Obtaining Polytene, Meiotic, and Mitotic Chromosomes from Mosquitoes for Cytogenetic Analysis
- 1Department of Entomology, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24060, USA
- 2Department of Genetics and Cell Biology, Tomsk State University, Tomsk 634050, Russia
- 3Laboratory of Evolutionary Genomics of Insects, Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Novosibirsk 630090, Russia
- ↵4Correspondence: msharakh{at}vt.edu
Abstract
Chromosome visualization is a key step for developing cytogenetic maps and idiograms, analyzing inversion polymorphisms, and identifying mosquito species. Three types of chromosomes—polytene, mitotic, and meiotic—are used in cytogenetic studies of mosquitoes. Here, we describe a detailed method for obtaining high-quality polytene chromosome preparations from the salivary glands of larvae and the ovaries of females for Anopheles mosquitoes. We also describe how to obtain mitotic chromosomes from imaginal discs of fourth-instar larvae and meiotic chromosomes from the testes of male pupae for all mosquitoes. These chromosomes can be used for fluorescence in situ hybridization (FISH), a fundamental technique in cytogenetic research that is used for physical genome mapping, detecting chromosomal rearrangements, and studying chromosome organization.
Footnotes
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From the Mosquitoes collection, edited by Laura B. Duvall and Benjamin J. Matthews.
