Biomedical subjects

T A Lovick

Publications and source records attributed to T A Lovick.

At least 19 recordsLinked to original sources

PubMed2006

Neuronal excitability in the periaqueductal grey matter during the estrous cycle in female Wistar rats.

Extracellular recordings were made from output neurons in the dorsal half of the periaqueductal gray matter (dPAG) in urethane-anesthetized female Wistar rats. All the neurons were quiescent. A basal level of firing was therefore induced by continuous iontophoretic application of D,L-homocysteic acid (DLH). In the presence of the GABA(A) receptor antagonist bicuculline methiodide (BIC 0-30 nA) the DLH-induced firing increased further, revealing the presence of ongoing GABAergic inhibitory tone on the recorded neurons. The BIC-induced increase in firing rate was significantly greater in neurons recorded during estrus (Est) and late diestrus (LD) compared with proestrus (Pro) and early diestrus (ED) suggesting that GABAergic tone was lower in Est and LD. I.v. injection of the panicogenic cholecystokinin (CCK)(B) receptor agonist pentagastrin (PG, 40 microg kg(-1)) produced an increase in firing rate in 12/17 (70%) of neurons tested in the dPAG. Iontophoretic application of PG (10-30 nA) also produced a current-related increase in firing rate in 73.6% of the neurons tested. The excitatory response was reduced during application of the selective CCK(B) receptor antagonist beta-[2-([2-(8-azaspiro[4.5]dec-8-ylcarbonyl)-4,6-dimethylphenyl]amino)-2-oxoethyl]-(R)-napthalenepropanoic acid (CR2945) (60 nA, n=6). The PG-evoked increase in firing rate was significantly greater in neurons recorded during Est and LD compared with during Pro and ED. Juxtacellular labeling with neurobiotin in eight neurons revealed multipolar cells 12-44 microm diameter with up to six primary dendrites. In three of eight neurons, a filled axon was present and coursed without branching toward the perimeter of the periaqueductal gray matter (PAG). The estrous cycle-related change in responsiveness to BIC and PG suggests that the panic circuitry in the PAG may become more responsive to panicogenic agents during estrus and late diestrus as a consequence of a decrease in the intrinsic level of inhibitory GABAergic tone. The findings may have implications for understanding the neural processes that underlie the development of premenstrual dysphorias in women.

Anesthesia↗

PubMed2006

Plasticity of GABAA receptor subunit expression during the oestrous cycle of the rat: implications for premenstrual syndrome in women.

Many women experience psychological changes during the luteal phase of their menstrual cycle. The late luteal (premenstrual) phase, when symptoms become most severe, is characterized by declining levels of ovarian progesterone. In female rats, withdrawal from prolonged dosing with progesterone leads to upregulation of alpha4 and delta subunits of the GABAA receptor in several brain regions. During the oestrous cycle of the rat, the natural fall in progesterone that occurs in late dioestrus is associated with a parallel increase in expression of alpha4, beta1 and delta GABAA receptor subunits in neurones in the periaqueductal grey matter (PAG), suggesting that new receptors of the alpha4beta1delta composition have been formed. Recombinant alpha4beta1delta receptors display a low EC50 for GABA, which is consistent with activation by extracellular levels of GABA. They are also likely to be located extrasynaptically and to carry tonic currents. In the PAG, a region involved in mediating panic-like anxiety, alpha4, beta1 and delta GABAA receptor subunits are located principally on GABAergic interneurones. On-going GABAergic neuronal activity normally limits and controls the excitability of the panic circuitry. During late dioestrus, when expression of alpha4, beta1 and delta subunits on GABAergic interneurones is upregulated, the increase in tonic current would be expected to lead to a reduction in the activity of the GABAergic population. Thus the panic circuitry would become intrinsically more excitable. It is suggested that during the menstrual cycle in women, plasticity of GABAA receptor subunit expression in brain regions such as the PAG, which are involved in mediating anxiety behaviour, may underlie some of the changes in mood that occur during the premenstrual period.

Animals↗

PubMed2006

Cardiovascular and respiratory responses to a panicogenic agent in anaesthetised female Wistar rats at different stages of the oestrous cycle.

In urethane-anaesthetised female Wistar rats, intravenous injection of the panicogenic CCK(B) receptor agonist pentagastrin (0.002-80 microg/kg) evoked a dose-related increase in blood pressure, heart rate and ventilation. The response was blocked in the presence of the selective CCK(B) receptor antagonist CR2945 (1 mg/kg i.v.). The same pattern of cardiovascular and respiratory changes was evoked by microinjection of pentagastrin (0.3 nmol in 250 nL) into the dorsal half of the periaqueductal grey matter (PAG). The effect of intra-PAG administration of pentagastrin was also abolished following injection of CR2945 (1 mg/kg, i.v.). Responsiveness to systemically administered pentagastrin was enhanced in rats in late dioestrus. At the highest dose tested (80 microg/kg), the pressor response, tachycardia and tachypnoea evoked in rats in late dioestrus was significantly higher than rats in proestrus. For rats in oestrus, the pressor response and tachycardia but not tachypnoea were also significantly larger than the response evoked in rats in early dioestrus. The results suggest that the dorsal half of the PAG (dPAG) plays a key role in mediating the cardiovascular and respiratory responses evoked by systemically administered CCK(B) agonists. The enhanced responsiveness to panicogenic agents during late dioestrus may be related to changes in the functional responsiveness of gamma-aminobutyric acid (GABA)ergic circuitry in the dPAG due to plasticity of GABA(A) receptor subunit expression as a consequence of falling progesterone levels.

Animals↗

PubMed2005

GABAergic neurones in the rat periaqueductal grey matter express alpha4, beta1 and delta GABAA receptor subunits: plasticity of expression during the estrous cycle.

Immunoreactivity for alpha4, beta1 and delta GABAA receptor subunits on neurones in the periaqueductal gray matter was investigated at different stages of the estrous cycle in Wistar rats. Immunostaining for alpha4, beta1 and delta GABAA receptor subunits was present on neurones throughout the periaqueductal gray matter. The numbers of subunit-immunoreactive neurones remained constant during the early phases of the estrous cycle (proestrus to early diestrus) but increased significantly in late diestrus. Dual immunolabeling for the GABA synthesizing enzyme glutamic acid decarboxylase revealed that almost 90% of the subunit-positive cells contained immunoreactivity for glutamic acid decarboxylase. During the early phases of the estrous cycle (proestrus to early diestrus), approximately one third of the glutamic acid decarboxylase-positive population co-localized alpha4, beta1 and delta GABAA receptor subunits. When the number of subunit positive cells increased during late diestrus, the proportion of the glutamic acid decarboxylase-containing population that expressed alpha4, beta1 and delta GABAA receptor subunits almost doubled. We propose that GABAA receptors with the alpha4beta1delta configuration are expressed by GABAergic neurones in the periaqueductal gray matter and that the numbers of cells expressing these subunits are increased in late diestrus in line with falling plasma progesterone levels. Changes in GABAA receptor expression may lead to changes in the excitability of the neural circuitry in the periaqueductal gray matter.

Animals↗

PubMed2005

Changes in GABA(A) receptor subunit expression in the midbrain during the oestrous cycle in Wistar rats.

In women, the late luteal phase or "premenstrual" period is commonly associated with psychological disturbances, which include mood changes and increased aggression. The underlying cause is unknown but one possibility is that fluctuations in levels of neuroactive steroids precipitate changes in expression of GABA(A) receptor subunits that result in functional changes in inhibitory control systems. The present study investigated the levels of expression of alpha4, beta1 and delta GABA(A) receptor subunits in the periaqueductal gray matter (PAG) in rats and whether plasticity occurs during the oestrous cycle in females. In male rats alpha4, beta1 and delta subunit immunoreactive neurones were present throughout the PAG in similar numbers. In female rats in proestrus, oestrus and early dioestrus, the density of alpha4, beta1 and delta subunit immunoreactive cells was similar to males. However, in late dioestrus, the numbers increased significantly, especially in the dorsolateral PAG, a region which is particularly rich in GABAergic interneurones. These parallel changes may reflect an increase in expression of the alpha4beta1delta GABA(A) receptor subtype. Recombinant alpha4beta1delta receptors, expressed in Xenopus oocytes, exhibited and EC(50) for GABA an order of magnitude lower (2.02+/-0.33 microM; mean+/-S.E.M.) than that found for the most ubiquitous alpha1beta2gamma2 GABA(A) receptor (32.8+/-2.5 microM). Increased expression of alpha4beta1delta GABA(A) receptors in the interneurones of the PAG could render the panic circuitry abnormally excitable by disinhibiting the ongoing GABAergic inhibition. Similar changes in neuronal excitability within the PAG in women consequent to falling steroid levels in the late luteal phase of the menstrual cycle could contribute to the development of pre-menstrual dysphoria.

Animals↗

PubMed2004

Neuronal activity-related coupling in cortical arterioles: involvement of astrocyte-derived factors.

Neuronal activity-evoked dilatation was investigated in cortical arterioles in brain slices from mature rats maintained in vitro at 31-33 degrees C. In the presence of the thromboxane A2 agonist U46619 (75 nM) to preconstrict vessels, internal diameter decreased by 14.2% and rhythmic contractile activity (vasomotion) developed. Addition of the epoxygenase inhibitor miconazole (20 microm) produced a further decrease in diameter and increase in the frequency of vasomotion, suggesting that tonic release of epoxygenase products maintains a level of cerebrovascular dilator tone. Addition of 1 mum AMPA for 5 min evoked a 15.4 +/- 3.7% increase in diameter and the frequency of vasomotion decreased by -6.7 +/- 1.4 contractions min(-1). The response persisted in the presence of 1 mum TTX, indicating that it was independent of neuronal activity and thus likely to have been evoked by activation of AMPA receptors on astrocytes rather than neurones. The response to the brief (5 min) application of AMPA remained unchanged in the presence of miconazole (20 microm). Prolonged (30 min) application of AMPA produced a +12.1 +/- 1.5% increase in internal diameter and reduction in vasomotion (-8.4 +/- 1.7 contractions min(-1)) that were sustained throughout the stimulation period. However, when AMPA was applied in the presence of miconazole (20 microm) it evoked only a transient increase in diameter (+9.8 +/- 3.1%) and decrease in vasomotion (-6.6 +/- 1.5 contractions min(-1)) that lasted for less than 10 min despite continued application of AMPA. The results suggest that products of epoxygenase activity, probably epoxyeicosatrienoic acids (EETs) are involved in activity-related dilatation in cortical arterioles. Whilst epoxygenase activity is not required to initiate dilatation, it appears to be involved in sustaining the response. Thus EETs released from membrane stores could contribute to the initial stages, but once these have been depleted de novo synthesis of EETs is required to maintain the effect.

Action Potentials↗

PubMed2002

Co-localization of 5-HT 2A -receptor- and GABA-immunoreactivity in neurones in the periaqueductal grey matter of the rat.

The dorsal half of the midbrain periaqueductal grey matter (dPAG) functions as a midbrain aversive system. Serotonin exerts anti-aversive effects in the dPAG that are mediated by actions at 5-hydroxytryptamine (5-HT)(1A)- and 5-HT(2A)-receptors. Since at the cellular level, 5-HT(2A)-receptor activation has been shown to evoke excitatory responses in the dPAG, it is possible that anti-aversive 5-HT(2A)-mediated effects are mediated indirectly by activation of inhibitory gamma-aminobutyric acid (GABA)ergic interneurones. In rats, immunoreactivity for 5-HT(2A)-receptors was present on the soma and dendrites of neurones throughout the PAG. Co-localization studies revealed that the majority (>90%) of 5-HT(2A)-receptor-labelled cells also showed immunoreactivity for GABA. These findings may provide an anatomical substrate for 5-HT(2A)-mediated anti-aversive effects in the dPAG.

Animals↗

PubMed2002

Inhibition of vasomotion in hippocampal cerebral arterioles during increases in neuronal activity.

The activity of small arterioles, internal diameter 9.9 +/- 0.8 microm (SEM), was investigated in the CA1 region of hippocampal slices maintained in vitro at 34 degrees C. Under resting conditions, the vessels were quiescent. However, in the presence of the thromboxane A2 agonist U46619 (75-100 nM), rhythmic contractile activity (vasomotion, 1.1-9.9 min(-1), mean 4.1 +/- 0.7 min(-1) SEM) developed in the smooth muscle cells of the vessel walls. Electrical stimulation of the Schaffer collateral fibre pathway was used to evoke increases in neuronal activity in CA1 in the vicinity of the vessels under investigation. A 3-min period of electrical stimulation of the Schaffer collateral fibre pathway produced a significant reduction in vasomotion in 8/8 vessels. During stimulation, vasomotion either ceased completely (n = 5) or the frequency decreased from 7.1, 3.3 and 3.2 min(-1) to 1.2, 0.4 and 0.6 min(-1), respectively (n = 3). In addition, the amplitude of the residual contractions was reduced by 66%, 12% and 52%. In the presence of 1 microM tetrodotoxin (TTX) (n = 4) to block the generation of action potentials, vasomotion was still present. However, the inhibition of vasomotion evoked by increased neuronal activity was blocked concomitant with the abolition of the field potentials recorded in CA1 in response to the stimulation of the Schaffer collaterals. These findings suggest that a reduction in vasomotion may contribute to the local hyperaemia, which accompanies increases in synaptic activity in the brain.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗

PubMed2001

Involvement of GABA in medullary raphe-evoked modulation of neuronal activity in the periaqueductal grey matter in the rat.

Experiments were carried out in urethane-anaesthetised rats to investigate whether GABA is involved in mediating inhibition of neuronal activity in the dorsal half of the periaqueductal grey matter (PAG) after stimulation of the serotonin-containing projection to the PAG from nucleus raphe obscurus (NRO). Multibarrelled micropipettes were used to make recordings from 42 neurones in the dorsal half of the PAG. Most (n = 36) cells were quiescent. Their firing rate was therefore raised to 10-16 Hz by continuous iontophoretic application of DL-homocysteic acid (DLH) in order to facilitate the study of inhibitory events. Iontophoretic application of GABA (0-10 nA) silenced every neurone tested (n = 42), and the effect was blocked by the GABAA antagonist bicuculline (BIC, 10-20 nA; 15/15 cells). BIC also produced an increase in ongoing activity in 14 of 15 cells, indicating the presence of inhibitory GABAergic tone. Iontophoretically applied serotonin (5-HT; 10-70 nA) also inhibited ongoing activity in 9 of 11 cells. The effect of 5-HT was not blocked by BIC. In six of seven cells, microinjection of 100-200 nl DLH into NRO produced a 72.3 +/- 9.4% decrease in neuronal firing rate which was maximal 112 +/- 18 s after the start of the injection and lasted for a total of 313 +/- 63 s. In five of six cells, the raphe-evoked inhibition was blocked by BIC. It is suggested that activation of the serotonergic projection to the PAG from NRO engages GABA-containing interneurones within the PAG which mediate the inhibitory effects of raphe stimulation.

Action Potentials↗

PubMed2000

Fluorescent imaging of nitric oxide production in neuronal varicosities associated with intraparenchymal arterioles in rat hippocampal slices.

The fluorescent indicator 4,5-diaminofluorescein (DAF-2) has been used to investigate the production of nitric oxide in the vicinity of intraparenchymal cerebral blood vessels. Slices of rat hippocampus 300-350 microm thick, were loaded with 5 microM DAF-2 diacetate. On exposure to light of 450-490 nm wavelength, point sources of fluorescence, 1.8+/-0.2 microm in diameter (mean+/-SEM), were observed in close apposition to the outer surface of the vascular smooth muscle wall of 10/15 arterioles. In fixed slices, resectioned and processed for nicotinamide adenine dinucleotide phosphate-dependent diaphorase, stained varicose fibres were also seen in close association with the smooth muscle wall of small arterioles. These findings suggest that tonic activity in perivascular nitrergic nerve fibres lying in close proximity to intraparenchymal microvessels may be a source of dilator tone within the parenchyma.

Animals↗

PubMed2000

Serotonergic transmission in the periaqueductal gray matter in relation to aversive behaviour: morphological evidence for direct modulatory effects on identified output neurons.

Intracellular recordings were made from 21 cells in the dorsolateral periaqueductal gray matter in coronal midbrain slices. In the majority (n = 20) bath application of 5-hydroxytryptamine (30 or 150 mM) evoked either hyperpolarizing (n = 11) or depolarizing (n = 9) responses. Reconstructions of 11 neurons in the dorsolateral periaqueductal gray matter after filling with biocytin revealed a population of output neurons whose axons followed a dorsolateral trajectory towards the perimeter of the ipsilateral periaqueductal gray matter. In seven cells, the axon could be followed into the adjacent mesencephalic reticular formation. At the light microscopic level, immunostaining for 5-hydroxytryptamine revealed immunoreactive processes throughout the dorsolateral periaqueductal gray matter but no labelled somata or dendrites. Close associations (i.e. no discernible gap) were observed between serotonergic profiles and the somata and dendrites of biocytin-filled cells. At the ultrastructural level, serial sections through 21 appositions on to biocytin-filled dendrites in three slices revealed 19 true appositions (i.e. having closely parallel plasma membranes with no intervening glial cell profiles) with the biocytin-filled dendrite. Only four of the appositions (21%) showed evidence of synaptic specializations which included aggregations of synaptic vesicles, and some thickening of the apposing membrane. The dense reaction product in the biocytin-filled cells precluded identification of the ultrastructure of postsynaptic elements. However, examination of contacts between 5-hydroxytryptamine-immunoreactive profiles and unlabelled elements in material taken from the contralateral side of the periaqueductal gray matter (i.e. no biocytin present) or in material taken from perfusion-fixed whole brain, in which ultrastructural preservation was superior compared with slices, revealed a similar incidence (21% and 23%, respectively) of synaptic specializations. The data indicate that serotonergic transmission on to output neurons in the dorsolateral periaqueductal gray matter is largely mediated by non-junctional contacts, suggesting that the actions of 5-hydroxytryptamine on these cells are mediated predominantly by volume rather than wiring transmission.

Animals↗

PubMed1999

Bio-imaging of nitric oxide-producing neurones in slices of rat brain using 4,5-diaminofluorescein.

4,5-Diaminofluorescein (DAF-2) was used to identify individual nitric oxide (NO)-producing neurones in brain slices in vitro. Coronal slices of midbrain or hippocampus, 300 microm thick from young adult rats, were incubated for 30 min in 1 microM DAF-2 diacetate (DAF-2 DA) and maintained in ACSF at 33 degrees C. Illumination at 450-490 nm revealed punctate fluorescence in neurones in the lateral tegmental nucleus, dorsal raphe nucleus, dorsolateral periaqueductal grey matter, deep collicular layers and cortical areas. Neurones in the hippocampal pyramidal cell layer, molecular layer of the dentate gyrus and the hilus fluoresced also. The fluorescence was abolished by pre-incubation of slices with L-NAME (100 microM-1 mM), the inhibitor of constitutive nitric oxide synthase (NOS), but not by D-NAME (100 microM) or L-NIL (5-50 microM), an inhibitor of inducible NOS. In some superficially located arterioles, there were small regions of bright fluorescence close to the outer smooth muscle wall and diffuse fluorescence within the adjacent smooth muscle cells. A diffuse fluorescence was also seen in some superficially located capillaries. Basal production of NO was not seen within deeper blood vessels. DAF-2 DA offers a sensitive indicator for visualising basal production of NO with high spatial resolution and could provide a means of identifying NOS-containing neurones in brain slices in vitro prior to neurophysiological study.

Animals↗

PubMed1999

Co-localization of GABA with nicotinamide adenine dinucleotide phosphate-dependent diaphorase in neurones in the dorsolateral periaqueductal grey matter of the rat.

Co-localization of GABA and nitric oxide synthase (NOS) has been investigated in neurones in the dorsolateral sector of the periaqueductal grey matter (PAG) in rats. GABA-immunoreactive neurones were found throughout the PAG although their density was greatest in the dorsolateral sector. Neurones containing nicotinamide adenine dinucleotide phosphate-dependent diaphorase, used to indicate NOS, were confined to the dorsolateral sector. GABA immunoreactivity was detected in 26.9% of 3009 diaphorase-reactive neurones counted at all rostro-caudal levels of the PAG in material from three rats. This double-labelled population may act as a neuronal gain control system which sets the level of excitability and responsiveness of effector systems in other sectors of the PAG.

Animals↗

PubMed1999

Neurones in the dorsolateral periaqueductal grey matter in coronal slices of rat midbrain: electrophysiological and morphological characteristics.

Intracellular recordings using the whole-cell patch-clamp configuration were made from 87 neurones in the dorsolateral "aversive" region of the periaqueductal grey matter (PAG) in coronal slices of rat midbrain. Camera lucida reconstructions made of 28 of the cells, which had been filled with biocytin, revealed round, triangular or oval cell bodies 11-40 microm in diameter. Between two and seven primary dendrites were present, which branched further, often becoming varicose. The dendritic tree was always contained within the dorsal half of the PAG. Biocytin-filled axons could be followed for 177-2315 microm from the soma. The axons typically travelled to the perimeter of the dorsal half of the ipsilateral PAG, before either turning ventrolaterally to run parallel to the tecto-bulbopsinal fibres or continuing their trajectory into the deep collicular layers or the mesencephalic reticular formation. Electrophysiologically, two functional categories of cells could be distinguished: type-A cells (30%) showed inward rectification at membrane voltages in excess of -77 mV and had short action potentials (1.6+/-0.07 ms), which were followed by a biphasic afterpolarisation characterised by an initial fast and a later slow phase. Only 18.5% of the type-A cells were spontaneously active (<4 Hz). The second category of cell (type B, 70% of the recorded population) did not show rectification and had longer-duration action potentials (2.1+/-0.07 ms), which showed a smooth decay of the afterhyperpolarisation phase. Approximately one third (37%) of the type-B cells fired spontaneously (<4 Hz). The gross morphology of the two types of cells was similar. However, in type-A cells (n=6), the axons could be seen to originate from the cell soma, whereas, in the majority of the type-B population (10/13), the axon arose from a primary dendrite. The results show that coronal slices of midbrain contain a viable population of "output" or "projection" neurones, which are accessible to electrophysiological and pharmacological investigation. In conscious animals, efferent output from this part of the PAG is concerned with mediating the autonomic and somatomotor changes which are characteristic components of aversive emotional behaviour. The output neurones should therefore reflect the net level of excitability in the PAG in relation to its functional activity.

Action Potentials↗

PubMed1999

Neurovascular relationships in hippocampal slices: physiological and anatomical studies of mechanisms underlying flow-metabolism coupling in intraparenchymal microvessels.

Experiments were carried out to investigate the functional and anatomical relationships between neuronal elements and cerebral microvessels in 300-350-microm thick coronal hippocampal slices maintained at 33-35 degrees C, obtained from 150-200 g male Wistar rats. Cerebral arterioles (9-22 microm in diameter) were visualized in situ and pre-constricted by 22.0+/-6.6% by the addition of the thromboxane A2 agonist U46619 (75 nM), to the bathing medium. The glutamate agonist N-methyl-D-aspartate (0.01-1 mM) produced a dose-related increase in luminal diameter of pre-constricted vessels. In the presence of 4 microM haemoglobin to scavenge nitric oxide from the extravascular environment of the slice, the increase in diameter evoked by 0.1 mM N-methyl-D-aspartate was significantly reduced from 17.5+/-4.6% to 4.8+/-1.7% indicating that N-methyl-D-aspartate-induced vasodilatation of cerebral microvessels is mediated via a mechanism which involves neuronally-derived nitric oxide. In a parallel anatomical study, beta-nicotinamide adenine dinucleotide phosphate-dependent diaphorase staining was used to reveal the enzyme nitric oxide synthase in vascular endothelium and neurons in slices. A small subpopulation (< 11 cells per slice) of darkly-stained multipolar neurons, 21-32 microm in diameter was observed to give rise to a dense network of fine diaphorase-reactive nerve fibres that ramified throughout the whole of the hippocampus and appeared to come into close apposition with arterioles. Morphometric analysis of the relationship between cerebral microvessels, beta-nicotinamide adenine dinucleotide phosphate, reduced form-dependent diaphorase-reactive neuronal elements and individual pyramidal layer neurons, identified by filling with biocytin, revealed that for a given point on a pyramidal layer neuron, the proximity of the nearest diaphorase-reactive nerve fibre was less than 10 microm, whilst the distance to the nearest arteriole (the smallest functional unit for controlling blood flow) was in excess of 70 microm. Such a distance would probably preclude diffusion of vasoactive metabolites in effective concentrations from the area of increased neuronal activity. We therefore propose that the diaphorase-reactive nerve network constitutes the functional link. It is possible that during periods of increased neuronal activity, spillover of glutamate from synapses may activate the diaphorase-reactive network. Release of nitric oxide from the network in the vicinity of local cerebral arterioles may then produce relaxation of the vascular smooth muscle, enabling increased blood flow into the capillary network supplying the region of increased metabolic activity. This study has shown that the process whereby increases in neuronal activity elicit a local change in cerebral blood flow remains functionally intact in hippocampal slice preparations. Nitric oxide of neuronal origin appears to be involved in mediating the coupling between neurons and cerebral arterioles. Stereological analysis of the relationship between neuronal and vascular elements within hippocampal slices suggested that a small subpopulation of nitric oxide synthase-containing neurons which give rise to a diffuse network of fine nitric oxide synthase-containing nerve fibres that lie in close apposition to cerebral arterioles may provide the anatomical substrate for coupling of blood flow to metabolism.

Animals↗

PubMed1997

Projections from dorsal raphe nucleus to the periaqueductal grey matter: studies in slices of rat midbrain maintained in vitro.

In coronal slices of rat midbrain localised injections of FluoroGold or DiO into the dorsolateral periaqueductal grey matter (PAG) labelled cells retrogradely in the dorsal half of the dorsal raphe nucleus (DRN) and the ipsilateral ventrolateral PAG. Low intensity electrical stimulation in the dorsolateral PAG (22.9 +/- 1.9 microA) evoked antidromic responses in neurones recorded intracellularly in the dorsal subnucleus of the DRN. Antidromic responses could also be evoked in neurones in the ventral DRN and its wings but only at much higher currents (40.9 +/- 2.5 microA) which likely spread to activate axons in the ventrolateral PAG that originated from perikarya in the ventral DRN. The findings are discussed in relation to modulation of the excitability of the aversive system in the dorsolateral PAG by the DRN.

Animals↗

PubMed1997

Role of nitric oxide and serotonin in modulation of the cardiovascular defence response evoked by stimulation in the periaqueductal grey matter in rats.

In rats anaesthetised with alphaxalone/alphadolone, electrical stimulation (10 s trains of 1 ms pulses at 80 Hz, 40-80 microA) in the dorsolateral and lateral periaqueductal grey matter (PAG), the midbrain defence area, evoked a pressor response with tachycardia and vasodilatation in the hindlimb. Microinjection of 200 nl 0.66 mM 5HT, but not 200 nl 165 mM NaCl, at the site of stimulation attenuated the components of the PAG-evoked response by 75-98%. The effect of 5HT was significantly reduced by prior intracerebroventricular injection of 100 microg N-nitro-L-arginine methyl ester (an inhibitor of nitric oxide synthase) but not N-nitro-D-arginine methyl ester. Resting cardiovascular parameters did not change significantly following any of these manipulations. The results suggest that serotonin exerts an inhibitory modulation on the excitability of the midbrain defence area by a mechanism which involves nitric oxide.

Animals↗

PubMed1997

Involvement of nitric oxide and serotonin in modulation of antinociception and pressor responses evoked by stimulation in the dorsolateral region of the periaqueductal gray matter in the rat.

In rats anaesthetized with alphaxalone/alphadolone, electrical stimulation in the periaqueductal gray matter in the region lying lateral and dorsolateral to the aqueduct produced a pressor response and an increase in the latency of the tail flick response to noxious heat applied to the tail. The antinociception and the pressor response were significantly attenuated following microinjection of 15 nmol 5-hydroxytryptamine at the site of stimulation in the periaqueductal gray matter. Microinjection of an equal volume of 165 mM saline had no effect. The inhibitory effects of 5-hydroxytryptamine were blocked by prior intracerebroventricular administration of 100 microg of the nitric oxide synthase inhibitor L-nitroarginine methyl ester. Neither 5-hydroxytryptamine or L-nitroarginine methyl ester had any effect on resting arterial pressure or on the baseline latency of the tail flick reflex. It is suggested that the inhibitory effects of 5-hydroxytryptamine in the dorsolateral periaqueductal gray matter are normally dependent on the functional integrity of local nitric oxide synthase-containing interneurons. Nitric oxide may act in association with 5-hydroxytryptamine to control the excitability of the aversive system in the midbrain.

Animals↗