Experimental techniques are the life blood of science. The better the methodology is, the more reliable and accurate the results will be. Ultimately, this will lead to a clearer interpretation of those results and firmer conclusions from any set of experiments. Experimental methodology in the area of cardiovascular biochemistry and molecular biology has advanced considerably in the last decade. Because of these factors, it was thought that a focused issue of Molecular and Cellular Biochemistry dedicated to the novel, latest technological advances in the field was warranted. We must thank Dr Naranjan S. Dhalla, Editor-in-Chief of Molecular and CellularBiochemistry, for his willingness to publish an issue with such a focus. We have attracted some of the leaders in the field of cardiovascular biology to submit articles describing some of the most novel, significant techniques currently in use in their laboratories. The purpose of the manuscripts was not to describe the recent experimental findings from each laboratory as is done in most conventional manuscripts. Instead, the purpose of the articles found within this focused volume of Molecular and Cellular Biochemistry was to describe how the technique is performed on the laboratory bench so that others less familiar with the technique may be able to use it in their own labs. The subjects described in this volume can be generally subdivided into three categories: molecular biology, cell biology and basic biochemistry. The methods cover wide areas including various DNA and RNA expression technologies, transfection techniques, quantification of ion flux movement, measurements of lipid metabolism, advances in the culture of specific cardiovascular cell populations, and the use of confocal microscopy to examine cell structure and function. We thank all of the authors who have contributed so much of their time and efforts and, most importantly, shared the `secrets' of these valuable techniques with the rest of the cardiovascular research community.
Author(s): Michael J. Mann, Ryuichi Morishita (auth.), Grant N. Pierce, William C. Claycomb (eds.)
Series: Developments in Molecular and Cellular Biochemistry 20
Edition: 1
Publisher: Springer US
Year: 1997
Language: English
Pages: 241
Tags: Biochemistry, general
Front Matter....Pages i-1
DNA transfer into vascular smooth muscle using fusigenic Sendai virus (HJV)-liposomes....Pages 3-12
Adenovirus mediated — gene transfer into cardiomyocytes....Pages 13-21
Retroviral techniques for studying organogenesis with a focus on heart development....Pages 23-35
Novel methods for adenovirus-mediated gene transfer to blood vessels in vivo ....Pages 37-46
Use of a hammerhead ribozyme with cationic liposomes to reduce leukocyte type 12-lipoxygenase expression in vascular smooth muscle....Pages 47-57
Immunodetection of activated mitogen-activated protein kinase in vascular tissues....Pages 59-66
Identification of proteins that interact with a protein of interest: Applications of the yeast two-hybrid system....Pages 67-79
Construction of a human heart cDNA library and identification of cardiovascular based genes (CVBest)....Pages 81-87
Competitive RT-PCR for studying gene expression in micro biopsies....Pages 89-95
The nuclear membrane integrity assay....Pages 97-102
Transfer of macromolecules into living adult cardiomyocytes by microinjection....Pages 103-109
Differential display: Identifying genes involved in cardiomyocyte proliferation....Pages 111-120
Analysis of inositol phosphates in heart tissue using anion-exchange high-performance liquid chromatography....Pages 121-127
Analysis of phospholipid molecular species....Pages 129-136
Measurements of fatty acid and carbohydrate metabolism in the isolated working rat heart....Pages 137-147
Fura-2 fluorescent technique for the assessment of Ca 2+ homeostasis in cardiomyocytes....Pages 149-157
Spectroscopic determination of sarcoplasmic reticulum Ca 2+ uptake and Ca 2+ release....Pages 159-170
The use of confocal microscopy in the investigation of cell structure and function in the heart, vascular endothelium and smooth muscle cells....Pages 171-194
A simple method for preparation of cultured cardiac fibroblasts from adult human ventricular tissue....Pages 195-198
DNA antisense strategies in the study of receptors for vasoactive peptides, and of growth and wound-healing factors....Pages 199-211
Potent and selective gene inhibition using antisense oligodeoxynucleotides....Pages 213-225
Immunohistochemical analysis of the adaptation of adult guinea-pig cardiomyocytes in long-term cultures and in cocultures with cardiac neurons: A novel model for studies of myocardial function....Pages 227-238
Back Matter....Pages 239-241
