RESUMEN
Objective To establish an animal behavioral model of tinnitus with step -down test ,and make use of the model to analyze the tinnitus genesis site in the salicylate-rats .Methods Forty wistar rats were includscl in this study ,they were randonly into 2 groups(experinent group and constrot group ,20 rats each group) .Ascertain that the animals treated with sodium salicylate(SS) have the sensation of tinnitus with the method of step -down test behavioral model .After having established a behavioral conditioned reflex ,subjects responded correctly to jump onto a climbing pole after hearing the sound .Then animals were under bilateral auditory nerves section ,to test whether the ablated-animals could still jump onto the pole after being treated with SS .Results Animals with the ab‐lating surgery could seldom jump onto the pole ,only reaching 1 .59 ± 0 .12 times average per 10 tests .Compared with con‐trol group ,there were significant differences with t-Test (P<0 .05) ,suggesting that subjects could not hear the tinnitus after the surgery .Conclusion Sodium salicylate could not make the animals with bilateral auditory nerves section to have the sensation of tinnitus .The peripheral auditory system plays an extremely important role in salicylate-induced tinnitus .
RESUMEN
OBJECTIVE@#To explore the methods of culture, identification and label of embryonic rat neural stem cells.@*METHOD@#The cells isolated from fetal rat hippocampus were identified with nestin immunocytochemical fluorescent staining. The cellular multiplication was observed by immunocytochemical fluorescence co-label after accession of BrDU. The neural stem cells (NSCs) were marked with fluorescent dye, bisbenzimide (Hoechest33342) and induced to differentiate. The differentiated cells were detected with Neuron Specific Enolase (NSE) and Glial Fibrillary Acidic Protein (GFAP) immunocytochemical fluorescent staining respectively.@*RESULT@#Nest-like clusters of neural stem cells were obtained in suspension and the cells could be differentiated into neurons and astrocytes which maintaining the main characteristics of NSCs after 8 passages of culture. The label efficiency of cells with Hoechest33342 was 97% and no attenuation of fluorescent brightness was observed after 8 passages of culture. The cellular fluorescence was observed in the NSCs and the differentiated cells.@*CONCLUSION@#The cells from embryonic rat hippocampus possessed the abilities of division, multiplication and self-renew, which were believed to be the main characteristics of NSCs of the central nervous system. The cells could be efficiently labeled with fluorescent dye and could be used as donor cells in experimental research on NSCs transplantation.
Asunto(s)
Animales , Femenino , Embarazo , Ratas , Técnicas de Cultivo de Célula , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Células Madre Embrionarias , Biología Celular , Células Madre Multipotentes , Células-Madre Neurales , Biología Celular , Neuronas , Biología Celular , Ratas Sprague-DawleyRESUMEN
AIM: To establish recombination plasmid pEGFP-NGB and to investigate the expression of pEGFP-NGB in culture neuroglia cells. METHODS: The NGB cds was isolated by using RT-PCR method with total RNA extracted from fetal Kunming mouse brain, then the NGB cds was cloned into the eukaryotic expression vector pEGFP-C1 of EGFP reported green fluorescence protein. The expression vector of recombinant plasmid pEGFP-NGB was successfully constructed. GeneJamer transfection reagent was used to transfer recombinant plasmid pEGFP-NGB into culture neuroglial cells. The mRNA and protein expression of pEGFP-NGB in culture neuroglial cells were investigated. RESULTS: The positive clone sequencing was consistent with the sequence of Genbank. The NGB mRNA and protein expression of pEGFP-NGB in culture neuroglial cells were detected at high levels. The high expression of green fluorescence protein was observed by fluorescence microscope in culture neuroglial cells. CONCLUSION: The expression vector of recombinant plasmid pEGFP-NGB was successfully constructed and green fluorescence protein was expressed in cultured neuroglial cells.