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A new co-culture method for identifying synaptic adhesion molecules involved in synapse formation

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Biophysics Reports 2019年第2期5卷 91-97页

作者： Wei Jiang Jihong Gong Yi Rong Xiaofei Yang Key Laboratory of Cognitive Science Hubei Key Laboratory of Medical Information Analysis and Tumor Diagnosis& Treatment Laboratory of Membrane Ion Channels and Medicine College of Biomedical Engineering SouthCentral University for Nationalities College of Biology Hunan University Key Laboratory of Molecular Biophysics of Ministry of Education College of Life Science and Technology Huazhong University of Science and Technology

The proper formation of synapses is essential for brain function. Synaptic cell adhesion molecules(CAMs) are thought to play essential roles in the initiation of the synapse formation process. The artificial synapse formation assay, in which non-neuronal cells and neurons are co-cultured, has been shown to be a powerful system for screening CAMs. However, controlling a large number of cell pools in co-culture is complicated, creating a potential barrier for high-throughput screening. This protocol describes a new co-culture method in which cDNA plasmid is transfected into human embryonic kidney 293T cells using polyetherimide 24 h after cells were mixed with neurons, and immunostaining and confocal imaging are employed for analyzing synaptogenesis. This optimized method is simpler and easier to perform than the traditional method for the examination of the synaptogenic activities of individual cell-surface proteins in isolation, and provides an unbiased screening platform for synaptogenic proteins.

关键词： Co-culture assay synapse formation Synaptic cell adhesion molecules Polyetherimide

The importance of fasciculation and elongation protein zeta-1 in neural circuit establishment and neurological disorders

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Neural Regeneration Research 2022年第6期17卷 1165-1171页

作者： Rafhanah Banu Bte Abdul Razar Yinghua Qu Saravanan Gunaseelan John Jia En Chua Department of Physiology Yong Loo Lin School of MedicineNational University of SingaporeSingaporeSingapore LSI Neurobiology Programme National University of SingaporeSingaporeSingapore Institute for Health Innovation and Technology National University of SingaporeSingaporeSingapore Healthy Longevity Translational Research Program Yong Loo Lin School of MedicineNational University of SingaporeSingaporeSingapore Institute of Molecular and Cell Biology Agency for ScienceTechnology and Research(A*STAR)SingaporeSingapore

The human brain contains an estimated 100 billion neurons that must be systematically organized into functional neural circuits for it to function properly.These circuits range from short-range local signaling networks between neighboring neurons to long-range networks formed between various brain regions.Compelling converging evidence indicates that alterations in neural circuits arising from abnormalities during early neuronal development or neurodegeneration contribute significantly to the etiology of neurological disorders.Supporting this notion,efforts to identify genetic causes of these disorders have uncovered an over-representation of genes encoding proteins involved in the processes of neuronal differentiation,maturation,synaptogenesis and synaptic function.Fasciculation and elongation protein zeta-1,a Kinesin-1 adapter,has emerged as a key central player involved in many of these processes.Fasciculation and elongation protein zeta-1-dependent transport of synaptic cargoes and mitochondria is essential for neuronal development and synapse establishment.Furthermore,it acts downstream of guidance cue pathways to regulate axo-dendritic development.Significantly,perturbing its function causes abnormalities in neuronal development and synapse formation both in the brain as well as the peripheral nervous system.Mutations and deletions of the fasciculation and elongation protein zeta-1 gene are linked to neurodevelopmental disorders.Moreover,altered phosphorylation of the protein contributes to neurodegenerative disorders.Together,these findings strongly implicate the importance of fasciculation and elongation protein zeta-1 in the establishment of neuronal circuits and its maintenance.

关键词： fasciculation and elongation protein zeta-1 neurological disorder neuronal development neuronal differentiation neuronal networks synapse formation synaptic function

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Glial cells in neuronal development:recent advances and insights from Drosophila melanogaster

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Neuroscience Bulletin 2014年第4期30卷 584-594页

作者： Jiayao Ou Yijing He Xi Xiao Tian-Ming Yu Changyan Chen Zongbao Gao Margaret S.Ho Department of Anatomy and Neurobiology School of MedicineTongji University School of Life Sciences Tongji University

Gila outnumber neurons and are the most abundant cell type in the nervous system. Whereas neurons are the major carriers, transducers, and processors of information, glial cells, once considered mainly to play a passive supporting role, are now recognized for their active contributions to almost every aspect of nervous system development. Recently, insights from the invertebrate organism Drosophila melanogaster have advanced our knowledge of glial cell biology. In particular, findings on neuron-glia interactions via intrinsic and extrinsic mechanisms have shed light on the importance of gtia during different stages of neuronal development. Here, we summarize recent advances in understanding the functions of Drosophila glia, which resemble their mammalian counterparts in morphology and function, neural stem-cell conversion, synapse formation, and developmental axon pruning. These discoveries reinforce the idea that glia are substantial players in the developing nervous system and further advance the understanding of mechanisms leading to neurodegeneration.

关键词： glia neuronal development Gcm neurodegeneration neural stem cell synapse formation axon pruning

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ntercellular protein-protein interactions at synapses

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Protein & Cell 2014年第6期5卷 420-444页

作者： Xiaofei Yang Dongmei Hou Wei Jiang Chen Zhang Key Laboratory of Cognitive Science Laboratory of Membrane Ion Channels and Medicine College of Biomedical Engineering South-Central University for Nationalities Wuhan 430074 China State Key Laboratory of Biomembrane and Membrane Biotechnology College of Life Sciences Peking University Beijing 100871 China PKU-IDG/McGovern Institute for Brain Research Peking University Beijing 100871 China

Chemical synapses are asymmetric intercellular junc. tions through which neurons send nerve impulses to communicate with other neurons or excitable cells. The appropriate formation of synapses, both spatially and temporally, is essential for brain function and depends on the intercellular protein-protein interactions of cell adhesion molecules （CAMs） at synaptic clefts. The CAM proteins link pre- and post-synaptic sites, and play essential roles in promoting synapse formation and maturation, maintaining synapse number and type, accumulating neurotransmitter receptors and ion chan- nels, controlling neuronal differentiation, and even regulating synaptic plasticity directly. Alteration of the interactions of CAMs leads to structural and functional impairments, which results in many neurological disorders, such as autism, Alzheimer＇s disease and schizophrenia. Therefore, it is crucial to understand the functions of CAMs during development and in the mature neural system, as well as in the pathogenesis of some neurological disorders. Here, we review the function of the major classes of CAMs, and how dysfunction of CAMs relates to several neurological disorders.

关键词： synapse formation cell-cell adhesion,synaptic plasticity neurological disorders protein-protein interaction cell adhesion molecules

Reduced npcl in nerve terminals produces defects in synaptic exocytosis and retrieval

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Reduced npcl in nerve terminals produces defects in synaptic...