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3005 Pseudorabies virus mutant as a transneuronal tracer
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3003
SIMULTANEOUS OPTICAL RECORDING OF CALCIUM AND VOLTAGE TRANSIENTS IN THE ORGANOTYPIC HIPPOCAMPAL CULTURES IN RAT Photon Medical Research Center, Hamamatsu University l, Department of Neurosurgery, Hamamatsu University2 Shinji Matsumura’ , Seiji Yamamoto2, Takashi Sakura? , Atsuo Miyakawa’ We tried to ‘simultaneously monitor the distribution and time course of electrical potential changes and calcium transients in organotypic hippocampal cultures in the rat. To detect voltage transients, we measured absorbance changes (at 700f60 nm) in the slice stained with the voltage-sensitive dye RH482 by using a 16x 16 photodiode array (PDA). We employed an ICCD camera to detect calcium transients by measuring fluorescence changes of the dye Calcium Green-l. The excitation light was allowed to pass through a bandpass filter (450490nm) to the preparation and the emitted light to pass through a bandpass filter (530flOnm) for [Ca”+]i measurement. A dichroic mirror (580nm) was used in order to distinguish calcium signals form voltage signals. Through the 10x objective lens, individual photo diodes detected the light from an area of tissue measuring 44pmx44pm. Image frames were sampled at 0.5 msec intervals on PDA and at 66msec intervals on ICCD camera.
3004
Brain function imaging using a three dimensional magnetometer-spatial
filter array.
Department of Physiology, Kyoto Prefectural University of Medicine’, Department of Neurology, Kyoto Prefectural University of Medicine 2, Technology Research Laboratory, Shiiadzu Co. LTD3 Kenji Yoshikawa’ , Jin XueLong’ , Keisuke Toyama l, Yoshiaki Takanashi2, Yasushi Kondo3, Yoshikazu Yoshida3 Magnetoencephalography (MEG) is characterized by high temporal resolution and poor spatial resolution compared with other noninvasive means of brain functional mapping. We attempted to overcome the difficulty of the MEG by introducing a spatial filter array combined with 129 channel three dimensional (3D) magnetometer system which transforms the magnetic fields into the local brain currents. Simulation and phatom experiments demonstrated that this system was capable of imaging the local brain currents with a spatial resolution (half width: 10 mm) almost comparable to that for PET and fMFU. In the human experiments where thumb and little fingers were stimulated, this system could image the local brain current sources localized in the corresponding primary somatosensory areas. These results indicate that the spatial filtering combined with 3D-MEG is useful means of the noninvasive functional imaging of the human brain.
3005 Dept. Anat’,
Pseudorabies virus mutant as a transneuronal
tracer
Dept. Surg, Wakayama Medical College’
Emiko Senba’ , Hiroki Imbe2, Misako Saika-Doi’ Fusion gene of glycoprotein gG and 1acZ was constructed and pseudorabies virus (PrV) mutants were produced. Characteristics of thii mutant virus as a transneuronal tracer was investigated by injecting it into masseter of SD rats. Injection of PrV solution with titer of 1.0 x lO’p.f.u./ml was most suitable for tracing. Numerous labeled neurons were observed in motor trigeminal n. neurons, but they were very few in the mesencephalic trigeminal n. Second- or third-order neurons were identified in the locus coeruleus, raphe magnus, medullary reticular formation, Edmger-Westphal n., tuberal magnocellular n., lateral hypothalamic n. and paraventricular hypothalamic n. (PVH). Injection of this virus into stomach labeled neurons in the motor dorsal vagal n., n. tractus solitarius, n. ambiguus, A5 and PVH. These findings indicate that this virus preferentially infecct somatic and visceral motoneurons and is transneuronally transfered to the CNS neurons.
3003
SIMULTANEOUS OPTICAL RECORDING OF CALCIUM AND VOLTAGE TRANSIENTS IN THE ORGANOTYPIC HIPPOCAMPAL CULTURES IN RAT Photon Medical Research Center, Hamamatsu University l, Department of Neurosurgery, Hamamatsu University2 Shinji Matsumura’ , Seiji Yamamoto2, Takashi Sakura? , Atsuo Miyakawa’ We tried to ‘simultaneously monitor the distribution and time course of electrical potential changes and calcium transients in organotypic hippocampal cultures in the rat. To detect voltage transients, we measured absorbance changes (at 700f60 nm) in the slice stained with the voltage-sensitive dye RH482 by using a 16x 16 photodiode array (PDA). We employed an ICCD camera to detect calcium transients by measuring fluorescence changes of the dye Calcium Green-l. The excitation light was allowed to pass through a bandpass filter (450490nm) to the preparation and the emitted light to pass through a bandpass filter (530flOnm) for [Ca”+]i measurement. A dichroic mirror (580nm) was used in order to distinguish calcium signals form voltage signals. Through the 10x objective lens, individual photo diodes detected the light from an area of tissue measuring 44pmx44pm. Image frames were sampled at 0.5 msec intervals on PDA and at 66msec intervals on ICCD camera.
3004
Brain function imaging using a three dimensional magnetometer-spatial
filter array.
Department of Physiology, Kyoto Prefectural University of Medicine’, Department of Neurology, Kyoto Prefectural University of Medicine 2, Technology Research Laboratory, Shiiadzu Co. LTD3 Kenji Yoshikawa’ , Jin XueLong’ , Keisuke Toyama l, Yoshiaki Takanashi2, Yasushi Kondo3, Yoshikazu Yoshida3 Magnetoencephalography (MEG) is characterized by high temporal resolution and poor spatial resolution compared with other noninvasive means of brain functional mapping. We attempted to overcome the difficulty of the MEG by introducing a spatial filter array combined with 129 channel three dimensional (3D) magnetometer system which transforms the magnetic fields into the local brain currents. Simulation and phatom experiments demonstrated that this system was capable of imaging the local brain currents with a spatial resolution (half width: 10 mm) almost comparable to that for PET and fMFU. In the human experiments where thumb and little fingers were stimulated, this system could image the local brain current sources localized in the corresponding primary somatosensory areas. These results indicate that the spatial filtering combined with 3D-MEG is useful means of the noninvasive functional imaging of the human brain.
3005 Dept. Anat’,
Pseudorabies virus mutant as a transneuronal
tracer
Dept. Surg, Wakayama Medical College’
Emiko Senba’ , Hiroki Imbe2, Misako Saika-Doi’ Fusion gene of glycoprotein gG and 1acZ was constructed and pseudorabies virus (PrV) mutants were produced. Characteristics of thii mutant virus as a transneuronal tracer was investigated by injecting it into masseter of SD rats. Injection of PrV solution with titer of 1.0 x lO’p.f.u./ml was most suitable for tracing. Numerous labeled neurons were observed in motor trigeminal n. neurons, but they were very few in the mesencephalic trigeminal n. Second- or third-order neurons were identified in the locus coeruleus, raphe magnus, medullary reticular formation, Edmger-Westphal n., tuberal magnocellular n., lateral hypothalamic n. and paraventricular hypothalamic n. (PVH). Injection of this virus into stomach labeled neurons in the motor dorsal vagal n., n. tractus solitarius, n. ambiguus, A5 and PVH. These findings indicate that this virus preferentially infecct somatic and visceral motoneurons and is transneuronally transfered to the CNS neurons.