G. Sandner

Induction of Fos immunoreactivity in the brain by exposure to the elevated plus-maze

The Fos protein immunohistochemistry technique was used to map the brain areas activated by a 15-min exposure of rats to the elevated plus maze, an ethologically based animal model of anxiety. Two hours after the test, labeling was found mainly in the piriform and entorhinal cortices, amygdala, midline thalamic nuclei, several medial hypothalamic nuclei, periaqueductal gray matter, superior and inferior colliculus, cuneiform nucleus, dorsal raphe nucleus and locus coeruleus. These results support a participation of these structures in anxiety.

Effect of propofol on affective state as assessed by place conditioning paradigm in rats.

Background Whether propofol produces a pleasant affective state remains unclear from clinical studies. In the current study, the effect on affective state of subanesthetic and anesthetic doses of propofol was assessed at a preclinical level with rats in a place conditioning paradigm. Propofol was compared with methohexital. Methods In the place conditioning paradigm, propofol-induced effect was repeatedly paired with one of two distinguishable compartments of the apparatus, whereas the vehicle-induced effect was repeatedly paired with the other compartment. During a subsequent free-choice test, a preference for the drug-paired compartment over the vehicle-paired compartment would be indicative of pleasant state induced by the drug. For all experiments, the conditioning session lasted 8 days and consisted of four pairings of the drug with one compartment and four pairings of the equivalent volume of vehicle with the other compartment. In experiment 1A, four groups of rats were designated according to the dose of propofol that they received intraperitoneally: 0, 30, 60, or 90 mg/kg. In experiment 1B, the same procedure was used with subanesthetic doses of intraperitoneal methohexital: 0, 10, 20, or 30 mg/kg. In experiment 2, the rats were conditioned during the recovery period from short-term anesthesia. For one group, anesthesia was induced by propofol (100 mg/kg) whereas for the other group, anesthesia was induced by an equivalent anesthetic dose of methohexital (40 mg/kg). Results In experiment 1A, the 30-mg/kg, 60-mg/kg, and 90-mg/kg groups showed a place preference for the drug-paired compartment, but only the group conditioned with 60 mg/kg propofol significantly differed from the 0-mg/kg group. In experiment 1B, the groups conditioned with methohexital showed no place preference for the drug-paired compartment. In experiment 2, the rats showed a place preference for the compartment in which they recovered from propofol-induced anesthesia but no place preference for the compartment in which they recovered from methohexital-induced anesthesia. Conclusions Propofol, but not methohexital, induced a pleasant affective state in rats at subanesthetic doses as well as during recovery from an anesthetic dose.

C-fos immunoreactivity in the brain following unilateral electrical stimulation of the dorsal periaqueductal gray in freely moving rats

C-fos immunoreactivity was used to reveal brain areas in which neurons were influenced by electrical stimulations applied to the dorsal periaqueductal gray. These stimulations were applied in freely moving rats so that the resulting behaviors could be observed. Shortly afterwards, the brains of the rats were processed for C-fos immunoreactivity. In order to determine the specificity of the brain areas thus labeled, control stimulations were applied to the ventral tegmental area of other rats. Immunoreactive cells were found surrounding the tip of the stimulation electrode within a radius of 0.5 mm. This labeled area extended further along the rostro-caudal axis than along the medio-lateral or dorso-ventral axis in the periaqueductal gray. Distally, clusters of labeled cells were found ipsilaterally in the caudal periaqueductal gray extending to the nucleus cuneiformis, and bilaterally in the locus coeruleus and supramamillary decussation. More widespread labeling was found in most hypothalamic subareas and in the lateral habenula. The labeled brain areas following ventral tegmental area stimulations were totally distinct, and comprised the medial forebrain bundle, the nucleus accumbens, the vertical limb of the diagonal band and the medial septum. The pattern of labeling produced by periaqueductal gray stimulations was therefore specific, and provided information about brain structures involved in the motivational and behavioral effects of such stimulations.

Conditioned place aversion produced by microinjections of semicarbazide into the periaqueductal gray of the rat

Previous studies have shown that the blockade of GABA-ergic neurotransmission in the periaqueductal gray (PAG) of the rat induce flight reactions. The present study examined whether a negative affective state was produced by such a blockade. Microinjections of semicarbazide, a GABA synthesis inhibitor, into the PAG were found to produce a conditioned place aversion. In a second experiment, it was found that the potent GABA agonist muscimol antagonized the effects of semicarbazide, without producing a conditioned place preference or aversion by itself. These results suggest that the blockade of the tonic inhibition exerted by GABAergic terminals in the PAG results in both an aversive experience and an overt flight reaction.

Escape and approach induced by brain stimulation:A parametric analysis

The study reported pursued two general aims: (a) to find out whether a brain-stimulation-induced escape response results from the bringing into play of similar mechanisms irrespective of whether the stimulation is applied to a site located in the mesencephalic central gray (CG), in the medial hypothalamus (MH) or in the lateral hypothalamus (LH); and (b) to verify whether escape and approach that can be induced by stimulating one and the same hypothalamic site result from a combined activation of two distinct neuronal systems, by means of specifying some of their functional characteristics that would possibly allow differentiation between them. Three experiments were carried out using a situation (shuttle-box) that allowed the measurement, for each given brain site, of both the time for which the rat underwent the stimulation before interrupting it (stimulation time, st., corresponding to an escape latency) and the time for which the animal remained non-stimulated before eventually restarting the stimulation himself (non-stimulation time, nst, corresponding to an approach latency). The following stimulation parameters were varied: the pulse duration (D), the stimulation intensity (I) and the interpulse interval (IPI = 1/stimulation frequency), i.e. the time interval that separated the beginning of a stimulating pulse from the beginning of the next one. The results obtained suggest the following. (1) Some of the mechanisms implicated in the elaboration of an escape response are similar irrespective of whether escape is induced by stimulating a site located in the CG, in the MH or in the LH.

Mapping of jumping, rearing, squealing and switch-off behaviors elicited by periaqueductal gray stimulation in the rat

Rats readily learn to escape from a stimulation applied to most mesencephalic periaqueductal gray (PAG) sites. In the present study, we tried to find out to what extent the differential effects induced by such stimulations actually reflect the existence of intraPAG functional subdivisions. To that end, a row of five electrodes was implanted into the PAG of each of 29 rats. Two kinds of effects were analyzed, the stimulation-elicited overt behaviors and the generalization of switch-off responding from one stimulation site to the others. Further, switch-off latency versus interpulse interval (IPI) relationships were established and both the threshold IPIs and the ceiling switch-off latencies were determined. The most commonly elicited behaviors (jumping, rearing and squealing) as well as the threshold IPIs and the ceiling switch-off latencies were mapped within the PAG. Switch-off behavior was elicited from all the stimulation sites studied. However, in the dorsal PAG the switch-off latency was found to decrease more steeply with decreasing IPI than it did in the ventral PAG. Switch-off generalization was less frequently observed between dorsally located stimulation sites. Jumps were most often elicited from dorsally and rostrally located PAG sites while squeals were more frequently elicited from the caudal part of the PAG and rearings from PAG subareas surrounding the aqueduct.

Central gray and medial hypothalamic stimulation: correlation between escape behavior and unit activity

A chronic experiment and an acute one were carried out in the same rat in order to get information on the neuronal events correlated with the escape latency (EL) induced by an electrical stimulation in the mesencephalic central gray (CG) or in the medial hypothalamus (MH). Escape latencies as well as unitary MH and CG neuronal responses (104 units in 25 rats) were studied as functions of the intensity and the pulse duration of a 50 pulses/sec stimulation train. The CG and/or the MH neuronal firing rate was either synchronized or diffusely altered, either inhibited or activated, by the CG or the MH stimulations. Many neuronal firing rate alterations were highly correlated with the escape speed (ES = 1/EL) induced by the same stimulation. Intensity duration trade-off functions were computed from both the EL and the unit recordings. Chronaxie determinations were performed from these data: the behaviorally determined chronaxies did not differ from the unitary ones (limit values: 0.05-0.42 msec). The discussion bear on the possible role of the observed neuronal alterations induced by CG or MH stimulations in relation to their behavioral effects.

Effect of 5,7-dihydroxytryptamine lesion on mianserin-induced conditioned place aversion and on 5-hydroxytryptamine1C receptors in the rat brain.

The lesion of serotonergic neurons (by an intraventricular injection of 5,7-dihydroxytryptamine) potentiated the conditioned place aversion induced by the 5-hydroxytryptamine1C/5-hydroxytryptamine2 antagonist mianserin in rats. This effect was selective for mianserin as the same lesion suppressed the conditioned place aversion induced by the benzodiazepine inverse agonist FG-7142. Previous results had shown the involvement of the 5-hydroxytryptamine1C receptors in the conditioned place aversion induced by mianserin [Rocha et al. (1993) Behav. Pharmac. 4, 101-106]. It was thus of interest to investigate the effect of the lesion on these receptor binding sites. Autoradiographic binding studies showed that the lesion significantly increased the concentration of the 5-hydroxytryptamine1C binding sites in various brain regions, including the amygdala, the hippocampus and the nucleus accumbens. Contrastingly, in these same brain regions, in situ hybridization histochemistry did not reveal an alteration of the level of messenger RNA coding for these receptors. On the one hand, correlating potentiation of the aversive effects of mianserin and increase of 5-hydroxytryptamine1C binding sites in the limbic system represent an interesting step in the comprehension of the molecular and motivational effects of serotonergic drugs. On the other hand, showing a dissociation between the expression of 5-hydroxytryptamine1C receptors and their corresponding messenger RNA, suggest that post-transcriptional mechanisms are involved in the regulation of these receptors.

Effects of dorsal raphe stimulation on escape induced by medial hypothalamic or central gray stimulation

Electrical stimulation of the ventral part of the mesencephalic central gray affects escape responding induced by stimulation of either the medial hypothalamus (MH) or dorsal central gray (CG) in the rat. The sign and magnitude of the effect appeared to depend on the following factors: (1) The location of the site stimulated in the ventral central gray and the intensity of the stimulation. A stimulation of the dorsal raphe nucleus (DR) which induced a marked rewarding and a low aversive effect reliably suppressed MH or CG induced escape. Stimulation of sites located at the boundary between DR and central gray enhanced escape at low intensities and suppressed it at higher intensities. When located more laterally in the ventral central gray, the stimulation induced less rewarding and more marked aversive effects and it reliably enhanced MH or CG induced escape. (2) The kind of stimulation applied (monopolar or bipolar). (3) The location of the site (MH or CG) from which the stimulation-induced escape responses were obtained.

Unit activity alterations induced in the mesencephalic periaqueductal gray by local electrical stimulation

In the framework of a series of investigations concerning the neural substrate of aversion, electrophysiological methods were used in order to specify, within the rats periaqueductal gray (PAG), functional properties, viz. conduction velocity and refractory period, of PAG neurons already assessed in previous studies by means of behavioral methods, and to gather data on their local synaptic relationships. Unit activities were recorded from the periaqueductal gray with the aim of analyzing those alterations that would be induced by locally applying an electrical stimulation with parameters shown to elicit escape behavior. An implanted row of electrodes allowed the application of a stimulation to several sites aligned along a mediolateral or a rostrocaudal axis through the periaqueductal gray. The results indicate that an electrical stimulation applied to the periaqueductal gray may induce its effects through the activation of a number of dendrites and many slow conducting fibers running in a great variety of directions and branching within the periaqueductal gray. Their refractory period was surprisingly low (0.6 ms) for slow conducting fibers (below 1 m/s). The local circuitry appears to include many inhibitory connections. Their organization is assumed to be partly recurrent. Stimulation-induced inhibition becomes predominant when the stimulation is moved away from the recorded neuron along the mediolateral axis, but not along the rostrocaudal axis.