实验仪器使用技术

CO2 fixation by the nervous system has been demonstrated with the use of 14C-labeled bicarbonate in the retina (Crane and Ball, 1951), in the brain of 1-d-old mice (Moldave et al., 1953), and in the adult cat brain (Berl et al., 1962a,b). When cat brain was perfused with labeled bicarbonate and the specific radi ...

The successful measurement of cerebral high-energy phosphates (ATP, ADP, AMP, and phosphocreatine) by the enzymatic techniques to be described in this chapter is dependent on: (1) adequate tissue fixation, (2) preparation, dissection, and weighing of samples to be analyzed, and (3) fluor ...

The purpose of this chapter is to describe the various methods that are presently available for the proper fixation of the brain in the investigation of cerebral energy metabolism. The advantages, disadvantages, and procedures of each method will be described in detail for those investiga ...

Under normal physiological conditions, cerebral function requires a continuous supply of glucose for its energetic and biosynthetic needs. Pyruvate oxidation via the pyruvate dehydrogenase enzyme complex (PDHC) represents a step of key importance in cerebral glucose utiliz ...

The present review focuses on the methodological aspects of glycolytic, tricarboxylic acid (TCA) cycle, and related enzymes, and on a discussion of how the methodologies can be usefully exploited in different experimental settings. Because of this primary focus and space limitation, t ...

The postulate that useful information about the activity of cerebral tissue can be deduced from a measure of energy metabolism, as reflected by the cerebral metabolic rate of glucose (CMRglc), is supported by a considerable body of data. Because there are adequate reviews of this subject (e.g., S ...

The brain is a complex, heterogeneous organ composed of many anatomical and functional components with markedly different levels of functional activity that vary independently with time and function. Other tissues are generally far more homogeneous, with most of their cells functi ...

The mammalian brain has very small reserves of carbohydrates, which under normal circumstances are the preferred sources of energy in vivo. Thus, the cerebral cortex contains glucose at l–l.5 μmol/g fresh weight and a little more of the equivalent amount of glucose stored as glycogen (McIlwain ...

Mammalian brain slices are used to study a broad range of cellular neural functions, and their use as a neuroscience tool has greatly expanded in the past decade, especially in the realm of electrophysiological determinations. Warburg (1923) initially developed tissue slice techniques ...

Until the mid-1960s, studies on the metabolism of brain mitochondria were hampered by the lack of suitable methodologies for brain mitochondrial isolation. In the majority of the studies up to this time, crude mitochondrial preparations were isolated from mammalian brain homogenates ...

The synapse plays pivotal roles in various neuronal functions, with its efficacy determining the opening and closing of neuronal circuits. Among the parameters that determine synaptic efficacy, presynaptic factors are of crucial importance. However, these factors are among those ...

Since late 1980s, the whole-cell patch-clamp technique has been used as a powerful tool for analyzing local circuits of the central nervous system in a brain slice preparation in which the fundamental architecture of local circuits is mostly maintained. When combined with intracellular s ...

The practical details of the “blind” patch-clamp recording method and its usefulness and applications to the analysis of synaptic transmission and plasticity in brain slice preparations are described. The blind patch-clamp method displays its greatest power in the experiments in wh ...

Patch-clamp is the best method for analyzing single-channel current activities today. The method is not good, however, at measuring the total number of channels expressed on the cell membrane that are responsible for the current of concern. In contrast, noise analysis has some advantage over ...

Analysis of data from single-channel studies can provide us with insights into the detailed physicochemical features of channel proteins, such as ion permeation rate, ion selectivity, and gating. However, single-channel analysis, particularly gating analysis, is sometimes time ...

The patch-clamp technique has revolutionized the study of membrane physiology, enabling unprecedented resolution in recording cellular electrical responses and underlying mechanisms. The perforated-patch variant of whole-cell patch-clamp recording was developed to ...

With the whole-cell mode of patch-clamp the membrane current is recorded from a cell under the voltage-clamp. The membrane potential response of a cell is also measured and manipulated under a current-clamp. The method enables one to correlate the macroscopic aspects of ion channels with their ...

The patch-clamp technique, invented by Erwin Neher and Bert Sakmann in 1976, was originally designed to detect the activity of single-ion channel proteins in the cell membrane. The technique is now widely used in the field of ion channel research for many purposes, such as to monitor changes in the tot ...

Solutions used in patch-clamp can be divided into two general categories by the side of the patch membrane with which the solution is in contact. The compositions of extracellular solutions are usually similar to those of the natural extracellular fluids, whereas intracellular solutio ...

Patch-clamp data, including channel currents and membrane voltage, are usually recorded to a computer via a process known as analog-to-digital conversion. This chapter gives essential information on how to avoid pitfalls and obtain error-free data with optimal resolution.