Abstract
The basidiomycete fungus Coprinus cinereus has naturally synchronous meiosis and is amenable to analysis using an array of well-developed genetic and molecular tools. In this chapter, we explain in detail the two methods most commonly employed for C. cinereus, staining of intact gill segments and chromosome spreads, with an example of the application of each. We describe iron-hematoxylin staining of intact gill segments for the brightfield examination of meiotic progression, and the use of surface spreads and fluorescence in situ hybridization (FISH) to investigate meiotic chromosome pairing. Gill segments can alternatively be stained with DAPI for the determination of meiotic stage, or propidium iodide for the quantitation of nuclear DNA content, and the chromosome fixation and spreading techniques used for FISH are also suitable for immunolocalization studies of chromosomal proteins.
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References
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Acknowledgments
The authors would like to dedicate this chapter to Dr. Ben Lu, who introduced each of them to C. cinereus cytology. He first provided P.J.P. with some aged ferric acetate so she could use the optimal iron-hematoxylin method right away. (In turn, she would be happy to supply aged ferric acetate to anyone who would like to try this method.) He also taught the techniques of chromosome spreading and silver staining to two generations of Zolan lab students and post-docs, who bestowed on him the title of Zolan-lab “fairy godfather.” M.E.Z. also thanks Sasha Savytskyy for help on this manuscript and for the data presented in Fig. 8.2, and Heather Palmerini, Felicia Kennedy, Claire Burns, and Elizabeth Sierra Potchanant for helpful comments on the manuscript. The work in our laboratories is supported by NSF grant EF-0412016 (to P.J.P.) and NIH grant GM43930 (to M.E.Z.).
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Zolan, M.E., Pukkila, P.J. (2009). Meiotic Cytogenetics in Coprinus cinereus . In: Keeney, S. (eds) Meiosis. Methods in Molecular Biology, vol 558. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-103-5_8
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DOI: https://doi.org/10.1007/978-1-60761-103-5_8
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