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L Dufy-Barbe

Publications and source records attributed to L Dufy-Barbe.

At least 19 recordsLinked to original sources
PubMed1992

Calcium-activated chloride conductance of lactotrophs: comparison of activation in normal and tumoral cells during thyrotropin-releasing-hormone stimulation.

We studied a chloride (Cl-) conductance activated by calcium (Ca2+) in normal rat lactotrophs and compared its activation during TRH stimulation in normal rat lactotrophs and in GH3 tumoral lactosomatotrophs cells, using the whole-cell configuration of the patch-clamp technique. The Cl- specificity of the conductance was assessed by manipulation of internal and external Cl- concentrations. The reversal potentials were in agreement with those predicted by the Nernst equation. Ca2+ ionophore A23187 and membrane depolarizations activated the Cl- conductance. However, a feedback effect of Cl- gradient modifications on Ca2+ movements was also observed in normal lactotrophs. In the latter, TRH (100 nM) mobilization of intracellular Ca2+ activated this Cl- conductance together with the potassium (K+) conductance when both ions were present in the intracellular medium (IM) or alone when K+ was absent. Chloride conductance was not activated in the GH3 cells, where mobilization of intracellular Ca2+ by TRH (100 nM) activated only Ca2(+)-dependent K+ conductance. It seems likely that the activation of Cl- conductance in these two different cell types involves different mechanisms.

Animals↗
PubMed1992

Calcium homeostasis in growth hormone (GH)-secreting adenoma cells: effect of GH-releasing factor.

Human GH-secreting tumors are heterogenous regarding their basal secretory activity and response to GH-releasing factor (GRF). We have investigated whether such different secretory properties could be accounted for by alterations of intracellular mechanisms occurring at the calcium level. Basal free intracellular calcium concentrations ([Ca2+]i) and Ca2+ responses to GRF were studied in single cells cultured from fragments of five GH-secreting pituitary adenomas. We used the microspectrofluorimetric method and indo-1 as the fluorescent probe. The cell populations cultured from the tumors of patients A and C showed increased hormone secretion in response to GRF in vitro, whereas cultures from patients B, D, and E were unresponsive to the peptide. Basal [Ca2+]i measured in the five cell populations ranged from 82 +/- 18 to 118 +/- 27 nM. A 10-sec application of 10 nM GRF induced an increase in [Ca2+]i in 60% and 54% of A and C cells, respectively. In the nonresponsive cell populations, the number of calcium responses to GRF was lower, 26% (B cells), 5% (D cells), and 10% (E cells). Two principal responses types were observed: 1) an initial increase in [Ca2+]i, followed by a sustained plateau phase lasting for more than 200 sec; and 2) a monophasic peak of increased [Ca2+]i lasting approximately 1 min before returning to baseline levels. GRF responses were totally suppressed in the absence of Ca2+ ions in the external medium. Sixteen to 30% of the cells cultured from four of the five tumors showed spontaneous fluctuations of [Ca2+]i. These spontaneous Ca2+ transients were suppressed in Ca(2+)-free medium. The number of cells exhibiting such Ca2+ transients decreased with time in culture. Basal hormone secretion was higher in cultures from patient D, in which no spontaneous Ca2+ transients were observed in any of the 72 studied cells, and in cultures from patients E, in which only 16% of cells were spontaneously active. We conclude that 1) in human responsive somatotrophs, the involvement of Ca2+ in GRF stimulus-secretion coupling mechanisms is apparently similar to that described in somatotrophs of other species; 2) the lack of a secretory response to GRF observed in some tumors may result from impairment of Ca2+ responsiveness in either cell recruitment or response amplitude and/or duration; and 3) spontaneous rhythmic Ca2+ activity is apparently dissociated from basal hormone secretion in some of these tumor cells.

Adenoma↗
PubMed1991

The gonadotropin-releasing hormone associated peptide reduces calcium entry in prolactin-secreting cells.

The precursor molecule to the GnRH contains a peptide named GnRH-associated peptide (GAP) with PRL-inhibiting properties. In this work, we have studied the electrophysiological properties and responses to GAP of three different types of PRL-secreting cells: 1) the rat tumor cell line GH3, 2) normal rat pituitary cells in primary culture, and 3) human PRL-secreting adenoma cells. Using different but complementary techniques we show that GAP reduces intracellular Ca++ levels, [Ca++]i, and inhibits Ca++ transients in these cells. This reduction of [Ca++]i results from coordinate actions of GAP on K+ and Ca++ conductances and may explain the inhibitory effect of GAP on hormonal secretion by PRL-secreting cells.

Adenoma↗
PubMed1991

Growth hormone-releasing factor stimulates calcium entry in the GH3 pituitary cell line.

The GH3 pituitary cell line has been extensively used to study various aspects of the stimulus secretion coupling process. It is known that GH3 cells release PRL and GH in the basal state and in response to various secretagogues. However, this cell line was considered unsuitable as a model for studying the effects of GHRF since the neuropeptide did not affect GH secretion or gene expression. This suggested that the GH3 cells may lack GHRF receptors. The present study investigates the effect of GHRF on free intracellular Ca2+ concentrations in GH3 cells. Cytosolic free calcium concentrations ([Ca2+]i) were monitored in individual cells by microspectrofluorimetry using the fluorescent dye indo 1. When the cells were challenged with a brief application of GHRF (100 nM; 15 sec), 36 out of 59 of these cells responded within a few seconds by a marked increase in [Ca2+]i. GHRF enhanced the frequency of [Ca2+]i oscillations in spontaneously active cells or triggered [Ca2+]i oscillations in inactive cells. The response to GHRF was totally blocked by external Ca2+ free solutions and Ca2+ channel blockers. Combined electrophysiological and fluorescent experiments were carried out in 16 cells. Eleven responded to GHRF. In all cases, the Ca2+ transients triggered by GHRF were associated with action potentials. The Ca2+ responses observed in our experiments clearly show that GH3 cells possess membrane receptors to GHRF. Thus, it is likely that the lack of secretory response observed in GH3 cells does not result from the absence of binding sites to the peptide. It is more likely to be related to alterations of transduction mechanisms resulting in uncoupling between stimulation and secretion.

Calcium↗
PubMed1991

[Stimulation of calcium intake by growth-hormone-releasing-hormone in GH3 cells].

The effect of GH-RH in the intra-cytosolic free Ca2+ concentration was studied in GH3 cells. To this end, we have used microspectrofluorimetry performed on single cells. We show that 60% of cells respond to a brief application of 100 nM GH-RH by an increase of their [Ca2+]i (mean increase 100% over basal values). This response which is blocked by calcium channel inhibitors results from an increased influx of Ca2+ ions from the external medium.

Animals↗
PubMed1991

[Electrophysiological study of the action mechanism of somatoliberin (GH-RH) on hypophyseal GH3 tumor cells].

We have investigated the electrical response of patched GH3 cells to Growth-Hormone Releasing-Hormone (GH-RH). GH-RH (100 nM) enhanced firing frequency of action potentials. This is accompanied by membrane depolarization (5-10 mV) and conductance increase. Voltage clamp studies reveal that GH-RH potentiates calcium inward currents and a calcium-dependent chloride current; transient outward current is diminished. These changes in membrane conductance account for the cytosolic free calcium rise shown by Indo-1 fluorescence measurements.

Action Potentials↗
PubMed1990

Electrophysiological response to thyrotropin-releasing hormone of rat lactotrophs in primary culture.

The response of rat pituitary cells to thyrotropin-releasing hormone (TRH) in primary culture was studied in the whole-cell configuration with the patch-clamp technique. Prolactin (PRL)-containing cells were identified in the culture with a peroxidase-antiperoxidase immunocytochemical method. The cells were cultured from the pituitaries of diestrous (D) and lactating (L) female rats. Membrane electrophysiological properties (resting potential and input resistance) of pituitary cells in primary culture varied widely. Under the recording conditions reported here, the mean resting potential of lactotrophs was about -30 mV. There were spontaneous fluctuations in membrane resting potential (10-15 mV) as well as of membrane input resistance, making these parameters difficult to evaluate accurately. Most of the cells exhibited spontaneous firing activity that was shown to be mainly calcium-dependent. There was no difference between L and D cells in resting membrane electrophysiological properties. TRH (10(-7) M) induced a transient hyperpolarization of the membrane similar to that previously described in the GH3 clonal pituitary cell line. Voltage-clamp studies showed that this hyperpolarization resulted from activation of an outward current, the reversal potential of which ranged from -48 to -86.5 mV. Experimental manipulations of the ionic composition of internal and external recording media suggested that both K+ and Cl- were involved. This hyperpolarizing response was observed both in D and L cells, although L cells had larger and faster responses than D cells. This observation may be of physiological significance because lactotrophs have been reported to exist in various subtypes.

Action Potentials↗
PubMed1988

The electrophysiological effects of thyrotropin-releasing hormone are similar in human TSH- and prolactin-secreting pituitary cells.

We studied the electrophysiological properties of individually characterized TSH-secreting cells cultured from pituitary fragments surgically removed from three patients, two who had primary TSH-secreting adenomas and one who had chronic TSH hypersecretion (hyperplasia) secondary to primary hypothyroidism. The TSH-secreting cells were excitable and had calcium-dependent action potentials. More than 80% of the cells cultured from the two patients with TSH-secreting adenomas were spontaneously active, whereas fewer cells (20%) cultured from the hypothyroid patient were spontaneously active. TRH (50 nmol/L) induced a complex pattern of electrical changes. The initial response was transient hyperpolarization (activation of potassium conductance), followed by increased low amplitude voltage fluctuations occasionally leading to action potentials. These TRH-induced electrophysiological changes were similar to those reported in rat and human PRL-secreting adenoma cells. These results suggest that TRH may have an identical mode of action in tumoral PRL and TSH cells. In the cells from the hypothyroid patient, the initial response to TRH cells was similar, but the second phase response was greater. The findings that the cells cultured from these patients behaved differently with regard to their electrophysiological characteristics (action potentials) and responses to TRH may reflect the different clinical conditions from which they were derived.

Action Potentials↗
PubMed1986

Anti-prolactin cell-surface immunoreactivity identifies a subpopulation of lactotrophs from the rat anterior pituitary.

Suspensions of cells dissociated from the anterior pituitary of the adult rat include many that contain intracellular PRL. After fixation, these cells can be identified and the distribution of their PRL determined by immunocytochemistry with anti-PRL antibodies. We have found that approximately 50% of the cells that contain intracellular PRL also have PRL or PRL-like immunoreactive material on the outer cell surface. Cell-surface PRL can be detected on unfixed anterior pituitary cells using anti-PRL antibodies and either fluorescence microscopy or fluorescence-activated cell sorting. Cell surface labeling by anti-PRL antibodies was restricted to PRL-containing cells; 85-97% of the labeled cells contained detectable intracellular PRL, while fewer than 2% contained detectable GH, ACTH, LH, or TSH. Subpopulations of live anterior pituitary cells could also be labeled on the cell surface by antibodies against GH, ACTH, LH, or TSH. This suggests that the presence of hormone at the cell surface may be a characteristic and identifying feature of different anterior pituitary cell types.

Animals↗
PubMed1986

[LHRH and LH release in the red fox Vulpes vulpes L].

Pituitary responsiveness to exogenous LHRH was studied in vivo and in vitro in the female red fox, a mono-oestrous species. In vivo, the ability of the pituitary to release LH in response to a single injection of LHRH (2 micrograms/kg) was determined at various stages of the reproductive cycle. The greatest responsiveness is observed during the preovulatory period, the lowest during the luteal phase. During the anoestrus phase, the responsiveness is reduced by more than 50% in lactating females compared to non lactating females. In vitro, dispersed fox anterior pituitary cells were exposed four times to LHRH (10(-9) M), hourly, for 8 min. Pituitary cells were taken from lactating and non lactating females. The cells are not sensitive to LHRH in lactating females but become more and more sensitive after weaning. It is suggested the inhibitory influence of lactation could be the result of prolactin-ovarian steroids-gonadotrophins interactions.

Animals↗
PubMed1979

[Electrophysiological study of hypothalamic neurons and gonadotropin regulation in rhesus monkey (author's transl)].

The electrical activity of various hypothalamic areas was recorded in anesthetized (sodium pentobartital) (n = 3) and unanesthetized (n = 3) ovariectomized rhesus monkeys. The plasma gonadotropin levels of the animals during the recording sessions were obtained by a chronic cardiac catheter. 1. Sodium pentobarbital greatly affected the electrical activity of most hypothalamic neurons (84%). The mean spontaneous electrical unit activity of neurons in the unanesthetized animals (n = 30) was 1.5 spike/sec. This value decreased to 0.1 spike/sec in the anesthetized group (n = 30). However, a small number of hypothalamic neurons were insensitive to the anaesthetic. 2. The multiunit electrical activity recorded in the median eminence showed marked periodic increases, about 200 sec before LH was observed to rise in the plasma (10/12). This increased firing lasted about 10 min (Fig. 1A). 3. In a few cases (2/12), however, increased firing was not associated with LH release. Since, in this area, multiunit activity represents the summation of a large number of axon terminals, the release of a hypothalamic hormone other than Gn-RH is put foreward as a possible hypothesis. 4. Of 51 neurons recorded in various hypothalamic areas, 3 showed periodic increases of electrical unit activity which corresponded to a rise in plasma LH levels (Fig. 1 B). These neurons are located in the arcuate area. 5. These results are consistent with the hypothesis of a relation between the electrical activity of some hypothalamic neurons and the periodical LH discharges observed in the ovariectomized rhesus monkey.

Animals↗
PubMed1978

Effects of protein synthesis inhibitors on the negative feed-back effect of estrogen on LH release.

Ovariectomized female rabbits were pretreated with actinomycin D (80 microgram/100 g body weight) cycloheximide (300 microgram/100 g body weight) or chloramphenicol (300 microgram/100 g body weight) 4 h before an intravenous injection of 25 microgram of estradiol benzoate. Actinomycin D, cycloheximide and chloramphenicol decreased basal gonadotropin levels, but actinomycin D was less active than chloramphenicol and cycloheximide. Despite this effect on basal levels, such treatment did not prevent the effect of estradiol benzoate on LH release. Moreover, no significant impairment of the LH response to LH-RH was found. The interpretation of experiments using drugs such actinomycin D, cycloheximide or chloramphenicol requires caution; however, one cannot disregard the possibility that the negative feed-back effect of estrogen, in contrast to the positive effect, may not be mediated by RNA-dependent DNA and the protein-synthesizing systems.

Animals↗
PubMed1976

Effects of estrogen on the electrical activity of identified and unidentified hypothalamic units.

Experiments performed on unanesthetized ovariectomized female rabbits demonstrated the effects of estradiol benzoate (EB; 20 microng i.v.) on the electrical activity of hypothalamic units which send their axons to the median eminence. Of a total of 1,840 cells recorded in hypothalamic and preoptic areas, 46 (2.5%) were antidromically activated by stimulating the median eminence. Under the present experimental conditions, EB induced a progressive diminution in the mean firing rate of these cells observed throughout the recording period (30-120 min). In addition to cells projecting to the median eminence, neurons which could not be antidromically invaded using our techniques were observed to be sensitive to estrogen. Estrogen administration produced a long-lasting inhibition of antidromically activated cells and a depression of much shorter duration (15-20 min) of unidentified nonstimulated units. These data suggest the existence of two types of hypothalamic neurons sensitive to estrogen.

Animals↗
PubMed1975

Effect of ovariectomy and estradiol benzoate treatment on LH-RH induced LH and FSH release in the female rabbit.

Synthetic LH-RH (0.75 and 2.5 mug/animal) was administered to mature intact and castrated female rabbits. The animals were injected for 3 consecutive days with either 40 mug estradiol benzoate (EB) dissolved in 0.5 ml propylene glycol 60%, water 40% or with the solvent alone. LH-RH was injected i.v. on the 4th day. In intact rabbits EB priming increased the pituitary LH response while LH basal levels were lowered. In the unprimed castrated rabbits, basal FSH and LH levels and the cumulative responses to LH-RH were higher than in intact control animals; EB priming lowered basal LH levels and cumulative responses to the values of intact control rabbits while both basal FSH levels and the cumulative responses were less influenced by the treatment. It is concluded that estrogen acts at both hypothalamic and pituitary levels to modify the LH responses to endogenous LH-RH, and that the complete inhibition of FSH hypersecretion in the ovariectomized rabbit probably needs another ovarian factor.

Animals↗