Techniques in the neurosciences are evolving rapidly. There are currently very few volumes dedicated to the methodology - ployed by neuroscientists, and those that are available often seem either out of date or limited in scope. This series is about the methods most widely used by modern-day neuroscientists and is written by their colleagues who are practicing experts. Volume 1 will be useful to all neuroscientists since it concerns those procedures used routmely across the widest range of s- disciplines. Collecting these general techniques together in a single volume strikes us not only as a service, but will no doubt prove of exceptional utilitarian value as well. Volumes 2 and 3 describe all current procedures for the analyses of amines and theirmetabolites and of amino acids, respectively. These collections will clearly be of value to all neurosclentists working m or contemplating research in these fields. Similar reasons exist for Volume 4 on receptor binding techniques, since experimental details are provided for all types of ligand-receptor binding, including chapters on general principles, drug discovery and development, and a most useful appendix on computer programs for Scatchard, nonlinear, and competitive d- placement analyses. Volume 5 provides procedures for the asse- ment of enzymes involved in biogenic amine synthesis and catabolism. Volumes in the NEUROMETHODS series will be useful to neurochemists, -pharmacologists, -physiologists, -anatomlsts, psychopharmacologists, psychiatrists, neurologists, and chemists (organic, analytical, pharmaceutical, medicinal); in fact, everyone involved in the neurosciences, both basic and clinical.

Leading neuroscience researchers offer a fresh perspective on neuronal function by examining all its many components-including their pertubation during major disease states-and relate each element to neuronal demands. Topics range from the dependency of neurons on metabolic supply, as well as on both ion and transmitter homeostasis, to their close interaction with the myelin sheath. Also addressed are the astrocytic signaling system that controls synaptic transmission, the extracellular matrix and space as communication systems, the role of blood flow regulation in neuronal demand and in blood-brain barrier function, and inflammation and the neuroimmune system. Insightful and integrative, The Neuronal Environment: Brain Homeostasis in Health and Disease demonstrates a clear new understanding that neurons do not work in isolation, that they need constant interactions with other brain components to process information, and that they are not the only information processing system in the brain.

Continuing the research of the best-selling first edition, this second edition collects three more years of research in the ever-expanding study of the cell membrane. It covers the latest developments in the "traditional" patch techniques. This authoritative second edition updates the standard techniques while introducing three brand new, cutting-edge technical advances in the field.

Thorough and timely, this edition is an invaluable resource.

There is a perception in the scientific community that the discipline of Physiology is in crisis, or at least, in a phase of profound transition and change. At the root of the problem is confusion between objectives (the biological questions to be solved) and the methods and technologies to be applied. Traditionally, ever since Claude Bernard's concept of the "milieu interieur," Physiology was an integrative science with the prime concern of studying regulatory mechanisms leading to adaptation and homeostasis in the presence of challenges from a dynamic internal and external environment. This study of control mechanisms can be applied on any level of fu- tion whether subcellular, cellular, and organ, but reaches its highest level of complexity with the functioning of the body as a whole and its interaction with the external environment. This involves the determination of the interaction of genetic with environmental factors and the resulting integrated body adaptation. It might seem obvious that in the pursuit of these questions any appropriate combination of techniques on any organizational level could be used. Yet the advent of molecular techniques has resulted in a preoccupation with the problems and challenges inherent in these techniques, sometimes at the expense of the original perspectives and concepts. The many new mechanisms that have been discovered at the molecular level, as well as their economical exploitation, have contributed to a climate of reductionism.

Continuing the research of the best-selling first edition, this second edition collects three more years of research in the ever-expanding study of the cell membrane. It covers the latest developments in the "traditional" patch techniques. This authoritative second edition updates the standard techniques while introducing three brand new, cutting-edge technical advances in the field.

Thorough and timely, this edition is an invaluable resource.

There is a perception in the scientific community that the discipline of Physiology is in crisis, or at least, in a phase of profound transition and change. At the root of the problem is confusion between objectives (the biological questions to be solved) and the methods and technologies to be applied. Traditionally, ever since Claude Bernard's concept of the "milieu interieur," Physiology was an integrative science with the prime concern of studying regulatory mechanisms leading to adaptation and homeostasis in the presence of challenges from a dynamic internal and external environment. This study of control mechanisms can be applied on any level of fu- tion whether subcellular, cellular, and organ, but reaches its highest level of complexity with the functioning of the body as a whole and its interaction with the external environment. This involves the determination of the interaction of genetic with environmental factors and the resulting integrated body adaptation. It might seem obvious that in the pursuit of these questions any appropriate combination of techniques on any organizational level could be used. Yet the advent of molecular techniques has resulted in a preoccupation with the problems and challenges inherent in these techniques, sometimes at the expense of the original perspectives and concepts. The many new mechanisms that have been discovered at the molecular level, as well as their economical exploitation, have contributed to a climate of reductionism.

Leading neuroscience researchers offer a fresh perspective on neuronal function by examining all its many components-including their pertubation during major disease states-and relate each element to neuronal demands. Topics range from the dependency of neurons on metabolic supply, as well as on both ion and transmitter homeostasis, to their close interaction with the myelin sheath. Also addressed are the astrocytic signaling system that controls synaptic transmission, the extracellular matrix and space as communication systems, the role of blood flow regulation in neuronal demand and in blood-brain barrier function, and inflammation and the neuroimmune system. Insightful and integrative, The Neuronal Environment: Brain Homeostasis in Health and Disease demonstrates a clear new understanding that neurons do not work in isolation, that they need constant interactions with other brain components to process information, and that they are not the only information processing system in the brain.

E. Neher and B. Sakman were the first to monitor the opening and closing of single ion channels and membranes by conductance measurements. In 1976, they used firepolished micropipets with a tip diameter of 3-5 pm to record currents from a small patch of the membranbe of sk- etal muscles, thereby decreasing background membrane noise. In order to reduce the dominant source of background noise-the leakage shunt under the pipet rim between m- the muscle membrane had to be treated brane and gla- enzymatically. Despite these early limitations, a new te- nique was born -the patch-clamp technique. The final bre- through came in 1981 when the same authors, in collaboration with 0. P. Hamill, A. Marty, and F. J. Sigworth, developed the gigaohm seal. Not only did this improve the quality of recordings, it was now possible to gently pull the membrane patch with the attached pipet off the cell and study its trapped ion channels in isolation. Another offshoot of the gigaohm seal technique was the whole-cell patch-clamp technique, in which the patch is ruptured without breaking the seal. This technique is really a sophisticated voltage-clamp technique and also allows for the altering of cytoplasmic constituents if the experimenter so wishes. The first part of Patch-Clamp Applications and Protocols presents modern developments associated with the techn- ogy of patch-clamp electrodes, of cell-free ion channel reco- ing, and of the whole-cell patch-clamp technique.

Most cells will survive removal from the natural mic- environment of their in vivo tissue and placement into a sterile culture dish under optimal conditions. Not only do they survive, but they also multiply and express differen- ated properties in such a culture dish. A few cells do this in suspension, but most will need some kind of mechanical support substituting for their natural connections with other cells. The surface of a culture dish that might have to be coated is usually sufficient. The recent trend to standa- ization of conditions and the existence of commercial ent- prises with adequate funds and specializing in the needs of scientists were responsible for the tremendous proliferation of cell culture techniques in all fields of research in the last 20 years. No longer does a scientist have to concentrate all his/her efforts on that technology; the new trends make it feasible to employ cell culture techniques as only one of the many methods available in a small corner of a larger research laboratory. Some areas of research depend more heavily than others on cell culture techniques. Neuroscience is one of the areas that has developed hand in hand with the prol- eration of cell culture methodology. Molecular biological aspects, cell differentiation and development, neurophy- ological and neurochemical studies, as well as investigations into the nature of various diseases are now to a large extent dependent on the use of cell cultures.

Most cells will survive removal from the natural mic- environment of their in vivo tissue and placement into a sterile culture dish under optimal conditions. Not only do they survive, but they also multiply and express differen- ated properties in such a culture dish. A few cells do this in suspension, but most will need some kind of mechanical support substituting for their natural connections with other cells. The surface of a culture dish that might have to be coated is usually sufficient. The recent trend to standa- ization of conditions and the existence of commercial ent- prises with adequate funds and specializing in the needs of scientists were responsible for the tremendous proliferation of cell culture techniques in all fields of research in the last 20 years. No longer does a scientist have to concentrate all his/her efforts on that technology; the new trends make it feasible to employ cell culture techniques as only one of the many methods available in a small corner of a larger research laboratory. Some areas of research depend more heavily than others on cell culture techniques. Neuroscience is one of the areas that has developed hand in hand with the prol- eration of cell culture methodology. Molecular biological aspects, cell differentiation and development, neurophy- ological and neurochemical studies, as well as investigations into the nature of various diseases are now to a large extent dependent on the use of cell cultures.

This volume explores major light microscopic imaging modalities that can be used to view nervous tissue, and discusses the steps needed to use each of them, and ways to interpret the data. The chapters in this book cover topics such as atlasing of insect brain; neuroanatomical tracing through fluorochrome expression; fluorescent probes for amyloids; or optical clearing for ultramicroscopy of GFP- expressing tissues. In the Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory. Authoritative and cutting-edge, Neurohistology and Imaging Techniques is a valuable resource for both expert and novice users of major light microscopic imaging techniques, and those interested in exploring alternate imaging tools.

E. Neher and B. Sakman were the first to monitor the opening and closing of single ion channels and membranes by conductance measurements. In 1976, they used firepolished micropipets with a tip diameter of 3-5 pm to record currents from a small patch of the membranbe of sk- etal muscles, thereby decreasing background membrane noise. In order to reduce the dominant source of background noise-the leakage shunt under the pipet rim between m- the muscle membrane had to be treated brane and gla- enzymatically. Despite these early limitations, a new te- nique was born -the patch-clamp technique. The final bre- through came in 1981 when the same authors, in collaboration with 0. P. Hamill, A. Marty, and F. J. Sigworth, developed the gigaohm seal. Not only did this improve the quality of recordings, it was now possible to gently pull the membrane patch with the attached pipet off the cell and study its trapped ion channels in isolation. Another offshoot of the gigaohm seal technique was the whole-cell patch-clamp technique, in which the patch is ruptured without breaking the seal. This technique is really a sophisticated voltage-clamp technique and also allows for the altering of cytoplasmic constituents if the experimenter so wishes. The first part of Patch-Clamp Applications and Protocols presents modern developments associated with the techn- ogy of patch-clamp electrodes, of cell-free ion channel reco- ing, and of the whole-cell patch-clamp technique.

The human brain represents about 2% of the body weight, yet it accounts for approximately 20% of aerobic metabolism. This high dependency on energy-consuming processes is mainly caused by the active transport of ions, which is necessary to compensate for the transmembrane ion currents that are part ofthe complex signaling processes in the brain. Ninety-five percent ofthe brain's ATP is derived from mitochondrial oxidative phosphorylation. Since that organ' s storage capacity for oxygen is minimal, any interruption of oxygen delivery to brain cells willlead to changes in membrane excitability and, there fore, to disruption of neuronal signaling within seconds. It seems that mamma lian brain is especially vulnerable to such an interruption, since oxygen deprivation leads to activation of ion channel mechanisms in neurons that impair their communications. Thus, the function of the brain as a coordinator of vital homeostatic reflexes, and complex body reactions to external challenges, depends critically on the rate of oxygen delivery and oxygen consumption. Oxygen delivery depends on two variables described in the Fick relation ship: volume flow rate ofblood and the arterial oxygen content. A reduction in either of these two variables will have serious effects on vital brain func tions. Reduction of arterial blood flow to the brain can be caused by cardiac arrest, shock, carotid occlusion, Of hypotension (global ischemia). Oxygen content is progressively decreased in asphyxia (including drowning)."

The human brain represents about 2% of the body weight, yet it accounts for approximately 20% of aerobic metabolism. This high dependency on energy-consuming processes is mainly caused by the active transport of ions, which is necessary to compensate for the transmembrane ion currents that are part ofthe complex signaling processes in the brain. Ninety-five percent ofthe brain's ATP is derived from mitochondrial oxidative phosphorylation. Since that organ' s storage capacity for oxygen is minimal, any interruption of oxygen delivery to brain cells willlead to changes in membrane excitability and, there fore, to disruption of neuronal signaling within seconds. It seems that mamma lian brain is especially vulnerable to such an interruption, since oxygen deprivation leads to activation of ion channel mechanisms in neurons that impair their communications. Thus, the function of the brain as a coordinator of vital homeostatic reflexes, and complex body reactions to external challenges, depends critically on the rate of oxygen delivery and oxygen consumption. Oxygen delivery depends on two variables described in the Fick relation ship: volume flow rate ofblood and the arterial oxygen content. A reduction in either of these two variables will have serious effects on vital brain func tions. Reduction of arterial blood flow to the brain can be caused by cardiac arrest, shock, carotid occlusion, Of hypotension (global ischemia). Oxygen content is progressively decreased in asphyxia (including drowning)."

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