By Paul N. Riccio, Odyssé Michos (auth.), Odyssé Michos (eds.)
Over the decade the advance of latest molecular biology instruments, complicated microscopy, reside imaging and platforms biology methods have revolutionized our perception of the way embryonic improvement proceeds. One primary point of improvement biology is the idea that of morphogenesis: figuring out how a gaggle of multipotent cells arrange and differentiate right into a advanced organ. In Kidney improvement: equipment and Protocols, professional researchers within the box element assorted methods to take on kidney improvement. those methods comprise tradition and reside imaging facets of kidney improvement, examining the three-d features of branching morphogenesis in addition to nephrogenesis, manipulation of the gene/protein expression in the course of kidney improvement in addition to within the grownup kidney, and the way to evaluate kidney malformation and ailment. Written within the hugely winning Methods in Molecular Biology™ sequence structure, chapters comprise introductions to their respective themes, lists of the required fabrics and reagents, step by step, without problems reproducible laboratory protocols, and key tips about troubleshooting and warding off recognized pitfalls.
Authoritative and practical, Kidney improvement: equipment and Protocols seeks to assist scientists within the extra research of the method of morphogenesis that's primary very important not just for learning developmental biology but in addition for regenerative medicine.
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Additional info for Kidney Development: Methods and Protocols
Live imaging of embryos can be performed on several species. The two major limitations are embryo size and embryo viability under the microscope. As the embryo gets bigger, the depth covered by confocal imaging becomes limiting. The depth of clear focal planes one can reach within a tissue depends primarily on the transparency of the embryo and the working distance of the objective. In an optimized confocal system, depths of up to 200 μm can be imaged (3). 5 mouse embryo but becomes progressively limiting for more central structures past this stage.
6. Rat serum: SD MALE. 7. DMEM/F12. 8. Antibiotic: Penicillin and streptomycin. 5 Organotypic Culture of the Urogenital Tract 47 9. Culture dishes with glass bottom (MatTeK corporation, P35G0-20-C); the day before experiment–coat the inside glass bottom of the dishes with the fibronectin (200 μg/mL solution in PBS), keep the plate at 4°C. 10. 4 μm culture membrane (Millipore, PICMORG50). 11. GDNF (R&D system). 12. HGF (R&D system). 13. FGF7 (R&D system). 14. Mineral oil (Sigma, M8410). 1. Medium Composition 1.
Kispert A, Vainio S, McMahon AP (1998) Wnt-4 is a mesenchymal signal for epithelial transformation of metanephric mesenchyme in the developing kidney. Development 125:4225–4234 6. Grobstein B (1955) Inductive interaction in the development of the mouse metanephros. J Exp Zool 130:319–340 7. Hedgepeth CM, Conrad LJ, Zhang J et al (1997) Activation of Wnt signalling pathway: a molecular mechanism for lithium activation. Dev Biol 185:82–91 8. Davies JA, Garrod DR (1995) Induction of early stages of kidney tubule differentiation by lithium ions.