Interactions between environmental aversiveness and the anxiolytic effects of enhanced cannabinoid signaling by FAAH inhibition in rats.

RATIONALE: Since the discovery of endogenous cannabinoid signaling, the number of studies exploring its role in health and disease has increased exponentially. Fatty acid amide hydrolase (FAAH), the enzyme responsible for degradation of the endocannabinoid anandamide, has emerged as a promising target for anxiety-related disorders. FAAH inhibitors (e.g., URB597) increase brain levels of anandamide and induce anxiolytic-like effects in rodents. Recent findings, however, questioned the efficacy of URB597 as an anxiolytic. OBJECTIVES: We tested here the hypothesis that conflicting findings are due to variations in the stressfulness of experimental conditions employed in various studies. RESULTS: We found that URB597 (0.1-0.3 mg/kg) did not produce anxiolytic effects when the aversiveness of testing procedures was minimized by handling rats daily before experimentation, by habituating them to the experimental room, or by employing low illumination during testing. In contrast, URB597 had robust anxiolytic effects when the aversiveness of the testing environment was increased by eliminating habituation to the experimental room or by employing bright lighting conditions. Unlike URB597, the benzodiazepine chlordiazepoxide (5 mg/kg) had anxiolytic effects under all testing conditions. The anxiolytic effects of URB597 were abolished by the cannabinoid CB1-receptor antagonist AM251, showing that they were mediated by CB1 receptors. Close inspection of experimental conditions employed in earlier reports suggests that conflicting findings with URB597 can be explained by different testing conditions, such as those manipulated in the present study. CONCLUSIONS: Our findings show that FAAH inhibition does not affect anxiety under mildly stressful circumstances but protects against the anxiogenic effects of aversive stimuli.

Medium-frequency oscillations dominate the inspiratory nerve discharge of anesthetized newborn rats.

In this study we examined the synchronization of the discharge of phrenic and recurrent laryngeal motoneurons in anesthetized rat pups 14 to 36 days of age and kittens, 14-15 days old. We found that the inspiratory nerve activity consisted of synchronized bursts separated by 20-35 ms, corresponding to medium-frequency oscillations (MFO). Accordingly, the autospectra of the neurograms had two peaks, one at the respiratory rate and the other between 22. 8-43.0 Hz. No significant coherence was found between MFOs in the discharges of different nerves. High-frequency oscillations (HFO) characteristic for the adult inspiratory nerve activity were not present in the newborn rats. These findings demonstrate that phrenic nerve discharge of rat pups, like that of kittens and piglets, is in the MFO range, and suggest that MFO activity is an index of an early developmental stage of the respiratory system.

Spinal segments communicating resting sympathetic activity to postganglionic nerves of the stellate ganglion.

It has been shown earlier using sympathetic reflexes and anatomic techniques that preganglionic neurons controlling different effectors occupy wide and overlapping ranges of adjacent segments in the spinal cord (cardiac: T1-T7, vertebral: T2-T8). Because, however, the majority of preganglionic neurons are silent at resting states, the present study was designed to estimate the segmental map of subsets of these neurons including only those active at rest using simultaneous recordings from the inferior cardiac and vertebral nerves, under chloralose-urethan or urethan anesthesia. In 22 cats, thoracic white rami T1-T8 were cut in a sequential manner. Three-minute-long data segments were recorded between sectionings and analyzed in the frequency domain using the fast Fourier transform. We found that cardiac and vertebral active maps involved segments T3-T5 and T4-T8, respectively. In individual experiments, however, most of the power of rhythmic activity originated from only one or two segments and the dominant segments for the two nerves never overlapped. Moreover, the separation between dominant segments generating cardiac and vertebral nerve discharges was wider and the distribution of tonically active preganglionic neurons projecting to each nerve was narrower under urethan than chloralose-urethan anesthesia. We conclude that the proportion of active to quiescent preganglionic neurons regulating cardiac and vertebral nerve discharges varies from spinal segment to segment and that active neurons projecting to these nerves are nonoverlapping.

Spinal cord segments mediating tonic sympathetic nerve discharge to the kidney in the anesthetized cat.

According to anatomical data, preganglionic neurons projecting to the kidney via sympathetic ganglia occupy a wide range of adjacent segments in the thoracolumbar spinal cord, from Th7 to L2. Since, however, the majority of preganglionic neurons is silent at resting states, the active segments indeed transmitting sympathetic activity, at rest, may be different. In the present experiments, the spontaneous sympathetic activity was recorded before and after the sympathetic trunk and white rami (WR) Th8-L3 were cut in a sequential manner. The step-by-step changes in the power of renal nerve discharge were estimated and used for mapping tonic renal outflow to the spinal cord. We found that powerful activity comprising 70-95% of the power of control recordings remained after eliminating the input from Th1-Th12, indicating that thoracic spinal cord including segments that contain the largest number of cells projecting to renal postganglionic neurons contributes relatively weakly to tonic renal nerve activity. It appeared that resting sympathetic nerve discharge was predominantly maintained by the caudalmost division of the renal preganglionic neuron population since the largest decrease in nerve power occurred after severing WR Th13, L1, and L2. These findings suggest that the 'active segmental map' of preganglionic neurons controlling a certain organ at rest does not necessarily match the distribution of the total population of neurons projecting to the same effector.

Power spectral analysis of inspiratory nerve activity in the anesthetized rat: uncorrelated fast oscillations in different inspiratory nerves.

The spectral composition of the inspiratory nerve discharge was studied in spontaneously breathing Sprague-Dawley rats under urethane (n = 7) or barbiturate (n = 10). Left phrenic nerve activity was recorded simultaneously with right phrenic or left recurrent laryngeal nerves. We found that all neurograms showed prominent fast oscillators at common frequencies in the high frequency (HFO) range. Concurrent medium frequency oscillations (MFO) were present in inspiratory nerve discharges of four rats anesthetized with Nembutal. Significant coherences between nerves were uncommon (n = 2) and were only found between HFOs. Thus although nerve autospectra were dominated by HFO, weak correlations indicated a relatively weak system HFO in th central pattern generator, in the anesthetized rat.

Differential sympathetic reactions during cerebral ischaemia in cats: the role of desynchronized nerve discharge.

1. Sympathetic nerve discharge (SND) of three postganglionic nerves with different functions and anatomical locations was simultaneously recorded at rest and during severe cerebral ischaemia (Cushing reaction). The three nerves, controlling the heart (inferior cardiac nerve), visceral (renal nerve) and skeletal muscle circulation (vertebral nerve), were selected with the assumption that their activity pattern will represent the differential central autonomic command to the major players of the circulatory response to cerebral ischaemia. 2. Changes in the power density spectra of the nerve signals, and in the pairwise coherence functions, elicited by the cerebral ischaemia, were evaluated separately for the rhythmic (R-SND, i.e. between 0 and 6 Hz) and high-frequency (HF-SND, i.e. between 12 and 100 Hz) components of the nerve signals. 3. The sympathetic nerve response to cerebral ischaemia developed in two phases. Phase 1 was a massive R-SND reaction and phase 2 was characterized by SND desynchronization and by the emergence of HF-SND. The power of HF-SND occupied a wide band between 12 and 80 Hz with maximum between 20 and 30 Hz. All three nerves were involved in the Cushing response but the magnitude and character of the reactions were specific for each nerve. In the cardiac nerve, the power of the rhythmic component of the discharge increased almost twice the control and remained dominant during the whole reaction, strongly modulating HF-SND during the second phase. In the vasomotor nerves, R-SND was suppressed during phase 2 and HF-SND occupied 65% of the total power of the signal. Near equal R- to HF-SND proportions, however, were reached on different activity levels in renal and vertebral nerves. Whereas total renal SND did not change, the power of the vertebral SND increased more than twice. In addition, desynchronization in the vertebral SND was preceded by a massive R-SND reaction during phase 1, which was missing in the renal nerve. 4. For all possible nerve pairs, R-SND was highly coherent before the reaction and remained so during intracranial pressure elevation, regardless of the direction and magnitude of the changes in absolute and/or relative power of this component in different nerves. On the other hand, HF-SND never correlated between any of the nerve pairs indicating that this component in each nerve originated from specific sources of regional sympathetic activity.(ABSTRACT TRUNCATED AT 400 WORDS)