The monophyletic order Osteoglossiformes represents one of the most ancestral groups of teleosts and has at least 1 representative in all continents of the southern hemisphere, with the exception of Antarctica. However, despite its phylogenetic and biogeographical importance, cytogenetic data in Osteoglossiformes are scarce. Here, karyotype and chromosomal characteristics of the lower Niger River population of the African butterfly fish Pantodon buchholzi, the sole species of the family Pantodontidae (Osteoglossiformes), were examined using conventional and molecular cytogenetic approaches. All specimens examined had 2n = 46 chromosomes, with a karyotype composed of 5 pairs of metacentric, 5 pairs of submetacentric, and 13 pairs of acrocentric chromosomes in both sexes. No morphologically differentiated sex chromosomes were identified. C-bands were located in the centromeric/pericentromeric region of all chromosomes and were associated with the single AgNOR site. FISH with ribosomal DNA probes revealed that both 5S and 18S rDNA were present in only 1 pair of chromosomes each, but did not colocalize. CMA3+ bands were observed near the telomeres in several chromosome pairs and also at the 18S rDNA sites. The mapping of di- and trinucleotide repeat motifs, Rex6 transposable element, and U2 snRNA showed a scattered distribution over most of the chromosomes, but for some microsatellites and the U2 snRNA also a preferential accumulation at telomeric regions. This study presents the first detailed cytogenetic analysis in the African butterfly fish by both conventional and molecular cytogenetic protocols. This is the first of a series of further cytogenetic and cytogenomic studies on osteoglossiforms, aiming to comprehensively examine the chromosomal evolution in this phylogenetically important fish order.

1.
Amemiya CT, Gold JR: Chromomycin A3 stains nucleolus organizer regions of fish chromosomes. Copeia 1:226-231 (1986).
2.
Arai R: Fish Karyotypes. A Check List (Springer, Tokyo 2011).
3.
Bănărescu P: Zoogeography of Fresh Waters. General Distribution and Dispersal of Freshwater Animals (Aula-Verlag, Wiesbaden 1990).
4.
Berra TM: Freshwater Fish Distribution (University of Chicago Press, Chicago 2007).
5.
Bertollo LAC, Cioffi MB, Moreira-Filho O: Direct chromosome preparation from freshwater teleost fishes, in Ozouf-Costaz C, Pisano E, Foresti F, Almeida Toledo LF (eds): Fish Cytogenetic Techniques (Ray-Fin Fishes and Chondrichthyans), pp 21-26 (CRC Press, Enfield 2015).
6.
Cioffi MB, Bertollo LAC: Chromosomal distribution and evolution of repetitive DNAs in fish, in Garrido-Ramos MA (ed): Repetitive DNA. Genome Dynamics, vol 7, pp 197-221 (Karger, Basel 2012).
7.
Cioffi MB, Martins C, Bertollo LAC: Comparative chromosome mapping of repetitive sequences. Implications for genomic evolution in the fish, Hoplias malabaricus. BMC Genet 10:34 (2009).
8.
Cross I, Merlo A, Manchado M, Ifante C, Canavate JP, Rebordinos L: Cytogenetic characterization of the sole Solea senegalensis (Teleostei: Pleuronectiformes: Soleidae): Ag-NOR, (GATA)n, (TTAGGG)n and ribosomal genes by one-color and two-color FISH. Genetica 128:253-259 (2006).
9.
da Rosa R, Rubert M, Caetano-Filho M, Giuliano-Caetano L: Conserved cytogenetic features in the Amazonian Arapaima, Arapaima gigas (Schinz 1822) from Jamari River, Rondônia-Brazil. Open Biol J 2:91-94 (2009).
10.
Donsakul T, Magtoon W: A Chromosome Study on Three Species of Notopterid Fishes, Notopterus notopterus, N. chitala and N. blanci from Thailand (Tech Pap Agric Biol, Srinakharinwirot University 1990).
11.
Ferreira DC, Porto-Foresti F, Oliveira C, Foresti F: Transposable elements as a potential source for understanding the fish genome. Mob Genet Elements 1:112-117 (2011).
12.
Gornung E: Twenty years of physical mapping of major ribosomal RNA genes across the teleosts: a review of research. Cytogenet Genome Res 141:90-102 (2013).
13.
Greenwood PH, Thomson KS: The pectoral anatomy of Pantodon buchholzi Peters (a freshwater flying fish) and the related Osteoglossidae. J Zool 135:283-301 (1960).
14.
Greenwood PH, Wilson MVH: Bonytongues and their allies, in Paxton JR, Eschmeyer WN (eds): Encyclopedia of Fishes, ed 2, pp 80-84 (Academic Press, San Diego 1998).
15.
Greenwood PH, Rosen DE, Weitzman SH, Myers GS: Phyletic studies teleostean fishes, with a provisional classification of living forms. Bull Am Mus Nat Hist 131:339-456 (1966).
16.
Hilton EJ: Comparative osteology and phylogenetic systematics of fossil and living bony-tongue fishes (Actinopterygii, Teleostei, Osteoglossomorpha). Zool J Linn Soc 137:1-100 (2003).
17.
Hirata J, Urushido T: Karyotypes and DNA content in the Osteoglossiformes (in Japanese with English abstract). Sci Rep Res Inst Evol Biol 9:83-90 (2000).
18.
Howell WM, Black DA: Controlled silver-staining of nucleolus organizer regions with a protective colloidal developer: a 1-step method. Experientia 36:1014-1915 (1980).
19.
Hua-Van A, Le Rouzic A, Maisonhaute C, Capy P: Abundance, distribution and dynamics of retrotransposable elements and transposons: similarities and differences. Cytogenet Genome Res 110:426-440 (2005).
20.
Inoue JG, Kumazawa Y, Miya M, Nishida M: The historical biogeography of the freshwater knifefishes using mitogenomic approaches: a Mesozoic origin of the Asian notopterids (Actinopterygii: Osteoglossomorpha). Mol Phylogenet Evol 51:486-499 (2009).
21.
Jurka J, Kapitonov VV, Pavlicek A, Klonowski P, Kohany O, Walichiewicz J: Repbase update, a database of eukaryotic repetitive elements. Cytogenet Genome Res 110:462-467 (2005).
22.
Kershaw DR: The cranial osteology of the ‘Butterfly Fish', Pantodon buchholzi Peters. Zool J Linn Soc 49:5-19 (1970).
23.
Kidwell MG: Transposable elements and the evolution of genome size in eukaryotes. Genetica 115:49-63 (2002).
24.
Krysanov EY, Golubtsov AS: Karyotypes of four fish species from the Nile and Omo-Turkana basins in Ethiopia, East Africa. J Ichthyol 54:889-992 (2014).
25.
Kubát Z, Hobza R, Vyskot B, Kejnovský E: Microsatellite accumulation on the Y chromosome in Silene latifolia. Genome 51:350-356 (2008).
26.
Lavoué S: Was Gondwanan breakup the cause of the intercontinental distribution of Osteoglossiformes? A time-calibrated phylogenetic test combining molecular, morphological, and paleontological evidence. Mol Phylogenet Evol 99:34-43 (2016).
27.
Lavoué S, Sullivan JP: Simultaneous analysis of five molecular markers provides a well-supported phylogenetic hypothesis for the living bony-tongue fishes (Osteoglossomorpha: Teleostei). Mol Phylogenet Evol 33:171-185 (2004).
28.
Lavoué S, Miya M, Arnegard ME, McIntyre PB, Mamonekene V, Nishida M: Remarkable morphological stasis in an extant vertebrate despite tens of millions of years of divergence. Proc Biol Sci 278:1003-1008 (2011).
29.
Levan A, Fredga K, Sandberg AA: Nomenclature for centromeric position on chromosomes. Hereditas 52:201-220 (1964).
30.
Li GQ, Wilson MVH: Phylogeny of Osteoglossomorpha, in Stiassny ML, Parenti LR, Johnson JD (eds): Interrelationships of Fishes, pp 163-174 (Academic Press, New York 1996).
31.
Marques DK, Venere PC, Galetti PM Jr: Chromosomal characterization of the bonytongue Arapaima gigas (Osteoglossiformes: Arapaimidae). Neotrop Ichthyol 4:215-218 (2006).
32.
Mayr B, Ráb P, Kalat M: Localization of NORs and counterstain-enhanced fluorescence studies in Perca fluviatilis (Pisces, Percidae). Genetica 67:51-56 (1985).
33.
Mu XD, Wang XJ, Song HM, Yang YX, Luo D, et al: Mitochondrial DNA as effective molecular markers for the genetic variation and phylogeny of the family Osteoglossidae. Gene 511:320-325 (2012).
34.
Nayyar RP: Karyotype studies in the genus Notopterus (Lacepede). The occurrence and fate of univalent chromosomes in spermatocytes of N. chitala. Genetica 36:398-406 (1965).
35.
Near TJ, Eytan RI, Dornburg A, Kuhn KL, Moore JA, et al: Resolution of ray-finned fish phylogeny and timing of diversification. Proc Natl Acad Sci USA 109:13698-13703 (2012).
36.
Nelson JS: Fishes of the World, ed 4 (John Wiley & Sons, Inc., Hoboken 2006).
37.
Ozouf-Costaz C, Coutanceau JP, Bonillo C, Belkadi L, Fermon Y, et al: First insights into karyotype evolution within the family Mormyridae. Cybium 39:227-236 (2015).
38.
Pendás AM, Morán 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).
39.
Pinkel D, Straume T, Gray J: Cytogenetic analysis using quantitative, high sensitivity, fluorescence hybridization. Proc Natl Acad Sci USA 83:2934-2938 (1986).
40.
Ráb P, Rábová M, Reed KM, Phillips RB: Chromosomal characteristics of ribosomal DNA in the primitive semionotiform fish, longnose gar Lepisosteus osseus. Chromosome Res 7:475-480 (1999).
41.
Rishi KK, Singh J: Chromosomes of Notopterus notopterus (Pallas) (Notopteridae: Clupeiformes). Chromosome Inform Service 34:10-11 (1983).
42.
Saidel WM, Strain GF, Fornari SK: Characterization of the aerial escape response of the African butterfly fish, Pantodon buchholzi Peters. Env Biol Fish 71:63-72 (2004).
43.
Sallan LC: Major issues in the origins of ray-finned fish (Actinopterygii) biodiversity. Biol Rev 89:950-971 (2014).
44.
Sambrook JH, Russell DW: Molecular Cloning (Cold Spring Harbor Laboratory Press, New York 2001).
45.
Schmid M: Chromosome banding in Amphibia. IV. Differentiation of GC- and AT-rich chromosome regions in Anura. Chromosoma 77:83-103 (1980).
46.
Schmid M, Guttenbach M: Evolutionary diversity of reverse (R) fluorescent chromosome bands in vertebrates. Chromosoma 97:101-114 (1988).
47.
Shen XY, Kwan HY, Thevasagayam NM, Prakki SRS, Kuznetsova IS, et al: The first transcriptome and genetic linkage map for Asian arowana. Mol Ecol Resour 14:622-635 (2014).
48.
Silawong K, Aoki S, Supiwong W, Tanomtong A, Khakhong S, et al: The first chromosomal characteristics of nucleolar organizer regions (NORs) in grey featherback fish, Notopterus notopterus (Osteoglossiformes, Notopteridae) by conventional and Ag-NOR staining techniques. Cytologia 77:279-285 (2012).
49.
Srivastava MD: The structure and behavior of chromosomes in six freshwater teleosts. Cellule 65:93-98 (1964).
50.
Stewart DJ: Re-description of Arapaima agassizii (Valenciennes), a rare fish from Brazil (Osteoglossomorpha: Osteoglossidae). Copeia 2013:38-51 (2013a).
51.
Stewart DJ: A new species of Arapaima (Osteoglossomorpha: Osteoglossidae) from the Solimões River, Amazonas State, Brazil. Copeia 2013:470-476 (2013b).
52.
Sumner AT: Simple technique for demonstrating centromeric heterochromatin. Exp Cell Res 75:304-306 (1972).
53.
Supiwong W, Tanomtong A, Khakhong S, Silawong K, Aoki S, Sanoamuang L: The first chromosomal characteristics of nucleolar organizer regions and karyological analysis of clown knife fish, Chitala ornata (Osteoglossiformes, Notopteridae) by T-lymphocyte cell culture. Cytologia 77:393-399 (2012).
54.
Suzuki A, Taki Y, Urushido T: Karyotypes of two species of arowana, Osteoglossum bicirrhosum and O. ferreirai. Jpn J Ichthyol 29:220-222 (1982).
55.
Takai A, Ojima Y: C-banded karyotype and nucleolus organizer regions of a notopterid fish, Notopterus chitala (Notopteridae, Osteoglossiformes). Chromosome Sci 2:35-38 (1998).
56.
Taverne L: Les Ostéoglossomorphes marins de l'Eocène du Monte Bolca (Italie): Monopteros Volta 1796, Thrissopterus Heckel, 1856 et Foreyichthys Taverne, 1979. Considérations sur la phylogénie des Téléostéens ostéoglossomorphes. Studi e Ricerche sui Giacimenti Terziari di Bolca (Museo Civico di Storia Naturale, Verona 1998).
57.
Teugels GG: Pantodontidae, in Paugy D, Lévêque C, Teugels GG (eds): The Fresh and Brackish Water Fishes of West Africa, vol 1, pp 148-150 (IRD Editions, Paris 2003).
58.
Úbeda-Manzanaro M, Merlo MA, Palazón JL, Cross I, Serasquete C, Rebordinos L: Chromosomal mapping of the major and minor ribosomal genes, (GATA)n and U2 snRNA gene by double-colour FISH in species of the Batrachoididae family. Genetica 138:787-794 (2010).
59.
Urushido T, Takahashi E, Taki Y: Karyotypes of three species of fishes in the order Osteoglossiformes. Chromosome Inform Service 18:20-22 (1975).
60.
Uyeno T: A comparative study of chromosomes in the teleostean fish order Osteoglossiformes. Jpn J Ichthyol 20:211-217 (1973).
61.
Volff JN, Korting C, Sweeney K, Schartl M: The non-LTR retrotransposon Rex3 from the fish Xiphophorus is widespread among teleosts. Mol Biol Evol 16:1427-1438 (1999).
62.
Volff JN, Bouneau L, Ozouf-Costaz C, Fischer C: Diversity of retrotransposable elements in compact pufferfish genomes. Trends Genet 19:674-678 (2003).
63.
Völker M, Ráb P: Direct chromosome preparation from regenerating fish fin tissue, in Ozouf-Costaz C, Pisano E, Foresti F, Almeida-Toledo LF (eds): Fish Cytogenetic Techniques (Ray-Fin Fishes and Chondrichthyans), pp 37-41 (CRC Press, Enfield 2015).
64.
Wilson MVH, Murray AM: Osteoglossomorpha: phylogeny, biogeography, and fossil record and the significance of key African and Chinese fossil taxa, in Cavin L, Longbottom A, Richter M (eds): Fishes and the Break-up of Pangaea, pp 185-219 (Geological Society of London, London 2008).
65.
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).
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.