Visualization of serotonin 1A receptor trafficking in neuron-like PC12 cells / 基础医学与临床

Objective To explore the mechanisms of trafficking and signaling of serotonin 1A receptor(5-HT_(1A))and its spatiotemporal distribution in living cells.Methods The mouse 5-HT_(1A) gene amplified by RT-PCR was recombined into pEGFP-N1 vector and the EGFP coding sequence was located in-frame at the C-terminal end of the 5-HT_(1A) receptor.The 5-HT_(1A)-EGFP was transfected into neuron-like PC12 cells as well as HEK293.The transfected cells were visualized using confocal microscopy,the mobility of 5-HT_(1A)-EGFP was monitored by live measurements and fluorescence recovery after photobleaching.Results The 5-HT_(1A) gene was identitical with the published gene sequence NM_008308.4 and a 5-HT_(1A)-EGFP fusion construct was created.After stable transfection of the plasimd into a PC12 cell line and analysis with a confocal laser scanning microscopy,the EGFP-tagged 5-HT_(1A) was predominantly associated with the plasma membrane,but some intracellular vesicles in the perinuclear region also contained the fusion protein.The predominant localization of 5-HT_(1A)-EGFP at the plasma membrane was confirmed in transiently transfected HEK293 cells.Bleached fluorescence was partialy recovered in 100 seconds,indicating that the 5-HT_(1A)-EGFP was mobiled on the membrane.Conclusion Spatiotemporal distribution and mobility of 5-HT_(1A) tagged with EGFP can be monitored in the 5-HT_(1A)-EGFP stable PC12 cell line,which could be an excellent neuron-like experimental cell model for research of 5-HT_(1A) trafficking and signaling.

Repair sciatic nerve gap of the rats with novel artifical nerve guide / 中国康复理论与实践

@# ObjectiveTo explore the possibility of repairing sciatic nerve gap of rats with artifical nerve graft.MethodsA novel artifical nerve guide was developed and used to suture the 15 milimeter long right sciatic nerve gap of 10 rats, other 7 rats were the control with the right sciatic nerve gap alone.2 and 4 monthes after operation, immunohistochemistry, Osmium staining, Bodian staining,motor end plate staining,WGA-HRP stain tracing have been done to observe the effect of repairing.Results2 months after operations, the sciatic nerve gap were repaired by the regeneration nerve.There was not evident inflammation in the defects.ConclusionsThe artifical nerve graft can induce the nerve to regenerate.

THE EXPRESSION OF AGRIN IN RAT CORTEX DURING THE POSTNATAL TIME / 解剖学报

Objective To observe the expression of heparan sulfate proteoglycan(HSPG)-Agrin in rat cortex of postnatal time in order to understand the relation between the development of cortex and agrin. Methods We performed immunohistochemical analysis of agrin expression in the postnatal rat cortex with agrin-specific polyclonal antibody. Results Agrin expression was shown to have significant differences in the different periods of postnatal time.It increased from postnatal 2 day(p2) to postnatal day 6(p6),then declined steadily after p6 and reached adult levels by 4w.In this periold of time,agrin has different expression peak following to the cortex layer development.Conclusion:Agrin is related to the development of cortex and may play an important role in the developing of central nervous system.;