实验仪器使用技术

The pancreatic beta cells are highly sensitive to the glucose concentration, and the minute to minute regulation of insulin secretion by glucose occurs at the level of exocytosis of insulin, transcriptional rate of the insulin gene, translation of the mRNA, and processing of the proinsulin to ...

Regulated exocytosis is the process of secretion in specialized cells. Decades of intensive research have defined the molecules that drive the process of membrane fusion, as well as a plethora of accessory factors that shape and modulate the exocytotic function. As regulated exocytosis ...

Understanding how signaling molecules are released from cells is essential for furthering our knowledge of the basic biological mechanisms controlling many significant biological pathways. These molecules, including neurotransmitters, hormones, growth factors, and p ...

Exocytosis proceeds through distinct stages. Based on morphological and functional criteria, they can be classified as pre- and hemifusion, fusion, and postfusion. During the prefusion stage, plasma and vesicle membranes approach each other. During the hemifusion stage, the outer m ...

Neurons use Ca2+-triggered exocytosis of synaptic vesicles for interneuronal communication. SNARE proteins and fusion regulatory factors play key roles in the exocytosis. In vitro fusion assays using reconstituted proteoliposomes have been widely used for studying the funct ...

We describe here the potential of viral-mediated gene transfer for the modeling and treatment of Huntington’s disease, focusing in particular on strategies for the tissue-specific targeting of various CNS cells. The protocols described here cover the design of lentiviral vectors, st ...

The use of viral-mediated gene transfer in animal behavioral experiments has become very popular over the last decade. Altering gene regulation and assessing subsequent behavior can be a useful tool in unraveling a specific gene’s contributions to complex behavioral processes such as ...

Gene therapy is a promising future tool for treatment of Parkinson’s disease (PD) and several different strategies are currently being evaluated. Although many of these strategies have shown promising results in animal models of PD and parkinsonian patients, some have been less effecti ...

Although our understanding of the mechanisms that underlie drug addiction have advanced significantly over the past few decades, the lack of efficacious treatments for this debilitating disorder suggest that there is much work still to be done to clarify the role of specific cells and circ ...

The ability to identify neurons that are involved in the acquisition and encoding of a specific memory can facilitate efforts to understand the neuronal and circuit mechanisms that underlie memory formation. In this chapter, we outline a method whereby a population of neurons in the lateral a ...

Neurons display strong plasticity of their cellular properties, especially by modulation of their synaptic inputs. The development of viral vectors as biological tools has been extremely useful to molecularly manipulate protein expression in the rodent brain for a better underst ...

Alphavirus vectors have been developed for transient gene expression. The most common approach has been to generate replication-deficient vectors based on Semliki Forest virus, Sindbis virus, or Venezuelan equine encephalitis virus. All these alphavirus vectors have demonst ...

Herpes simplex virus type 1 (HSV-1) is a neurotropic virus displaying several outstanding adaptations to the nervous system, and each of them can be rationally exploited in the design of gene therapy vectors with regard to neurological applications. Replication-competent attenuat ...

Viral vectors based on recombinant adeno-associated virus have gained increasing interest over the last two decades as promising delivery vehicles in gene therapy. This enthusiasm is based on their ability to infect a broad range of tissues including proliferative and quiescent cell ...

Recombinant adenoviruses have been extensively applied in basic research and gene therapy. Recently, there is an increase in neurobiological studies using recombinant adenovirus technology to manipulate gene expression in neurological systems. Recombinant adenovirus ...

Viral vectors are now common in contemporary neuroscience research and their use as gene transfer tools for the central nervous system has seen an enormous growth in the last 2 decades. This chapter discusses about designing, production, and use of lentiviral vectors (LVs), one of the most popul ...

Motor neuron diseases such as amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA) are characterized by the progressive loss of motor neurons in the spinal cord and primary motor cortex. Subsequent paralysis of skeletal muscles leads to variable degrees of motor impai ...

Loss of function is a standard approach to elucidate the function of a specific protein. Among these multiple strategies for silencing genes in living animals, genetic knockout in mice have been so far most frequently used. However, short hairpin RNAs (shRNAs) and microRNAs (miRNAs) deliver ...

Neural stem/precursor cells (NPCs) are defined according to their ability to self-renew and differentiate into all the major cellular types of the central nervous system (CNS). They can survive and proliferate in defined culture conditions as spherical aggregates (neurospheres) for a ...

Optogenetics is a technical methodology that allows direct light-based manipulation of genetically specified cells. Optogenetic methods have provided novel insights into the role of defined neuronal populations in brain function and animal behavior. An expanding palette of s ...