Hyung Cheul Shin

A micromachined silicon depth probe for multichannel neural recording

A process of making a new type of silicon depth-probe microelectrode array is described using a combination of plasma and wet etch. The plasma etch, which is done using a low temperature oxide (LTO) mask, enables probe thickness to be controlled over a range from 5 to 90 /spl mu/. Bending tests show that the probes mechanical strength depends largely on shank thickness. More force can he applied to thicker shanks while thinner shanks are more flexible. One can then choose a thickness and corresponding mechanical strength using the process developed. The entire probe shaping process is performed only at low temperature, and thus is consistent with the standard CMOS fabrication. Using the probe in recording from rats somatosensory cortex, the authors obtained four channel simultaneous recordings which showed clear independence among channels with a signal-to-noise ratio performance comparable with that obtained using other devices.

A Novel Neuroprotective Mechanism of Riluzole: Direct Inhibition of Protein Kinase C

In addition to its antiexcitotoxic action, the anti-amyotrophic lateral sclerosis (ALS) neuroprotectant riluzole protects against nonexcitotoxic oxidative neuronal injury. In light of evidence that protein kinase C (PKC) mediates oxidative stress in cortical culture, we examined the possibility that riluzoles antioxidative neuroprotection involves PKC inhibition. Riluzole (30 microM) blocked phorbol 12-myristate 13-acetate (PMA)-induced increases in membrane PKC activity in cultured cortical cells. Suggesting a direct action, riluzole also inhibited the activity of purified PKC. Consistently, both PKC depletion and oxidative neuronal death induced by PMA were markedly attenuated by riluzole. The site of action of riluzole on PKC was not likely the diacylglycerol binding site but the catalytic domain, since riluzole did not alter radiolabeled phorbol-12,13-dibutyrate binding, but inhibited PKM, the catalytic domain of PKC. However, increasing ATP concentrations did not alter the inhibition of PKC by riluzole, making it unlikely that riluzole is a competitive inhibitor of ATP binding at PKM. Present results have demonstrated that riluzole directly inhibits PKC, which action may contribute to its antioxidative neuroprotective effects. In addition, it appears possible that PKC inhibition may be able to explain some of its well-known channel inhibitory and neuroprotective effects. Combined with findings that PKC activity is increased in ALS, the present results suggest that PKC may be a potential therapeutic target in ALS.

Voltage‐gated calcium channels play crucial roles in the glutamate‐induced phase shifts of the rat suprachiasmatic circadian clock

The resetting of the circadian clock based on photic cues delivered by the glutamatergic retinohypothalamic tract is an important process helping mammals to function adaptively to the daily light–dark cycle. To see if the photic resetting relies on voltage‐gated Ca2+ channels (VGCCs), we examined the effects of VGCC blockers on the glutamate‐induced phase shifts of circadian firing activity rhythms of suprachiasmatic nucleus (SCN) neurons in hypothalamic slices. First, we found that a cocktail of amiloride, nimodipine and ω‐conotoxin MVIIC (T‐, L‐ and NPQ‐type VGCC antagonists, respectively) completely blocked both phase delays and advances, which were, respectively, induced by glutamate application in early and late night. Next, we discovered that: (i) amiloride and another T‐type VGCC antagonist, mibefradil, completely obstructed the delays without affecting the advances; (ii) nimodipine completely blocked the advances while having less impact on delays; and (iii) ω‐conotoxin MVIIC blocked largely, if not entirely, both delays and advances. Subsequent whole‐cell recordings revealed that T‐type Ca2+ currents in neurons in the ventrolateral, not dorsomedial, region of the SCN were larger during early than late night, whereas L‐type Ca2+ currents did not differ from early to late night in both regions. These results indicate that VGCCs play important roles in glutamate‐induced phase shifts, T‐type being more important for phase delays and L‐type being so for phase advances. Moreover, the results point to the possibility that a nocturnal modulation of T‐type Ca2+ current in retinorecipient neurons is related to the differential involvement of T‐type VGCC in phase delays and advances.

Hemodynamic Responses of Rat Brain Measured by Near-infrared Spectroscopy During Various Whisker Stimulations

NIRS (Near-infrared spectroscopy) is a relatively, new, non-invasive, and non-ionizing method of measuring hemodynamic responses in thick biological tissues such as the cerebral cortex. In this study, we measured the hemodynamic responses of the rat barrel cortex to whisker stimulation by using a frequency-domain NIRS system. We designed multiple optical probes comprising multi-mode optical fibers and manipulating arms, both of which can be easily applied to small animals. Various electrical stimulations were applied to rat whiskers at different voltage levels and stimulation frequencies. Our results show that the hemodynamic responses are highly dependent on the stimulation voltage level, and not so much on stimulation frequency. This paper suggests that NIRS technology is highly suitable for the study of small animal brains.

Neuronal Responses in the Globus Pallidus during Subthalamic Nucleus Electrical Stimulation in Normal and Parkinson's Disease Model Rats

Deep brain stimulation (DBS) of the subthalamic nucleus (STN) has been widely used as a treatment for the movement disturbances caused by Parkinsons disease (PD). Despite successful application of DBS, its mechanism of therapeutic effect is not clearly understood. Because PD results from the degeneration of dopamine neurons that affect the basal ganglia (BG) network, investigation of neuronal responses of BG neurons during STN DBS can provide informative insights for the understanding of the mechanism of therapeutic effect. However, it is difficult to observe neuronal activity during DBS because of large stimulation artifacts. Here, we report the observation of neuronal activities of the globus pallidus (GP) in normal and PD model rats during electrical stimulation of the STN. A custom artifact removal technique was devised to enable monitoring of neural activity during stimulation. We investigated how GP neurons responded to STN stimulation at various stimulation frequencies (10, 50, 90 and 130 Hz). It was observed that activities of GP neurons were modulated by stimulation frequency of the STN and significantly inhibited by high frequency stimulation above 50 Hz. These findings suggest that GP neuronal activity is effectively modulated by STN stimulation and strongly dependent on the frequency of stimulation.

A quantitative comparison of basal ganglia neuronal activities of normal and Parkinson's disease model rats

The purpose of this study was to identify consistent characteristic changes of neuronal activity in basal ganglia (BG) nuclei associated with Parkinsons disease (PD) so that a reliable index of PD can be derived. A simple algorithm for automatic identification of firing patterns was devised as an essential tool to achieve this goal. A detailed quantitative analysis of firing patterns as well as firing rate was performed in three BG nuclei: the subthalamic nucleus (STN), the substantia nigra pars reticulate (SNpr), and the globus pallidus (GP). The results showed that the firing rate of STN neurons was not significantly altered in PD model rats. We also did not find a significant alteration in firing rates in the SNpr and GP between normal and PD model rats. In contrast, consistent changes of firing patterns were observed in all three BG nuclei in that the percentage of neurons with a regular firing pattern decreased whereas those with irregular, mixed, or burst patterns increased. This enables a simple algorithm based on burst detection and the shape of the interspike interval histogram to identify whether the neuronal activity is from normal or PD model rats.

Activity-dependent conduction velocity changes of Aδ fibers in a rat model of neuropathy

ACTIVITY-DEPENDENT changes of conduction velocity (CV) and conduction block in single Aδ fibers of primary afferent neurons were characterized in a rat model of neuropathy (NP). Injured dorsal root (DR) fiber in NP rats exhibited profoundly greater decreases of CV following impulse activity than did DR fiber in normal rats. Activity-dependent conduction block was absent up to 100 Hz of activity rate in DR fiber of NP rats, but was present above 25 Hz in normal rats. Profiles of activity dependence in sciatic fibers were similar in both NP and normal rats. These results suggest that nerve injury may alter activity-dependent hypoexcitability of Aδ DR fibers. Furthermore, this excitability change may be responsible for the elevated pain perception in neuropathy.

Bombesin-induced changes of sensory transmission in the dorsal column nuclei

Bombesin was intracisternally administered to determine the effects on the neuronal activities of the dorsal column nuclei (DCN) of anesthetized rats. Although averaged afferent somatosensory transmission through 22 DCN neurons did not appear to be altered by bombesin (-1.11 +/- 3.5%), 12 of them were actually augmented (+17.15 +/- 2.7%) and 10 of them were suppressed (-26.15 +/- 4.9%) during 40 min after bombesin (0.01 microgram) administration. Pretreatment of a bombesin antagonist ([Leu13,Psi(CH2NH)Leu14]bombesin, 0.1 microgram) blocked the effects of bombesin. Lower dose (0.001 microgram) of bombesin did not exert any influences. These results suggest that bombesin may heterogeneously influence afferent somatosensory information in the brain stem of the rat.

Increased immunoreactivity of c‑Fos in the spinal cord of the aged mouse and dog

Expression of c‑Fos in the spinal cord following nociceptive stimulation is considered to be a neurotoxic biomarker. In the present study, the immunoreactivity of c‑Fos in the spinal cord was compared between young adult (2‑3 years in dogs and 6 months in mice) and aged (10‑12 years in dogs and 24 months in mice) Beagle dogs and C57BL/6J mice. In addition, changes to neuronal distribution and damage to the spinal cord were also investigated. There were no significant differences in neuronal loss or degeneration of the spinal neurons observed in either the aged dogs or mice. Weak c‑Fos immunoreactivity was observed in the spinal neurons of the young adult animals; however, c‑Fos immunoreactivity was markedly increased in the nuclei of spinal neurons in the aged dogs and mice, as compared with that of the young adults. In conclusion, c‑Fos immunoreactivity was significantly increased without any accompanying neuronal loss in the aged spinal cord of mice and dogs, as compared with the spinal cords of the young adult animals.

Manipulation of Rat Movement via Nigrostriatal Stimulation Controlled by Human Visually Evoked Potentials

Here, we report that the development of a brain-to-brain interface (BBI) system that enables a human user to manipulate rat movement without any previous training. In our model, the remotely-guided rats (known as ratbots) successfully navigated a T-maze via contralateral turning behaviour induced by electrical stimulation of the nigrostriatal (NS) pathway by a brain- computer interface (BCI) based on the human controller’s steady-state visually evoked potentials (SSVEPs). The system allowed human participants to manipulate rat movement with an average success rate of 82.2% and at an average rat speed of approximately 1.9 m/min. The ratbots had no directional preference, showing average success rates of 81.1% and 83.3% for the left- and right-turning task, respectively. This is the first study to demonstrate the use of NS stimulation for developing a highly stable ratbot that does not require previous training, and is the first instance of a training-free BBI for rat navigation. The results of this study will facilitate the development of borderless communication between human and untrained animals, which could not only improve the understanding of animals in humans, but also allow untrained animals to more effectively provide humans with information obtained with their superior perception.