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Increasing the resolution of chromosome analysis using pyrido[1,2-a]benzimidazoles

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Abstract

We studied the influence of three derivatives of pyrido[1,2-a]benzimidazoles (PBIs), which have DNA-intercalating properties, on plant mitotic chromosome condensation, in order to increase the resolution of chromosome analysis. The efficiency of the influence of these agents was assessed using the median chromosome length on chromosome slides, as well as by the number and size of chromosome DAPI bands. We used the third chromosome of Linum grandiflorum Desf. in these experiments. The chromosome was identified on the slides using its DAPI band pattern and a molecular marker, viz., the 5S rDNA site, which is located in the proximal region of the long arm of the chromosome. The influence of the well-known 9-aminoacridine (9-AMA) DNA intercalator, which is widely used in karyotype studies of short-chromosome organisms, was used as a control in all of the experiments. It was found that the influence of each of the three PBIs in the study on the root meristem of L. grandiflorum resulted in an increase in the median length of the third chromosome, the linear centromeric DAPI band size, and the number of intercalary DAPI bands. All three PBIs acted more efficiently than 9-AMA. The median chromosome length was increased by 15–40% and the number of intercalary bands increased by 1.5–3 times after PBI treatment, as compared to 9-AMA treatment. At the same time, 7-CF3-PBI, in a similar manner to 9-AMA, did not change the relative size of the centromeric DAPI band, while 7-NH2-PBI and 9-NH2-7-CF3-PBI gradually increased this parameter. It is concluded that these substances can be used as intercalating agents in cytogenetic studies in order to increase the resolution of chromosome analysis.

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References

  1. Caspersson, T., Zech, L., Johansson, C., and Modest, E.J., Identification of Human Chromosomes by DNA-Binding Fluorescent Agents, Chromosoma, 1970, vol. 30, pp. 215–227.

    Article  PubMed  CAS  Google Scholar 

  2. Arrighi, F.E. and Hsu, T.C., Localization of Heterochromatin in Human Chromosomes, Cytogenetics, 1971, vol. 10, pp. 81–86.

    Article  PubMed  CAS  Google Scholar 

  3. Samner, A.T., Evans, H.J., and Buckland, R.A., New Technique for Distinguishing between Human Chromosomes, Nature, 1971, vol. 232, pp. 31–32.

    Google Scholar 

  4. Muravenko, O.V. and Zelenin, A.V., Chromosomal Organization of the Genomes of Small-Chromosome Plants, Russ. J. Genet., 2009, vol. 45, no. 11, pp. 1338–1350.

    Article  CAS  Google Scholar 

  5. Zakharov, A.F. and Egolina, N.A., Differential Spiralization along Mammalian Mitotic Chromosomes: 1. BUDR-Revealed Differentiation in Chinese Hamster Chromosomes, Chromosoma, 1972, vol. 38, pp. 341–365.

    Article  PubMed  CAS  Google Scholar 

  6. Ikeuchi, T., Inhibitory Effect of Ethidium Bromide on Mitotic Chromosome Condensation and Its Application to High-Resolution Chromosome Banding, Cytogenet. Cell Genet., 1984, vol. 38, pp. 42–56.

    Article  Google Scholar 

  7. Lerman, L.S., Structural Consideration on Interaction of DNA with Acridines, J. Mol. Biol., 1961, vol. 3, pp. 18–30.

    Article  PubMed  CAS  Google Scholar 

  8. Zelenin, A.V., Acridine Orange as a Probe for Cell and Molecular Biology, Fluorescent and Luminescent Probes for Biological Activity, Maison, W.T., Ed., London: Academic Press, 1999. pp. 117–135.

    Chapter  Google Scholar 

  9. Ronne, M., Chromosome Preparation and High Resolution Banding Technique, J. Dairy Sci., 1989, vol. 72, no. 5, pp. 1363–1377.

    Article  PubMed  CAS  Google Scholar 

  10. Zakharov, A.F., Baranovskaya, L.I., Demintseva, V.S., and Ibraimov, A.I., Differentiation of Human Chromosomes according to Length and the Problem of Their Identification: 1. The General Pattern of Differentiation as Revealed by 5-Bromodeoxyuridine, Tsitologiya, 1973, vol. 15, no. 5, pp. 508–518.

    CAS  Google Scholar 

  11. Zakharov, A.F., Benyush, V., Kuleshov, N.P., and Baranovskaya, L.I., Khromosomy cheloveka: Atlas (Human Chromosomes: Atlas), Moscow: Meditsina, 1982.

    Google Scholar 

  12. Geoffrey, H., Bourne, G., and Chapman, P., Plant Chromosome Ultrastructure, Int. Rev. Cytol., 1985, vol. 94, pp. 119–126.

    Google Scholar 

  13. Zelenin, A.V., Vzaimodeistvie proizvodnykh aminoakridina s kletkoi (Interaction of Aminoacridine Derivatives with Cells), Moscow: Nauka, 1971.

    Google Scholar 

  14. Muravenko, O.V., Amosova, A.V., Samatadze, T.E., et al., 9-Aminoacridine: An Efficient Reagent to Improve Human and Plant Chromosome Banding Patterns and to Standardize Chromosome Image Analysis, Cytometry, 2003, vol. 51A, no. 1, pp. 52–57.

    Article  Google Scholar 

  15. Popov, K.V., Muravenko, O.V., Samatadze, T.E., et al., Peculiarity of the Analysis of Heterochromatic Regions in Small Chromosomes of Plants, Dokl. Akad. Nauk, 2001, vol. 381, no. 1–6, pp. 543–546.

    CAS  Google Scholar 

  16. Muravenko, O.V., Amosova, A.V., Samatadze, T.E., et al., Improvement of Human and Plant Chromosome High Resolution Banding Patterns Using 9-Aminoacridine for the Standardization of Chromosome Image Analysis, Cytometry, 2000, no. 10, pp. 163–164.

  17. Muravenko, O.V., Samatadze, T.E., and Popov, K.V., Comparative Genome Analysis in Two Flax Species by C-Banding Patters, Russ. J. Genet., 2001, vol. 37, no. 3, pp. 253–256.

    Article  CAS  Google Scholar 

  18. Samatadze, T.E., Muravenko, O.V., Popov, K.V., and Zelenin, A.V., Genome Comparison of the Matricaria chamomilla L. Varieties by the Chromosome C- and OR-Banding Patterns, Caryologia, 2001, vol. 54, no. 4, pp. 299–306.

    Google Scholar 

  19. Pastor, J., Siro, J., Garcia-Navio, J.L., et al., Synthesis of New Azino Fused Benzimidazolium Salts: A New Family of DNA Intercalating Agents, Bioorg. Med. Chem. Lett., 1995, vol. 5, pp. 3043–3048.

    Article  Google Scholar 

  20. Ryzvanovich, G.A., Begunov, R.S., Rachinskaya, O.A., et al., Synthesis and Intercalating Activity of New Pyrido[1.2-a]benzimidazoles, Khim.-Farm. Zh., 2011, vol. 43, no. 3, pp. 100–102.

    Google Scholar 

  21. Semenova, O.Yu., Samatadze, T.E., Zelenin, A.V., and Muravenko, O.V., Comparative Study of the Species of Adenolinum and Stellerolinum Sections by Means of FISH Technique, Biol. Membrany, 2006, vol. 23, no. 6, pp. 453–460.

    Article  CAS  Google Scholar 

  22. Muravenko, O.V., Samatadze, T.E., Popov, K.V., et al., Comparative Genome Analysis in Two Flax Species by C-Banding Patterns, Russ. J. Genet., 2001, vol. 37, no. 3, pp. 253–256.

    Article  CAS  Google Scholar 

  23. Muravenko, O.V., Yurkevich, O.Yu., Bolsheva, N.L., et al., Comparison of Eight Species of Sections Linum and Adenolinum from the Genus Linum Based on Chromosome Banding, Moleculars and RAPD Analysis, Genetics, 2009, vol. 135, pp. 245–255.

    CAS  Google Scholar 

  24. Samatadze T.E., Muravenko O.V., and Bolsheva, N.L., Investigation of Chromosomes in Varieties and Translocation Lines of Pea Pisum sativum L. by FISH, Ag-NOR, and Differential DAPI Staining, Russ. J. Genet., 2005, no. 12, pp. 1381–1389.

  25. Sakore, T.D., Jain, S.C., Tsai, C.-C., and Sobell, H.M., Mutagen-Nucleic Acid Intercalative Binding: Structure of a 9-Aminoacridine: 5-Iodocitidylyl(3′-5′)Guanosine Crystalline Complex, Proc. Natl. Acad. Sci. USA, 1977, vol. 74, no. 1, pp. 188–192.

    Article  PubMed  CAS  Google Scholar 

  26. Lerman, L.S., The Structure of the DNA-Acridine Complex, Proc. Nat Acad. Sci. USA, 1963, vol. 49, pp. 94–102.

    Article  PubMed  CAS  Google Scholar 

  27. Vershinin, A.V., Schwarzacher, T., and Heslop-Harrison, J.S., The Large-Scale Genomic Organization of Repetitive DNA Families at the Telomeres of Rye Chromosomes, Plant Cell, 1995, vol. 7, pp. 1823–1833.

    PubMed  CAS  Google Scholar 

  28. Vosa, C.G., Heterochromatin Recogniting and Analysis of Chromosome Variation in Scilla sibirica, Chromosoma, 1973, vol. 43, no. 3, pp. 269–278.

    Article  Google Scholar 

  29. Linde-Laursen, J., Giemsa C-Banding of Barley Chromosomes: 1. Bending Pattern Polymorphism, Hereditas, 1978, vol. 88, no. 1, pp. 55–64.

    Article  Google Scholar 

  30. Muravenko, O.V., Badaev, N.S., Borisov, J.M., et al., Karyotype Formation in Relative Barley Varieties, Russ. J. Genet., 1991, vol. 27, no. 3, pp. 2109–2118.

    Google Scholar 

  31. Badaeva, E.D., Sozinova, L.F., Badaev, N.S., et al., “Chromosomal Passport” of Triticum aestivum L. em. Thell. cv. Chinese Spring and Standardization of Chromosomal Analysis of Cereals, Cer. Res. Commun., 1990, vol. 18, no. 4, pp. 273–281.

    Google Scholar 

  32. Muravenko, O.V., Fedotov, A.R., Punina, E.O., et al., Comparison of Chromosome BrdU-Hoechst-Giemsa Banding Patterns of the A1 and (AD)2 Genomes of Cotton, Genome, 1998, vol. 41, pp. 616–625.

    CAS  Google Scholar 

  33. Samatadze, T.E., Zelenin, A.V., Suslina, S.N., et al., Comparative Cytogenetic Study of the Forms of Macleaya cordata (Willd.) R. Br. from Different Localities, Russ. J. Genet., 2012, vol. 48, no. 1, pp. 63–69.

    Article  CAS  Google Scholar 

  34. Rachinskaya, O.A., Lemesh, V.A., Muravenko, O.V., et al., Genetic Polymorphism of Flax Linum usitatissimum Based on the Use of Molecular Cytogenetic Markers, Russ. J. Genet., 2011, vol. 47, no. 1, pp. 56–65.

    Article  CAS  Google Scholar 

  35. Siro, J., Pastor, J., Garcia-Navio, J.L., and Juan, J., Unexpected N-C Bond Fission of Fused N-Alkylbenzidazolium Salts: A New Approach to Pyrido[1,2-α]or Pyridazino[1,6-α]benzimidazoles, Tetrahedron, 1998, vol. 54, pp. 1929–1936.

    Article  CAS  Google Scholar 

  36. Brana, M.F., Castellano, J.M., Keilhauer, G., et al., Benzimidazo[1,2-c]quinazolines: A New Class Antitumor Compounds, Anti-Cancer Drug Des., 1994, vol. 9, pp. 527–538.

    CAS  Google Scholar 

  37. Weinkauf, R.L., Chen, A.Y., Yu, C., et al., Antineoplastic Activity of Benzimidazo[1,2-b]isoquinolines, Indolo[2,3-b]quinolines and Pyridocarbazoles, Bioorg. Med. Chem., 1994, vol. 2, no. 8, pp. 781–786.

    Article  PubMed  CAS  Google Scholar 

  38. Deady, L.W., Rodemman, T., Finlay, G.J., et al., Synthesis and Cytotoxic Activity of Carboxamide Derivatives of Benzimidazo[2,1-a]isoquinoline and Pyrido[3′,2′:4.5]imidazo[2,1-a]isoquinoline, Anti-Cancer Drug Des., 2000, vol. 15, no. 5, pp. 339–346.

    CAS  Google Scholar 

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Correspondence to A. V. Zelenin.

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Original Russian Text © O.A. Rachinskaya, K.V. Popov, G.A. Ryzvanovich, N.L. Bol’sheva, R.S. Begunov, O.Yu. Yurkevich, A.V. Zelenin, O.V. Muravenko, 2012, published in Genetika, 2012, Vol. 48, No. 10, pp. 1228–1236.

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Rachinskaya, O.A., Popov, K.V., Ryzvanovich, G.A. et al. Increasing the resolution of chromosome analysis using pyrido[1,2-a]benzimidazoles. Russ J Genet 48, 1055–1062 (2012). https://doi.org/10.1134/S1022795412100080

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