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Karyotypic characterization and genomic organization of the 5S rDNA in Polypterus senegalus (Osteichthyes, Polypteridae)

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Abstract

Polypteridae (Cladistia) is a family of archaic fishes, confined to African freshwaters. On account of their primitiveness in anatomical and morphological characters and mosaic relationships among lower Osteichthyans fishes, they constitute an important subject for the study of evolution in vertebrates. Very little is known about the karyological structure of these species. In this article, a cytogenetic analysis on twenty specimens of Polypterus senegalus (Cuvier, 1829) was performed using both classical and molecular techniques. Karyotype (2n = 36; FN = 72), chromosome location of telomeric sequences (TTAGGG) n , (GATA)7 repeats and ribosomal 5S and 18S rRNA genes were examined by using Ag-NOR, classical C-banding, CMA3 staining and FISH. Staining with Ag-NOR showed the presence of two GC rich NORs on the p arm of the chromosome pair no. 1. CMA3 marked all centromerical and some (no. 1 and no. 14) telomeric regions. FISH with 5S rDNA marked the subtelomeric region of the q arm of the chromosome pair no. 14. FISH with 18S rDNA marked the telomeric region of the p arm of the chromosome pair no. 1, previously marked by Ag-NOR. (GATA)7 repeats marked the subtelomeric regions of all chromosome pairs, with the exclusion of the no. 1, 3 and 14. Hybridization with telomeric probes (TTAGGG) n showed bright signals at the end of all chromosomes. After cloning, the 5SrDNA alignment revealed an organization of sequences made up of two different classes of tandem arrays (5S type I and 5S type II) of different lengths.

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References

  • Almeida-Toledo LF, Ozouf-Costaz C, Foresti F, Bonillo C, Porto-Foresti F, Daniel-Silva M (2002) Conservation of the 5S-bearing chromosome pair and co-localization with major rDNA clusters in five species of Astyanax (Pisces, Characidae). Cytogenet Genome Res 97:229–233

    Article  PubMed  CAS  Google Scholar 

  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410

    PubMed  CAS  Google Scholar 

  • Arnason U, Gulberg A, Janke A (2001) Molecular phylogenetics of gnatosthomous (jawed) fishes: old bones, new cartilage. Zool Scr 30:249–259

    Article  Google Scholar 

  • Arratia G, Cloutier R (1996) Reassessment of the morphology of Cheirolepis canadensis (Actinopterygii). In: Shultze HP, Cloutier R (eds) Devonia fishes and plants of Miguasha Quebec, Canada. Verlag Dr. Pfeil. Munchen, pp 165–197

  • Bartsch P, Gemballa S, Piotrowski T (1997) The embryonic and larval development of Polypterus senegalus Cuvier, 1829: ist staging with reference to external and skeletal features, behaviour and locomotory habits. Acta Zool 78:309–328

    Article  Google Scholar 

  • Bi CP, Benham CJ (2004) WebSIDD: server for predicting stress-induced duplex destabilized (SIDD) sites in superhelical DNA. Bioinformatics 20(9):1477–1479

    Article  PubMed  CAS  Google Scholar 

  • Britz R, Bartsch P (2003) The myth of dorsal ribs in gnathostome vertebrates. Proc R Soc Lond B 270(Suppl):S1–S4

    Article  Google Scholar 

  • Britz R, Johnson GD (2003) On the homology of the posterior most gill arch in polypterids (Cladistia, Actinopterygii). Zool J Linn Soc 138(4):495–530

    Article  Google Scholar 

  • Cao Y, Waddel PJ, Okada N, Hasegawa M (1998) The complete mitochondrial sequence of the shark Mustelus manazo: evaluating rooting contradictions to living bony vertebrates. Mol Biol Evol 15:1637–1646

    PubMed  CAS  Google Scholar 

  • Capanna E, Cataudella S (1973) The chromosomes of Calamoichthys calabaricus (Pisces, Polypteriformes). Experientia 29:491–492

    Article  PubMed  CAS  Google Scholar 

  • Cataudella S, Sola L, Capanna E (1978) Remarks on the karyotype of the Polypteriformes. The chromosomes of Polypterus delhezi, P. endlicheri congicus and P. palmas. Experientia 34(8):999–1000

    Article  PubMed  CAS  Google Scholar 

  • Chiu C, Dewar K, Wagner GP, Takahashi K, Ruddle F, Ledje C, Bartsh P, Scemama JL, Stellwag E, Fried C, Prohaska SJ, Stadler PF, Amemiya CT (2006) Bichir HoxA cluster sequence reveals surprising trends in ray-finned fish genomic evolution. Genome Res 14:11–17

    Article  CAS  Google Scholar 

  • Cross I, Diaz E, Sanchez I, Rebordinos L (2005) Molecular and cytogenetic characterization of Crassostrea angulata chromosomes. Acquaculture 247:135–144

    Article  CAS  Google Scholar 

  • Cross I., Merlo A, Manchado M, Infante C, Canavate JP, Rebordinos L (2006) 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

    Article  PubMed  Google Scholar 

  • Daget J, Gayet M, Meunier FJ, Sire JY (2001) Major discoveries on the dermal skeleton of fossil and recent polypteriforms: a review. Fish Fisheries 2:113–122

    Article  Google Scholar 

  • Deiana AM, Cau A, Salvadori S (2000) Major and 5S ribosomal sequences of the largemouth bass Micropterus salmoides (Perciformes, Centrarchidae) are localized in GC-rich regions of the genome. Chromosome Res 8:213–218

    Article  PubMed  CAS  Google Scholar 

  • Denton TE, Howell WM (1973) Chromosomes of the African Polypterid fishes, Polypterus palmas and Calamoichthys calabaricus (Pisces: Brachiopterygii). Experientia 29:122–124

    Article  Google Scholar 

  • Fontana F, Tagliavini J, Congiu L (2001) Sturgeon genetics and citogenetics: recent advancements and perspectives. Genetica 111:359–373

    Article  PubMed  CAS  Google Scholar 

  • Gallardo-Escàrate C, Alvarez-Borrego J, Del Rio-Portilla MA, Cross I, Merlo MA, Rebordinos L (2005) Fluorescence in situ hybridization of rDNA, telomeric (TTAGGG)n and (GATA)n repeats in the red abalone Haliotis rufescens (Archaeogasteropoda: Haliotidae). Hereditas 142:73–79

    Article  PubMed  Google Scholar 

  • Gardiner B (1984) The relationships of the palaeoniscid fishes, a review based on new specimens of Mimia and Moythomasia from the Upper Devonia of Western Australia. Bulletin of the British Museum (Natural History). Geology 37:175–428

    Google Scholar 

  • Gardiner B, Schaeffer B, Masserie J (2004) A review of the lower actinopterygian phylogeny. Zool J Linn Soc 144:511–525

    Article  Google Scholar 

  • Gayet M, Meunier F, Werner C (2002) Diversification in Polypteriformes and special comparison with the Lepisosteiformes. Palaeontology 45:361–376

    Article  Google Scholar 

  • Gemballa S, Hagen K, Roder K, Rolf M, Treiber K (2003) Structure and evolution of the horizontal septum in vertebrates. J Evol Biol 16:966–975

    Article  PubMed  CAS  Google Scholar 

  • Gosse JP (1984) Polypteridae. In: Daget J, Gosse JP, Thys van den Audernaerde DFE (eds) Check-list of the freshwater fishes of Africa. Cloffa I, Paris, ORSTOM Tervuren MRAC, pp 18–29

  • Greenwood PH (1984) Polypterus and Erpetoichtys. Anachronistic Osteichthyans. In: Eldredge N, Stanley S (eds) Living fossils. Springer – Verlag, Berlin, pp 143–147

    Google Scholar 

  • Hoegg S, Brinkmann H, Taylor JS, Meyer A, (2004) Phylogenetic timing of the fish-specific Genome Duplication correlates with the diversification of Teleost fish. J Mol Evol 59:190–203

    Article  PubMed  CAS  Google Scholar 

  • Howell WM, Black DA (1980) Controlled silver staining of nucleolus organizer regions with a protective colloidal developer: a 1-step method. Experientia 36:1014–1015

    Article  PubMed  CAS  Google Scholar 

  • Inafuku J, Nabeyama M, Kikuma Y, Saitoh J, Kubota S, Kohno S (2000) Chromosomal location and nucleotide sequences of 5S ribosomal DNA of two cyprinid species (Osteichthyes, Pisces). Chromosome Res 8:193–199

    Article  PubMed  CAS  Google Scholar 

  • Kikugawa K, Katoh K, Kuraku S, Sakurai H, Ishida O, Iwabe N, Miyata T (2004) Basal jawed vertebrate phylogeny inferred from multiple nuclear DNA-coded genes. BMC Biol 2(3):1–11

    Google Scholar 

  • Lund R (2000) The new Actinopterygian order Guildayichthyiformes from the Lower Carboniferous of Montana (USA). Geodiversitas 22:171–206

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbour Laboratory Press, New York

    Google Scholar 

  • Martins C, Galetti PM Jr (1999) Chromosomal localization of 5S rDNA genes in Leporinus fish (Anostomidae, Characiformes). Chromosome Res 7:363–367

    Article  PubMed  CAS  Google Scholar 

  • Martins C, Galetti PM Jr (2000) Conservative distribution of 5S rDNA loci in Schizodon (Pisces, Anostomidae) chromosomes. Chromosome Res 8:353–355

    Article  CAS  Google Scholar 

  • Martins C, Galetti PM Jr (2001a) Organization of 5S rDNA in species of the fish Leporinus: two different genomic locations are characterized by distinct nontranscribed spacers. Genome 44:903–910

    Article  PubMed  CAS  Google Scholar 

  • Martins C, Galetti PM Jr (2001b) Two 5S array in Neotropical fish species: is it a general rule for fishes? Genetica 111:439–446

    Article  PubMed  CAS  Google Scholar 

  • Martins C, Wasko AP, Oliveira C (2002) Dynamics of 5S rDNA in the tilapia (Oreochromis niloticus) genome: repeat units, inverted sequences, pseudogenes and chromosome loci. Cytogenet Gen Res 98:78–85

    Article  CAS  Google Scholar 

  • Martins C, Wasko AP, Oliveira C, Wright JM (2000) Nucleotide sequence of 5S rDNA and localization of ribosomal RNA genes to metaphase chromosomes of the tilapiine cichlid fish, Oreochromis niloticus. Hereditas 133:39–46

    Article  PubMed  CAS  Google Scholar 

  • Mayne J, Baker RJ, Hobart HH, Hsu TC, Ryder OA, Ward OG, Wiley JE, Wurster-Hill DH, Yates TL, Moyzis RK (1990) Distribution of non-telomeric sites of the (TTAGGG)n telomeric sequence in vertebrate chromosomes. Chromosoma 99(1):3–10

    Article  Google Scholar 

  • Messias LH, Ferreira DC, Wasko AP, Oliveira C, Foresti F, Martins C (2003) 5S rDNA organization in the fish Synbrancus marmoratus (Symbranchidae; Symbranchiformes). Hereditas 139(3):228–231

    Article  PubMed  Google Scholar 

  • Mezzanotte R, Ferrucci L, Vanni R, Bianchi U (1983) Selective digestion of human metaphase chromosomes by AluI restriction endonuclease. J Histochem Cytochem 31:553–556

    PubMed  CAS  Google Scholar 

  • Morescalchi MA, Rocco L, Stingo V (2001) New karyological and molecular data in three species of ancient Osteichthyes. 14th International Chromosome Conference, 4–8 Sept. 2001, Wurzburg, Germany, p 73

  • Morescalchi MA, Rocco L, Stingo V (2002) Cytogenetic and molecular studies in a lungfish, Protopterus annectens (Osteichthyes, Dipnoi). Gene 295:279–287

    Article  PubMed  CAS  Google Scholar 

  • Morescalchi MA (2005) Karyotype, molecular characterization and phylogenetic implications in the Sarcopterygian and Polypteriformes fishes: a review. Recent Res Dev Genes Genomes 1:35–60

    Google Scholar 

  • Morescalchi MA, Liguori I, Rocco L, Stingo V (2006) Karyotypic characterization and genomic organization of the 5S rDNA in Erpetoichthys calabaricus (Osteichthyes, Polypteridae). Genetica, Nov 30, ISSN 0016–6707

  • Negi MS, Rajagopal J, Chauhan N, Cronn R, Lakshmikumaran M (2002) Length and sequence heterogeneity in 5S rDNA of Populus deltoides. Genome 45:1181–1188

    Article  PubMed  CAS  Google Scholar 

  • Ozouf-Costaz C, Pisano E, Bonillo C, Williams R (1997) Ribosomal RNA location in the Antarctic fish Champsocephalus gunnari (Notothenioidei, Channichthyidae) using banding and fluorescence in situ hybridization. Chromosome Res 4(8):557–561

    Article  Google Scholar 

  • Pasolini P, Costagliola D, Rocco L, Tinti F (2006) Molecular organization of 5S rDNAs in Rajidae (Chondrichthyes): structural features and evolution of piscine 5S rRNA genes and nontrascribed intergenic spacers. J Mol Evol 62:564–574

    Article  PubMed  CAS  Google Scholar 

  • Patterson C (1982) Morphology and interrelationships of primitive actinopterygian fishes. Am Zool 22:9759–9774

    Google Scholar 

  • Pendas AM, Moran P, Freije JP, Garcia-Vasquez E (1994) Chromosomal mapping and nucleotide sequence of two tandem repeats of Atlantic salmos 5S rDNA. Cytogenet Cell Genet 67:31–36

    PubMed  CAS  Google Scholar 

  • Ràb P, Rabova M, Reed KM, Phillips RB (1999) Chromosomal characteristics of ribosomal DNA in the primitive semionotiform fish, longnose gar, Lepisosteus osseus. Chromosome Res 7:475–480

    Article  PubMed  Google Scholar 

  • Rasmussen AS, Arnason U (1999a) Phylogenetic studies of complete mitochondrial molecules place cartilaginous fishes within the tree of bony fishes. J Mol Evol 48:118–123

    Article  PubMed  CAS  Google Scholar 

  • Rasmussen AS, Arnason U (1999b) Molecular studies suggest that cartilaginous fishes have a terminal position in the piscine tree. Proc Natl Acad Sci USA 96:2177–2182

    Article  PubMed  CAS  Google Scholar 

  • Saitoh K, Hayashizaki K, Yokoyama Y, Asahida T, Toyohara H, Yamashita Y (2000) Complete nucleotide sequence of Japanese flounder (Paralichthys olivaceus) mitochondrial genome: structural properties and cue for resolving teleostean relationships. J Hered 91(4):271–278

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Russel DW (2001) Molecular cloning. A laboratory manual, 3rd edn. Cold Spring Harbour Laboratory Press

  • Schafer F (2004) Polypterus, Flosselhechte Bichirs. Verlag A.C.S. GmbH (Aqualog.)

  • Sola L, Rossi AR, Iaselli V, Rasch EM, Monaco PG (1992) Cytogenetics of bisexual/unisexual species of Poecilia II. Analysis of heterochromatin and nucleolar organizer regions in Poecilia mexicana maxicana by C-banding and DAPI, quinacrine, chromomycin A3 and silver staining. Cytogenet Cell Genet 60:229–235

    PubMed  CAS  Google Scholar 

  • Sola L, Rossi AR, Annesi F, Gornung E (2003b) Cytogenetic studies in Sparus auratus (Pisces, Perciformes); molecular organization of 5S rDNA and chromosomal mapping of 5S and 45S ribosomal genes and of telomeric repeats. Hereditas 139(3):232–236

    Article  PubMed  CAS  Google Scholar 

  • Subramanian S, Mishra RK, Singh L (2003) Genome-wide analysis of Bkm sequences (GATA repeats): predominant association with sex chromosomes and potential role in higher order chromatin organization and function. Bioinformatics 19(6):681–615

    Article  PubMed  CAS  Google Scholar 

  • Tiersh TR, Simco BA, Davis KB, Watchel SS (1992) Molecular genetics of sex determination in channel catfish: studies on SRY, ZFY, Bkm and human telomeric repeats. Biol Reprod 47(2):185–192

    Article  Google Scholar 

  • Tigano C, Rocco L, Ferrito V, Costagliola D, Pappalardo AM, Stingo V (2004) Chromosomal mapping and molecular characterization of ribosomal RNA genes in Lebias fasciata (Teleostei, Cyprinodontidae). Genetica 121:95–100

    Article  PubMed  CAS  Google Scholar 

  • Urushido T, Takahashi Y, Taki Y, Kondo N (1977) A karyotype study of polypterid fishes, with notes on their phyletic relationships. Proc Jpn Acad 53:95–98

    Article  Google Scholar 

  • Venkatesh B, Ning Y, Brenner S (1999) Late changes in spliceosomal introns define clades in vertebrate evolution. Proc Nat Acad Sci USA 96:10267–10271

    Article  PubMed  CAS  Google Scholar 

  • Venkatesh B, Erdmann MV, Brenner S (2001) Molecular synapomorphyes resolve evolutionary relationships of extant jawed vertebrates. Proc Nat Acad Sci USA 98:11382–11387

    Article  PubMed  CAS  Google Scholar 

  • Vervoort A (1980a) Tetraploidy in Protopterus (Dipnoi). Experientia 36:294–296

    Article  Google Scholar 

  • Vervoort A (1980b) Karyotypes and nuclear DNA contents of Polypteridae (Osteichthyes). Experientia 36(6):646–647

    Article  PubMed  CAS  Google Scholar 

  • Volff JN, Schartl M (2001) Variability of genetic sex determination in poeciliid fishes. Genetica 111:101–110

    Article  PubMed  CAS  Google Scholar 

  • Zardoya R, Meyer A (1996) Evolutionary relationships of the coelacanth, lungfishes and tetrapods based on the 28s ribosomal gene. Proc Natl Acad Sci USA 93:5449–5454

    Article  PubMed  CAS  Google Scholar 

  • Zardoya R, Meyer A (2001) Vertebrate phylogeny: limits of inference of mitochondrial genome and nuclear rDNA sequence data due to an adverse phylogenetic signal/noise ratio. In: Ahlberg PE (ed) Major events in early Vertebrate evolution. Taylor & Francis, London, pp 135–155

    Google Scholar 

  • Zardoya R, Cao Y, Hasegawa M, Meyer A (1998) Searching for the closest living relative(s) of tetrapods through evolutionary analyses of mitochondrial and nuclear data. Mol Biol Evol 15:506–517

    PubMed  CAS  Google Scholar 

  • Wachtel S, Demas S, Tiersh T, Pechan P, Shapiro D (1991) Bkm satellite DNA and ZFY in the coral reef fish Anthias squamipinnis. Genome 34(4):612–617

    PubMed  CAS  Google Scholar 

  • Wasko AP, Martins C, Wright JM, Galetti PM (2001) Molecular organization of 5S rDNA in fishes of the genus Brycon. Genome 44:893–902

    Article  PubMed  CAS  Google Scholar 

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Morescalchi, M.A., Liguori, I., Rocco, L. et al. Karyotypic characterization and genomic organization of the 5S rDNA in Polypterus senegalus (Osteichthyes, Polypteridae). Genetica 132, 179–186 (2008). https://doi.org/10.1007/s10709-007-9160-7

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