Lipolysis involves the sequential breakdown of triglycerides into free fatty acids and glycerol. The extent of lipolysis is therefore a key determinant of the energy status of an individual and also dictates insulin resistance. Here, we describe a protocol for estimating lipolysis in mu ...

Glucose is the main metabolic fuel in mammalian cells. Glucose entry into cells is facilitated by a family of ubiquitously expressed glucose transporter proteins. Typically, glucose transporters are localized on the plasma membrane. One notable exception is the glucose transporter ...

Adipogenesis is an important component of adipose tissue development and growth. Alterations in adipogenesis may promote adipose tissue insulin resistance and inflammation. The ability of preadipocytes to differentiate into mature adipocytes depends on the activation of ph ...

Adipose tissue plays a critical role in energy homeostasis, not only in storing triglycerides, but also in responding to nutrient, neural, and hormonal signals, and secreting adipokines that control feeding, thermogenesis, immunity, and neuroendocrine function. It is conceivable ...

In vivo adipose tissue quantification is an important tool to characterize phenotypes of obesity, especially in the human. The amount and distribution of adipose tissue is associated with many of the adverse consequences of obesity. Recent studies suggest that adipose tissue is not a sing ...

Brown and white adipose tissue have recently gained prominence as key players in obesity and related health problems, such as type-2 diabetes and cardiovascular disease. Brown adipose tissue-dependent nonshivering thermogenesis significantly affects the body's energy bala ...

Angiogenesis is required for the growth and expansion of both healthy and pathological tissues. The plasticity of the adipose tissue is reflected by its remarkable ability to expand or to reduce in size throughout the adult lifespan. We, and others, have recently shown that expansion of fat mass is ...

Regulation of blood flow in tissues such as skeletal muscle, liver, and adipose tissue is needed to meet the changing local metabolic and physiological demands under varying conditions. In healthy individuals, adipose tissue blood flow (ATBF) is remarkably responsive to meal ingestion, ...

Brown and white adipose tissues in mammals have a number of similar properties, such as lipid storage and adipokine production, but also distinctive properties. The energy-storing white adipose tissue has few mitochondria and low oxidative capacity. The heat-producing brown adipose ...

Role of specific reactive lipids as well as amino acids in control of insulin signalling in adipose tissue is well recognized. Since it is practically impossible to measure the levels of all metabolites in the biological sample simultaneously with a single analytical platform, we utilize mu ...

The spinal cord represents a complex system that serves in the encoding of sensory information and organization of autonomic and somatomotor outflow. As such, it has become a target of investigation for subjects ranging from pain to hypertension to motor spasticity. A primary method in such i ...

The electrical signal associated with nerve cells, mainly as a result of changes in the membrane potential during functional activity, can be recorded extracellularly to study central mechanisms underlying sensory processing. The secondary neurons in the spinal trigeminal comp ...

The dorsal horn of the spinal cord is a key relay in the transmission of sensory information to the brain. Furthermore, this circuitry of spinal-cord neurons, and hence the spinal processing of sensory information, is subject to a great deal of plasticity, both pharmacological and physiologic ...

This chapter focuses on in vivo and in vitro recording setups of extracellular single-unit recordings of peripheral sensory nerve or dorsal root fibers in rodents. Extracellular single-unit recording methods have been used to obtain a wealth of data about the properties of peripheral ne ...

We describe methods for labeling and identifying neurons within the central nervous system involved in the transmission of nociceptive stimuli. The most reliable methods are physiological identification followed by intracellular injection or immunocytochemical detec ...

There is increasing use of isolated sensory neuronal preparations to examine the cellular mechanisms involved in pain signaling. Indeed, these in viro preparations have several advantages that make them beneficial for examining physiological and/or pathological processes a ...

Generating gene-expression profiles from laser-captured cells requires the successful combination of laser-capture microdissection, RNA extraction, RNA amplification, and microarray analysis. To permit single-cell gene-expression profiling, the RNA amplificat ...

Primary cultures of neurons provide opportunities to study the cell biology of neurons under controlled conditions. Because differences exist in cellular properties among populations of neurons in the brain, survival requirements for neurons among these regions differ as well. T ...

In this chapter we cover what we have found to be a “best practice” for real-time polymerase chain reaction (PCR) for relative mRNA quantification. We describe our techniques for tissue-sample collection and freezing, sample handling for quick and reproducible extraction of total RNA, fir ...

Hybridization array technology makes it possible to compare global gene-expression patterns in any experimental context for which good-quality RNA can be generated. To date, DNA arrays have been used as a tool to compare functional genomic changes (differences in wholesale gene expre ...