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Morphological study on neurotrophic action of gliostatin on cortical neurons in vitro
s199 12-13
THREE DIMENSIONAL CELL AGGREGATION EVOKES STRONG RESPONSE TO NGF IN HIDENORI HORIE AND YOSHIKO AKAHORI. Department. ADULT SENSORY NEURONS. 9-f pqsCq_~~_~~,~~Schqpl of Medicine, Yokohama CiLUniversityl__3-_9 Fukuura _L-_- Kana~_~_ _- ____ sayaku,Yokohama, 236,_Jap_ag, Dorsal root ganglion (DRG) explants and neurons in vitro have been thought to be receptive to the growth promoting activity of the NGF only during enhancement of a restricted period of their early embryonic life. However, neurite regeneration by NGF was clearly seen in aged DRG explants without nerve fibers. Collagenase-trypsin treatment further enhanced the response to regenerate many neurites all around the explants like embryonic ones. These results indicate that NGF could strongly activate capacity of neuronal regeneration in After dissociation with enzymes we aggreadult sensory neurons in the explant. gated highly purifjed isolated adult DRG neurons into thin thylindrical space in collagen gel. Three dimensional cell assembly evoked strong response to NGF to regenerate many neurites and promote neurite growth similar to the enzyme treated explants. This result suggests that three dimensional interaction among cells revives strong reaction to NCF in adult sensory neurons.
12-14 MORPHOLOGICAL STUDY ON NEUROTROPHIC ACTION OF GLIOSTATIN ON CORTICAL NEURONS IN VITRO. TAKATOSHI UEKI., KIYOFUMI ASAI AND TAIJI ment of BioreauJ_&.ion Research. Naaov!itv Universitv Medical School. 1 Kawasd Mizuho-Cho. Mizuho-Ku. Naaova 467Jam Gliostatin is a polypeptide factor, which is of an apparent Mr=lOO k with a homodimeric structure comprising two 50 kDa subunits, acting on cortical neurons (neurotrophic action) as well as astrocytic cells (growth inhibition). Under the coculture system of cerebral cortical neurons and astrocytes from fetal rats (El5 or E16), the neurotrophic action of gliostatin was examined immunocytochemically Inununostaining by an anti-neurofilament (NF) and electron-microscopically. monoclonal antibody visualized a marked neurite-outgrowth and interconnecting bundles of neuritic processes induced by gliostatin in the coculture system. Neurons stimulated by gliostatin were densely aggregated in clumps, while neurons in control cocultures spread out. Gliostatin has also shown survival-promoting effects on neurons. The electron-microscopic examination further substantiated the immunocytochemical observations. These suggest that gliostatin may play physiological roles on neuronal development through its neurotrophic action.
12-15 Pharmacolow,
KAINIC ACID INDUCES MASABUMI MINAMI, Facultv of Pharmaceutical
LEUKEMIA INHIBITORY FACTOR mRNA IN THE RAT BRAIN. KAZUKI YABUUCHI, MASAMICHI SATOH, Deuartment @ Sciences. Kyoto Universitv, Kvoto 606-01, Japan
Leukemia inhibitory factor (LIF) is a cytokine which has been reported to act on neuronal cells as a cholinergic differentiation factor. In this study, we examined the expression of LIF mRNA in the brain of both normal and kainic acid (KA)-treated rats. Male Sprague-Dawley rats were injected with KA (12 mg/kg i.p.) and with diazepam (10 mg/kg i.p.), in most cases, 4 hr after that to suppress extensive seizure activity. The rats were decapitated 2,4,8,24,48,72 hr after the injection of KA. The expression of LIF mRNA was examined using both northern blot analysis and in situ hybridization technique, but not detected in any regions of the normal rat brain. The northern blot analysis showed that the expression of LIF mRNA was observed highly in the hippocampus, moderately in the cerebral cortex (CC), thalamus (THL) and hypothalamus (HT), weakly in the striatum at 8 hr after the injection of KA. The expression of LIF mRNA was able to be detected at 2 hr, reached to the maximum at 8 or 24 hr and reduced to the undetectable levels at 72 hr. Although pretreatment with diazepam at the same dose 10 min before that of KA suppressed both seizure and the induction of LIF mRNA at 8 hr, the injection of diazepam 4 hr after that of KA hardly inhibited the expression of LIF mRNA. In situ hybridization histochemistry revealed that intense expression of LIF mRNA was observed in the lateral septum, hippocampal dentate gyrus, amygdala and meninges. Moderate signals were observed in the olfactory bulb, CC, THL and HT. These findings suggest that LIF might be produced in the brain.
THREE DIMENSIONAL CELL AGGREGATION EVOKES STRONG RESPONSE TO NGF IN HIDENORI HORIE AND YOSHIKO AKAHORI. Department. ADULT SENSORY NEURONS. 9-f pqsCq_~~_~~,~~Schqpl of Medicine, Yokohama CiLUniversityl__3-_9 Fukuura _L-_- Kana~_~_ _- ____ sayaku,Yokohama, 236,_Jap_ag, Dorsal root ganglion (DRG) explants and neurons in vitro have been thought to be receptive to the growth promoting activity of the NGF only during enhancement of a restricted period of their early embryonic life. However, neurite regeneration by NGF was clearly seen in aged DRG explants without nerve fibers. Collagenase-trypsin treatment further enhanced the response to regenerate many neurites all around the explants like embryonic ones. These results indicate that NGF could strongly activate capacity of neuronal regeneration in After dissociation with enzymes we aggreadult sensory neurons in the explant. gated highly purifjed isolated adult DRG neurons into thin thylindrical space in collagen gel. Three dimensional cell assembly evoked strong response to NGF to regenerate many neurites and promote neurite growth similar to the enzyme treated explants. This result suggests that three dimensional interaction among cells revives strong reaction to NCF in adult sensory neurons.
12-14 MORPHOLOGICAL STUDY ON NEUROTROPHIC ACTION OF GLIOSTATIN ON CORTICAL NEURONS IN VITRO. TAKATOSHI UEKI., KIYOFUMI ASAI AND TAIJI ment of BioreauJ_&.ion Research. Naaov!itv Universitv Medical School. 1 Kawasd Mizuho-Cho. Mizuho-Ku. Naaova 467Jam Gliostatin is a polypeptide factor, which is of an apparent Mr=lOO k with a homodimeric structure comprising two 50 kDa subunits, acting on cortical neurons (neurotrophic action) as well as astrocytic cells (growth inhibition). Under the coculture system of cerebral cortical neurons and astrocytes from fetal rats (El5 or E16), the neurotrophic action of gliostatin was examined immunocytochemically Inununostaining by an anti-neurofilament (NF) and electron-microscopically. monoclonal antibody visualized a marked neurite-outgrowth and interconnecting bundles of neuritic processes induced by gliostatin in the coculture system. Neurons stimulated by gliostatin were densely aggregated in clumps, while neurons in control cocultures spread out. Gliostatin has also shown survival-promoting effects on neurons. The electron-microscopic examination further substantiated the immunocytochemical observations. These suggest that gliostatin may play physiological roles on neuronal development through its neurotrophic action.
12-15 Pharmacolow,
KAINIC ACID INDUCES MASABUMI MINAMI, Facultv of Pharmaceutical
LEUKEMIA INHIBITORY FACTOR mRNA IN THE RAT BRAIN. KAZUKI YABUUCHI, MASAMICHI SATOH, Deuartment @ Sciences. Kyoto Universitv, Kvoto 606-01, Japan
Leukemia inhibitory factor (LIF) is a cytokine which has been reported to act on neuronal cells as a cholinergic differentiation factor. In this study, we examined the expression of LIF mRNA in the brain of both normal and kainic acid (KA)-treated rats. Male Sprague-Dawley rats were injected with KA (12 mg/kg i.p.) and with diazepam (10 mg/kg i.p.), in most cases, 4 hr after that to suppress extensive seizure activity. The rats were decapitated 2,4,8,24,48,72 hr after the injection of KA. The expression of LIF mRNA was examined using both northern blot analysis and in situ hybridization technique, but not detected in any regions of the normal rat brain. The northern blot analysis showed that the expression of LIF mRNA was observed highly in the hippocampus, moderately in the cerebral cortex (CC), thalamus (THL) and hypothalamus (HT), weakly in the striatum at 8 hr after the injection of KA. The expression of LIF mRNA was able to be detected at 2 hr, reached to the maximum at 8 or 24 hr and reduced to the undetectable levels at 72 hr. Although pretreatment with diazepam at the same dose 10 min before that of KA suppressed both seizure and the induction of LIF mRNA at 8 hr, the injection of diazepam 4 hr after that of KA hardly inhibited the expression of LIF mRNA. In situ hybridization histochemistry revealed that intense expression of LIF mRNA was observed in the lateral septum, hippocampal dentate gyrus, amygdala and meninges. Moderate signals were observed in the olfactory bulb, CC, THL and HT. These findings suggest that LIF might be produced in the brain.