实验仪器使用技术

Neuroproteomic technologies are crucial for a deeper understanding of molecular mechanisms underlying neuronal plasticity and pathologies of the central nervous system. For a comprehensive neuroproteomic analysis, high-resolution high-throughput techniques are i ...

Parkinson’s disease (PD) is characterized by the progressive loss of dopamine neurons. Here, we describe how to use human SH-SY5Y neuroblastoma cells as an in vitro cell model to study the effects of candidate PD susceptibility genes on dopamine neuron differentiation and viability. This ce ...

The covalent attachment of the small protein ubiquitin to other proteins is known to control a host of biological pathways and is emerging as an important regulatory factor in various processes specific to the nervous system. Ubiquitination is also tightly linked to most neurodegenerat ...

Recent progress in mass spectrometry allowed the identification of numerous proteins in complex soluble mixtures; however, the analysis of membranous subproteomes is still a challenging task. Integral membrane proteins, in particular those with more than one transmembrane dom ...

Protein phosphorylation plays an important role in biological process such as cell differentiation, cell cycle control, metabolism, and apoptosis. Toward global analysis of the phosphoproteome, enrichment is an essential step to overcome analytical challenges associated w ...

The 1D-PAGE/LC-ESI MS/MS approach is widely used for the qualitative and quantitative analysis of proteomes ranging from low to high complexity. As the first dimension of separation is based on SDS-PAGE, this method is compatible with the analysis of all classes of proteins including hydrop ...

The proteomic analysis at the peptide level is increasingly becoming a method of choice for complex samples. The success, however, depends on the development of attractive peptide fractionation methodologies to decrease the sample complexity prior to mass analysis. Recently, the OF ...

Various proteomic approaches are being applied in brain tumor proteomics with regard to targeted proteins of interest, to discover phenotype specific markers which could facilitate diagnosis as well as potential antitumor drug targets. iTRAQ technology is a multiplexing protein ...

Blue native polyacrylamide gel electrophoresis (BN-PAGE) is a powerful strategy for the separation of native multiprotein complex (MPC). Combined with other techniques, such as SDS–PAGE, immunoblotting, mass spectrometry, and antibody-shift assay, BN-PAGE has been widely used in ...

Neurons are highly polarized cells characterized by subcellular microdomains: the synapses. These compartments are specialized structures and are, for certain cellular pathways, independent from the cell body. To achieve such a functional specificity, including local mRNA tr ...

Plasma membrane (PM) proteins take center-stage in most of the fundamental processes of the nervous system. They impart specificity to the formation of neuronal circuits, and determine the mode of neurotransmission. To accomplish these tasks, they demonstrate spatially and tempora ...

A protocol presents a purification of postsynaptic density (PSD), from rat brain by subcellular fractionation using solubilization of membrane with Triton X-100 and sucrose density centrifugation. The protocol also includes purification of other synapse subdomains such as syn ...

Accurate annotation of protein identifications in organellar proteomics highly depends on the sample quality with special respect to contaminations from other subcellular compartments. In this respect, Free-flow electrophoresis (FFE) offers a valuable alternative to cla ...

Dissection of brain tissue is an important step in sample preparation for (subcellular) proteomics studies. In this chapter, brain removal and separate dissection of multiple brain regions from a single brain are described in step-by-step protocol. This concerns dissection from fresh ...

Neuroproteomics is a branch of proteomics that specifically studies qualitatively and/or quantitatively the tissue/organelle proteomes of the nervous system. This chapter introduces the various aspects of neuroproteomics, and outlines the range of methods that are commonly ...

Modeling human disease in animals has traditionally been used to elucidate pathogenesis and test promising new approaches to treatment. This approach has now identified a plethora of interventions that ameliorate functional deficits associated with spinal cord injury (SCI) in ro ...

The cat can recover a robust pattern of hindlimb locomotion following partial or complete spinal cord injuries. As a result, it has been instrumental in our understanding of spinal networks controlling and regulating locomotor activity. Thanks to our feline friend, spinal cord-injured ...

This chapter gives reasons for the choice of rabbit in experimental neurobiology and includes descriptions of both the spinal cord injury models and behavioral neuroscience methods. The models provide mechanisms for hypothesis generation that can be investigated at molecular and ...

Most animals of contused, compressed, or transected spinal cord injury require that a laminectomy is performed. However, increasing evidence suggests that laminectomies may induce undesirable effects including neuropathic pain, spinal instabilities, longer anesthetic a ...

Dorsal root injury provides opportunities for highly reproducible lesions and for detailed anatomical, physiological, and behavioral outcome assessment with high precision and validity. Dorsal root injury models are used to several aspects of relevance to spinal cord injury re ...