研究神经元分枝投射的荧光素双标记法

近代神经解剖学研究中,由于应用轴浆运输原理,开展了辣根过氧化物酶(HRP)及放射性同位素自显影(ARG)的技术后,进行了大量的工作并取得了许多成果。但对一个神经元的轴突分支投射到不同脑部的问题仍难以解决。

Hub Genes of Astrocyte Involved in Glaucoma with Ocular Hypertension by Integrated Bioinformatics Analysis

This study was conducted to elucidate the potential key candidate genes and pathways in role of astrocyte involved in glaucoma with ocular hypertension.Methods Expression profiles GSE2378 and GSE758 including 27 reactive optic nerve head astrocytes(ONHAs)by hypertensions and 26 normal controls,were integrated and deeply analyzed.Differentially expressed genes(DEGs)were sorted and candidate genes and pathways enrichment were analyzed.DEGs-associated protein-protein interaction network(PPI)was performed.Results A total of 119 consistently expressed genes were identified from 281 commonly changed DEGs,including 68 up-regulated genes and 51 down-regulated genes.PPI network complex filtered 75 DEGs(43 up-regulated and 32 down-regulated genes)of the 119 consistently altered DEGs and developed 117 edges,and 10 hub genes were identified.The most significant 3 modules were filtered from PPI,pathway enrichment analysis showed that module 1 was associated with extracellular exosome.Module 2 was mainly associated with antibody-dependent cellular cytotoxicity(ADCC)and module 3 was mainly associated with Hippo signaling pathway.Conclusion Taken above,using integrated bioinformatical analysis,we have identified DEGs candidate genes and pathways in role of astrocyte involved in glaucoma with ocular hypertension,which could improve our understanding of the cause and underlying molecular events,and these candidate genes and pathways could be therapeutic targets for glaucoma.

Neurogenesis by Activation of Inherent Neural Stem Cells in the Rat Hippocampus after Cerebral Infarction

Objective To investigate the changes of neural stem cells (NSCs) in the rat hippocampus after cerebral infarction (CI) and to evaluate the neurogenesis caused by the activation of NSCs. Methods CI models of rats were made and rats were assigned to 6 groups: sham-operated, 1 day, 3 days, 7 days, 14 days, and 28 days after CI. The dynamic expression of bromodeoxyuridine (BrdU), polysialylated neural cell adhesion molecule (PSA-NCAM), glial fibrillary acidic protein (GFAP), and neuronal nuclear antigen (NeuN) were determined by immunohistochemistry and immunofluorescence staining. BrdU was used to mark the proliferated NSCs. PSA-NCAM was used to mark the plasticity of activated NSCs. GFAP and NeuN were used to mark the differentiated NSCs. Results Compared with the controls, the number of BrdU+ cells in the hippocampus increased significantly at 1 day after CI (P<0.05), reached peak at 7 days after CI (P<0.05), decreased but still elevated compared with the controls at 14 days after CI (P<0.05), and nearly unchanged at 28 days after CI. The number of BrdU+/PSA-NCAM+ cells increased significantly at 7 days after CI (P<0.05), reached peak at 14 days after CI (P<0.05), and decreased but still elevated compared with the controls at 28 days after CI (P<0.05). The number of BrdU+/PSA-NCAM+ cells was equal to 60% of the number of BrdU+ cells in all the same period. The number of BrdU+/NeuN+ cells in the hippocampus increased significantly at 14 days after CI (P<0.05) and reached peak at 28 day after CI (P<0.05). The number of BrdU+/GFAP+cells in the hippocampus nearly unchanged after CI. Conclusion CI can stimulate the proliferation of inherent NSCs, and most proliferated NSCs may differentiate into neurons and represent neural plasticity.

The role of microglia in thalamic reticular nucleus in acupuncture regulating cognitive deficits in insomnia rats

Objective:To explore the mechanism of acupuncture in regulating cognitive deficits in insomnia rats by observing the effect of acupuncture on microglia in thalamic reticular nucleus(TRN).Methods:Thirty rats were randomly divided into a control group,a model group and an acupuncture group,with 10 rats in each group.The insomnia model was established by intraperitoneal injection of para-chlorophenylalanine(PCPA)once a day for 2 d.Rats in the control group were intraperitoneally injected with the same amount of normal saline.Rats in the acupuncture group received acupuncture at Neiguan(PC 6)and Zusanli(ST 36)for 5 consecutive days.The CLOCKLAB 2 data acquisition system was used to dynamically observe the sleep of the rats throughout the experiment.The cognition of rats was evaluated by event-related potentials(ERPs).After intervention,brain tissue was extracted.Immunofluorescence was used to test the fluorescence expression in TRN region.The concentrations of interleukin(IL)-1βand tumor necrosis factor(TNF)-αwere detected by enzyme-linked immunosorbent assay.Results:After intraperitoneal injection of PCPA suspension,the spontaneous activity in light period of rats in the model group and acupuncture group increased significantly compared with the control group(both P<0.01).After acupuncture treatment,the rats in the acupuncture group had much less spontaneous activity during the light period than those in the model group(P<0.01),and the results indicated that acupuncture could effectively improve the sleep quality of insomnia rats.Compared with the control group,rats in the model group showed that the P3 latency,the average optical density of microglia,and the concentrations of IL-1βand TNF-αincreased significantly(all P<0.05),and the P3 amplitude decreased significantly(P<0.01).Compared with the model group,rats in the acupuncture group presented that the P3 latency,the average optical density of microglia,and the concentrations of IL-1βand TNF-αwere significantly decreased(all P<0.05),and the amplitude of P3 was significantly increased(P<0.05).Conclusion:Acupuncture possesses an ability to improve the cognitive state in insomnia rats.The mechanism may be related to inhibiting the microglial activation,diminishing the levels of pro-inflammatory mediators like IL-1βand TNF-α,and promoting the recovery of central nervous system function.

Expression and role of PAK6 after spinal cord injury in adult rat

Objective： To observe p21-activated kinase 6 （PAK6） expression and its possible role after spinal cord injury （SCI） in adult rat.Methods： Sprague-Dawley rats were subjected to spinal cord injury. To explore the pathological and physiological significance of PAK6, the expression patterns and distribution of PAK6 were observed by Western blot, immunohistochemistry and immunofluorescence.Results： Western blot analysis showed PAK6 protein level was significantly up-regulated on day 2 and day 4,then reduced and had no up-regulation till day 14. Immunohistochemistry analysis showed that the expression of PAK6 was significantly increased on day 4 compared with the control group. Besides, double immunofluorescence staining showed PAK6 was primarily expressed in the neurons and astrocytes in the control group. While after injury, the expression of PAK6 was increased significantly in the astrocytes and neurons, and the astrocytes were largely proliferated. We also examined the expression of proliferating cell nuclear antigen （PCNA） and found its change was correlated with the expression of PAK6. Importantly, double immunofluorescence staining revealed that cell proliferation evaluated by PCNA appeared in many PAK6-expressing cells on day 4 after injury.Conclusion： The up-regulation of PAK6 in the injured spinal cord may be associated with glial proliferation.