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J J Nordmann

Publications and source records attributed to J J Nordmann.

At least 19 recordsLinked to original sources
PubMed1992

Endothelin regulation of neuropeptide release from nerve endings of the posterior pituitary.

We have investigated the role of endothelin (ET) in the stimulus-secretion coupling mechanism in the posterior pituitary. We report that isolated nerve endings contain immunoreactive endothelin, the level of which is regulated by homeostatic mechanisms involved in control of water balance. ET-1 and ET-3 potentiate vasopressin release induced by depolarization through interaction with specific receptors of the ETA subtype and this response is antagonized by sarafotoxin S6b. The second messenger for this effect, however, remains unknown since the potentiation of depolarization-induced vasopressin release occurs in the absence of an increase in cellular calcium.

Animals↗
PubMed1992

Intracellular calcium and hormone release from nerve endings of the neurohypophysis in the presence of opioid agonists and antagonists.

Rat neural lobes and isolated nerve terminals from the neurohypophysis were stimulated in the presence of different opioid agonists and antagonists. The secretion of arginine vasopressin and oxytocin and rise in cytoplasmic calcium induced by depolarization were analyzed by radioimmunoassay and the fluorescent probe fura-2, respectively. The kappa-agonists dynorphin A(1-13) and dynorphin A(1-8) did not affect electrically evoked release of vasopressin, although oxytocin release was slightly reduced. U-50 488, a relatively specific kappa-receptor agonist, had no effect on the amount of vasopressin or oxytocin secreted, although it significantly reduced K(+)-evoked changes in [Ca2+]i in isolated nerve endings. Two kappa-receptor antagonists, MR 2266 and diprenorphin, alone had no effect on vasopressin and oxytocin secretion from isolated nerve endings depolarized with potassium. Opioid agonists less selective for the kappa receptors, etorphin and ethylketocyclazocin, were found to inhibit the release of both vasopressin and oxytocin significantly. Naloxone, a nonselective opiate receptor antagonist, alone had no effect on vasopressin release but potentiated the electrically evoked release of oxytocin. Naloxone also could overcome the inhibitory effect of etorphin on oxytocin and vasopressin release observed after electrical stimulation of the neural lobe. A number of inconsistencies therefore exist between the effects of opioid agonists and antagonists on neuropeptide release and on the evoked changes in [Ca2+]i. In view of these inconsistencies and the high concentrations of opioid agonists and antagonists necessary to modify release, we conclude that it is doubtful that opioid molecules have a physiological role in controlling neurohypophysial secretion.

Animals↗
PubMed1992

G-proteins mediate inhibition and activation of Ca(2+)-induced exocytosis from SLO-permeabilized peptidergic nerve endings.

In SLO-permeabilized isolated nerve endings from the rat neurohypophysis, GTP, guanosine 5'[y-thio]triphosphate (GTPyS) and guanosine 5'(beta y-imido]triphosphate (GMPPNP) inhibit the Ca(2+)-evoked vasopressin release. Pretreatment with pertussis toxin enhances the inhibitory effects of both GTP-analogues. Omission of Mg2+ overcomes the effect of GMPPNP and reverses the inhibitory effect of GTP and GTPyS. In the absence of Mg2+, GTP and GTPyS now potentiate Ca(2+)-evoked secretion.

Animals↗
PubMed1992

Exo-endocytosis in isolated peptidergic nerve terminals occurs in the sub-second range.

Exo- and endocytotic processes induced by depolarization of isolated neurosecretory nerve terminals show a close temporal correlation, which suggests a short time of integration of the neurosecretory granule membrane with the plasma membrane. In order to determine minimal time requirements for exocytosis-coupled endocytosis to occur, we have analyzed by electron microscopy uptake of horseradish peroxidase (HRP) as a fluid phase marker at the onset of depolarization. We have applied rapid mixing and sampling (quenched flow) to assess events in sub-second time periods after stimulation. A significant number of labelled endocytotic vacuoles was observed during the first second of depolarization. This number then further increased by a factor of about 2 (within 5 s) and 4 (within 50 s). Thus, as for exocytosis, the rate of endocytosis decreased considerably during prolonged stimulation. These data indicate i) that a substantial proportion of secretory granules undergoes exocytosis very shortly after stimulation, and ii) that, following exocytosis, the minimal time required for consecutive membrane retrieval is in the sub-second range.

Animals↗
PubMed1992

The light chain of tetanus toxin inhibits calcium-dependent vasopressin release from permeabilized nerve endings.

The effects of tetanus toxin and its light and heavy chain subunits on vasopressin release were investigated in digitonin-permeabilized neurosecretory nerve terminals isolated from the neural lobe of the rat pituitary gland. Exocytosis was induced by challenging the permeabilized nerve endings with micromolar calcium concentrations. Tetanus toxin inhibited vasopressin release only in the presence of the reducing agent dithiothreitol. This effect was irreversible. The purified light chain of tetanus toxin strongly inhibited exocytosis in a dose-dependent manner with half-maximal effect at c. 10 nM. The action of the light chain was observed after only 2.5 min of preincubation. Separated heavy chain subunit had no effect on hormone secretion. Inhibition of vasopressin release could be prevented by preincubating the light chain of tetanus toxin with an immune serum against tetanus toxin. The data clearly demonstrate that in mammalian neurosecretory nerve endings tetanus toxin acts at a step downstream from the activation by Ca2+ of the exocytotic machinery and that the functional domain of this toxin is confined to its light chain.

Animals↗
PubMed1992

Membrane retrieval following exocytosis in isolated neurosecretory nerve endings.

In the neurosecretory nerve endings of the neurohypophysis depolarization-induced exocytosis is followed by endocytosis of vacuole-like structures with diameter similar to that of neurosecretory granules. However, it remains unknown whether the membrane of the endocytotic vacuoles is comprised primarily of retrieved secretory granule membrane, plasma membrane or of a mixture of the granule and plasma membrane. In the present paper membrane retrieval following depolarization-induced exocytosis has been studied in isolated neurosecretory nerve endings from the rat neurohypophysis. The origin of the retrieved membrane was assessed by pre-labeling the plasma membrane with an antibody against neural cell adhesion molecule, a plasma membrane specific protein. Horseradish peroxidase was used as an index of fluid endocytosis and secretion of vasopressin was measured by radioimmunoassay. Following potassium-induced depolarization, endocytotic vacuoles showed labeling with the fluid phase marker horseradish peroxidase but never showed significant neural cell adhesion molecule labeling. The time-course of endocytosis following closely that of exocytosis as endocytotic vacuoles labeled with horseradish peroxidase were only observed when the fluid phase marker was present in the extracellular medium during the period of evoked exocytosis. Our results are consistent with a model in which in neurosecretory nerve endings, after transient exocytotic fusion of the granule membrane with the plasma membrane, the granule membrane is rapidly and selectively retrieved into the nerve endings in the form of vacuoles similar in size to that of the neurosecretory granules.

Animals↗
PubMed1992

Possible role during exocytosis of a Ca(2+)-activated channel in neurohypophysial granules.

Ion channels from bovine neurohypophysial granules were incorporated into artificial lipid bilayers. The larger amplitude channel is permeable to cations and exhibits multiple conductances. The channel opens only in the presence of free Ca2+, but is inhibited by relatively high Ca2+ concentrations. Release of vasopressin from permeabilized neurohypophysial terminals also shows a similar biphasic dependence on Ca2+. Release is selectively inhibited by low concentrations of the long-chain alcohol octanol, but not by high concentrations of ethanol, as is the neurosecretory granule Ca(2+)-activated cation channel. Furthermore, Ca(2+)-evoked release and channel activity are both inhibited by the long-chain tetraethylammonium analogs decamethonium and decyl-triethyl ammonium bromide. The close correlation between channel and release properties lead us to conclude that the Ca(2+)-activated channel is involved in peptide secretion.

Alcohols↗
PubMed1992

Depolarization, intracellular calcium and exocytosis in single vertebrate nerve endings.

We have investigated the temporal relationship between depolarization, elevation of [Ca2+]i and exocytosis in single vertebrate neuroendocrine nerve terminals. The change of [Ca2+]i and vasopressin release were measured with a time resolution of less than 1 s in response to K(+)-induced depolarization. Exocytosis was also monitored in the whole-terminal patch-clamp configuration by time resolved capacitance measurements while [Ca2+]i was simultaneously followed by fura-2 fluorescence measurements. In intact as well as patch-clamped nerve terminals sustained depolarization leads to a sustained rise of [Ca2+]i. The rate of vasopressin release from intact nerve terminals rises in parallel with [Ca2+]i but then declines rapidly towards basal (t1/2 approximately 15 s) despite the maintained high [Ca2+]i indicating that only a limited number of exocytotic vesicles can be released. We demonstrate that in nerve terminals exocytosis can be followed during step depolarization by capacitance measurements. The capacitance increase starts instantaneously whereas [Ca2+]i rises with a half time of several hundred milliseconds. An instantaneous steep capacitance increase is followed by a slow increase with a slope of 25-50 fF/s indicating the sequential fusion of predocked and cytoplasmic vesicles. During depolarization the capacitance slope declines to zero with a similar time course as the vasopressin release indicating a decrease in exocytotic activity. Depolarization per se in the absence of a sufficient rise of [Ca2+]i does not induce exocytosis but elevation of [Ca2+]i in the absence of depolarization is as effective as in its presence. The experiments suggest that a rapid rise of [Ca2+]i in a narrow region beneath the plasma membrane induces a burst of exocytotic activity preceding the elevation of bulk [Ca2+]i in the whole nerve terminal.

Animals↗
PubMed1991

Ca(2+)-independent regulation of neurosecretion by intracellular Na+.

While secretion from nerve endings is strictly controlled by an increase in cytoplasmic free calcium several reports suggest intracellular sodium may serve a regulatory role. Whether sodium acts directly to modulate secretion or indirectly by influencing cytoplasmic calcium dynamics is unknown. This study shows, based on parallel experiments studying [Na+]i, [Ca2+]i and vasopressin secretion, that sodium acts directly to regulate secretion in isolated nerve endings from the rat neurohypophysis. The elevation in secretion that develops is dose-dependently related to the [Na+]i and can occur in the absence of changes in [Ca2+]i.

Animals↗
PubMed1991

Ethanol reduces vasopressin release by inhibiting calcium currents in nerve terminals.

Ingestion of ethanol (EtOH) is known to result in a reduction of plasma arginine-vasopressin (AVP) levels in mammals. We examined the basis for this effect using a combination of biochemical and electrophysiological techniques. Release of AVP from nerve terminals isolated from the rat neurohypophysis was very sensitive to EtOH, with significant reductions in AVP release evident in 10 mM EtOH. However, EtOH did not affect the release of AVP from terminals which had been permeabilized with digitonin, suggesting that voltage-gated calcium channels might be the target of EtOH's actions. Patch clamping of these terminals indicated that both inactivating and long-lasting calcium currents were reduced in EtOH, but the long-lasting currents were more sensitive (significant reductions in 10 mM EtOH). EtOH-induced decreases in plasma AVP levels can be explained by EtOH's inhibition of calcium currents in the nerve terminals.

Animals↗
PubMed1991

Exocytosis in neurohypophysial nerve terminals is not coupled to protein kinase C translocation.

Protein kinase C (PKC) has been implicated in the mechanism of exocytosis, although various studies have been unable to pinpoint actual translocation or activation of PKC during exocytosis. We have studied, in neurohypophysial nerve endings, intracellular Ca2+ levels, secretion of neuropeptides and PKC translocation. Neurohormone secretion was triggered by K(+)-induced or electrically induced depolarization in both the absence and the presence of phorbol esters. PKC was translocated from the cytosol to the membrane on electrical stimulation or K+ depolarization, but not to the extent obtained with phorbol ester. Data are presented clearly demonstrating that the translocation of PKC from cytosol to membrane is not required for exocytosis, nor does it alter in any way neuropeptide release from neurohypophysial nerve terminals.

Alkaloids↗
PubMed1991

Effect of sodium and calcium on basal secretory activity of rat neurohypophysial peptidergic nerve terminals.

1. The release of arginine vasopressin (AVP) from a perifused preparation of peptidergic nerve terminals isolated from rat neurohypophyses was studied during manipulations of the external sodium and calcium concentrations. Intracellular concentrations of these two ions were manipulated by use of ouabain and a calcium ionophore, respectively. 2. Removal of extracellular Na+ caused, in a concentration-dependent manner, a significant decrease of secretory activity. Conversely, graded addition of Na+ to a Na(+)-free perifusion medium increased secretion. Half-maximal activation of secretory activity was attained at ca 75 mM [Na+]o. 3. Manipulations of extracellular Ca2+ did not affect the level of hormonal secretion in the absence of extracellular Na+. However, when Na+ was present in the perifusion medium, removal of extracellular Ca2+ induced an increase of secretory activity. 4. The effects of manipulations of [Na+]o were not dependent on the presence of Ca2+ in the perifusion medium nor on the nature of the Na+ replacement used (i.e. choline or mannitol). 5. Ouabain (0.1 mM) increased the basal secretory activity and potentiated the secretory response to removal of Ca2+ from the perifusion medium. 6. The Ca2+ ionophore A23187 stimulated, in a concentration-dependent fashion, the secretory activity of the peptidergic nerve terminals and this stimulation was strictly dependent on the presence of Ca2+ in the perifusion medium. 7. These results show that basal secretion is directly dependent on [Na]o and indicate that intracellular Na+ is an important factor in the control of secretory mechanisms. Evidence is presented in regard to a possible antagonistic effect of extracellular Ca2+ and Na+ on secretion.

Animals↗
PubMed1991

Calcium currents and peptide release from neurohypophysial terminals are inhibited by ethanol.

The effects of EtOH on peptide release and on high-threshold, voltage-activated calcium (Ca++) channels were examined in acutely dissociated rat neurohypophysial terminals. These terminals release the peptide hormones, arginine vasopressin (AVP) and oxytocin. Release of AVP from isolated intact neurohypophyses, induced by either electrical stimulation or elevated potassium, was inhibited by clinically relevant concentrations of EtOH. "Whole-cell" patch-clamp recording methods were used to study the effects of EtOH on voltage-activated Ca++ currents (ICa) in the peptidergic nerve terminals. Amplitudes of both fast-inactivating ICa and long-lasting ICa were reduced in EtOH, and the reduction in ICa did not result from a shift in its current-voltage or steady-state inactivation relationships. Only the fast-inactivating component recovered after removal of EtOH. The effects of EtOH on ICa could not be attributed to changes in osmolarity. In contrast to ICa, the fast, transient K+ current was insensitive to EtOH. These results suggest that EtOH-induced reduction of ICa in the peptidergic nerve terminals produces a decrease in AVP release, resulting in lowered plasma AVP levels.

Animals↗
PubMed1990

Release of vasopressin from isolated permeabilized neurosecretory nerve terminals is blocked by the light chain of botulinum A toxin.

The intracellular action on exocytosis of botulinum A toxin and constituent chains was studied using permeabilized isolated nerve endings from the rat neural lobe. The release of the neuropeptide vasopressin was measured by radioimmunoassay. In the presence of the reducing agent dithiothreitol, the two-chain form of botulinum A toxin inhibited vasopressin release induced by 10 microM free calcium. Half maximal inhibition was obtained with 15 nM botulinum A toxin. In the absence of the heavy chain the light chain of the toxin strongly inhibited exocytosis with a half maximal effect of 2.5 nM. The inhibitory effects on secretion could be prevented by incubating the light chain with an immune serum against botulinum A toxin. The heavy chain of botulinum A toxin did not affect vasopressin release. However, it prevented the inhibitory effects of the light chain on stimulated exocytosis. It is concluded that botulinum A toxin inhibits the calcium-dependent step leading to exocytosis by interfering with a target present in the isolated and permeabilized nerve terminals. The functional domain of this neurotoxin, which is responsible for the inhibition of vasopressin release, is present in its light chain.

Animals↗
PubMed1989

Chloride and magnesium dependence of vasopressin release from rat permeabilized neurohypophysial nerve endings.

The role of Cl- and Mg+ ions has been studied on the secretory mechanism leading to the release of vasopressin from digitonin permeabilized nerve endings isolated from the rat neurohypophysis. Secretion was triggered by challenging the permeabilized nerve endings with 1.1 microM free Ca2+. Magnesium enhances secretion and its maximal effect occurred at a concentration of about 2 mM. Further increase of this divalent cation concentration however led to an inhibition of secretion. Chloride ions are necessary for the final steps in exocytosis and this effect of Cl- was inhibited by the chloride channel antagonist N144. It is concluded that in neurosecretory nerve endings magnesium and chloride ions are crucial components for exocytosis to occur.

Animals↗