Abstract
Polypterids are a group of Osteichthyan fish whose evolutionary relationships with closer basal ray-finned and lobe-finned fish have been disputed since their discovery. Very little is known about the evolutive karyology in the whole Polypteriformes group. In order to fill this gap, a cytogenetic analysis of Erpetoichthys calabaricus species was performed, using both classical and molecular techniques. Karyotype structure (2n = 36; FN = 72), chromosome location of telomeric sequences (TTAGGG) n and ribosomal 5S and 18S rRNA genes were examined in twenty specimens of E. calabaricus by using Ag-NOR, classical C-banding, sequential CMA3/4′,6-diaminidino-2-phenylindole (DAPI) staining and fluorescent in situ hybridization (FISH). CMA3 marked all centromerical and some (n°1 and n°15) telomeric regions. Staining with Ag-NOR and CMA3 showed the presence of two NORs on the p arm of the chromosome pair n°1. Hybridization with telomeric probes (TTAGGG) n showed signals at the end of all chromosomes. 5S rDNA was cloned and sequenced. After the alignment, the 5S rRNA sequences revealed an organization made up of two different classes of tandem arrays (type I and type II). FISH with 5S rDNA marked the telomeric regions of the small chromosome pair n°15, while FISH with 18S rDNA marked the telomeric region of the pair n°1. The results obtained were compared with cariological data on closer species now available in literature.
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References
Albuin M, Martinez P, Sanchez L (1996) Localization of the repetitive telomeric sequences (TTAGGG) n in four salmonid species. Genome 39:1035–1038
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Molec Biol 215:403–410
Birstein VJ, Hanner R, Desalle R (1997) Phylogeny of the Acipenseriformes: cytogenetic and molecular approaches. Env Biol Fishes 48:127–156
Capanna E, Cataudella S (1973) The chromosomes of Calamoichthys calabaricus (Pisces, Polypteriformes). Experientia 29:491–492
Carrol RL (1988) Vertebrate paleontology and evolution. Freedman, New York
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
Chew JS, Oliveira C, Wright JM, Dobson MJ (2002) Molecular and cytogenetic analysis of the telomeric (TTAGGG) n repetitive sequences in the Nile tilapia, Oreochromis niloticus (Teleostei: Cichlidae). Chromosoma 111(1):45–52
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
Denton TE, Howell WM (1973) Chromosomes of the African Polypterid fishes, Polypterus palmas and Calamoichthys calabaricus (Pisces: Brachiopterygii). Experientia 29:122–124
Fajkus J, Sykorova E, Leitch AR (2005) Telomeres in evolution and evolution of telomeres. Chromosome Res 13(5):469–479
Ferro DA, Neo M, Moreira-Filho O, Bertollo LAC (2001) Nucleolar organizing regions, 18S and 5S rDNA in Astyanax scabripinnis (Pisces, Characidae): populations distribution and functional diversità. Genetica 110:55–62
Fontana F, Bruch RM, Binkowski FP, Lanfredi M, Chicca M, Feltrami N, Congiu L (2004) Karyotype characterization of the lake strurgeon, Acipenser fulvescens (Rafinesque, 1817) by chromosome banding and fluorescent in situ hybridization. Genome 47(4):742–746
Fontana F, Tagliavini J, Congiu L (1998) Karyotypic characterization of the great sturgeon, Huso huso, by multiple staining techniques and fluorescent in-situ hybridization. Mar Biol 132:495–501
Gardiner BG, Schaeffer AB (1989) Interrelationships of lower actinopterygian fishes. Zool J Linn Soc 97:135–187
Gardiner BG, Shaeffer AB, Masserie J (2005) A review of the lower actinopterigian phylogeny. Zool J Linn Soc 144:511–525
Garrido-Ramos MA, de la Herran R, Rejon CR, Rejon MR (1998) A satellite DNA of the sparidae family (pisces, perciformes) associated with telomeric sequences. Cytogenet Cell Genet 83:3–9
Gayet M, Meunier F (1991) Polypteridae (Pisces, Cladistia, Polypteriformes) from the late Cretaceous and early Paleocene of Bolivia. In: Suarez-Soruco R (ed) Fosiles Y Facies de Bolivia. Revista Tecnica de Yacimientos Petroliferos Fiscales Bolivianos, pp 495–498
Gayet M, Meunier F (1992) Polypteriformes (Pisces, Cladistia) du Maastrichtien et du Paleocene de Bolivie. Geobios 14:159–168
Gayet M, Meunier F, Werner C (2002) Diversification in Polypteriformes and special comparison with the Lepisosteiformes. Palaeontology 45:361–376
Gosse JP (1984) Polypteridae. In: Daget J, Gosse P, Thys van den Audernaerde DFE (eds) Check-list of the freshwater fishes of Africa. Cloffa I, Paris, ORSTOM Tervuren MRAC, pp 18–29
Gosse JP (1988) Revision systematique de deux especes du genre Polypterus (Pisces, Polypteridae). Cybium 12:239–245
Hanssens MM, Tegels GG, Thys van den Audenaerde DFE (1995) Subspecies in the Polypterus palmas complex (Brachiopterygii: Polypteridae) from west and central Africa. Copeia 3:694–705
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
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). Chrom Res 8:193–199
Lauder GV, Liem KF (1983) The evolution and interrelationships of the actinopterygian fishes. Bull Mus Comp Zool 150:95–197
Mandrioli M, Manicardi GC, Machella N, Caputo V (2000) Molecular and cytogenetic analysis of the Gobius niger (Teleostei, Gobiidae). Genetica 110(1):73–78
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbour Laboratory Press, New York
Martins C, Galetti PM Jr (1999) Chromosomal localization of 5S rDNA genes in Leporinus fish (Anostomidae, Characiformes). Chrom Res 7:363–367
Martins C, Galetti PM Jr (2000) Conservative distribuition of 5S rDNA loci in Schizodon (Pisces, Anostomidae) chromosomes. Chrom Res 8:353–355
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
Martins C, Galetti PM Jr (2001b) Two 5S array in Neotropical fish species: is it a general rule for fishes? Genetica 111:439–446
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
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
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
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
Moran P, Martinez JL, Garcia-Vasquez E, Pendas AM (1996) Sex chromosome linkage of 5S rDNA in rainbow trout (Oncorhyncus mykiss). Cytogenet Cell Genet 75:145–150
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
Nelson JS (1994) Fishes of the world. John Wiley, NY
Ocalewicz K, Babiak I, Dobosz S, Nowaczyk J, Goryczko K (2004b) The stability of telomereless chromosome fragments in adult androgenetic raibow trout. J Exp Biol 207:2229–2236
Ocalewicz K, Sliwinska A, Jankun M (2004a) Autosomal localization OD interstitial telomeric sites (ITS) in brook trout, Salvelinus fontinalis (Pisces, Salmonidae). Cytogenet Genome Res 105(1):79–82
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
Pendas AM, Moran P, Freije JP, Garcia-Vazquez E (1994) Chromosomal mapping and nucleotide sequence of two tandem repeats of Atlantic salmos 5S rDNA. Cytogenet Cell Genet 67:31–36
Perez J, Moran P, Garcia-vazquez E (1999) Cloning and physical mapping of Atlantic salmon (Salmo salar L.) telomeric sequences. Heredity 82:409–414
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
Rabova M, Rab P, Ozouf-Costaz C (2001) Extensive polymorphism and chromosomal characteristics of ribosomal DNA in a loach fish, Cobitis vardarensis (Ostariophysi, Cobitidae) detected by different banding techniques and fluorescence in situ hybridization (FISH). Genetica 111:413–422
Reed KM, Phillips RB (1995) Molecular cytogenetic analysis of the double-CMA3 chromosome of lake trout, Salvelinus namaycush. Cytogenet Cell Genet 70:104–107
Rocco L, Costagliola D, Stingo V (2001) (TTAGGG) n telomeric sequences in selachian chromosomes. Heredity 87:583–588
Rocco L, Morescalchi MA, Costagliola D, Stingo V (2002) Karyotype and genome characterization in four cartilaginous fishes. Gene 7:295(2):289–298
Sajidak SL, Reed KM, Phillips RB (1998) Intraindividual and interspecies variation in the 5S rDNA of coregonid fish. J Mol Evol 46:680–688
Salvadori S, Deiana AM, Cosuccia E, Floridia G, Rossi E, Zuffardi O (1995) Colocalization of (TTAGGG) n telomeric sequences and ribosomal genes in Atlantic eels. Chromosome Res 3:54–58
Sambrook J, Russel DW (2001) Molecular cloning. A laboratory manual, 3rd edn. Cold Spring Harbour Laboratory Press
Schmid M, Guttenbach M (1988) Evolutionary diversity of reverse (R) fluorescent bands in vertebrates. Chromosoma 97:327–344
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
Sola L, Gornung E, Naoi H, Gunji R, Sato C, Kawamura K, Arai R, Ueda T (2003a) FISH-mapping of 18S ribosomal RNA genes and telomeric sequences in the Japanese bitterlings Rhodeus ocellatus kurumeus and Tanakia limbata (Pisces, Cyprinidae) reveals significant cytogenetic differences in morphologically similar karyotypes. Genetica 119(1):99–106
Sola L, Rossi AR, Annesi F, Gornung E (2003b) Cytogenetic srudies 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
Tagliavini J, Williot P, Congiu L, Chicca M, Lanfredi M, Rossi R, Fontana F (1999) Molecular cytogenetic analysis of the karyotype of the European Atlantic sturgeon, Acipenser sturio. Heredity 83:520–525
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
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
Vervoort A (1980a) Tetraploidy in Protopterus (Dipnoi) Experientia 36:294–296
Vervoort A (1980b) Karyotypes and nuclear DNA contents of Polypteridae (Osteichthyes). Experientia 36(6):646–647
Volff JN, Schartl M (2001) Variability of genetic sex determination in poeciliid fishes. Genetica 111:101–110
Wasko AP, Martins C, Wright JM, Galetti PM (2001) Molecular organization of 5S rDNA in fishes of the genus Brycon. Genome 44:893–902
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Morescalchi, M.A., Liguori, I., Rocco, L. et al. Karyotypic characterization and genomic organization of the 5S rDNA in Erpetoichthys calabaricus (Osteichthyes, Polypteridae). Genetica 131, 209–216 (2007). https://doi.org/10.1007/s10709-006-9119-0
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DOI: https://doi.org/10.1007/s10709-006-9119-0