Analysis of recordings of single-unit firing and population activity in the dorsal subiculum of unrestrained,freely moving rats

We examined neuronal activity in the dorsal subiculum of unrestrained, male adult Wistar rats, which were implanted with a moveable eight-electrode microdrive. The subiculum is the primary hippocampal formation output area and is comparatively uninvestigated neurophysiologically. We compared subicular unit activity and the subicular EEG while rats occupied a small, restricted environment and also correlated neuronal activity with the ongoing behavior of the animal. Units were separated using their electrophysiological characteristics into bursting units, regular spiking units, theta-modulated units, and fast spiking units. The bursting and regular spiking unit classes are similar to hippocampal CA1 units, whereas the fast spiking units appear to be interneurons. Bursting units were variable in their behavior: some units bursted regularly, and others bursted only occasionally. Theta-modulated units have not been described before; these were similar to regular spiking units in all respects except that they increased their firing significantly when theta oscillations were present in the simultaneous EEG record. Subicular EEG was similar to hippocampal EEG, with theta oscillations dominating "alert, moving" behaviors, while large amplitude irregular activity (LIA), which included sharp waves, predominated when theta oscillations were not present, mainly during "alert, still" and "quiet" behaviors. A relatively small proportion of subicular recordings (approximately 32%) were phase-locked to theta; this is a smaller proportion than in areas from which the subiculum takes major inputs. The relative lack of entrainment of subicular neurons by this important intrinsic rhythm is suggestive of a limit to which theta might be capable of affecting both subicular and hippocampal information processing more generally.     

Physiological and behavioural responses to duodenal pain in freely moving rats

Luminal distention of the intestine can be aversive in humans and laboratory animals, and hypersensitivity to distention is found in functional gastrointestinal disorders. Current animal models either require anaesthesia or acute balloon intubation or use implanted balloons of irritant materials, for which the aversive quality of distention and physiological responses have not been well characterised. We report here that silicone balloon catheters implanted in the duodenum via the stomach have long patency without obvious tissue damage. Balloon inflation in freely moving rats caused passive avoidance learning and classic 'pain' postures, as well as graded cardiovascular responses which can be recorded telemetrically. The method should make long-lasting studies of pharmacological and environmental effects on visceral sensitivity more feasible.     

Oscillatory entrainment of thalamic neurons by theta rhythm in freely moving rats

The anterior thalamic nuclei are assumed to support episodic memory with anterior thalamic dysfunction a core feature of diencephalic amnesia. To date, the electrophysiological characterization of this region in behaving rodents has been restricted to the anterodorsal nucleus. Here we compared single-unit spikes with population activity in the anteroventral nucleus (AV) of freely moving rats during foraging and during naturally occurring sleep. We identified AV units that synchronize their bursting activity in the 6-11 Hz range. We show for the first time in freely moving rats that a subgroup of AV neurons is strongly entrained by theta oscillations. This feature together with their firing properties and spike shape suggests they be classified as "theta" units. To prove the selectivity of AV theta cells for theta rhythm, we compared the relation of spiking rhythmicity to local field potentials during theta and non-theta periods. The most distinguishable non-theta oscillations in rodent anterior thalamus are sleep spindles. We therefore compared the firing properties of AV units during theta and spindle periods. We found that theta and spindle oscillations differ in their spatial distribution within AV, suggesting separate cellular sources for these oscillations. While theta-bursting neurons were related to the distribution of local field theta power, spindle amplitude was independent of the theta units' position. Slow- and fast-spiking bursting units that are selectively entrained to theta rhythm comprise 23.7% of AV neurons. Our results provide a framework for electrophysiological classification of AV neurons as part of theta limbic circuitry.     

Electric activity in the neocortex of freely moving young and aged rats

Electroencephalographic activity of the neocortex was evaluated in young (5-7 months) and aged (26-28 months) rats. All animals in the aged group showed behavioral impairment in a spatial task (water maze). A neocortical electroencephalogram was derived simultaneously from 16 different neocortical locations and was subjected to spectral analysis. The frequency of occurrence and duration of high-voltage spindles was determined in two sessions, each involving a total of 30 min alert immobility. Changes in spectral characteristics and high-voltage spindles in response to scopolamine administration were also evaluated. The power of high-frequency activity (8-20 Hz) was significantly reduced in the aged subjects. This was greatest in the temporo-occipital regions, while no significant changes were seen in the mediofrontal region. Scopolamine resulted in a large power increase in all frequency bands, but the increase in the higher-frequency range (8-20 Hz) was significantly less in the aged group. The incidence of high-voltage spindles was 6 times higher and their total duration was 9 times longer in aged rats, with virtually no overlap with the young group. In young rats, scopolamine increased the incidence and total duration of high-voltage spindles, while it decreased both parameters in the aged subjects. Cholinergic neurons in the nucleus basalis appeared shrunken in the aged animals. These findings demonstrate that reliable electroencephalographic changes are present in the neocortex of the aged rat, and that some of the physiological alterations may be due to the pathological changes in the cholinergic nucleus basalis.     

Calpain inhibitor I retards seizure offset in the hippocampus of freely moving rats

The implications of increased calpain-mediated proteolysis during epileptic seizures are still unclear, and in this study we investigate the effect of the continuous perfusion of calpain inhibitor I on picrotoxin-induced seizures in chronic freely moving rats. Continuous intrahippocampal microperfusion of 500 microM calpain inhibitor I had no effect on basal EEG, but doubled (P < 0.05) average seizure duration, and increased more than five-fold (P < 0.01) the total seizure time and three-fold (P < 0.01) the seizure offset time compared to picrotoxin alone, in each individual rat. However, seizure type and onset time were not modified by calpain inhibitor I. These results indicate that a calpain-mediated mechanism may be responsible for seizure offset, probably through AMPA glutamate receptors internalization and further degradation.     

Eye-movement recording in freely moving animals

A new method is described for precise recording of eye movements in freely moving animals using Hall-effect devices. This inexpensive system, of small size and low weight, allows the analysis of horizontal and vertical components of saccadic eye movements, optokinetic nystagmus, slow tracking movements, eye vergence, etc., in unrestrained animals. A set of Hall-effect devices mounted in the skull is used to sense variations in the position of high-power miniature magnets fixed to the eye sclera. The output of the Hall-effect devices is amplified by operational amplifiers and collected through an analog-to-digital converter to be displayed on-line in a personal computer and stored for later analysis by specific software. Some examples of simultaneous body- and eye-movement recordings obtained in freely moving goldfish in different experimental situations are presented. This method would be useful in the recording of eye and gaze movements under natural conditions and for behavioural studies in freely moving animals.     

Acetyl-L-carnitine enhances acetylcholine release in the striatum and hippocampus of awake freely moving rats

The effect of acetyl-L-carnitine (ALC) on the spontaneous release of acetylcholine (ACh) in the striatum and hippocampus of freely moving rats was investigated using brain microdialysis coupled with HPLC-electrochemical detection. Systemic administration of ALC, in a dose-dependent manner, stimulated ACh release in both areas, while the D-enantiomer was substantially ineffective. The effect of ALC was strongly Ca2+ dependent and tetrodotoxin (TTX) sensitive. These features of an exocytotic and impulse flow-dependent mechanism suggest that the increase in ACh release is the result of ALC activation of a physiological mechanism in cholinergic neurons.     

Circadian rhythm of histamine release from the hypothalamus of freely moving rats.

Using an in vivo microdialysis technique coupled with HPLC-fluorometry, the release of neuronal histamine from the anterior hypothalamic area was monitored continuously in conscious, freely moving rats under a 12:12 h light:dark cycle. Spontaneous locomotor activity of the rats was measured simultaneously using a locomotor activity counter. Histamine release gradually increased in the second half of the light period (1400-2000) and the average histamine release during the dark period (2000-0800, 0.20 +/- 0.02 pmol/30 min) was significantly higher than that during the light period (0.12 +/- 0.01 pmol/30 min). This clear circadian change in the release suggests that the central histaminergic system is related to the circadian rhythm of rats.     

Thioperamide and cimetidine modulate acetylcholine release from the amygdala of freely moving rats

Inflammation Research -     

Involvement of corticosterone in cardiovascular responses to an open-field novelty stressor in freely moving rats

The aim of the present study was to investigate the modulatory action of different concentrations of circulating corticosterone occupying either predominantly mineralocorticoid receptors (MR) or both MR and glucocorticoid receptors (GR) in control of cardiovascular responses to a novelty stressor. Six groups of rats were instrumented with radiotelemetry transmitters: sham-operated controls, adrenalectomised (ADX) controls, ADX with chronic implantation of a 20-mg corticosterone pellet, ADX with chronic implantation of a 100-mg corticosterone pellet, ADX receiving acute bolus injection of 0.25 mg/kg of corticosterone, and ADX with both implantation of a 20-mg corticosterone pellet and bolus treatment. Exposure to the novelty of an open field caused an increase in blood pressure, heart rate, body temperature and exploratory locomotor activity. The pressor response was dose-dependently increased in ADX rats implanted with a corticosterone pellet. Bolus injection of corticosterone at 10 min prior to novelty had no effect. The tachycardia was reduced in ADX rats compared to sham-operated controls, and this effect was restored by implantation of a 20-mg, but not 100-mg, corticosterone pellet. Bolus injection of corticosterone facilitated the return of heart rate towards baseline levels. The increase in body temperature was reduced in ADX rats, a deficit that was normalised by implantation of either corticosterone dose but not by acute bolus treatment. Locomotor activity was not different between the groups except for a slightly more rapid decline of locomotor activity in both groups treated with a bolus injection of corticosterone. These data show an important role of putative brain MR in maintaining adequate cardiovascular and behavioural responsiveness to a mild psychological stressor, while additional acute or chronic occupation of GR has further differential and sometimes opposing effects.     

Somatostatin inhibits the centrally mediated hypertensive response to clonidine in freely moving rats.

The effect of somatostatin on the hypertensive response induced by intracerebroventricular (i.c.v.) injection of clonidine was investigated in freely moving rats. A 10 micrograms i.c.v. injection of clonidine produced a marked pressor response and a decrease in heart rate. No depressor response was induced by clonidine. The i.c.v. pretreatment with somatostatin (2 and 5 micrograms) dose-dependently inhibited the pressor response to i.c.v. injected clonidine (10 micrograms), and a long-lasting depressor response was observed. Systemic (i.v.) treatment with somatostatin had no such effect. These results suggest that brain somatostatin modulates the centrally-mediated pressor response to clonidine.     

A novel carbon fiber implantation assembly for cerebral voltammetric measurements in freely moving rats

In vivo voltammetry has been widely employed to monitor the effects of drugs on dopamine metabolism. In the present study, we report on the use of a newly designed assembly for implantation of carbon fiber microelectrodes for voltammetric measurements in freely moving animals. The assembly consists of an electrode holder and a fixed support (implanted stereotaxically) which is cemented to the rat skull. The working carbon fiber electrode is cemented to the electrode holder, and can be lowered into the fixed support for implantation into the brain parenchyma. This implantation assembly has been used to study the effects of tail-pinch on dopamine metabolism (extracellular DOPAC levels) in the nucleus accumbens and ventral tegmental area of the awake rat. Using this implantation assembly and 3 carbon fiber electrodes, oxidation peaks for ascorbic acid (-100 mV) and DOPAC (+ 100 mV) were recorded in both brain structures. Mild tail pressure for 7 min led to an increase in extracellular DOPAC levels in the nucleus accumbens. A smaller increase was observed in the ventral tegmental area. The DOPAC signals remained elevated for some time after removal of the stressor. This implantation assembly represents a convenient system for implanting carbon fiber electrodes in the freely moving rat, which can be employed to investigate the effects of behavioral manipulation on dopaminergic neurons at the terminal as well as at the cell body level. This microdevice could also be usefully employed for other in vivo electrochemical studies.     

Regulatory role of nitric oxide over extracellular taurine in the hippocampus of freely moving rats

We have studied the effects of drugs which manipulate nitric oxide (NO) levels as well the effect of N-methyl-d-aspartate (NMDA) infusion on extracellular taurine in rat hippocampus using in vivo microdialysis. The NO donor S-nitroso-N-acetylpenicillamine (SNAP) increased dialysate taurine in a concentration-dependent manner, and this effect was blocked by the inhibitor of soluble guanylate cyclase1H-[1,2,4]oxadiazolo[4,3-alpha]quinoxalin-1-one (ODQ). NMDA (100 microM) increased hippocampal taurine release, an effect that was reversed by the NMDA receptor antagonist 2-amino-5-phosphonopentanoic acid (AP5; 10 microM). The non-selective nitric oxide synthase (NOS) inhibitor N-nitro-l-arginine methyl ester (L-NAME; 100 microM and 1.0 mM) increased extracellular taurine in a concentration-dependent manner while 7-nitroindazole (7-NI), a relatively selective neuronal NOS (nNOS) inhibitor, at the same concentrations decreased extracellular taurine. L-NAME (1.0 mM) infused prior to NMDA did not alter the effect of NMDA on extracellular taurine having an effect essentially identical to that seen with L-NAME infused alone. In contrast, when 7-NI was infused for 30 min prior to NMDA, taurine levels were no longer increased above basal. This suggests to us that taurine efflux is mediated by two different mechanisms: an NMDA-evoked, 7-NI-sensitive pathway which may be dependent on cyclic guanosine monophosphate formation, and an L-NAME-modulated mechanism which presumably involves other members of the NOS group of enzymes than nNOS alone.     

Possible contribution of central gamma-aminobutyric acid receptors to resting vascular tone in freely moving rats

Previous studies have shown that central administration of GABA (gamma -aminobutyric acid), an inhibitory neurotransmitter, preferentially reduces hindquarters and carotid vascular resistances but not renal and coeliac vascular resistances in conscious rats. This study tested the hypothesis that these preferential actions of central GABA receptors are related to differences between vessels in resting autonomic vascular tone in freely moving rats. Rats were chronically implanted with intracisternal cannulas and/or electromagnetic probes to measure regional blood flows. In response to GABA administration, the changes in vascular resistance (arterial blood pressure/regional blood flow) of the hindquarters (n = 23) and carotid (n = 12) vascular beds were significantly and negatively correlated with basal vascular resistance. No such relationship was found for the renal (n = 21), coeliac (n = 13) and superior mesenteric (n = 23) vascular beds. This finding indicates that the responsiveness to GABA of brainstem pathways controlling the hindquarters and carotid vascular beds co-varies with resting resistance in hindquarters and carotid vessels. A similar analysis was performed, correlating the ongoing vascular resistance of each vessel with its response to ganglionic blockade by chlorisondamine. In this case, a significant negative correlation was also found for the hindquarters (n = 26) and carotid (n = 15) vascular beds, but not for the coeliac (n = 17) or superior mesenteric (n = 19) vessels. Together, these findings suggest that central GABA receptors accessible from the cisterna magna preferentially affect two vascular beds which, in the freely moving rat, show resting autonomic vascular tone.     

Single unit and population responses during inhibitory gating of striatal activity in freely moving rats

The striatum is thought to be an essential region for integrating diverse information in the brain. Rapid inhibitory gating (IG) of sensory input is most likely an early factor necessary for appropriate integration to be completed. Gating is currently evaluated in clinical settings and is dramatically altered in a variety of psychiatric illnesses. Basic neuroscience research using animals has revealed specific neural sites involved in IG including the hippocampus, thalamus, brainstem, amygdala and medial prefrontal cortex. The present study investigated local IG in the basal ganglia structure of the striatum using chronic recording microwires. We obtained both single unit activations and local field potentials (LFPs) in awake behaving rats from each wire during the standard two-tone paradigm. Single units responded with different types of activations including a phasic and sustained excitation, an inhibitory response and a combination response that contained both excitatory and inhibitory components. IG was observed in all the response types; however, non-gating was observed in a large proportion of responses as well. Positive wave field potentials at 50-60 ms post-stimulus (P60) showed consistent gating across the wire arrays. No significant correlations were found between single unit and LFP measures of gating during the initial baseline session. Gating was strengthened (Tamp/Camp ratios approaching 0) following acute stress (saline injection) at both the single unit and LFP level due to the reduction in the response to the second tone. Alterations in sensory responding reflected by changes in the neural response to the initial tone were primarily observed following long-term internal state deviation (food deprivation) and during general locomotion. Overall, our results support local IG by single neurons in striatum but also suggest that rapid inhibition is not the dominant activation profile observed in other brain regions.     

5-HT3 receptors control dopamine release in the nucleus accumbens of freely moving rats

ICS 205-930, a selective and potent 5-HT3 receptor antagonist applied either systemically, or locally into the ventral tegmental area, antagonized the stimulation of dopamine release in the nucleus accumbens, induced by the subcutaneous administration of morphine. These findings, obtained by the use of brain microdialysis in awake freely-moving rats, demonstrate in vivo a functional role of 5-HT3 receptors in the brain. Since stimulation of dopamine release in the nucleus accumbens is a prerequisite for the expression of the rewarding properties of morphine, its suppression by ICS 205-930 suggests a possible application of 5-HT3 receptor antagonists in the treatment of addiction.