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Optical recording of synaptic potential in the embryonic chick brain stem slice preparation using a voltage-sensitive dye
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OPTICAL RECORDING OF SYNAPTIC POTENTIAL IN THE EMBRYONIC CHICK BRAIN STEM SLICE PREPARATION USING A VOLTAGE-SENSITIVE DYE. HITOSHI KOMURO, YOKO MOMOSE, TETSURO SAKAI, AKIHIKO HIROTA, KOHTARO KAMINO, Department of Phvsioloqv , Tokyo Ikx
Gku
School of Medicine, l-5-45 Yushima, Bunkvo-ku, Tokyo 113, Japan.
Using an optical method for monitoring electrical activity, we have recorded synaptic potential from the embryonic chick brain stem. Vaguslbrain stems were isolated from 6 to lo-day old chick embryos and then the slice preparations with both the right and left vagus nerve fibers were prepared by transverse sectioning. The slice preparations were stained with a voltage-sensitive merocyanine-rhodanine dye (NK2761). Voltage-related optical absorbance changes evoked by vagus nerve stimulation with depolarizing pulses using a suction electrode were tried to record simultaneously from 127 separated adjacent loci in the preparation using a 12 x 12-element photodiode array. Optical responses were detected from limited area near the dorsal surface of the stimulated side. When relatively large strengths of stimulating current were applied, optical changes related to postsynaptic potential were detected. The postsynaptic potentialrelated optical changes appeared to be concentrated in a smaller region within the vagal response area, and they were completely eliminated in the presence of kynurenic acid which is a blocker of glutamatergic synaptic transmission.
I~ISTCCHEMIC4L
LOCALIZATICN OF NERVE GROWTH FACTOR RECEPTOR CONTAINING NERVE IN THE RAT CENTRAL NERVOUS SYSTEM MIWAKO SEKITANI', MIKAKO IKEDA’ , YASUHIDE LEE2, SADAO SHIOSAKA', HIROSHI HATANAKAS AND MASAYA TCHYAMA2,Department of Neuroanatomy,BiomedicaI Research TERMINALS
Center', 2nd Department of Anatomy' and Protein Research Center3,0saka University Medical School.3-57-4 Nakanosiima Kitaku. Osaka.530 Iaoan. The -distribution of’nerve growth factor recepior (NGF-R) ” containing anerve terminals in the rat central nervous system was imnunohistochemically analysed using a monoclonal antibody against rat NGF-R, MC1921gG. Dense nerve positive terminals were found in the olfactory glomerular layer, suprachiasmatic nucleus, 01 ivary pretectal nucleus, lateral geniculate nucleus, super ior colliculus, solitary tract nucleus, trigeminal spinal tract nucleus, gracile nucleus, cuneate nucleus and dorsal horn of spinal cord. A double-labeling of retrograde tracer technique and an imnunofluorescnt technique for NGF-R showed that the positive nerve terminals in the 01 factory glomerular layer originated from NGF-R neurons in the diagonal band and substantia Innominata.
REGULATORY ACTION OF ACIDIC FIBROBLAST GROWTH FACTOR ON FOOD INTAKE. SAKI'. KENJI SUZUKI"', TADASHI MUTO*'. KAZUMITSU HANAIX3. IKUO T,OYA,:~~AH~~~~~';;,~~~ll" for Wakan-Yaku, “Scientific Instrument Center, Toyama Medical and Research Institute Pharmaceutical University, Sugitani, Toyama 930-01, jDepartment of Biology, Faculty of Science, Kyushu University, Fukuoka 812, and 41nstitute of Molecular Neurobiology, Shiga University of Medical Science, Ootsu, Shiga 520-21, Japan. Acidic fibroblast growth factor (aFGF) has been isolated from neural tissue including hypothalamus as well as retina. Although this factor has mitogenic activity for glial, endothelial and vascular smooth muscle cells in vitro, the role of aFGF in the in viva central nervous system remains obscure. Recently, we have reported that the level of aFGF in cerebrospinal fluid of rats increases after feeding or after intraperitoneal injection of glucose. In the present study, we infused glucose into third ventricle with a dose of 0.2 mg/lO ul to investigate a direct effect of glucose on the release of aFGF in cerebrospinsl fluid. The level of aFGF was determined by a bioassay system in which aFGF specifically depressed the Hydra japonica feeding response elicited by S-methylglutathione. The contents of s.FGF in cerebrospinal fluid increased 1000 times greater than that of the pi-e-infusionperiod 2 hr after glucose infusions and it returned to pre-infusion levels 5 hr after infusions. Saline or mannitol infusions had no effect on aFGF levels. When the central action of aFGF or its antibody on food intake was examined, intracersbroventricular administrations of aFGF dose-dependently decreased food intake in rats, whereas bilateral infusions of antibody of aFGF into lateral hypothalamus increased food intake. These results suggest that aFGF is one of the endogenous substances which centrally regulate food intake.
OPTICAL RECORDING OF SYNAPTIC POTENTIAL IN THE EMBRYONIC CHICK BRAIN STEM SLICE PREPARATION USING A VOLTAGE-SENSITIVE DYE. HITOSHI KOMURO, YOKO MOMOSE, TETSURO SAKAI, AKIHIKO HIROTA, KOHTARO KAMINO, Department of Phvsioloqv , Tokyo Ikx
Gku
School of Medicine, l-5-45 Yushima, Bunkvo-ku, Tokyo 113, Japan.
Using an optical method for monitoring electrical activity, we have recorded synaptic potential from the embryonic chick brain stem. Vaguslbrain stems were isolated from 6 to lo-day old chick embryos and then the slice preparations with both the right and left vagus nerve fibers were prepared by transverse sectioning. The slice preparations were stained with a voltage-sensitive merocyanine-rhodanine dye (NK2761). Voltage-related optical absorbance changes evoked by vagus nerve stimulation with depolarizing pulses using a suction electrode were tried to record simultaneously from 127 separated adjacent loci in the preparation using a 12 x 12-element photodiode array. Optical responses were detected from limited area near the dorsal surface of the stimulated side. When relatively large strengths of stimulating current were applied, optical changes related to postsynaptic potential were detected. The postsynaptic potentialrelated optical changes appeared to be concentrated in a smaller region within the vagal response area, and they were completely eliminated in the presence of kynurenic acid which is a blocker of glutamatergic synaptic transmission.
I~ISTCCHEMIC4L
LOCALIZATICN OF NERVE GROWTH FACTOR RECEPTOR CONTAINING NERVE IN THE RAT CENTRAL NERVOUS SYSTEM MIWAKO SEKITANI', MIKAKO IKEDA’ , YASUHIDE LEE2, SADAO SHIOSAKA', HIROSHI HATANAKAS AND MASAYA TCHYAMA2,Department of Neuroanatomy,BiomedicaI Research TERMINALS
Center', 2nd Department of Anatomy' and Protein Research Center3,0saka University Medical School.3-57-4 Nakanosiima Kitaku. Osaka.530 Iaoan. The -distribution of’nerve growth factor recepior (NGF-R) ” containing anerve terminals in the rat central nervous system was imnunohistochemically analysed using a monoclonal antibody against rat NGF-R, MC1921gG. Dense nerve positive terminals were found in the olfactory glomerular layer, suprachiasmatic nucleus, 01 ivary pretectal nucleus, lateral geniculate nucleus, super ior colliculus, solitary tract nucleus, trigeminal spinal tract nucleus, gracile nucleus, cuneate nucleus and dorsal horn of spinal cord. A double-labeling of retrograde tracer technique and an imnunofluorescnt technique for NGF-R showed that the positive nerve terminals in the 01 factory glomerular layer originated from NGF-R neurons in the diagonal band and substantia Innominata.
REGULATORY ACTION OF ACIDIC FIBROBLAST GROWTH FACTOR ON FOOD INTAKE. SAKI'. KENJI SUZUKI"', TADASHI MUTO*'. KAZUMITSU HANAIX3. IKUO T,OYA,:~~AH~~~~~';;,~~~ll" for Wakan-Yaku, “Scientific Instrument Center, Toyama Medical and Research Institute Pharmaceutical University, Sugitani, Toyama 930-01, jDepartment of Biology, Faculty of Science, Kyushu University, Fukuoka 812, and 41nstitute of Molecular Neurobiology, Shiga University of Medical Science, Ootsu, Shiga 520-21, Japan. Acidic fibroblast growth factor (aFGF) has been isolated from neural tissue including hypothalamus as well as retina. Although this factor has mitogenic activity for glial, endothelial and vascular smooth muscle cells in vitro, the role of aFGF in the in viva central nervous system remains obscure. Recently, we have reported that the level of aFGF in cerebrospinal fluid of rats increases after feeding or after intraperitoneal injection of glucose. In the present study, we infused glucose into third ventricle with a dose of 0.2 mg/lO ul to investigate a direct effect of glucose on the release of aFGF in cerebrospinsl fluid. The level of aFGF was determined by a bioassay system in which aFGF specifically depressed the Hydra japonica feeding response elicited by S-methylglutathione. The contents of s.FGF in cerebrospinal fluid increased 1000 times greater than that of the pi-e-infusionperiod 2 hr after glucose infusions and it returned to pre-infusion levels 5 hr after infusions. Saline or mannitol infusions had no effect on aFGF levels. When the central action of aFGF or its antibody on food intake was examined, intracersbroventricular administrations of aFGF dose-dependently decreased food intake in rats, whereas bilateral infusions of antibody of aFGF into lateral hypothalamus increased food intake. These results suggest that aFGF is one of the endogenous substances which centrally regulate food intake.