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Nigrostriatal dopamine neurons form wide and highly dense axon terminals in the striatum of rat: Single-cell study
S170
Abstracts / Neuroscience Research 58S (2007) S1–S244
1 Differences between single neuronal responses in P2-h0
4 Novel hydrolase AHO-3 is required for associative P2-h0
anterior and posterior insula during multi-trial reward schedules
learning between cultivation temperature and feeding state in C. elegans
Takashi Mizuhiki 1,2 , Barry J. Richmond 3 , Munetaka Shidara 1,2 1 Graduate School of Comprehensive Human Science, University of Tsukuba, Japan; 2 Neuroscience Research Institute, AIST, Tsukuba, Ibaraki, Japan; 3 Laboratory of Neuropsychology, NIMH, Bethesda, USA The anterior (AI) and posterior insula (PI) have anatomically different neuronal connections with other brain areas, suggesting the functional subdivision between them. We previously showed that many of single neurons in monkey AI reacted to immediately forthcoming reward during multi-trial reward schedules. To examine whether PI neurons also show reward-expectancy-related responses, we recorded 38 PI neurons using the same task. Though 21/120 AI neurons reacted to possible reward, preliminary results show that none of 38 PI neurons reacted to it. On the other hand, none of 120 AI neurons reacted to reward solenoid apparatus activation whereas 8/38 PI neurons reacted to it (possibly its activation sound). These differences in information processing might come from different anatomical connections that AI and PI have dense connections with the areas responsive to reward and sound, respectively.
Nana Nishio 1 , Akiko Mohri 2 , Eiji Kodama 1 , Kotaro Kimura 3 , Atsushi Kuhara 1 , Ikue Mori 1,4 1 Division of Biological Science, Nagoya University, Nagoya, Japan; 2 Present address: University of Texas, Health Science Centre, USA; 3 Present address: National Institute of Genetics, Japan; 4 CREST, Japan Animals exhibit many types of behavioral plasticity. To elucidate the regulation mechanism of behavioral plasticity, we are using the temperature-food associative learning paradigm in C. elegans: well-fed animals migrate to and starved animals avoid from their cultivation temperature on a temperature gradient. The aho-3 mutant has defect in this associative learning; aho-3 mutants showed defect in starvation-induced cultivation temperature avoidance, while they showed normal response in food recognition. We found that aho-3 gene encodes a predicted hydrolase that is a highly conserved protein throughout yeast to human. However, the molecular properties have not been characterized. aho-3 was expressed in several neurons required for thermotaxis, and the learning defect of aho-3 mutant was partially rescued by aho-3 expression in thermotaxis neural circuit. Currently, we are trying to dissect the molecular and cellular analysis of aho-3 gene further. Research fund: KAKENHI
2 Visualization of the convergence of entorhinal and hipP2-h0
5 P2-h0
pocampal CA3 input onto CA1 neurons
roaches
Riichi Kajiwara 1 , Floris G. Wouterlood 2 , Anupam Sah 2 , Amber J. enne T.G. Baks-te Bulte 2 , Menno P. Witter 2,3 Boekel 2 , Luci¨ 1 Neuroscience Research Institute, AIST, Tsukuba, Japan; 2 Department of Anatomy, VU Univ Med Ctr, Amsterdam, Netherlands; 3 Ctr Biol of Memory, NTNU, Trondheim, Norway The hippocampal area CA1 receives excitatory input from the entorhinal cortex through the perforant pathway, directly from layer III and indirectly from layer II through the dentate gyrus-CA3-Schaffer collateral circuit. Here we report anatomical evidence in support of such convergence in CA1 single neurons. In rats, anterograde tracers were injected into layer III of the entorhinal cortex and into CA3, to tag simultaneously the entorhinal fibers and the Schaffer collaterals with different Alexa FluorTM dyes (488 and 633). In 400 m thick slices of fixed brains we intracellularly filled the Alexa Fluor hydrazide 555 in CA1 pyramidal cells and interneurons. Thin sections of these slices were scanned in a confocal laser scanning microscope and with the use of 3D reconstruction algorithms we confirmed that boutons on the labeled input fibers from both entorhinal cortex and CA3 were in contact with single CA1 interneurons as well as pyramidal cells.
3 Establishment of in vivo gene transfer system into P2-h0 newly hatched chick brain Naoki Hirose 1 , Sachiko Katagiri 1 , Shinji Yamaguchi 1 , Yasuyuki Fujimoto 1 , Toshiya Matsushima 2 , Tatsuya Takano 1 , Koichi Homma 1 1 Faculty of Pharmaceutical Sciences, Teikyo University, Sagamihara-shi, Kanagawa, Japan; 2 Department of Biology, Faculty of Science, Hokkaido University, Hokkaido, Japan Newly hatched domestic chick serves an ideal model for studies of neural basis of behavioral plasticity, particularly for understanding the mechanism of learning such as filial imprinting. In order to elucidate the molecular basis and gene functions involved in learning, we developed an in vivo gene transfer system in the brain of a living chick using electroporation. When green fluorescent protein-encoding plasmids were transfected to a chick brain, green fluorescence was clearly observed, and expression at the protein level was confirmed by immunoblotting. Most of the transfected brain cells exhibited dendrites and were immunostained using anti-NeuN antibodies, suggesting that these cells were neuronal cells. This neuron-selective electroporation system will facilitate the analysis of gene function in the living chick brain and provide further clues as to the molecular mechanisms of avian learning.
Context-dependent olfactory conditioning in cock-
Chihiro Sato 1 , Yukihisa Matsumoto 1 , Midori Sakura 2 , Hidehiro Watanabe 1 , Makoto Mizunami 1 1 Graduate School of Life Sciences, Tohoku University, Sendai, Japan; 2 Research Institute for Electronic Science, Hokkaido University, Sapporo, Japan We demonstrated context-dependent olfactory learning by behavioral and electrophysiological methods. Cockroaches, Periplaneta americana, were trained with differential conditioning procedure in which the arrangement of conditioning stimuli (CS) and unconditioned stimuli (US) is the opposite between two light conditions (contexts); under context 1, odor A was paired with reward (US+) and odor B was paired with punishment (or presented without pairing, US−) and under context 2, the pairings were AUS− and B-US+. After training in the behavioral experiment, cockroaches selected the rewarded odor in each context. In the electrophysiological experiment, response of salivary neurons to rewarded odor was significantly greater than that to unrewarded odor after training in each context. We showed that (1) cockroaches are capable of selecting different odor according to the visual context and that (2) the contextual learning can be monitored by activities of salivary neurons.
6 Nigrostriatal dopamine neurons form wide and highly P2-h0 dense axon terminals in the striatum of rat: Single-cell study Wakoto Matsuda 1 , Takahiro Furuta 2 , Kouichi Nakamura 2 , Takeshi Kaneko 2 , Ryohachi Arai 1 1 Department of Anatomy, Shiga University of Medical Science, Otsu, Japan; 2 Department of Morphological Brain Science, Graduate School of Medicine, Kyoto University, Kyoto, Japan Dopamine (DA) neurons in the midbrain mediate a variety of brain functions, such as motor control, emotion, and reward. We studied the axonal arborization of DA neurons with Sindbis virus vectors that expressed membrane-targeted green fluorescent protein (GFP). After injections of the viral vectors into the rat substantia nigra compacta (SNc), infected single neurons were first examined their tyrosine hydroxylase immunoreactivity (TH-IR) and were then visualized completely by the inimmunoperoxidase method with an anti-GFP antibody. Of 70 infected neurons, 8 nigrostriatal neurons showing TH-IR were reconstructed by camera lucida and formed wide and highly dense bushes in the striatum, amounted to hundreds of thousands m in total length. The present results are considered to support the reinforcement-learning hypothesis of the basal ganglia.
Abstracts / Neuroscience Research 58S (2007) S1–S244
1 Differences between single neuronal responses in P2-h0
4 Novel hydrolase AHO-3 is required for associative P2-h0
anterior and posterior insula during multi-trial reward schedules
learning between cultivation temperature and feeding state in C. elegans
Takashi Mizuhiki 1,2 , Barry J. Richmond 3 , Munetaka Shidara 1,2 1 Graduate School of Comprehensive Human Science, University of Tsukuba, Japan; 2 Neuroscience Research Institute, AIST, Tsukuba, Ibaraki, Japan; 3 Laboratory of Neuropsychology, NIMH, Bethesda, USA The anterior (AI) and posterior insula (PI) have anatomically different neuronal connections with other brain areas, suggesting the functional subdivision between them. We previously showed that many of single neurons in monkey AI reacted to immediately forthcoming reward during multi-trial reward schedules. To examine whether PI neurons also show reward-expectancy-related responses, we recorded 38 PI neurons using the same task. Though 21/120 AI neurons reacted to possible reward, preliminary results show that none of 38 PI neurons reacted to it. On the other hand, none of 120 AI neurons reacted to reward solenoid apparatus activation whereas 8/38 PI neurons reacted to it (possibly its activation sound). These differences in information processing might come from different anatomical connections that AI and PI have dense connections with the areas responsive to reward and sound, respectively.
Nana Nishio 1 , Akiko Mohri 2 , Eiji Kodama 1 , Kotaro Kimura 3 , Atsushi Kuhara 1 , Ikue Mori 1,4 1 Division of Biological Science, Nagoya University, Nagoya, Japan; 2 Present address: University of Texas, Health Science Centre, USA; 3 Present address: National Institute of Genetics, Japan; 4 CREST, Japan Animals exhibit many types of behavioral plasticity. To elucidate the regulation mechanism of behavioral plasticity, we are using the temperature-food associative learning paradigm in C. elegans: well-fed animals migrate to and starved animals avoid from their cultivation temperature on a temperature gradient. The aho-3 mutant has defect in this associative learning; aho-3 mutants showed defect in starvation-induced cultivation temperature avoidance, while they showed normal response in food recognition. We found that aho-3 gene encodes a predicted hydrolase that is a highly conserved protein throughout yeast to human. However, the molecular properties have not been characterized. aho-3 was expressed in several neurons required for thermotaxis, and the learning defect of aho-3 mutant was partially rescued by aho-3 expression in thermotaxis neural circuit. Currently, we are trying to dissect the molecular and cellular analysis of aho-3 gene further. Research fund: KAKENHI
2 Visualization of the convergence of entorhinal and hipP2-h0
5 P2-h0
pocampal CA3 input onto CA1 neurons
roaches
Riichi Kajiwara 1 , Floris G. Wouterlood 2 , Anupam Sah 2 , Amber J. enne T.G. Baks-te Bulte 2 , Menno P. Witter 2,3 Boekel 2 , Luci¨ 1 Neuroscience Research Institute, AIST, Tsukuba, Japan; 2 Department of Anatomy, VU Univ Med Ctr, Amsterdam, Netherlands; 3 Ctr Biol of Memory, NTNU, Trondheim, Norway The hippocampal area CA1 receives excitatory input from the entorhinal cortex through the perforant pathway, directly from layer III and indirectly from layer II through the dentate gyrus-CA3-Schaffer collateral circuit. Here we report anatomical evidence in support of such convergence in CA1 single neurons. In rats, anterograde tracers were injected into layer III of the entorhinal cortex and into CA3, to tag simultaneously the entorhinal fibers and the Schaffer collaterals with different Alexa FluorTM dyes (488 and 633). In 400 m thick slices of fixed brains we intracellularly filled the Alexa Fluor hydrazide 555 in CA1 pyramidal cells and interneurons. Thin sections of these slices were scanned in a confocal laser scanning microscope and with the use of 3D reconstruction algorithms we confirmed that boutons on the labeled input fibers from both entorhinal cortex and CA3 were in contact with single CA1 interneurons as well as pyramidal cells.
3 Establishment of in vivo gene transfer system into P2-h0 newly hatched chick brain Naoki Hirose 1 , Sachiko Katagiri 1 , Shinji Yamaguchi 1 , Yasuyuki Fujimoto 1 , Toshiya Matsushima 2 , Tatsuya Takano 1 , Koichi Homma 1 1 Faculty of Pharmaceutical Sciences, Teikyo University, Sagamihara-shi, Kanagawa, Japan; 2 Department of Biology, Faculty of Science, Hokkaido University, Hokkaido, Japan Newly hatched domestic chick serves an ideal model for studies of neural basis of behavioral plasticity, particularly for understanding the mechanism of learning such as filial imprinting. In order to elucidate the molecular basis and gene functions involved in learning, we developed an in vivo gene transfer system in the brain of a living chick using electroporation. When green fluorescent protein-encoding plasmids were transfected to a chick brain, green fluorescence was clearly observed, and expression at the protein level was confirmed by immunoblotting. Most of the transfected brain cells exhibited dendrites and were immunostained using anti-NeuN antibodies, suggesting that these cells were neuronal cells. This neuron-selective electroporation system will facilitate the analysis of gene function in the living chick brain and provide further clues as to the molecular mechanisms of avian learning.
Context-dependent olfactory conditioning in cock-
Chihiro Sato 1 , Yukihisa Matsumoto 1 , Midori Sakura 2 , Hidehiro Watanabe 1 , Makoto Mizunami 1 1 Graduate School of Life Sciences, Tohoku University, Sendai, Japan; 2 Research Institute for Electronic Science, Hokkaido University, Sapporo, Japan We demonstrated context-dependent olfactory learning by behavioral and electrophysiological methods. Cockroaches, Periplaneta americana, were trained with differential conditioning procedure in which the arrangement of conditioning stimuli (CS) and unconditioned stimuli (US) is the opposite between two light conditions (contexts); under context 1, odor A was paired with reward (US+) and odor B was paired with punishment (or presented without pairing, US−) and under context 2, the pairings were AUS− and B-US+. After training in the behavioral experiment, cockroaches selected the rewarded odor in each context. In the electrophysiological experiment, response of salivary neurons to rewarded odor was significantly greater than that to unrewarded odor after training in each context. We showed that (1) cockroaches are capable of selecting different odor according to the visual context and that (2) the contextual learning can be monitored by activities of salivary neurons.
6 Nigrostriatal dopamine neurons form wide and highly P2-h0 dense axon terminals in the striatum of rat: Single-cell study Wakoto Matsuda 1 , Takahiro Furuta 2 , Kouichi Nakamura 2 , Takeshi Kaneko 2 , Ryohachi Arai 1 1 Department of Anatomy, Shiga University of Medical Science, Otsu, Japan; 2 Department of Morphological Brain Science, Graduate School of Medicine, Kyoto University, Kyoto, Japan Dopamine (DA) neurons in the midbrain mediate a variety of brain functions, such as motor control, emotion, and reward. We studied the axonal arborization of DA neurons with Sindbis virus vectors that expressed membrane-targeted green fluorescent protein (GFP). After injections of the viral vectors into the rat substantia nigra compacta (SNc), infected single neurons were first examined their tyrosine hydroxylase immunoreactivity (TH-IR) and were then visualized completely by the inimmunoperoxidase method with an anti-GFP antibody. Of 70 infected neurons, 8 nigrostriatal neurons showing TH-IR were reconstructed by camera lucida and formed wide and highly dense bushes in the striatum, amounted to hundreds of thousands m in total length. The present results are considered to support the reinforcement-learning hypothesis of the basal ganglia.