Different spatial expressions of c-Fos in the nucleus of the solitary tract following taste stimulation with sodium, potassium, and ammonium ions in rats.

Cation-specific epithelial receptors on the tongue have been well demonstrated. However, active regions along the nucleus of the solitary tract (NST) for cations Na(+), K(+), NH4(+) are still unclear, even though the best responses of NST neurons to taste stimuli vary depending on the cell. In the present study, the spatial distribution patterns of cation-specific active regions in the NST are investigated. The tongues of urethane-anesthetized Sprague-Dawley rats (n = 25) were stimulated with artificial saliva (control), 0.5 M NaCl, 1.0 M NaCl, 0.5 M KCl, and 0.3 M NH(4) Cl. Then, the three-dimensional positions of c-Fos-like-immunoreactive (cFLI) cells in the NST were generated. The spatial distributions of cFLI cells in the NST were compared among five taste stimulations. cFLI cells were observed throughout the NST, irrespective of the stimulus; however, the intermediate-medial central regions of the NST had higher numbers of cFLI cells than the other regions in all taste stimulations. Analysis of images revealed that the activated regions in the NST differed significantly depending on the cations. The intermediate-dorsal-central region and the caudal-ventral region were activated by a 0.5 M concentration of sodium, the rostral-ventral region and the intermediate-dorsal/ventral region were activated by a 1.0 M concentration of sodium, the intermediate-dorsal/ventral region was activated by potassium ions, and the rostral-ventral region and the intermediate-ventral central region were activated by ammonium ions. These results suggest that the responses of NST cells to cation salt ions are regulated differentially.

Changes in cytokine expression after electroacupuncture in neuropathic rats.

The production of proinflammatory cytokines including interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) plays a key role in chronic pain such as neuropathic pain. We investigated changes in cytokine expression in injured peripheral nerves and dorsal root ganglia (DRG) following electroacupuncture (EA) treatment. Neuropathic pain was induced by peripheral nerve injury to the left hind limb of Sprague-Dawley rats under pentobarbital anesthesia. Two weeks later, the nerve-injured rats were treated by EA for 10 minutes. The expression levels of IL-1ß, IL-6, and TNF-α in peripheral nerves and DRG of neuropathic rats were significantly increased in nerve-injured rats. However, after EA, the cytokine expression levels were noticeably decreased in peripheral nerves and DRG. These results suggest that EA stimulation can reduce the levels of proinflamtory cytokines elevated after nerve injury.

c-Fos Expression in the Nucleus of the Solitary Tract in Response to Salt Stimulation in Rats.

Salt signals in tongue are relayed to the nucleus of the solitary tract (NST). This signaling is very important to determine whether to swallow salt-related nutrition or not and suggests some implications in discrimination of salt concentration. Salt concentration-dependent electrical responses in the chorda tympani and the NST were well reported. But salt concentration-dependency and spatial distribution of c-Fos in the NST were not well established. In the present study, NaCl signaling in the NST was studied in urethane-anesthetized rats. The c-Fos immunoreactivity in the six different NST areas along the rostral-caudal axis and six subregions in each of bilateral NST were compared between applications of distilled water and different concentrations of NaCl to the tongue of experimental animals. From this study, salt stimulation with high concentration (1.0 M NaCl) induced significantly higher c-Fos expression in intermediate NST and dorsal-medial and dorsal-middle subregions of the NST compared to distilled water stimulation. The result represents the specific spatial distribution of salt taste perception in the NST.

Interferon-gamma alters electrical activity and clock gene expression in suprachiasmatic nucleus neurons.

The proinflammatory cytokine interferon (IFN-gamma) is an immunomodulatory molecule released by immune cells. It was originally described as an antiviral agent but can also affect functions in the nervous system including circadian activity of the principal mammalian circadian pacemaker, the suprachiasmatic nucleus. IFN-gamma and the synergistically acting cytokine tumor necrosis factor-alpha acutely decrease spontaneous excitatory postsynaptic activity and alter spiking activity in tissue preparations of the SCN. Because IFN-gamma can be released chronically during infections, the authors studied the long-term effects of IFN-gamma on SCN neurons by treating dispersed rat SCN cultures with IFN-gamma over a 4-week period. They analyzed the effect of the treatment on the spontaneous spiking pattern and rhythmic expression of the "clock gene," Period 1. They found that cytokine-treated cells exhibited a lower average spiking frequency and displayed a more irregular firing pattern when compared with controls. Furthermore, long-term treatment with IFN-gamma in cultures obtained from a transgenic Per1-luciferase rat significantly reduced the Per1-luc rhythm amplitude in individual SCN neurons. These results show that IFN-gamma can alter the electrical properties and circadian clock gene expression in SCN neurons. The authors hypothesize that IFN-gamma can modulate circadian output, which may be associated with sleep and rhythm disturbances observed in certain infections and in aging.

Fractal stochastic modeling of spiking activity in suprachiasmatic nucleus neurons.

Individual neurons in the suprachiasmatic nucleus (SCN), the master biological clock in mammals, autonomously produce highly complex patterns of spikes. We have shown that most (approximately 90%) SCN neurons exhibit truly stochastic interspike interval (ISI) patterns. The aim of this study was to understand the stochastic nature of the firing patterns in SCN neurons by analyzing the ISI sequences of 150 SCN neurons in hypothalamic slices. Fractal analysis, using the periodogram, Fano factor, and Allan factor, revealed the presence of a 1/f-type power-law (fractal) behavior in the ISI sequences. This fractal nature was persistent after the application of the GABAA receptor antagonist bicuculline, suggesting that the fractal stochastic activity is an intrinsic property of individual SCN neurons. Based on these physiological findings, we developed a computational model for the stochastic SCN neurons to find that their stochastic spiking activity was best described by a gamma point process whose mean firing rate was modulated by a fractal binomial noise. Taken together, we suggest that SCN neurons generate temporal spiking patterns using the fractal stochastic point process.

Dynamical heterogeneity of suprachiasmatic nucleus neurons based on regularity and determinism.

The suprachiasmatic nucleus (SCN) is known to be the master biological clock in mammals. Despite the periodic mean firing rate, interspike interval (ISI) patterns of SCN neurons are quite complex and irregular. The aim of the present study was to investigate the existence of nonlinear determinism in the complex ISI patterns of SCN neurons. ISI sequences were recorded from 173 neurons in rat hypothalamic slice preparations using a cell-attached patch recording technique. Their correlation dimensions (D2) were estimated, and were then compared with those of the randomly-shuffled surrogate data. We found that only 16 neurons (16/173) exhibited deterministic ISI patterns of spikes. In addition, clustering analysis revealed that SCN neurons could be divided into two subgroups of neurons each having distinct values of coefficient of variation (CV) and skewness (SK). Interestingly, most deterministic SCN neurons (14/16) belonged to the group of irregularly spiking neurons having large CV and SK values. To see if the neuronal coupling mediated by the gamma-aminobutyric acid (GABA), the major neurotransmitter in the SCN, contributed to the deterministic nature, we examined the effect of the GABAA receptor antagonist bicuculline on D2 values of 56 SCN neurons. 8 SCN neurons which were originally stochastic became to exhibit deterministic characteristics after the bicuculline application. This result suggests that the deterministic nature of the SCN neurons arises not from GABAergic synaptic interactions, but likely from properties inherent to neurons themselves.

A calcium flux is required for circadian rhythm generation in mammalian pacemaker neurons.

Generation of mammalian circadian rhythms involves molecular transcriptional and translational feedback loops. It is not clear how membrane events interact with the intracellular molecular clock or whether membrane activities are involved in the actual generation of the circadian rhythm. We examined the role of membrane potential and calcium (Ca2+) influx in the expression of the circadian rhythm of the clock gene Period 1 (Per1) within the rat suprachiasmatic nucleus (SCN), the master pacemaker controlling circadian rhythmicity. Membrane hyperpolarization, caused by lowering the extracellular concentration of potassium or blocking Ca2+ influx in SCN cultures by lowering [Ca2+], reversibly abolished the rhythmic expression of Per1. In addition, the amplitude of Per1 expression was markedly decreased by voltage-gated Ca2+ channel antagonists. A similar result was observed for mouse Per1 and PER2. Together, these results strongly suggest that a transmembrane Ca2+ flux is necessary for sustained molecular rhythmicity in the SCN. We propose that periodic Ca2+ influx, resulting from circadian variations in membrane potential, is a critical process for circadian pacemaker function.