Abstract
In this paper, we have developed a method to prepare spreads of mitotic chromosomes of Amoeba proteus and described the process of Amoeba proteus karyotyping. This protocol allows obtaining spread chromosomes with a characteristic pattern of chromomeres on individual chromosomes. It is shown that, in the metaphase of mitosis, amoebas of strain B (one of the type strains of A. proteus in the Amoebae Cultures Collection of the Institute of Cytology, Russian Academy of Sciences) contain 27 pairs of chromosomes. It is established that the pattern of chromomeres is a chromosome-specific feature. A typical karyogram and image bank of DAPI- and YoYo1-banded individual chromosomes of A. proteus, strain B composed of five different spreads of mitotic cells are presented.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Afon’kin, S.Yu., The content of DNA in nuclei of different Amoeba proteus strains according to cytofluorometry data, Tsitologiia, 1983, vol. 25, no. 7, pp. 771–776.
Afon’kin, S.Yu., Induced and spontaneous polyploidization in large Amoebae, Int. Rev. Cytol., 1989, vol. 115, pp. 231–266.
Barnitzke, S., Bullerdiek, J., and Schloot, W., Effects of hypotonic treatment on human metaphase chromosome length, Cytobios, 1981, vol. 31, pp. 75–80.
Chalkley, H.W. and Daniel, G.E., The relation between the form of the living cell and the nuclear phases of division in Amoeba proteus (Leidy), Physiol. Zool., 1933, vol. 6, pp. 593–619.
Claussen, U., Mazur, A., and Rubtsov, N., Chromosomes are highly elastic and can be stretched, Cytogenet. Cell Genet., 1994, vol. 66, pp. 120–125.
Comings, D.E., Mechanisms of chromosome banding and implication for chromosome structure, Ann. Rev. Genet., 1978, vol. 12, pp. 25–46.
Demin, S., Pleskach, N., Svetlova, M., and Solovyeva, L., High-resolution mapping of interstitial telomeric repeats in Syrian hamster metaphase chromosomes, Cytogenet. Genome Res., 2011, vol. 132, pp. 151–155.
Demin, S.Yu., Berdieva, M.A., Podlipaeva, Yu.I., Yudin, A.L., and Goodkov, A.V., Optical tomography analysis of Amoeba proteus chromatin organization at various cell cycle stages, Cell Tissue Biol., 2015, vol. 10, pp. 84–94.
George, P., Sharakhova, M.V., and Sharakhov, I.V., Highresolution cytogenetic map for the African malaria vector Anopheles gambiae, Insect Mol. Biol., 2010, vol. 19, pp. 675–682.
Goodkov, A.V., Yudin, A.L., and Podlipaeva, Yu.I., Collection of the proteus-type Amoebae at the Institute of Cytology, Russian Academy of Sciences, I. History, goals and research field, Protistology, 2014, vol. 8, no. 2, pp. 71–75.
Goodkov, A.V., Yudin, A.L., and Podlipaeva, Yu.I., Collection of the proteus-type Amoebae at the Institute of Cytology, Russian Academy of Sciences, II. Index of strains and list of publications, Protistology., 2015, vol. 9, no. 2, pp. 99–111.
Gromov, D.B., Ultrastructure of mitosis in Amoeba proteus, Protoplasma, 1985, vol. 126, pp. 130–139.
Hliscs, R., Muhlig, P., and Claussen, U., The spreading of metaphase is a slow process which leads to a stretching of chromosomes, Cytogenet. Cell Genet., 1997, vol. 76, pp. 167–171.
Kapuscinski, J., DAPI: a DNA-specific fluorescent probe, Biotech. Histochem., 1995, vol. 70, pp. 220–233.
Khodyuchenko, T., Gaginskaya, E., and Krasikova, A., Non-canonical Cajal bodies form in the nucleus of late stage avian oocytes lacking functional nucleolus, Histochem. Cell Biol., 2012, vol. 138, pp. 57–73.
Makhlin, E.E., Kudryavtseva, M.V., and Kudryavtsev, B.N., Peculiarities of changes in DNA content of Amoeba proteus nuclei during interphase, Exp. Cell Res., 1979, vol. 118, pp. 143–150.
Makhlin, E.E., The extra DNA synthesis in Amoeba proteus nuclei during the cell cycle, Tsitologiia, 1993, vol. 35, no. 3, pp. 109–121.
Makhlin, E.E., Variability of DNA quantity synthesized in Amoeba proteus nuclei during the cell cycle, Tsitologiia, 1987, vol. 29, no. 12, pp. 1379–1384.
Marakhova, N.V., Sopina, V.A., Skarlato, S.O., and Gromov, D.B., Separation of chromosome-sized DNA from Amoeba proteus by transverse alternating-field electrophoresis, Dokl. Akad. Nauk, 1993, vol. 330, no. 6, pp. 794–796.
Novikov, D.V., Kireev, I., and Belmont, A.S., High-pressure treatment of polytene chromosomes improves structural resolution, Nat. Methods, 2007, vol. 4, no. 6, pp. 483–485.
Ord, M.J., Chemical mutagens, in The Biology of Amoeba, New York: Acad. Press, 1973, pp. 349–370.
Ord, M.J., The synthesis of DNA through the cell cycle of Amoeba proteus, J. Cell Sci., 1968, vol. 3, pp. 483–491.
Podlipaeva, Yu., Demin, S., and Goodkov, A., New method for cell cycle synchronization in Amoeba proteus culture, Protistology, 2013, vol. 8, no. 1, pp. 3–7.
Prescott, D.M., and Carrier, R.F., Experimental procedures and cultural methods for Euplotes eurystomus and Amoeba proteus, in Methods in Cell Physiology, New York, London: Acad. Press, 1964, vol. 1, pp. 85–95.
Rogerson, A., Generation times and reproductive rates of Amoeba proteus (Leidy) as influenced by temperature and food concentration, Can. J. Zool., 1980, vol. 58, pp. 543–548.
Rønne, M., Andersen, O., and Erlandsen, M., Effect of colcemid exposure and methanol acetic fixation on human metaphase structure, Hereditas, 1979, vol. 90, pp. 195–201.
Rye, H.S., Yue, S., Wemmer, D.E., and Quesada, M.A. et, al., Stable fluorescent complexes of double-stranded DNA with bis-intercalating asymmetric cyanine dyes: properties and applications, Nucleic Acids Res., 1992, vol. 20, pp. 2803–2812.
Solovyeva, A.I., Stefanova, V.N., Podgornaya, O.I., and Demin, S.Iu., Karyotype features of trematode Himasthla elongata, Mol. Cytogenet., 2016, vol. 9, p. 34. doi 10.1186/s13039-016-0246-8
Sopina, V.A., The multiplication rate of Amoebae related to the cultivation temperature, J. Therm. Biol., 1976, vol. 1, 199–204.
Squarzoni, S., Cinti, C., Santi, S., Valmori, A., and Maraldi, N., Preparation of chromosome spreads for electron (TEM, SEM, STEM), light and confocal microscopy, Chromosoma, 1994, vol. 103, pp. 381–392.
Yudin, A.L., Amoeba and other Protozoa, in Animal Species for Developmental Studies, Invertebrates, New York, London: Consultants Bureau, 1990, vol. 1, pp. 1–11.
Yudin, A.L., Amoeba, in Ob”ekty biologii razvitiya (Objects of Developmental Biology), Moscow: Nauka, 1975, pp. 5–12.
Author information
Authors and Affiliations
Corresponding author
Additional information
Original Russian Text © S.Yu. Demin, M.A. Berdieva, Yu.I. Podlipaeva, A.L. Yudin, A.V. Goodkov, 2016, published in Tsitologiya, 2016, Vol. 58, No. 12, pp. 971–976.
Rights and permissions
About this article
Cite this article
Demin, S.Y., Berdieva, M.A., Podlipaeva, Y.I. et al. Karyotyping of Amoeba proteus . Cell Tiss. Biol. 11, 308–313 (2017). https://doi.org/10.1134/S1990519X17040046
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S1990519X17040046