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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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O P T I C A L R E C O R D I N G O F S Y N A P T I C P O T E N T I A L IN T H E E M B R Y O N I C C H I C K B R A I N S T E M S L I C E P R E P A R A T I O N USING A V O L T A G E - S E N S I T I V E DYE. HITOSHI KOMURO, YOKO MOMOSE, TETSURO S A K A I , A K I H I K O H I R O T A , K O H T A R O K A M I N O , D e p a r t m e n t of P h y s i o l o q y , T o k y o I k a - S h i k a D a i q a k u S c h o o l of M e d i c i n e , I - 5 - 4 5 Y u s h i m a , B u n k y o - k u , T o k y o 113, J a p a n . U s i n g a n o p t i c a l m e t h o d for m o n i t o r i n g e l e c t r i c a l a c t i v i t y , w e h a v e r e c o r d e d s y n a p t i c p o t e n t i a l f r o m t h e e m b r y o n i c c h i c k b r a i n stem. Vagus/brain stems were i s o l a t e d f r o m 6 t o 1 0 - d a y o l d c h i c k e m b r y o s a n d t h e n the s l i c e p r e p a r a t i o n s w i t h both the right and left vagus nerve fibers were prepared by transverse sectioning. The slice preparations were stained with a voltage-sensitive merocyanine-rhodanine d y e (NK2761). Voltage-related optical absorbance changes evoked by vagus nerve stimulation with depolarizing pulses using a suction e l e c t r o d e w e r e t r i e d t o r e c o r d s i m u l t a n e o u s l y f r o m 127 s e p a r a t e d a d j a c e n t l o c i in t h e p r e p a r a t i o n u s i n g a 12 x 1 2 - e l e m e n t p h o t o d i o d e a r r a y . Optical responses were d e t e c t e d f r o m l i m i t e d a r e a n e a r t h e d o r s a l s u r f a c e of the s t i m u l a t e d side. When r e l a t i v e l y l a r g e s t r e n g t h s of s t i m u l a t i n g c u r r e n t w e r e a p p l i e d , o p t i c a l c h a n g e s r e l a t e d to p o s t s y n a p t i c p o t e n t i a l w e r e d e t e c t e d . The postsynaptic potentialr e l a t e d o p t i c a l c h a n g e s a p p e a r e d t o b e c o n c e n t r a t e d in a s m a l l e r r e g i o n w i t h i n t h e v a g a l r e s p o n s e a r e a , a n d t h e y w e r e c o m p l e t e l y e l i m i n a t e d in t h e p r e s e n c e of k y n u r e n i c a c i d w h i c h is a b l o c k e r of g l u t a m a t e r g i c s y n a p t i c t r a n s m i s s i o n .
IMvlUNOHISTOO-IEMICAL LOCALIZATI(~ OF NERVE GROWI'H FACTOR RECEPTOR CONTAINING NERVE TERMINALS IN THE RAT CENTRAL NERVOUS SYSTEM MIWAKO SEKITANI 2, MIKAKO 1KEDA1 , YASUHIDE LEE 2 S A D A O SHIOSAKA 1 HIROSHI HATANAKA3 , AND M~SAYA TOHYAMA2,Department o f Neuroanatomy~Biomedical Research C e n t e r ~, 2nd Department of A n a t o m y 2, a nd Protein Research Center3,Osaka University Medical School,3-57-4 Nakanoshima Kitaku, Osaka,530 Japan. The distribution of n e r v e g r o w t h f a c t o r receptor (NGF-R) containing nerve terminals in the rat central nervous system was immunohistochemically analysed using a monoclonal antibody against r a t NGF-R, MC1921gG. D e n s e positive nerve terminals w e r e f o u n d in t h e o l f a c t o r y glomerular layer, suprachiasmatic nucleus, olivary pretectal nucleus, lateral geniculate nucleus, superior colliculus, solitary tract nucleus, trigeminal spinal tract nucleus, gracile nucleus, cuneate n u c l e u s and d o r s a l h o r n of s p i n a l c o r d . A double-labeling of retrograde tracer t e c h n i q u e a nd an immunofluorescnt technique for NGF-R s h o w e d t h a t t h e p o s i t i v e nerve terminals in the olfactory glomerular layer originated f r o m NGF-R n e u r o n s in the diagonal band and substantia innominata.
REGULATORY ACTION OF ACIDIC FIBROBLAST GROWTH FACTOR ON FOOD INTAKE. YUTAKA OOMURA I , KAZU0 SASAKI 2 , KENJI SUZUKI "I , TADASHI MUT0 "I . KAZUMITSU HANAI *3 . IKU0 T00YAMA4 . AND HIROSHI KIMURA4 . IResearch Institute for Wakan-Yaku, ZScientific Instrument Center, Toyama Medical and Pharmaceutical University, Sugitani, Toyama ~30-01, 3Department of Biology, Faculty of Science, Kyushu University r Fukuoka 8121 and 4Institute 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 vivo 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/10 ul to investigate a direct effect of glucose on the release of aFGF in cerebrospinal 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 aFGF in cerebrospinal fluid increased 1000 times greater than that of the pre-infusion period 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, intracerebroventricular administrations of aFOF 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.
O P T I C A L R E C O R D I N G O F S Y N A P T I C P O T E N T I A L IN T H E E M B R Y O N I C C H I C K B R A I N S T E M S L I C E P R E P A R A T I O N USING A V O L T A G E - S E N S I T I V E DYE. HITOSHI KOMURO, YOKO MOMOSE, TETSURO S A K A I , A K I H I K O H I R O T A , K O H T A R O K A M I N O , D e p a r t m e n t of P h y s i o l o q y , T o k y o I k a - S h i k a D a i q a k u S c h o o l of M e d i c i n e , I - 5 - 4 5 Y u s h i m a , B u n k y o - k u , T o k y o 113, J a p a n . U s i n g a n o p t i c a l m e t h o d for m o n i t o r i n g e l e c t r i c a l a c t i v i t y , w e h a v e r e c o r d e d s y n a p t i c p o t e n t i a l f r o m t h e e m b r y o n i c c h i c k b r a i n stem. Vagus/brain stems were i s o l a t e d f r o m 6 t o 1 0 - d a y o l d c h i c k e m b r y o s a n d t h e n the s l i c e p r e p a r a t i o n s w i t h both the right and left vagus nerve fibers were prepared by transverse sectioning. The slice preparations were stained with a voltage-sensitive merocyanine-rhodanine d y e (NK2761). Voltage-related optical absorbance changes evoked by vagus nerve stimulation with depolarizing pulses using a suction e l e c t r o d e w e r e t r i e d t o r e c o r d s i m u l t a n e o u s l y f r o m 127 s e p a r a t e d a d j a c e n t l o c i in t h e p r e p a r a t i o n u s i n g a 12 x 1 2 - e l e m e n t p h o t o d i o d e a r r a y . Optical responses were d e t e c t e d f r o m l i m i t e d a r e a n e a r t h e d o r s a l s u r f a c e of the s t i m u l a t e d side. When r e l a t i v e l y l a r g e s t r e n g t h s of s t i m u l a t i n g c u r r e n t w e r e a p p l i e d , o p t i c a l c h a n g e s r e l a t e d to p o s t s y n a p t i c p o t e n t i a l w e r e d e t e c t e d . The postsynaptic potentialr e l a t e d o p t i c a l c h a n g e s a p p e a r e d t o b e c o n c e n t r a t e d in a s m a l l e r r e g i o n w i t h i n t h e v a g a l r e s p o n s e a r e a , a n d t h e y w e r e c o m p l e t e l y e l i m i n a t e d in t h e p r e s e n c e of k y n u r e n i c a c i d w h i c h is a b l o c k e r of g l u t a m a t e r g i c s y n a p t i c t r a n s m i s s i o n .
IMvlUNOHISTOO-IEMICAL LOCALIZATI(~ OF NERVE GROWI'H FACTOR RECEPTOR CONTAINING NERVE TERMINALS IN THE RAT CENTRAL NERVOUS SYSTEM MIWAKO SEKITANI 2, MIKAKO 1KEDA1 , YASUHIDE LEE 2 S A D A O SHIOSAKA 1 HIROSHI HATANAKA3 , AND M~SAYA TOHYAMA2,Department o f Neuroanatomy~Biomedical Research C e n t e r ~, 2nd Department of A n a t o m y 2, a nd Protein Research Center3,Osaka University Medical School,3-57-4 Nakanoshima Kitaku, Osaka,530 Japan. The distribution of n e r v e g r o w t h f a c t o r receptor (NGF-R) containing nerve terminals in the rat central nervous system was immunohistochemically analysed using a monoclonal antibody against r a t NGF-R, MC1921gG. D e n s e positive nerve terminals w e r e f o u n d in t h e o l f a c t o r y glomerular layer, suprachiasmatic nucleus, olivary pretectal nucleus, lateral geniculate nucleus, superior colliculus, solitary tract nucleus, trigeminal spinal tract nucleus, gracile nucleus, cuneate n u c l e u s and d o r s a l h o r n of s p i n a l c o r d . A double-labeling of retrograde tracer t e c h n i q u e a nd an immunofluorescnt technique for NGF-R s h o w e d t h a t t h e p o s i t i v e nerve terminals in the olfactory glomerular layer originated f r o m NGF-R n e u r o n s in the diagonal band and substantia innominata.
REGULATORY ACTION OF ACIDIC FIBROBLAST GROWTH FACTOR ON FOOD INTAKE. YUTAKA OOMURA I , KAZU0 SASAKI 2 , KENJI SUZUKI "I , TADASHI MUT0 "I . KAZUMITSU HANAI *3 . IKU0 T00YAMA4 . AND HIROSHI KIMURA4 . IResearch Institute for Wakan-Yaku, ZScientific Instrument Center, Toyama Medical and Pharmaceutical University, Sugitani, Toyama ~30-01, 3Department of Biology, Faculty of Science, Kyushu University r Fukuoka 8121 and 4Institute 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 vivo 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/10 ul to investigate a direct effect of glucose on the release of aFGF in cerebrospinal 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 aFGF in cerebrospinal fluid increased 1000 times greater than that of the pre-infusion period 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, intracerebroventricular administrations of aFOF 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.