absolute amount of Prl released by TRH without suppressing the ability of TRH to stimulate Prl release.

5,8,11,14-Eicosatetraynoic Acid↗
PubMed1990

Two types of voltage-dependent calcium current in rat somatotrophs are reduced by somatostatin.

1. Somatotrophs were obtained from rat pituitary glands after dissociation, separation and enrichment on a continuous gradient of bovine serum albumin at unit gravity. Somatotrophs were enriched up to 85% in the heavy fractions (F8 and F9). 2. After identification by reverse hemolytic plaque assay, patch-clamp recording in the whole-cell mode was performed on somatotrophs. 3. Under voltage-clamp conditions, two types of Ca2+ currents were recorded. From a holding potential of -70 mV, depolarizing voltage steps to potentials more positive than -50 mV activated a current which rapidly inactivated and which was very sensitive to Ni2+ but not to Cd2+. This current corresponds to T-type current. Depolarizing steps to potentials more positive than -30 mV from a holding potential of -40 mV triggered a current which slowly inactivated and which was very sensitive to Cd2+ but not to Ni2+. This current corresponds to L-type current. 4. Application of somatostatin to the bath solution (10 nM) markedly reduced the amplitudes of both T- and L-type currents. Somatostatin decreased the conductance of L-type current without modifying its time- and voltage-dependent inactivation but its activation was not affected. However, somatostatin decreased the conductance of T-type currents, and also accelerated its time-dependent inactivation. Half-inactivation voltage of T-type current was shifted from -52 to -63 mV by somatostatin but no change was obtained in the current activation curve. 5. All these modifications in Ca2+ currents were abolished by a pre-treatment of the cultures with pertussis toxin (100 ng/ml, for 10 h). This pre-treatment also blocked the inhibitory effect of somatostatin on high-K(+)-stimulated growth hormone release. 6. Our results show that somatostatin acts on somatotrophs by attenuating the voltage-dependent Ca2+ currents. These effects may contribute to a somatostatin-induced reduction in [Ca2+]i and the subsequent decline in growth hormone release.

Animals↗
PubMed1990

Somatostatin increases voltage-dependent potassium currents in rat somatotrophs.

To study the modulatory effects of somatostatin on membrane K+ currents, whole cell voltage-clamp recordings were performed on identified rat somatotrophs in primary culture. In the presence of Co2+ (2 mM) and tetrodotoxin (1 microM) in the bath solution to block Ca2+ and Na+ inward currents, two types of voltage-activated K+ currents were identified on the basis of their kinetics and pharmacology. First, a delayed rectifier K+ current (IK) had a threshold of -20 mV, did not decay during voltage steps lasting 300 ms, and was markedly attenuated by extracellular application of tetraethylammonium (TEA, 10 mM). Second, a transient outward K+ current (IA) was activated at -40 mV (from a holding potential of -80 mV) and persisted despite the presence of TEA. This IA was blocked by 4-aminopyridine (2 mM). Somatostatin (10 nM) increased IK by 75% and IA by 45% without obvious effects on steady-state voltage dependency of activation or inactivation, and these effects were reversible. This increase in K+ currents may contribute in part to the inhibitory effect of somatostatin on growth hormone release.

4-Aminopyridine↗
PubMed1990

Plateau potentials recorded from lactating rat enriched lactotroph cells are triggered by thyrotropin releasing hormone and shortened by dopamine.

Enrichment of dispersed pituitary cells from normal lactating rats on a continuous BSA gradient permitted the isolation of two prolactin cell populations--light and heavy. Hormone release studies indicated that spontaneous prolactin (PRL) release of the heavy fraction cells was particularly sensitive to TRH stimulation (mean of 450%) and that this effect of TRH was totally inhibited by Ca2+ channel blockers (Co2+, Cd2+, Ni2+ and Mn2+). For this reason, the electrophysiological response to TRH was investigated on heavy fraction cells. Experiments performed on 264 cells after 4-12 days in culture showed that these cells could be divided into two groups. The first group, called high resting potential (HRP) cells, constituted 73% of the total and was characterized by a mean resting potential of -60 mV and a mean input membrane resistance of 700 M omega, and these cells displayed plateau potentials, which were triggered by application of brief (2 s), large (1 nA) depolarizing or hyperpolarizing current steps. The plateaux were characterized by a sustained depolarization at a potential near -20 mV and they were concomitant with an increase of the membrane conductance. The repolarization consisted of a slight, gradual hyperpolarization followed by a rapid return to the resting potential. The second group of cells (the remaining 27%; called low resting potential or LRP cells) was characterized by a mean resting potential of -45 mV and a mean input membrane resistance of 250 M omega. These cells were excitable, 30% of them displaying spontaneous activity but never showing plateaux. Electrophysiological experiments showed that the majority of the HRP cells (99%) responded to TRH but were insensitive to dopamine (DA) ejections (up to 10(-6) M) at resting potential. Ejection of TRH onto HRP cells induced a slow depolarization (10-15 mV) concomitant with a decrease of the membrane resistance. This led the cell membrane potential to a critical value (approximately -50 mV), at which the plateau response was triggered. The plateau lasted for about 10 s from potentials greater than -100 mV, and could reach 10 min from holding potentials close to the resting potential. The amplitude of the plateau varied according to the holding potential and the reversal potential was found to be -20 mV. Local application of tetraethylammonium chloride (TEA, 30 mM) only slightly affected the amplitude of the plateaux but they were shortened or totally blocked by Co2+, Cd2+ and Ni2+.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1990

Effects of dopamine on voltage-dependent potassium currents in identified rat lactotroph cells.

The effects of dopamine (DA) on voltage-dependent potassium currents were investigated in rat lactotrophs maintained in primary culture. Lactotroph cells were identified using the reverse hemolytic plaque assay. Membrane currents and potentials of lactotroph cells were recorded using the patch-clamp recording technique in the 'whole-cell' configuration. In the presence of cobalt (2 mM), two types of voltage-dependent K+ currents were recorded, a voltage-activated delayed K+ current (IK) and a voltage-activated transient K+ current (IA). The current IK was activated at membrane potentials varying from -20 to +40 mV and did not inactivate during prolonged voltage steps (up to 25 s); it was blocked by tetraethylammonium (10 mM). The current IA was activated at membrane potentials higher than -45 mV and showed a voltage-dependent inactivation between -110 and -40 mV; it was slightly inhibited by 4-aminopyridine (5 mM). Under current-clamp conditions, the majority of the cells (60%) showed spontaneous Ca2(+)-dependent action potentials (APs) while silent cells (40%) were excitable by depolarizing current pulses. Bath application of 10 nM DA evoked a hyperpolarizing response, blocked spontaneous APs and decrease the amplitude of evoked APs. Only the hyperpolarizing response faded during the course of the whole cell recording experiments. Under voltage-clamp conditions, DA induced a reversible increase in both voltage-dependent outward K+ currents, without modifying their thresholds. Steady-state inactivation of IA was not affected by DA. These DA-induced responses were dose-dependent and they involved D2 receptor activation. They were mimicked by the specific D2 receptor agonist bromocriptine (10 nM) and blocked by the specific D2 receptor antagonist sulpiride (100 nM), the D1 antagonist SCH 23390 being ineffective. The ability of DA to increase voltage-dependent K+ currents cannot be observed without GTP in the recording pipette. It was pertussis-toxin-sensitive but was affected neither by bath application of 1 mM forskolin nor by the presence of 500 microM cyclic AMP with 500 microM 3-isobutyl-1-methylxanthine in the pipette solutions. We conclude that in lactotroph cells DA specifically increases two voltage-dependent K+ currents via a pertussis-toxin-sensitive guanine nucleotide regulatory protein and appears to be independent of intracellular cyclic AMP. This effect leads to a decrease in the excitability of the cell, explaining in part the inhibitory effect of DA on prolactin release.

4-Aminopyridine↗
PubMed1990

Dopamine inhibits two characterized voltage-dependent calcium currents in identified rat lactotroph cells.

The effects of dopamine (DA) on voltage-dependent Ca2+ currents were investigated in cultured rat lactotroph cells using the patch clamp recording technique. Each recorded cell was identified by the reverse hemolytic plaque assay. In the whole-cell configuration, two types of Ca2+ currents, L and T, were characterized on the basis of their kinetics, voltage sensitivity, and pharmacology. The L component had a threshold of -25 mV, showed little inactivation during a 150-msec voltage step, and was maximal at +10 mV. Cadmium ions (100 microM) significantly reduced its amplitude (75%). The T component was activated at a membrane potential close to -50 mV, was maximal at -10 mV, and showed a voltage-dependent inactivation between -90 and -30 mV. It was quickly inactivated during a maintained depolarization (time constant, 27 ms at -30 mV) and was strongly reduced (80%) by nickel ions (100 microM). Bath application of DA (10 nM) caused a markedly general depression of inward Ca2+ currents, acting differently on the T- and L-type currents. DA application shifted the voltage-dependence of the L-type current activation toward depolarization values (8 mV) without modifying its time- and voltage-dependent inactivation. In contrast, DA enhanced the inactivation of the T-type current by accelerating its time-dependent inactivation (25% decrease in the time constant of inactivation) and by shifting the voltage-dependence of the T-type current inactivation toward hyperpolarizing values (-63 mV in control vs. -77 mV in the presence of DA). These effects of DA were dose-dependent and involved the activation of a D2 receptor type. They were mimicked by bromocriptine application (10 nM), whereas sulpiride (100 nM) blocked the DA-evoked response. The D1 antagonist SCH 23390 was ineffective up to 100 microM. All of these DA-induced modifications in Ca2+ currents were abolished using a GTP-free pipette solution or after pretreatment of cells with pertussis toxin, suggesting that DA can regulate the function of Ca2+ channels through GTP-binding proteins (G-proteins). Our results show that DA acts simultaneously by reducing both voltage-dependent Ca2+ currents on lactotroph cells. Thus, DA reduces the entry of Ca2+ ions across the surface membrane and thereby influences electrical activity and the cytosolic free Ca2+ concentration involved in both basal and evoked PRL release.

Animals↗
PubMed1989

Electrophysiological responses to somatostatin of rat hypophysial cells in somatotroph-enriched primary cultures.

1. Somatotroph cells were obtained from pituitaries of adult male rats by dissociation, separation and enrichment on a continuous gradient of bovine serum albumin at unit gravity. They were kept in culture for 7-15 days before electrophysiological experiments. 2. Immunofluorescent staining of the resulting gradient fractions (numbered F2 to F9) indicated that the majority of somatotrophs (75-85%) were located in the heavy fractions (F8 and F9). However, a small percentage (15-20%) of cells in these fractions were identified as lactotrophs. 3. Perifusion experiments indicated that on the one hand release of growth hormone from somatotroph-enriched fractions was stable at the level of 6 ng (2 min)-1 (10(6) cells)-1 and was markedly inhibited by somatostatin (1.9 ng (2 min)-1 (10(6) cells)-1) but not by dopamine. On the other hand, in the same cell preparations, basal prolactin release (1.6 ng (2 min)-1 (10(6) cells)-1) was significantly reduced by dopamine (0.08 ng (2 min)-1 (10(6) cells)-1) but remained unchanged by somatostatin treatment. 4. The inhibitory effect of somatostatin on growth hormone release was dose dependent. This effect was not abolished by tetraethylammonium (40 mM) or 4-aminopyridine (5 mM), but somatostatin decreased high-potassium-induced release. 5. In all the cells recorded (n = 187), 14% (n = 26) displayed a low resting potential (less than -30 mV) and poor membrane resistance (less than 50 M omega). The recording was unstable and resting potentials decreased regularly to 0 mV in less than 5 min. The other 86% of the cells displayed resting potentials varying from -45 to -65 mV and had a membrane resistance of more than 150 M omega. Only cells which displayed these membrane characteristics showed clear responses to somatostatin or dopamine, and were therefore chosen for experiments. 6. In all the cells selected for the experiments (n = 161), 78% (n = 126) showed either triggered or spontaneous action potentials. The action potentials remained insensitive to sodium-free bath solution, but were reversibly blocked by the calcium channel blockers cobalt (5 mM) or nickel (5 mM). 7. When the cells were at resting potential, somatostatin induced a hyperpolarizing response associated with a decrease of membrane resistance. During this response, spontaneous or triggered action potentials were inhibited. The hyperpolarizing response induced by somatostatin was dose-dependent.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1989

Electrophysiological responses of rat pituitary cells in somatotroph-enriched primary culture to human growth-hormone releasing factor.

The in vitro effects of human growth hormone releasing factor (hGRF, 1-44) were studied in somatotroph-enriched cultures (75-85%) obtained from adult male rat pituitaries. Cells perifused with hGRF showed an up to 800% increase in growth hormone (GH) release over basal values in a dose-dependent manner. Calcium current blockers (5 mM of Co2+, Ni2+ or Cd2+) completely inhibited this stimulating effect but sodium-free (choline) medium did not. Using a single-intracellular-electrode recording technique, it was found that hGRF induced a dose-dependent depolarizing response concomitant with a decrease in membrane resistance in 38% of the cells recorded (98 of 258 cells). The reversal potential of this response was close to -40 mV. This depolarizing response was recorded in both excitable and unexcitable cells with no marked difference. Co2+ and Ni2+ (5 mM) completely and reversibly inhibited the membrane response to hGRF. Application of hGRF and somatostatin (SRIF), a hormone that inhibits GH release, to the same cell cultures showed the existence of two subpopulations: one was responsive to both hGRF and SRIF (53%, n = 62), another was only responsive to SRIF (47%, n = 62). Human GRF did not affect prolactin release and did not modify the electrical properties of cells responding to dopamine and therefore considered as lactotrophs. These results suggest that (1) hGRF leads to an increase in growth hormone release and a modification of membrane electrical properties by means of an extracellular Ca2+-dependent pathway, and (2) according to their responses to SRIF and hGRF, there are at least two subpopulations of somatotrophs.

Animals↗
PubMed1987

Evidence of a direct action of triiodothyronine (T3) on the cell membrane of GH3 cells: an electrophysiological approach.

Electrophysiological experiments demonstrate that triiodothyronine (T3) exerts a direct effect on the membrane of a strain of cultured rat pituitary tumor cells, GH3/B6. These cells respond to pressure application of T3 (2-5 nl, concentration 1 X 10(-10) M) with an increase in the membrane resistance (Rm) and a hyperpolarization. Spontaneously firing cells become silent.

Action Potentials↗
PubMed1987

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↗
PubMed1987

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↗
PubMed1987

[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↗
PubMed1985

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↗
PubMed1983

Electrophysiological properties of normal somatotrophs in culture. An intracellular study.

Normal rat pituitary cells were isolated and enriched to obtain a relatively pure population of growth hormone (GH) secreting cells by sedimentation at unit gravity and subsequently grown in culture. The electrophysiological properties of these cells were studied with intracellular recording techniques. 4-6-day-old cells did not show active electrical properties. At 7-9 days the cells were able to generate evoked responses (fast responses) which were blocked by inhibitors of calcium flux and prolonged with tetraethylammonium. The resting potential of these cells was sensitive to external local application of substances known to act on the release of GH. External elevated K+ induced a strong depolarizing response which was associated with an increase of membrane conductance. Somatostatin induced a hyperpolarizing response associated with a decrease of membrane conductance. It is suggested that the fast Ca2+-dependent responses and the slow variations of the resting potential induced by substances which modify hormone release may play an important role in the control of the secretory process of the presumably GH-secreting cells.

Animals↗
PubMed1982

Lidocaine in the treatment of tinnitus aurium. A double-blind study.

Available reports suggest that lidocaine is capable of relieving symptoms of tinnitus, but no convincing data support this conclusion. By use of a placebo-controlled double-blind crossover study we found that 19 out of 26 patients with subjective tinnitus reported that their tinnitus was temporarily better or gone with intravenous (IV) lidocaine and that there was no change with IV saline.

Clinical Trials as Topic↗