Tracking of Neural Stem Cells in Rats with Intracerebral Hemorrhage by the Use of 3T MRI

OBJECTIVE: To access the feasibility of clinically available 3T MRI to detect the migration of labeled neural stem cells (NSCs) in intracerebral hemorrhage (ICH) in a rat model. MATERIALS AND METHODS: The ethics committee of our institution approved this study. ICH was induced by the injection of collagenase type IV into the right striatum of ten Sprague-Dawley rats. Human NSCs conjugated with Feridex (super-paramagnetic iron oxide: SPIO) were transplanted into the left striatum one week after ICH induction. MRI was performed on a 3T scanner during the first, second, third, fourth, and sixth weeks post-transplantation. MRI was obtained using coronal T2- and T2*-weighted sequences. Two rats were sacrificed every week after in vivo MRI in order to analyze the histological findings. RESULTS: ICH in the right striatum was detected by MRI one and two weeks after transplantation without migration of the NSCs. There was no migration of the NSCs as seen on the histological findings one week after transplantation. The histological findings two weeks after transplantation showed a small number of NSCs along the corpus callosum. On MRI three weeks after transplantation, there was a hypointense line along the corpus callosum and decreased signal intensity in the right periventricular region. Histological findings three weeks after transplantation confirmed the presence of the hypointense line representing SPIO-labeled NSCs. MRI four and six weeks after transplantation showed a hypointense spot in the right periventricular region. The histological findings four and six weeks after transplantation showed the presence of prominent NSCs in the right periventricular region. CONCLUSION: 3T MRI can detect the migration of NSCs in rats with ICH along the corpus callosum. Therefore, 3T MRI could be feasible for detecting the migration of NSCs in the clinical setting of stem cell therapy.

Effects of Phenylephrine on the Excitability of Medial Vestibular Nuclear Neurons in Rats

Coeruleo-vestibular pathway which connects locus coeruleus and vestibular nuclei is noradrenergic. This study was designed to elucidate the effects of phenylephrine on the spontaneous activity of acutely isolated medial vestibular nuclear neurons of rat by whole-cell patch-clamp technique. Sprague-Dawley rats, aged 14 to 16 days, were used. After enzymatic digestion, dissociated medial vestibular neurons were transferred to a recording chamber mounted on an inverted microscope, and spontaneous action potentials were recorded by standard patch-clamp techniques. In current-clamp mode, the frequency of spontaneous action potential of medial vestibular nuclear neurons was decreased by phenylephrine (n=15). Phenylephrine increased the amplitude of afterhyperpolarization without changes in the resting membrane potential and spike width. In voltage-clamp mode, the whole potassium currents of the medial vestibular nuclear neurons were increased by phenylephrine (n=12). These experimental results suggest that alpha-receptor mediates the inhibitory effects on the neuronal activity of the medial vestibular nuclear neuron.

Effects of a delta-opioid Agonist on the Brainstem Vestibular Nuclear Neuronal Activity of Rats

This study was undertaken to investigate the effects of [D-Ala2, D-Leu5]-enkephalin (DADLE) on the spontaneous activity of medial vestibular nuclear neurons of the rat. Sprague-Dawley rats, aged 14 to 16 days, were anesthetized with ether and decapitated. After enzymatic digestion, the brain stem portion of medial vestibular nuclear neuron was obtained by micropunching. The dissociated neurons were transferred to a recording chamber mounted on an inverted microscope, and spontaneous action potentials were recorded by standard patch-clamp techniques. The spontaneous action potentials were increased by DADLE in 12 cells and decreased in 3 cells. The spike frequency and resting membrane potential of these cells were increased by DADLE. The depth of afterhyperpolarization was not affected by DADLE. The potassium currents were decreased in 20 cells and increased in 5 cells. These results suggest that DADLE increases the neuronal activity of the medial vestibular nuclear neurons by altering resting membrane potential.

The Effect of Acetylcholine on the Potassium Currents and Length of Outer Hair Cell / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Inner hair cells (IHCs) of the organ of Corti change the external sound stimulus into the electrical signal and transmit this signal to the auditory cortex through afferent nerve fibers. Outer hair cells (OHCs) control the sound transmission function of IHC. OHCs respond with a somatic shape change to alterations in their membrane potential and this electromotile response is believed to provide mechanical feedback to the basilar membrane. Efferent nerve fibers which arise from the superior olivary nucleus in the midbrain and transmit to OHCs through medial olivocochlear bundle use acetylcholine (ACh) as a neurotransmitter. The cholinergic response of OHCs' alpha-9 nicotinic ACh receptor increase the Ca2+ influx, which control OHCs' electromotility by changing a membrane potential. In this research, the effect of ACh on the K+ current in OHC of guinea pig was studied, and the change of OHC length by ACh was studied. MATERIALS AND METHODS: Using the extracted OHC from a guinea pig potassium currents induced by ACh were recorded using the whole-cell patch clamp technique. The change of OHC length when ACh was applied was observed. RESULTS: 1) ACh increases voltage-dependent K+ current in OHC. 2) In the condition, which Ca2+-dependent K+ current is blocked by removing Ca2+ from intra-cellular fluid, ACh has no effect on K+ current in OHC. 3) ACh increases OHC length. CONCLUSION: These experimental results show that ACh from the medial olivocochlear efferent system regulates mobility of OHC, increases the Ca2+-dependent K+ currents in OHC.

Effects of Aminoglycoside Antibiotics on Acetylcholine-induced Potassium Currents in Guinea-pig Outer Hair Cell / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Aminoglycoside antibiotics are widely used for the treatment of infection caused by Gram-negative bacilli, but prolonged exposure to it can cause permanent hearing loss since are reported to interrupt the function of medial olivocochlear bundle. This study was designed to explore the effects of several aminoglycoside antibiotics on acetylcholine, the neurotransmitter of medial olivocochlear bundle, and how it affects potassium currents of the guinea pig outer hair cells. MATERIALS AND METHOD: Using extracted outer hair cells from Guinea pigs, potassium currents induced by acetylcholine were recorded using the whole-cell patch clamp technique. The effects of different aminoglycoside antibiotics on the potassium current were also studied. RESULTS: Acetylcholine increased the potassium currents of outer hair cells. All of aminoglycoside antibiotics tested such as neomycin, streptomycin, gentamicin, and amikacin decreased the acetylcholine-induced potassium current. CONCLUSION: These experimental results suggest that aminoglycoside antibiotics have effects on the membrane potential of outer hair cells regulated by acetylcholine, which is thought to be one of the mechanisms of aminoglycoside ototoxicity.

Inhibitory Effects of Norepinephrine on the Potassium Current of Rat Medial Vestibular Nuclear Neurons

BACKGROUND: This study aimed to explore the effects of norepinephrine on the potassium currents of rat medial vestibular nuclear neurons by using the whole-cell patch clamp technique. METHODS: Sprague-Dawley rats aged 14 to 16 days were anesthetized with ether and decapitated. After enzymatic digestion, the portion of the medial vestibular nucleus neurons were obtained by micropunching. The dissociated neurons were transferred into a recording chamber mounted on an inverted microscope and potassium currents were recorded by standard patch-clamp techniques under voltage-clamp modes. RESULTS: Norepinephrine inhibited the whole potassium currents of the medial vestibular nuclear neurons. Outward potassium currents were more suppressed in 4 mM 4-aminopyridine and norepinephrine added solutions than 4 mM 4-aminopyridine only, but were not suppressed in 10 mM tetraethylammonium and norepinephrine added solutions compared to 10 mM tetraethylammonium only. CONCLUSIONS: These results suggest that norepinephrine blocks the delayed rectifier potassium channels in medial vestibular nuclear neurons.

Excitatory Effects of 5-Hydroxytryptamine on the Resting Activity of Medial Vestibular Nuclear Neurons in the Rat / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: This study was aimed to investigate the effects of 5-Hydroxytryptamine (5-HT) on the spontaneous activity of medial vestibular nuclear neurons of the rat. MATERIALS AND METHOD: Sprague-Dawley rats aged 14 to 16 days were anesthetized with ether and decapitated. After enzymatic digestion, the brainstem portion of medial vestibular nuclear neuron was obtained by micropunching. The dissociated neurons were transferred into a recording chamber mounted on an inverted microscope and spontaneous action potentials were recorded by standard patch-clamp techniques. RESULTS: The response of medial vestibular nuclear neurons to 5-HT applied in the recording chamber was not uniform. The spontaneous action potentials were increased in 10 cells, decreased in 3 cells and not altered in 2 of 15 cells. The spike width of the neurons was not affected by 5-HT. The spike frequency and resting membrane potential of these cells were increased by 5-HT. The depth of afterhyperpolarization was decreased by 5-HT. CONCLUSION: These results suggest 5-HT increases spontaneous firing of the medial vestibular nuclear neurons by altering the afterhyperpolarization.

Effects of 5-Hydroxytryptamine on Potassium Currents of the Medial Vestibular Nuclear Neuron in the Rat / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: This study was aimed to explore the effects of the 5-hydroxytryptamine (5-HT) on potassium currents in rat vestibular nuclear neurons. MATERIALS AND METHOD: Sprague-Dawley rats aged 14 to 16 days were anesthetized with ether and decapitated. After enzymatic digestion, the portion of medial vestibular nucleus neuron was removed by micropunching and gently agitated. The dissociated neurons were transferred into a recording chamber mounted on an inverted microscope and whole-cell membrane currents were recorded at room temperature by using standard patch-clamp techniques. RESULTS: When cells were held at -70 mV and depolarized from -60 mV to +40 mV in 10 mV increments, sustained outward potassium currents were evoked. The response of medial vestibular nuclear neurons to 5-HT was not uniform. The outward potassium currents were increased in 17 of 40 cells and decreased in 23 of 40 cells. 5-carboxamidotryptamine, 5-HT1 agonist increased the outward potassium currents of the medial vestibular nuclear cell. alpha-methyl-5-hydroxytryptamine, 5-HT2 agonist decreased the outward potassium currents of the medial vestibular nuclear cell. CONCLUSION: These results suggest that 5-HT affects the potassium currents of the cell with different effects according to the receptor subtype on which it acts.