This chapter presents the histological assessment of cartilage and bone of tibial plateaus, by procedures that have been applied and validated in two animal models of osteoarthritis: meniscectomized rats and guinea pigs. It starts from bone sampling, followed by all the steps of sample pre ...

Most current approaches to the analysis of gene expression in arthritic tissue samples are based on RNA isolated either from cultured synovial cells or from synovial biopsies. However, this strategy does not distinguish between specific gene expression profiles of cells originating ...

The procedure described below is useful for extracting proteins, nucleic acids, and glycosaminoglycans from 5–40 mg of cartilage or tissue-engineered cartilage samples. This extraction method will generate samples compatible with Western blot, RNase protection, dimethyl me ...

Protein localization in cartilage sections by antibodies that specifically bind to epitopes of a protein is one of the most powerful technologies in modern cartilage research. Studies using two or more primary antibodies that recognize different protein epitopes allow the colocal ...

Cartilage functions as a low-friction, wear-resistant, load-bearing tissue. During a normal gait cycle, one cartilage surface rolls and slides against another, all the while being loaded and unloaded. The durability of the tissue also makes for an impressive material to study. However, wh ...

Genomic DNA in eukaryotic cells is packaged with histone and nonhistone proteins into chromatin structure. Both biochemical and genetic evidences indicate that chromatin structure imposes constraints on nuclear processes including transcription, replication, recomb ...

Thyroid hormone receptors (TRs) are capable of both activating and repressing transcription from genes bearing TR-binding elements (TREs). In general, TRs function as activators in the presence of thyroid hormone (T3) and repressors in the absence of T3 (1). The ability of TRs to regulate tran ...

Thyroid hormone receptors (TRs) play a major role in animal physiology. TRs are important and very interesting regulators of diverse aspects, including brain development, hearing, bone growth, morphogenesis, metabolism, intestine, and heart rate in vertebrates (Fig. 1). Aberrant fu ...

In mammals, thyroid hormones (TH) have been shown to control the postnatal development of many organs, such as brain, intestine and long bone, and to participate in the maintenance of homeostasis in adults by controlling basal metabolism, heart rate, and body temperature (1,2). To ensure this last ...

Thyroid hormone receptors (TRs) and retinoic acid receptors (RARs) are nuclear hormone receptors that play crucial roles in embryogenesis, cell proliferation, differentiation, and metabolism. TRs and RARs repress basal transcription in the absence of ligand and activate transc ...

Among the most dramatic actions of thyroid hormone are those exerted on brain development and function. In the adult human brain, a deficiency or excess of thyroid hormone may lead to various psychiatric manifestations, but it is during development when thyroid hormone exerts its most varied ...

The v-erbA oncogene is the most clear-cut case of an oncogene that acts by blocking a differentiation sequence (for a review, see ref. 1). It has been discovered as one of the two oncogenes carried by the avian erythroblastosis virus (AEV), a leukemia-inducing retrovirus. It is derived from the c-erbA pr ...

The transfection and overexpression of the cDNA encoding the thyroid hormone receptor (TR) in mammalian cells has shed light into several aspects of the function and biological characteristics of the TR in cells. Using this method, thyroid hormone and TR-responsive genes and response ele ...

Resistance to thyroid hormone (RTH) syndrome is an inherited human endocrine disease, which is manifested as a failure to respond properly to elevated circulating thyroid hormone (1-4). RTH syndrome behaves as an autosomal dominant trait, and has been mapped at the molecular level to a diverse ...

Thyroid hormone (T3) plays important roles during vertebrate development (1). In humans, T3 is detected in the embryonic plasma by 6 mo and rises to high levels around birth (2). During this postembryonic period, extensive tissue remodeling and organogenesis take place. T3 deficiency during ...

Recent molecular genetic studies in cardiac and skeletal muscle have revealed mutations in a battery of sarcomeric muscle-restricted genes that appear to be associated with various myopathies (1,2). In sharp contrast, no mutations in smooth muscle cell (SMC)-restricted genes have be ...

Protein kinases play pivotal roles in almost all signal transduction pathways in eukaryotic cells (1–4) and are implicated in most major human diseases, including atherosclerosis and associated vasculoproliferative disorders of arteries such as restenosis and graft stenosis ...

The development of new and effective techniques to study differential gene expression has revolutionized biomedical research during the last decade. Such techniques include differential display reverse transcription polymerase chain reaction (ddRTPCR) (see Chapter 9), f ...

The vast majority of past and present efforts in the molecular cloning of expressed sequences involve isolation of clones from cDNA libraries constructed in bacteriophage lambda (1,2). As discussed in Chapter 6, screening these cDNA libraries using labeled probes remains the most strai ...

The purpose of screening cDNA libraries is to isolate a particular cDNA clone encoding a mRNA and by implication, a protein, of interest. The screening is based on identification of the desired clone among a large number of recombinant clones within the library selected (1,2). As an example of both the u ...