By Donna Woulfe, Jing Yang, Nicolas Prevost, Peter O’Brien, Ryan Fortna (auth.), Jonathan M. Gibbins, Martyn P. Mahaut-Smith (eds.)
Platelets are a vital portion of the body's hemostasis process, and but, via their involvement in thrombosis, also are a tremendous reason behind morbidity and mortality. Platelets have a constrained lifespan and needs to be consistently synthetic by means of their precursor, the megakaryocyte. In Platelets and Megakaryocytes, quantity 2: views and methods prime specialist researchers-many accountable for seminal discoveries within the field-offer simple and complex thoughts for learning mobile signaling within the law and serve as of platelets and megakaryocytes, in addition to state of the art commentaries on our knowing of those cells. The equipment comprise ligand-binding assays, the learn of protein and lipid kinases and phosphatases, the research of lipid rafts within the rules of mobile signaling, and the dimension of intracellular calcium degrees. There also are options for the examine of electrophysiological occasions, nitric oxide signaling, venom proteins, and for the internalization of proteins into platelets via permeabilization. the appliance of many molecular and post-genomic how you can the learn of platelets and megakaryocytes is additionally defined. All protocols persist with the winning equipment in Molecular Biology™ sequence structure, each delivering step by step laboratory directions, an advent outlining the primary in the back of the procedure, lists of apparatus and reagents, and tips about troubleshooting and keeping off recognized pitfalls. A significant other quantity, Platelets and Megakaryocytes, quantity 1: practical Assays describes in step by step element easy and complex options for interpreting platelet and megakaryocyte functionality.
finished and state-of-the-art, the 2 volumes of Platelets and Megakaryocytes provide beginner and skilled mobile biologists, hematologists, and clinicians not just a benchmark survey of the sphere, but in addition a finished library of confirmed thoughts crucial for effective learn on platelets and megakaryocytes today.
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Platelets are an important component to the body's hemostasis method, and but, via their involvement in thrombosis, also are a massive explanation for morbidity and mortality. Platelets have a restricted lifespan and has to be constantly synthetic through their precursor, the megakaryocyte. In Platelets and Megakaryocytes, quantity 2: views and methods major specialist researchers-many accountable for seminal discoveries within the field-offer simple and complicated thoughts for learning mobile signaling within the law and serve as of platelets and megakaryocytes, in addition to cutting-edge commentaries on our figuring out of those cells.
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Extra resources for Platelets and Megakaryocytes: Volume 2: Perspectives and Techniques
It is difficult at the moment to predict exactly which genetic modifications will produce such a “knockdown” phenotype, although the insertion of the neoR selection cassette within an intron can often have such an effect. Alternatively, modification of certain promoter elements might produce a reduction in expression levels, as has been seen for the GATA-1 gene knockdown mutation in a promoter proximal regulatory element (41). Again, prediction of which strategies are likely to be effective in megakaryocytes will become easier as we learn more from this technology and discover more about regulatory mechanisms that control megakaryocyte-specific gene expression.
The feasibility of using recombinant adenoviruses to infect megakaryocytes has been shown, for example, by Faraday et al. (12), although there seems to be no obvious advantage to match the efficiency, specificity, and heritable integration that can be achieved with retroviral and lentiviral vectors. Finally, if it is necessary to introduce very large segments of CH02v2,33-56,24pgs 6/10/04 11:25 AM Page 40 40 Emambokus et al. DNA (20–100 kbp), then herpes simplex virus type I (HSV-1) could be a possible solution (see, for example, ref.
25, 444–447. 44. 44 Tsang, A. , Visvader, J. , Turner, C. , Weiss, M. , et al. (1997) FOG, a multiple zinc finger protein, acts as a cofactor for transcription factor GATA-1 in erythroid and megakaryocytic differentiation. Cell 90, 109–119. 45. Tsang, A. , Hom, D. , and Orkin, S. H. (1998) Failure of megakaryopoiesis and arrested erythropoiesis in mice lacking the GATA-1 transcriptional cofactor FOG. Genes & Dev. 12, 1176–1188. 46. , and Orkin, S. H. (2002) The zinc-finger proto-oncogene Gfi-1b is essential for development of the erythroid and megakaryocytic lineages.