scholarly journals Spatiotemporally resolved subcellular phosphoproteomics

2021 ◽  
Vol 118 (25) ◽  
pp. e2025299118
Author(s):  
Yanjun Liu ◽  
Ruxin Zeng ◽  
Ruixuan Wang ◽  
Yicheng Weng ◽  
Ruixiang Wang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Underlying Mechanism ◽  
Living Cells ◽  
Light Illumination ◽  
Primary Neuron ◽  
Quantitative Mass Spectrometry ◽  
Neuron Culture ◽  
Biotin Ligase ◽  
Primary Neuron Culture ◽  
Rapid Activation

Proteome-wide profiling of protein phosphorylation has been widely used to reveal the underlying mechanism of diverse cellular signaling events. Yet, characterizing subcellular phosphoproteome with high spatial–temporal resolution has remained challenging. Herein, we developed a subcellular-specific uncaging-assisted biotinylation and mapping of phosphoproteome (SubMAPP) strategy to monitor the phosphorylation dynamics of subcellular proteome in living cells and animals. Our method capitalizes on the genetically encoded bioorthogonal decaging strategy, which enables the rapid activation of subcellular localized proximity labeling biotin ligase through either light illumination or small-molecule triggers. By further adopting an integrated orthogonal pull-down strategy with quantitative mass spectrometry, SubMAPP allowed for the investigation of subcellular phosphoproteome dynamics, revealing the altered phosphorylation patterns of endoplasmic reticulum (ER) luminal proteins under ER stress. Finally, we further expanded the scope of the SubMAPP strategy to primary neuron culture and living mice.

Effects of hypoxic preconditioning on the expression of iron influx and efflux proteins in primary neuron culture

2012 ◽  
Vol 60 (4) ◽  
pp. 335-343 ◽  
Author(s):  
Fang Du ◽  
Ming Fan ◽  
Qi Gong ◽  
Ling Ling Zhu ◽  
Zhou-Jing Zhu ◽  
...  
Keyword(s):  
Hypoxic Preconditioning ◽  
Primary Neuron ◽  
Neuron Culture ◽  
Primary Neuron Culture ◽  
Efflux Proteins

scholarly journals Primary Neuron Culture for Nerve Growth and Axon Guidance Studies in Zebrafish (Danio rerio)

PLoS ONE ◽  
2013 ◽  
Vol 8 (3) ◽  
pp. e57539 ◽  
Author(s):  
Zheyan Chen ◽  
Han Lee ◽  
Steven J. Henle ◽  
Thomas R. Cheever ◽  
Stephen C. Ekker ◽  
...  
Keyword(s):  
Axon Guidance ◽  
Danio Rerio ◽  
Primary Neuron ◽  
Neuron Culture ◽  
Nerve Growth ◽  
Primary Neuron Culture

Neurosphere Development from Hippocampal and Cortical Embryonic Mixed Primary Neuron Culture: A Potential Platform for Screening Neurochemical Modulator

2018 ◽  
Vol 9 (11) ◽  
pp. 2870-2878 ◽  
Author(s):  
Juhee Khan ◽  
Gaurav Das ◽  
Varsha Gupta ◽  
Saswat Mohapatra ◽  
Subhajit Ghosh ◽  
...  
Keyword(s):  
Primary Neuron ◽  
Neuron Culture ◽  
Primary Neuron Culture

Toward a Merged Microfluidic/Microelectrode Array Device for Culture of Unidirectional Neural Network

2010 ◽  
Vol 1272 ◽  
Author(s):  
Alexander Mo ◽  
Sarah Heilshorn
Keyword(s):  
Neural Network ◽  
Soft Lithography ◽  
Microelectrode Array ◽  
Primary Neurons ◽  
Primary Neuron ◽  
Neuron Culture ◽  
Initial Validation ◽  
Primary Neuron Culture ◽  
Neural Cultures ◽  
Two Populations

AbstractTraditional neural cultures are formed from disassociating primary neurons that are sourced from animals. However by disassociating them, any larger organizational structure between the neurons is lost. In an effort to regain some of the lost organization a device that is capable of recreating and monitoring a unidirectional connection between two populations is proposed. Initial validation toward a fully functional device is presented. Using soft lithography techniques, a microfluidic chip has been designed and produced with a design conducive for facilitating such a neuron culture. Using a specialized adhesive method, this chip can be attached and removed from a commercially available microelectrode array. Finally, cell viability is demonstrated using the PC12 neuron-like cell line after exploring several device configurations. Further efforts will be focused on primary neuron culture and establishment of a unidirectional network.

scholarly journals A Primary Neuron Culture System for the Study of Herpes Simplex Virus Latency and Reactivation

10.3791/3823 ◽  
2012 ◽  
Author(s):  
Mariko Kobayashi ◽  
Ju-Youn Kim ◽  
Vladimir Camarena ◽  
Pamela C. Roehm ◽  
Moses V. Chao ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Culture System ◽  
Primary Neuron ◽  
Neuron Culture ◽  
Virus Latency ◽  
Primary Neuron Culture ◽  
Simplex Virus

scholarly journals Intracellular proteome compartmentalization: a biotin ligase-based proximity labeling approach

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Olanrewaju Ayodeji Durojaye
Keyword(s):  
Mass Spectrometry ◽  
Underlying Mechanism ◽  
Cellular Structures ◽  
Biological Processes ◽  
Interacting Proteins ◽  
Cellular Processes ◽  
Biotin Ligase ◽  
Labeling Approach ◽  
Cellular Compartments

AbstractSpecialized biological processes occur in different regions and organelles of the cell. Additionally, the function of proteins correlate greatly with their interactions and subcellular localization. Understanding the mechanism underlying the specialized functions of cellular structures therefore requires a detailed identification of proteins within spatially defined domains of the cell. Furthermore, the identification of interacting proteins is also crucial for the elucidation of the underlying mechanism of complex cellular processes. Mass spectrometry methods have been utilized systematically for the characterization of the proteome of isolated organelles and protein interactors purified through affinity pull-down or following crosslinking. However, the available methods of purification have limited these approaches, as it is difficult to derive intact organelles of high purity in many circumstances. Furthermore, contamination that leads to the identification of false positive is widespread even when purification is possible. Here, we present a highlight of the BioID proximity labeling approach which has been used to effectively characterize the proteomic composition of several cellular compartments. In addition, an observed limitation of this method based on proteomic spatiotemporal dynamics, was also discussed.

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