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Synchronous neuronal activity in the hippocampus (theta rhythm) can be elicited in urethanized rats with sensory stimulation as well as with electrical or pharmacological stimulation of different nuclei of the brainstem. It is known that two of these nuclei, the nucleus pontis oralis (RPO) and the pedunculopontine nucleus (PPN), play an important role in theta regulation, however, it is still unclear which of them is essential for expression of theta in the hippocampus. In the present study we investigated the effect of temporal inactivation of PPN on the hippocampal theta rhythm induced by electrical stimulation applied to RPO. The experiments were performed on 5 male Wistar rats in deep urethane anesthesia with its level monitored on the basis of breathing rate. Animals were implanted with bilateral recording electrodes into the dorsal hippocampus and stimulation electrode into the RPO. Hippocampal EEG was recorded during repeated electrical stimulation of RPO in control conditions and also following intra-PPN administration of procaine. In all animals electrical stimulation of the RPO (200 - 300 mA, 30 s) induced episodes of robust hippocampal theta rhythm in both hippocampi which lasted for the whole period of the electrical stimulation (30 s) with no latency. After temporal inactivation of the PPN by direct procaine microinjection (20% solution/0.5 μl), electrical stimulations of the RPO were not able to induce synchronous activity in the hippocampus. Neuronal activity within the RPO and PPN nuclei changes during sleep/wake cycle including paradoxical sleep, of which hippocampal theta rhythm is an important indicator. Regular theta rhythm in the hippocampus is also present during urethane anesthesia which was applied in our experiments. Our results indicate that undisturbed neuronal activity within the PPN is crucial for evoking hippocampal theta rhythm with electrical stimulation of RPO, which suggests superior role of the PPN.
The nucleus reticularis pontis oralis (RPO) is a reticular structure important for the regulation of paradoxical sleep (PS). However, the data concerning the relation between the RPO and the main tonic indicator of PS, hippocampal theta rhythm, are contradictory: although electrical or cholinergic stimulation of the RPO evoked well-synchronized theta activity, the electrolytic lesion of the structure had no effect on theta. In our experiment, the effect of procaine injections into different parts of the RPO on the electrical activity of the hippocampus, as well as on tail pinch-elicited hippocampal theta rhythm was assessed in urethanized rats. Power spectral analysis was performed using a Fast Fourier Transform routine in 1-Hz and 3-Hz bands between 0.6 and 12 Hz frequency. We have found that unilateral procaine inactivation of neurons in the caudal part of the RPO blocked the sensory-elicited theta rhythm. The same injection into the rostral RPO either had no effect or evoked long-lasting episodes of theta rhythm without sensory stimulation. These results suggest functional diversity of the parts of the RPO in mechanisms underlying production of hippocampal theta.
Our previous study indicated that microinjection of procaine or electrolytic lesion of the ventral tegmental area (VTA) suppressed hippocampal theta rhythm in urethane-anaesthetized rats. The aim of this study was to verify the hypothesis that electrical stimulation of the VTA induces hippocampal theta rhythm and to fi nd brain structures particularly active during this phenomenon and probably involved in its mechanism. The study was performed on urethane anaestethized male Wistar rats with an electrode implanted unilaterally in the VTA or zona incerta (ZI – control group). Stimulation was applied as 0.1-ms rectangular impulses of 50 Hz frequency and duration of 30 s at 10-min intervals. VTA stimulation within the current intensity range of 100–240 mA evoked hippocampal theta rhythm, manifested as synchronization of the EEG signal and an increase in the power at 3–6 Hz band. ZI stimulation did not elicit such effects. After VTA stimulation we also found induction of c-fos expression in brain regions connected to the VTA: nucleus accumbens, lateral septum, or engaged in the regulation of hippocampal theta rhythm: medial septum, midline thalamic nuclei, hypothalamic nuclei, pedunculopontine, laterodorsal and cuneiform tegmental nuclei. The results indicate that the VTA may be a part of the brainstem theta synchronizing system and may infl uence the hippocampal EEG through indirect pathway via hypothalamus and the medial septum, simultaneously increasing thalamic activity.
Ventral tegmental area (VTA) is thought to be an important component in the mesocorticolimbic system involved in the regulation of theta rhythm in the hippocampus. In this study we investigated the effect of pharmacological inactivation of the VTA on theta rhythm parameters in fear conditioned rats. Animals were implanted with bilateral recording electrodes into the dorsal hippocampus and bilateral injection cannula into the VTA. Hippocampal LFP was recorded throughout the experiment, in pre- and post-injection condition during freezing behavior and active locomotion (avoidance). We found that intra-VTA injection of procaine temporary suppressed avoidance response and decreased the power of hippocampal theta rhythm during freezing in comparison to control (water). Our results indicate that inactivation of neuronal activity in the VTA reduces hippocampal theta rhythm as well as avoidance response in fear conditioned animal. Supported by MNiSW Grant IP2011 034371.
INTRODUCTION: Two notable targets of the pedunculopontine tegmental nucleus (PPN) circuitry, the substantia nigra (SN) and the ventral tegmental area (VTA), are implicated in locomotion and reward processing. A dysfunction of these regions occurs in Parkinson’s and related disorders as well as in various psychiatric conditions, and over the course of normal aging AIM(S): In the present study, we were interested in understanding NMDA-receptors involvement in the interactions between the PPN and SN/VTA midbrain complex. In order to obtain more insight into this process, we analyzed the number and the distribution of midbrain tyrosine hydroxylase positive cells (TH+). METHOD(S): All rats were implanted with bilateral stimulating electrodes in the VTA and with bilateral guide cannulas for intracerebral injections into the PPN. Immunohistochemistry for TH+ was used to measure the number of active dopaminergic neurons in midbrain (VTA-SN) of rats subjected to unilateral VTA electrical stimulation and local injection of MK‑801 (5 μg) or NMDA (3 μg) to the contralateral or ipsilateral hemispheres into the PPN (4 experimental groups). The control brains were from rats in which only the 14‑day unilateral electrical VTA‑stimulation was performed (control group). RESULTS: Immunohistochemical analysis revealed a decrease in the number of TH+ cells in the midbrain. When the main subdivisions of the VTA/SN were subjected to a separate analysis, a significantly lower number of TH+ cells were found in all experimental groups in the PBP (parabrachial pigmented nucleus), PB (paranigral nucleus) and SNc (SN, pars compacta), as compared to the control group. CONCLUSIONS: The level of NMDA receptor arousal in the PPN regulates the activity of the midbrain dopaminergic cells. FINANCIAL SUPPORT: The research was funded by the Polish National Science Center; decision no: DEC‑2013/09/N/NZ4/02195 and by the Faculty of Biology, University of Gdansk, Poland.
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