细胞功能测定

Peripheral adipose tissue contains a population of clonogenic precursor cells referred to as adipose-derived stem cells (ASC) that retain the capacity to differentiate into multiple cell types including osteoblasts, adipocytes, chondrocytes, myocytes, and neuronal cells fo ...

The following chapter describes several methods involved in the detection of MAPK activities and phosphorylated proteins during early development of Xenopus laevis. The Xenopus embryo provides a powerful platform for biochemical studies. We describe here basic methods of embryo ...

The technique of single muscle-fiber cultures has already proven valuable in extending knowledge of myogenesis, stem cell heterogeneity, the stem cell niche in skeletal muscle, and satellite cell activation. This report reviews the background of the model and applications, and detai ...

Phase specificity, the temporal and tissue restriction of teratogen-induced defects during embryonic �development, is a poorly understood but common property of teratogens, an important source of human birth defects. Somite counting and somite units are novel chronometric too ...

Caenorhabditis elegans is a premier model genetic system for discovering new information about the assembly and maintenance of striated muscle. The localization of a protein within a nematode muscle cell can reveal important clues to its function. In C. elegans, proteins can be localized by ...

The Drosophila system has been invaluable in providing important insights into mesoderm specification, muscle specification, myoblast fusion, muscle differentiation, and myofibril assembly. Here, we present a series of Drosophila protocols that enable the researcher to vis ...

During myogenesis, cells gradually transition from mesodermal precursors to myoblasts, myocytes, and then to muscle fibers. The molecular characterization of this process requires the ability to identify each of these cell types and the factors that regulate the transitions betwe ...

Resistance loading provides an important tool for understanding skeletal muscle responses and adaptations to various perturbations. A model using anesthetized rodents provides the means to control the input parameters carefully, and to measure the output parameters of each mus ...

Plasmid DNA electrotransfer is a direct method of gene delivery to skeletal muscle commonly used to identify endogenous signaling pathways that mediate muscle remodeling or pathological states in adult rodents. When plasmids encoding a protein to be overexpressed are fused to a fluor ...

Skeletal muscle is a highly adaptive tissue that modifies its size in response to a variety of external stimuli. In adult mammals, skeletal muscle hypertrophy occurs primarily as a response to increases in external loading. Here, we describe the methods that should be used for a comprehensive as ...

Skeletal muscle mass is determined by the balance between rates of protein synthesis and degradation. Protein synthesis rates can be measured in vivo by administering an amino acid as a tracer that is labeled with an isotope (radioactive or stable) of C, H, or N. The rate at which the labeled amino acid is in ...

The mechanisms by which muscle gene expression is initiated and maintained are not fully understood. Muscle genes are regulated by combinatorial interactions between numerous transcription factors bound to enhancers and promoters, and their associated protein complexes. Am ...

Ca2+ signaling plays an essential role in several functions of cardiac myocytes. Transient rises and reductions of cytosolic Ca2+, permitted by the sarcoplasmic reticulum Ca2+ ATPase (SERCA2) and other proteins, control each cycle of contraction and relaxation. Here we provide a practi ...

Ca2+ sparks are the elementary units of Ca2+ signaling in striated muscle fibers that appear as highly localized Ca2+ release events through ryanodine receptor (RyR) Ca2+ release channels in the sarcoplasmic reticulum (SR). While these events are commonly observed in resting cardiac myo ...

Calcium transients elicited by IP3 receptors upon electrical stimulation of skeletal muscle cells (slow calcium signals) are often hard to visualize due to their relatively small amplitude compared to the large transient originated from ryanodine receptors associated to excit ...

The ryanodine receptor ion channels (RyRs) release Ca2+ from the endo/sarcoplasmic reticulum in a variety of nonvertebrate and vertebrate species including flies, crustaceans, birds, fish, and amphibians. They are most abundant in skeletal and cardiac muscle, where in response to an ac ...

Mass spectrometry-based proteomics had a major impact on the global characterization of skeletal muscles and has decisively enhanced the field of neuromuscular pathology. Proteomic profiling of x-linked muscular dystrophy has identified a large number of new signature molecu ...

The lipidome of skeletal muscles is a worthwhile target of research, as it affects a multitude of biological functions, and is, in turn, affected by factors such as diet, physical activity, and development. We present two methods for the analysis of the main lipid classes in skeletal muscles of humans ...

Gene profiling is an excellent tool to identify the genetic mechanisms, networks, and molecular pathways involved in skeletal muscle development and muscular disorders. Oligonucleotide or cDNA microarray can be the first step to identify the global gene expression in the study of inte ...

Gene inactivation has become the gold standard for determining gene function in the mouse. Many genes inactivated in the germ line cause early lethality that precludes phenotypic assessment at a later time point. Conditional gene inactivation using Cre recombinase expressed via a tiss ...