The speciose neotropical genus Characidium has proven to be a good model for cytogenetic exploration. Representatives of this genus often have a conserved diploid chromosome number; some species exhibit a highly differentiated ZZ/ZW sex chromosome system, while others do not show any sex-related chromosome heteromorphism. In this study, chromosome painting using a W-specific probe and comparative chromosome mapping of repetitive sequences, including ribosomal clusters and 4 microsatellite motifs - (CA)15, (GA)15, (CG)15, and (TTA)10 -, were performed in 6 Characidium species, 5 of which possessed a heteromorphic ZW sex chromosome system. The W-specific probe showed hybridization signals on the W chromosome of all analyzed species, indicating homology among the W chromosomes. Remarkably, a single major rDNA-bearing chromosome pair was found in all species. The 18S rDNA localized to the sex chromosomes in C. lanei, C. timbuiense and C. pterostictum, while the major rDNA localized to one autosome pair in C. vidali and C. gomesi. In contrast, the number of 5S rDNA-bearing chromosomes varied. Notably, minor ribosomal clusters were identified in the W chromosome of C. vidali. Microsatellites were widely distributed across almost all chromosomes of the karyotypes, with a greater accumulation in the subtelomeric regions. However, clear differences in the abundance of each motif were detected in each species. In addition, the Z and W chromosomes showed the differential accumulation of distinct motifs. Our results revealed variability in the distribution of repetitive DNA sequences and their possible association with sex chromosome diversification in Characidium species.

Keywords:
Chromosome painting, Crenuchidae, Fish cytogenetics, Karyotype diversification
1.
Buckup PA: Phylogenetic interrelationships and reductive evolution in neotropical characidiin fishes (Characiformes, Ostariophysi). Cladistics 9:305-341 (1993).
2.
Cioffi MB, Bertollo LAC: Initial steps in XY chromosome differentiation in Hoplias malabaricus and the origin of an X1X2Y sex chromosome system in this fish group. Heredity (Edinb) 105:554-561 (2010).
3.
Cioffi MB, Camacho JPM, Bertollo LAC: Repetitive DNAs and differentiation of sex chromosomes in Neotropical fishes. Cytogenet Genome Res 132:188-194 (2010).
4.
Cioffi MB, Kejnovský E, Bertollo LAC: The chromosomal distribution of microsatellite repeats in the genome of the wolf fish Hoplias malabaricus, focusing on the sex chromosomes. Cytogenet Genome Res 132:289-296 (2011).
5.
Cioffi Mde B, Kejnovský E, Marquioni V, Poltronieri J, Molina WF, et al: The key role of repeated DNAs in sex chromosome evolution in two fish species with ZW sex chromosome system. Mol Cytogenet 5:42 (2012).
6.
Dover GA: Evolution of genetics redundancy for advanced player. Curr Opin Genet Dev 3:902-910 (1993).
7.
Eschmeyer WN: Catalog of Fishes: Genera, Species, References. http://researcharchive.calacademy.org/research/ichthyology/catalog/fishcatmain.asp (accessed April 4, 2015).
8.
Hancock JM: Simple sequences and the expanding genome. Bioessays 18:421-425 (1996).
9.
Henning F, Trifonov V, Ferguson-Smith MA, de Almeida-Toledo LF: Non-homologous sex chromosomes in two species of the genus Eigenmannia (Teleostei: Gymnotiformes). Cytogenet Genome Res 121:55-58 (2008).
10.
Kejnovský E, Michalovova M, Steflova P, Kejnovska I, Manzano S, et al: Expansion of microsatellites on evolutionary young Y chromosome. PLoS One 8:e45519 (2013).
11.
Kubat Z, Hobza R, Vyskot B, Kejnovský E: Microsatellite accumulation on the Y chromosome of Silene latifolia. Genome 51:350-356 (2008).
12.
Levan A, Fredga K, Sandberg AA: Nomenclature for centromeric position on chromosomes. Hereditas 52:201-220 (1964).
13.
Lohe AR, Hilliker AJ, Roberts PA: Mapping simple repeated DNA sequences in heterochromatin of Drosophila melanogaster. Genetics 134:1149-1174 (1993).
14.
López-Flores I, Garrido-Ramos MA: The repetitive DNA content of eukaryotic genomes, in Garrido-Ramos MA (ed): Repetitive DNA. Genome Dyn, vol 7, pp 1-28 (Karger, Basel 2012).
15.
Machado TC, Pansonato-Alves JC, Pucci MB, Nogaroto V, Almeida MC, et al: Chromosomal painting and ZW sex chromosomes differentiation in Characidium (Characiformes, Crenuchidae). BMC Genet 12: 65 (2011).
16.
McMurray CT: Mechanisms of DNA expansion. Chromosoma 104:2-13 (1995).
17.
Milani D, Cabral-de-Mello DC: Microsatellite organization in the grasshopper Abracris flavolineata (Orthoptera: Acrididae) revealed by FISH mapping: remarkable spreading in the A and B chromosomes. PLoS One 9:e97956 (2014).
18.
Nanda I, Schories S, Tripathi N, Dreyer C, Haaf T, et al: Sex chromosome polymorphism in guppies. Chromosoma 123:373-383 (2014).
19.
Pansonato-Alves JC, Paiva LRS, Oliveira C, Foresti F: Interespecific chromosomal divergences in the genus Characidium (Teleostei: Characiformes: Crenuchidae). Neotrop Ichythyol 8:77-86 (2010).
20.
Pansonato-Alves JC, Vicari MR, Oliveira C, Foresti F: Chromosomal diversification in samples of Characidium cf. gomesi (Teleostei, Crenuchidae). J Fish Biol 78:183-194 (2011a).
21.
Pansonato-Alves JC, Oliveira C, Foresti F: Karyotypic conservatism in samples of Characidium cf. zebra (Teleostei, Characiformes, Crenuchidae): physical mapping of ribosomal genes and natural triploidy. Genet Mol Biol 34:208-213 (2011b).
22.
Pansonato-Alves JC, Serrano ÉA, Utsunomia R, Camacho JPM, Costa-Silva GJ, et al: Single origin of sex chromosomes and multiple origins of B chromosomes in fish genus Characidium. PLoS One 9:e107169 (2014).
23.
Pazian MF, Shimabukuro-Dias CK, Pansonato-Alves JC, Oliveira C, Foresti F: Chromosome painting of Z and W sex chromosomes in Characidium (Characiformes, Crenuchidae). Genetica 141:1-9 (2013).
24.
Pendás AM, Moran P, Freije JP, Garcia-Vazquez E: Chromosomal mapping and nucleotide sequence of two tandem repeats of Atlantic salmon 5S rDNA. Cytogenet Cell Genet 67:31-36 (1994).
25.
Pinkel D, Straume T, Gray JW: Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization. Proc Natl Acad Sci USA 83:2934-2938 (1986).
26.
Pokorná M, Kratochvíl L, Kejnovský E: Microsatellite distribution on sex chromosomes at different stages of heteromorphism and heterochromatinization in two lizard species (Squamata: Eublepharidae: Coleonyx elegans and Lacertidae: Eremias velox). BMC Genet 12:90 (2011).
27.
Poltronieri J, Marquioni V, Bertollo LAC, Kejnovský E, Molina WF, et al: Comparative chromosomal mapping of microsatellites in Leporinus species (Characiformes, Anostomidae): unequal accumulation on the W chromosomes. Cytogenet Genome Res 142:40-45 (2013).
28.
Pucci MB, Barbosa O, Nogaroto V, Almeira MC, Artoni RF, et al: Population differentiation and speciation in the genus Characidium (Characiformes: Crenuchidae): effects of reproductive and chromosomal barriers. Biol J Linn Soc 111:541-553 (2014).
29.
Ruiz-Ruano FJ, Cuadrado Á, Montiel EE, Camacho JPM, López-León MD: Next generation sequencing and FISH reveal uneven and nonrandom microsatellite distribution in two grasshopper genomes. Chromosoma 124:221-234 (2015).
30.
Scacchetti PC, Utsunomia R, Pansonato-Alves JC, Costa-Silva GJ, Oliveira C, Foresti F: Extensive spreading of interstitial telomeric sites on the chromosomes of Characidium (Teleostei, Characiformes). Genetica 143:263-270 (2015).
31.
Schemberger MO, Bellafronte E, Nogaroto V, Almeida MC, Schühli GS, et al: Differentiation of repetitive DNA sites and sex chromosome systems reveal closely related group in Parodontidae (Actinopterygii: Characiformes). Genetica 139:1499-1508 (2011).
32.
da Silva M, Matoso DA, Vicari MR, Almeida MC, Margarido VP, Artoni RF: Physical mapping of 5S rDNA in two species of Knifefishes: Gymnotus pantanal and Gymnotus paraguensis (Gymnotiformes). Cytogenet Genome Res 134:303-307 (2011).
33.
Silva DMZA, Pansonato-Alves JC, Utsunomia R, Daniel SN, Hashimoto DT, et al: Chromosomal organization of repetitive DNA sequences in Astyanax bockmanni (Teleostei, Characiformes): dispersive location, association and co-localization in the genome. Genetica 141:329-336 (2013).
34.
Sumner AT: A simple technique for demonstrating centromeric heterochromatin. Expl Cell Res 75:304-306 (1972).
35.
Terencio ML, Schneider CH, Gross MC, Vicari MR, Farias IP, et al: Evolutionary dynamics of repetitive DNA in Semaprochilodus (Characiformes, Prochilodontidae): a fish model for sex chromosome differentiation. Sex Dev 7:325-333 (2013).
36.
Tóth G, Gáspári Z, Jurka J: Microsatellites in different eukaryotic genomes survey and analysis. Genome Res 10:967-981 (2000).
37.
Vicari MR, Artoni RF, Moreira-Filho O, Bertollo LAC: Diversification of a ZZ/ZW sex chromosome system in Characidium fish (Crenuchidae, Characiformes). Genetica 134:311-317 (2008).
38.
Yano CF, Poltronieri J, Bertollo LAC, Artoni RF, Liehr T, Cioffi MB: Chromosomal mapping of repetitive DNAs in Triportheus trifurcatus (Characidae, Characiformes): insights into the differentiation of the Z and W chromosomes. PLoS One 9:e90946 (2014).
39.
White TJ, Bruns T, Lee S, Taylor L: Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics, in Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds): PCR Protocols: A Guide to Methods and Applications, pp 312-315 (Academic Press Inc, London/New York 1990).
40.
Ziemniczak K, Traldi JB, Nogaroto V, Almeida MC, Artoni RF, et al: In situ localization of (GATA)n and (TTAGGG)n repeat DNAs and W sex chromosome differentiation in Parodontidae (Actinopterygii: Characiformes). Cytogenet Genome Res 144:325-332 (2014).
© 2015 S. Karger AG, Basel
2015
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.