Biomedical subjects

Mike Ludwig

Publications and source records attributed to Mike Ludwig.

16 recordsLinked to original sources

PubMed2006

Histaminergic and glycinergic modulation of GABA release in the vestibular nuclei of normal and labyrinthectomised rats.

Vestibular compensation (the behavioural recovery that follows unilateral vestibular de-afferentation), is facilitated by histamine, and is associated with increased central histamine release and alterations in histamine H(3) receptor expression in the vestibular nuclei. However, little is known of the effects of histamine on neurotransmission in the vestibular nuclei, and the mechanisms by which histamine may influence compensation are unclear. Here we examined the modulatory effects of histaminergic agents on the release of amino acid neurotransmitters in slices of the medial vestibular nucleus (MVN) prepared from normal and labyrinthectomised rats. The release of GABA, but not glutamate, glycine or aspartate, was robustly and reproducibly evoked by a high-K(+) stimulus applied to normal MVN slices. Histamine inhibited the evoked release of GABA, both through a direct action on presynaptic H(3) receptors (presumably located on GABAergic terminals), and through a novel, indirect pathway that involved the increased release of glycine by activation of postsynaptic H(1)/H(2) receptors (presumably on glycinergic neurons). After unilateral labyrinthectomy (UL), the direct H(3) receptor-mediated inhibition of GABA release was profoundly downregulated in both ipsi-lesional and contra-lesional MVNs. This effect appeared within 25 h post-UL and persisted for at least 3 weeks post-UL. In addition, at 25 h post-UL the indirect glycinergic pathway caused a marked suppression of GABA release in the contra-lesional but not ipsi-lesional MVN, which was overcome by strychnine. Stimulation of histamine H(3) receptors at 25 h post-UL restored contra-lesional GABA release to normal, suggesting that acutely after UL H(3) receptors may strongly modulate glycinergic and GABAergic neurotransmission in the MVN. These findings are the first to demonstrate the modulatory actions of the histaminergic system on neurotransmission in the vestibular nuclei, and the changes that occur during vestibular system plasticity. During vestibular compensation, histaminergic modulation of glycine and GABA release may contribute to the rebalancing of neural activity in the vestibular nuclei of the lesioned and intact sides.

Amino Acids↗

PubMed2006

Endogenous activation of supraoptic nucleus kappa-opioid receptors terminates spontaneous phasic bursts in rat magnocellular neurosecretory cells.

Phasic activity in magnocellular neurosecretory vasopressin cells is characterized by alternating periods of activity (bursts) and silence. During phasic bursts, action potentials (spikes) are superimposed on plateau potentials that are generated by summation of depolarizing after-potentials (DAPs). Burst termination is believed to result from autocrine feedback inhibition of plateau potentials by the kappa-opioid peptide, dynorphin, which is copackaged in vasopressin neurosecretory vesicles and exocytosed from vasopressin cell dendrites during phasic bursts. Here we tested this hypothesis, using intracellular recording in vitro to show that kappa-opioid receptor antagonist administration enhanced plateau potential amplitude to increase postspike excitability during spontaneous phasic activity. The antagonist also increased postburst DAP amplitude in vitro, indicating that endogenous dynorphin probably reduces plateau potential amplitude by inhibiting the DAP mechanism. However, the kappa-opioid receptor antagonist did not affect the slow depolarization that follows burst termination, suggesting that recovery from endogenous kappa-opioid inhibition does not contribute to the slow depolarization. We also show, by extracellular single-unit recording, that that there is a strong random element in the timing of burst initiation and termination in vivo. Administration of a kappa-opioid receptor antagonist eliminated the random element of burst termination but did not alter the timing of burst initiation. We conclude that dendritic dynorphin release terminates phasic bursts by reducing the amplitude of plateau potentials to reduce the probability of spike firing as bursts progress. By contrast, dendritic dynorphin release does not greatly influence the membrane potential between bursts and evidently does not influence the timing of burst initiation.

Action Potentials↗

PubMed2006

Dendritic peptide release and peptide-dependent behaviours.

Neuropeptides that are released from dendrites, such as oxytocin and vasopressin, function as autocrine or paracrine signals at their site of origin, but can also act at distant brain targets to evoke long-lasting changes in behaviour. Oxytocin, for instance, has profound effects on social bonding that are exerted at sites that richly express oxytocin receptors, but which are innervated by few, if any, oxytocin-containing projections. How can a prolonged, diffuse signal have coherent behavioural consequences? The recently demonstrated ability of neuropeptides to prime vesicle stores for activity-dependent release could lead to a temporary functional reorganization of neuronal networks harbouring specific peptide receptors, providing a substrate for long-lasting effects.

Animals↗

PubMed2005

Regulation of activity-dependent dendritic vasopressin release from rat supraoptic neurones.

Magnocellular neurones of the hypothalamus release vasopressin and oxytocin from their dendrites and soma. Using a combination of electrophysiology, microdialysis, in vitro explants, and radioimmunoassay we assessed the involvement of intracellular Ca(2+) stores in the regulation of dendritic vasopressin release. Thapsigargin and cyclopiazonic acid, which mobilize Ca(2+) from intracellular stores of the endoplasmic reticulum, evoked vasopressin release from dendrites and somata of magnocellular neurones in the supraoptic nucleus. Thapsigargin also produced a dramatic potentiation of dendritic vasopressin release evoked by osmotic or high potassium stimulation. This effect is long lasting, time dependent, and specific to thapsigargin as caffeine and ryanodine had no effect. Furthermore, antidromic activation of electrical activity in the cell bodies released vasopressin from dendrites only after thapsigargin pretreatment. Thus, exposure to Ca(2+) mobilizers such as thapsigargin or cyclopiazonic acid primes the releasable pool of vasopressin in the dendrites, so that release can subsequently be evoked by electrical and depolarization-dependent activation. Vasopressin itself is effective in inducing dendritic vasopressin release, but it is ineffective in producing priming.

Animals↗

PubMed2004

Regulation of neuronal nitric oxide synthase mRNA expression in the rat magnocellular neurosecretory system.

We examined the activation of nNOS mRNA expression within the supraoptic and paraventricular nuclei (SON and PVN) of the hypothalamus. In salt-loaded rats nNOS mRNA expression was significantly increased in both nuclei. In rats given i.p. injections of 1.5 M NaCl (4 ml/kg), a small but significant increase in nNOS mRNA expression in the SON and PVN was found 6 h after injection; no change was detected 2 or 4 h after injection. In rats in which hyponatraemia had been induced experimentally, nNOS mRNA was downregulated in the SON, and expression levels were not increased within 4 h after intense acute osmotic stimuli. Finally, neurons of the SON were antidromically-activated by neural stalk stimulation for 2 h. No increase of nNOS mRNA expression in the SON was observed 2 h after stimulation. Thus, increased electrical activity is not directly coupled to rapidly increased expression of nNOS mRNA, and hence acute increases in nNOS mRNA expression are unlikely to play a role in short-term adaptation of the magnocellular system to osmotic stimulation.

Analysis of Variance↗

PubMed2004

Phasic spike patterning in rat supraoptic neurones in vivo and in vitro.

In vivo, most vasopressin cells of the hypothalamic supraoptic nucleus fire action potentials in a 'phasic' pattern when the systemic osmotic pressure is elevated, while most oxytocin cells fire continuously. The phasic firing pattern is believed to arise as a consequence of intrinsic activity-dependent changes in membrane potential, and these have been extensively studied in vitro. Here we analysed the discharge patterning of supraoptic nucleus neurones in vivo, to infer the characteristics of the post-spike sequence of hyperpolarization and depolarization from the observed spike patterning. We then compared patterning in phasic cells in vivo and in vitro, and we found systematic differences in the interspike interval distributions, and in other statistical parameters that characterized activity patterns within bursts. Analysis of hazard functions (probability of spike initiation as a function of time since the preceding spike) revealed that phasic firing in vitro appears consistent with a regenerative process arising from a relatively slow, late depolarizing afterpotential that approaches or exceeds spike threshold. By contrast, in vivo activity appears to be dominated by stochastic rather than deterministic mechanisms, and appears consistent with a relatively early and fast depolarizing afterpotential that modulates the probability that random synaptic input exceeds spike threshold. Despite superficial similarities in the phasic firing patterns observed in vivo and in vitro, there are thus fundamental differences in the underlying mechanisms.

Action Potentials↗

PubMed2004

TFF3 induced Fos protein expression in the magnocellular oxytocin neurons of the hypothalamus.

TFF3 is synthesized in magnocellular oxytocin neurons of the supraoptic (SON) and paraventricular nuclei (PVN) of the rat and human hypothalamus. Here we investigated whether intracerebroventricular (i.c.v.) injection of TFF3 stimulates oxytocin release into the blood and activates Fos protein immunoreactivity in oxytocin neurons of the SON and PVN in rats. The results show that plasma oxytocin concentrations were not altered after i.c.v. injection of TFF3 or vehicle. Fos protein expression was significantly increased in both the SON and PVN after TFF3 injections and double labeling studies showed that the Fos signal was predominantly in oxytocin neurons.

Animals↗

PubMed2004

The activity of the hypothalamo-neurohypophysial system in response to acute stressor exposure: neuroendocrine and electrophysiological observations.

The present mini review focuses on stress-induced alterations of the electrical and secretory activity of vasopressin (AVP) and oxytocin (OXT) neurones originating within the supraoptic nucleus (SON) and constituting the hypothalamo-neurohypophysial system (HNS) in the male rat. Previously, it was thought that SON neurones are predominantly activated by osmotic and reproductive stimuli. However, recent findings also suggest a selective activation of AVP and/or OXT neurones in response to specific stressors. Inhibitory amino acids seem to participate at the level of the SON in the control of HNS activity during stress. Taurine, probably of glial origin, selectively inhibits the secretory activity of AVP neurones. In contrast, GABA, probably of neuronal origin, interferes with the release of OXT both from axon terminals into blood and from somata/dendrites into the extracellular fluid of the SON. Depending upon whether a defined stressor triggers taurine and/or GABA release within the SON the secretion of AVP and/or OXT from HNS neurones will be inhibited. These observations shed new light on the neurone-neurone and glial-neurone interactions that ensure an appropriate neuroendocrine stress response.

Acute Disease↗

PubMed2004

GABA selectively controls the secretory activity of oxytocin neurons in the rat supraoptic nucleus.

Recently we reported that a single social defeat experience triggers the release of oxytocin (OXT) from somata and dendrites, but not axon terminals, of neurons of the hypothalamic-neurohypophysial system. To further investigate the regulatory mechanisms underlying this dissociated release, we exposed male Wistar rats to a 30-min social defeat and monitored release of the inhibitory amino acids gamma amino butyric acid (GABA) and taurine within the hypothalamic supraoptic nucleus (SON) using microdialysis. Social defeat caused a significant increase in the release of both GABA and taurine within the SON (up to 480%; P < 0.01 vs. prestress release). To reveal the physiological significance of centrally released GABA, the specific GABAA-receptor antagonist bicuculline (0.02 mm) was administered into the SON via retrodialysis. This approach caused a significant increase in the release of OXT both within the SON and into the blood under basal conditions and during stress (up to 300 and 200%, respectively; P < 0.05 vs. basal values), without affecting plasma vasopressin. Electrophysiological studies confirmed the selective action of bicuculline on the firing activity of OXT neurons in the SON. Taken together, our data demonstrate that GABA is released within the SON during emotional stress to act as a selective inhibitor of both central and peripheral OXT secretion.

Action Potentials↗

PubMed2004

Thapsigargin-induced mobilization of dendritic dense-cored vesicles in rat supraoptic neurons.

Dense-cored vesicles (DCVs) containing oxytocin or vasopressin are secreted from both the nerve terminals in the posterior pituitary and dendrites in the hypothalamus of magnocellular supraoptic neurons. Dendritic secretion can be enhanced (primed) by pretreatment with either thapsigargin or oxytocin for subsequent activity-dependent release. Here, we determined whether priming involves a translocation of DCV closer to the dendritic membrane. To reduce total vesicle content, rats were salt-loaded for 24 h before application of thapsigargin or vehicle onto the ventrally exposed surface of the supraoptic nucleus in vivo. Tissues were then prepared for quantitative electron microscopic analysis of the total incidence of DCVs within supraoptic dendritic cross-sections, and of the incidence and distance (within a 500-nm margin) of each DCV to the dendritic plasma membrane. Salt loading per se did not alter the frequency distribution or average proportion of DCVs found in the 500-nm margin but significantly decreased the average incidence of DCVs per dendrite by 30% (P < 0.05). However, thapsigargin treatment resulted in a significant increase in the total incidence of DCVs within the 500-nm margins and a higher incidence of DCVs within the first 200 nm of the plasma membrane (P < 0.05), indicating that the thapsigargin-induced priming involves a relocation of DCVs closer to sites of secretion.

Animals↗

PubMed2003

Neurosteroid regulation of oxytocin and vasopressin release from the rat supraoptic nucleus.

In adult rats somato-dendritic release of oxytocin and vasopressin from magnocellular neurones in the supraoptic nucleus of the hypothalamus has important autoregulatory actions on the neuronal electrical activity, and in neonatal rats it plays a role in the development of dendritic arborisation. In the adult, oxytocin effects are modulated by allopregnanolone via an interaction with inhibitory GABAA receptors. This study examined the effects of allopregnanolone, progesterone and 17beta-oestradiol on oxytocin and vasopressin release from intact isolated supraoptic nuclei and from the neurophypophyses in rats of differing ages. In supraoptic nuclei from rats of 3-4 weeks old or less, all three neurosteroids induced oxytocin release from the isolated supraoptic nucleus, but only allopregnanolone induced significant release of vasopressin. Surprisingly, in these very young rats, allopregnanolone-induced oxytocin release was inhibited by GABAA receptor antagonists as well as by an oxytocin receptor antagonist. By contrast, in supraoptic nuclei from adult rats allopregnanolone-induced oxytocin release was much smaller, and was enhanced in the presence of bicuculline. The GABAA receptor agonist muscimol also induced oxytocin release from supraoptic nuclei in young rats, but had no effect in adult rats. Oxytocin cells isolated from young rats showed an increase in [Ca2+]i in response to both allopregnanolone and muscimol. Allopregnanolone had no effect on [Ca2+]i or on the release of oxytocin or vasopressin from neurohypophysial axon terminals in either young or old rats. We conclude that, in very young rats, (i) neurosteroids induce oxytocin release from the supraoptic nucleus by a mechanism that partly depends on the presence of GABA, which in young rats is depolarising to oxytocin cells, and which also partly depends upon endogenous oxytocin, and (ii) the effect of allopregnanolone upon oxytocin release changes with age, as the functional activity of GABAA receptors changes from excitation to inhibition of oxytocin cells.

Aging↗

PubMed2003

Talking back: dendritic neurotransmitter release.

Classical transmitters and neuropeptides can be released from the dendrites of many neuronal populations, to act as retrograde signals that modulate synaptic transmission, electrical activity and, in some cases, morphology of the cell of origin. For the hypothalamic neuroendocrine cells that release vasopressin and oxytocin, the stimuli, mechanisms and physiological functions of dendritic release have been revealed in detail that is not yet available for other neurons. The regulation of dendritic transmitter release is complex and at least partially independent from axon terminal release. Here, we provide an overview of recent findings on the mechanisms and physiological consequences of dendritic neuropeptide release and place this in the context of discoveries of dendritic neurotransmitter release in other brain regions.

Animals↗

PubMed2003

The role of nitric oxide in morphine dependence and withdrawal excitation of rat oxytocin neurons.

Magnocellular oxytocin neurons develop morphine dependence after intracerebroventricular infusion for 5 days as revealed by their profound excitation following naloxone-induced withdrawal. Oxytocin neurons strongly express nitric oxide synthase (NOS) and nitric oxide (NO) inhibits their activity. This study investigated whether excitation of oxytocin neurons during morphine withdrawal involves reduced activity of NOS and NO. Neuron activity was measured in urethane-anaesthetized rats with blood sampling for oxytocin radioimmunoassay and extracellular single unit firing rate recording of supraoptic nucleus oxytocin neurons. To compare morphine-dependent and -naive rats oxytocin secretion was measured during stimulation by intravenous hypertonic saline infusion. Prior treatment with Nomega-nitro-l-arginine methyl ester, a NOS inhibitor, facilitated osmotically stimulated oxytocin secretion in both morphine-dependent and -naive rats. The facilitation was not different between these groups when corrected for the slower responses observed in morphine-dependent rats. Treatment of morphine-dependent rats with Nomega-nitro-l-arginine methyl ester also enhanced oxytocin secretion during naloxone-precipitated withdrawal. Oxytocin neurons excited by withdrawal were recorded during microdialysis application to the supraoptic nucleus of the NO donor sodium nitroprusside alone and in combination with the GABAA antagonist bicuculline. Sodium nitroprusside inhibited oxytocin neurons during naloxone-precipitated morphine withdrawal and, while bicuculline alone increased firing rate, it did not reduce the inhibition by sodium nitroprusside, in contrast with previous findings in naive rats. Together, these findings indicate that NO restraint of oxytocin secretion is not curtailed during morphine dependence and remains a potent inhibitor of withdrawal excitation despite reduced effectiveness on GABA innervation of the supraoptic nucleus. Hence there is no evidence that changes in NO regulation underlie excitation of oxytocin neurons during opiate withdrawal in morphine dependence.

Action Potentials↗

PubMed2002

Intracellular calcium stores regulate activity-dependent neuropeptide release from dendrites.

Information in neurons flows from synapses, through the dendrites and cell body (soma), and, finally, along the axon as spikes of electrical activity that will ultimately release neurotransmitters from the nerve terminals. However, the dendrites of many neurons also have a secretory role, transmitting information back to afferent nerve terminals. In some central nervous system neurons, spikes that originate at the soma can travel along dendrites as well as axons, and may thus elicit secretion from both compartments. Here, we show that in hypothalamic oxytocin neurons, agents that mobilize intracellular Ca(2+) induce oxytocin release from dendrites without increasing the electrical activity of the cell body, and without inducing secretion from the nerve terminals. Conversely, electrical activity in the cell bodies can cause the secretion of oxytocin from nerve terminals with little or no release from the dendrites. Finally, mobilization of intracellular Ca(2+) can also prime the releasable pool of oxytocin in the dendrites. This priming action makes dendritic oxytocin available for release in response to subsequent spike activity. Priming persists for a prolonged period, changing the nature of interactions between oxytocin neurons and their neighbours.

Animals↗

PubMed2002

The active role of dendrites in the regulation of magnocellular neurosecretory cell behavior.

The interactions of the dendritically released neuropeptides vasopressin and oxytocin with co-released neuroactive substances such as opioids and nitric oxide are reviewed. Endogenous opioids regulate magnocellular neurons at the level of the supraoptic nucleus and the relationship of dendritically released peptides and co-released opioids seems to be dependent on the stimulus given and the physiological state of the animal. Nitric oxide has a prominent inhibitory action on supraoptic neurons and these actions are predominantly mediated indirectly by GABA inputs. The role of these co-released neuroactive substances in differentially regulated release of neuropeptides from dendrites versus distant axon terminals has to be determined in more detail. A picture emerges in which release of vasopressin and oxytocin from different anatomical compartments of a single neuron may arise from different intracellular secretory pools and their preparation before release.

Animals↗

PubMed2002

Vasopressin from hypothalamic magnocellular neurons has opposite actions at the adenohypophysis and in the supraoptic nucleus on ACTH secretion.

Magnocellular vasopressinergic and oxytocinergic neurons of the hypothalamic supraoptic (SON) and paraventricular nuclei comprise the hypothalamic-neurohypophysial system, which is crucially involved in the regulation of body fluid and electrolyte homeostasis. However, still controversial is to what extent the same system influences the secretion of adrenocorticotropic hormone (ACTH) from the adenohypophysis. Therefore, we selectively stimulated magnocellular neurons of the SON of conscious male Wistar rats via retrodialysis. As expected, dialysis of the SON with hypertonic medium increased both the release of vasopressin within the SON and the secretion of vasopressin and oxytocin into the systemic blood circulation. This activation of the hypothalamic-neurohypophysial system was accompanied by a fivefold increase in plasma ACTH concentration. This effect was observed only if the tip of the microdialysis probe was within the SON. Intravenous infusion of the vasopressin V1 receptor antagonist d(CH2)5Tyr(Me)AVP significantly attenuated the effects of local osmotic stimulation of the SON on ACTH secretion. In contrast, administration of the same antagonist directly into the SON significantly enhanced the osmotically stimulated secretion of ACTH and corticosterone, primarily by delaying the restoration of the hormone secretion to prestimulation levels. We conclude from these findings that vasopressin from the hypothalamic-neurohypophysial system participates in the regulation of the hormonal stress response in a counterbalanced manner at the level of the SON and the adenohypophysis.

Adrenocorticotropic Hormone↗