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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 ...

In this post-genomic era, emphasis has shifted from identifying genes to understanding the physiological functions of gene products and their implications in human diseases. The use of transgenic mice is one of the key approaches in elucidating gene function and regulation. Transgenic ...

Models of acute nociception using a thermal stimulus are widely employed as screening methods for nociceptive properties of new drug compounds. In this chapter, detailed descriptions for conducting of two of the most commonly used models; the hot plate test and the “Hargreaves test,” are des ...

Gene targeting in mice by homologous recombination is a powerful approach to study the role of specific genes in vivo. This technology is now applied to pain-related genes to understand molecular mechanisms of nociceptive behaviors. In this chapter, we provide detailed methodological i ...

Pancreatic β-cells, responsible for the synthesis and secretion of insulin in response to a glucose challenge, are located in the islets of Langerhans. Islets are comprised of a heterogeneous population of endocrine cells, including insulin-producing β-cells (approx. 65–70%), gluca ...

The β-cell is receptive to intricate hormonal, neuronal and nutrient signaling which is key for normal physiology but complicates the study of specific effects of individual factors on β-cell function. To preserve the microenvironment of the β-cell, most studies of β-cell physiology have ...

Insulin is synthesized by the β cells of the pancreatic islets as part of a single 110-amino acid precursor, preproinsulin (see Fig. 1). Processing is initiated by removal of the amino terminal, 24-amino acid signal sequence (1). The resulting 86-amino acid product folds through the formation of th ...

The placenta is a specialized pregnancy-specific structure that develops concurrently with development of the embryo and fetus. From an evolutionary perspective, the placenta was the essential factor in permitting viviparity, a reproductive strategy in which fetal developme ...

This chapter describes methods used to investigate implantation in the common marmoset monkey, Callithrix jacchus. A reverse-transcriptase polymerase chain reaction-strategy with which to detect transcripts for steroid receptors and enzymes involved in estradiol biosy ...

For age- and sex-matched subjects, osteoporotic bone is more fragile than healthy bone. Vibrational infrared spectroscopy and in particular infrared microspectroscopic imaging is a useful tool for investigating and characterizing changes associated with metabolic bone dis ...

Analysis of X-chromosome inactivation patterns can be a useful tool in the identification of carriers of certain X-linked diseases and also for other investigations, such as gene mapping and clonality analysis. X inactivation is the process in females whereby one of the two X chromosomes of a ce ...

Every genetic laboratory diagnosis carries some degree of uncertainty, even if only the risk of laboratory error. Increasingly, diagnosis is based on direct tests that tell us whether or not the patient carries a given mutation. In these cases formal risk analysis is scarcely necessary—alt ...

The hemoglobinopathies are a diverse group of inherited recessive disorders consisting of the structural hemoglobin variants and the thalassemias. They can occur at very high carrier frequencies in the malarious regions of the world and are regionally specific, with each population ...

The culture of chondrocytes is one of the most powerful tool for exploring the intracellular and molecular features of chondrocyte differentiation and activation. However, chondrocytes tend to dedifferentiate to fibroblasts when they are subcultured, which is a major problem. This ...