The technique of needle microinjection has become a prominent experimental approach in biological research. Cellular organelles, DNA and RNA, enzymes, structural proteins, metabolites, ions and antibodies are just some of the molecular and cellular elements that have been transposed from test tubes into living cells by needle injection. This technique is broadly used as a valuable tool for the study of many different cell responses in a variety of systems. This text presents information on the technique of needle microinjection's principles, the required equipment, preparation of receiving cells and injected material and the analysis of the results. It provides detailed protocols for a wide range of important applications and different cellular systems from the fields of cell cycle regulation, signal transduction, study of transcriptional regulations, cytoskeletal functions, secretion and intracellular transport.
1. Needle Microinjection: A Brief History.- 2. Microinjection of Macromolecules into Mammalian Cells in Culture.- 3. The Use of Plasmid Microinjection to Study Specific Cell Cycle Phase Transitions.- 4. Combining Microinjection with Microanalytical Methods to Investigate Regulation of Metabolic Pathways.- 5. Import of Stably Folded Proteins into Peroxisomes.- 6. Study of the Function of Microtubule Proteins.- 7. Cytoskeleton Regulation by Rho Proteins.- 8. Cytoskeletal Regulation by Rho-p21 Associated Kinases: Analysis of ROK and PAK Function by Plasmid Microinjection.- 9. Use of Microinjection to Study Apoptosis and its Prevention.- 10. Role of the InsP3 Receptor in Intracellular Ca2+ Release and Ca2+ Entry.- 11. Microinjection of Xenopus laevis Oocytes: A Model System.- 12. Xenopus laevis Oocytes as a Model for Studying the Activation of Intracellular Kinases.- 13. Signal Transduction in Xenopus laevis Oocytes by Ras Oncoproteins and Lipid Metabolites.- 14. The Multiple Roles of Mos During Meiosis.- 15. Use of the Xenopus Oocyte System to Study RNA Transport.- 16. Functional Expression of G Protein-Coupled Receptors in Xenopus laevis Oocytes.- 17. Expression of Glucose Transporters in Xenopus laevis Oocytes: Applications for the Study of Membrane Transporter Function and Regulation.- 18. DNA Injection into Xenopus laevis Embryos as a Tool to Study Spatial Gene Activity.- 19. Transformation of Nematodes by Microinjection.- Guide to Solutions.- Guide to Protocols.- Troubleshooting Guide.
Series: Methods and Tools in Biosciences and Medicine
Number Of Pages: 248
Published: 1st May 1999
Publisher: SPRINGER VERLAG GMBH
Country of Publication: CH
Dimensions (cm): 25.04 x 18.77
Weight (kg): 0.5