Abstract
Karyotype and other chromosomal characteristics in the Adriatic brook lamprey Lampetra zanandreai, representative of one of the most ancestral group of vertebrates, were examined using conventional (Ag-staining, C-banding as well as CMA3 and DAPI fluorescence) and molecular (FISH with 18/28S rDNA and EcoRI satDNA as probes) protocols with metaphase chromosomes derived from whole blood cultures. The chromosome complement had a modal diploid chromosome number of 2n = 164, as in other petromyzontid lamprey species. Ag-staining and CMA3 fluorescence, as well as FISH with 18/28S rDNA probes, detected nucleolar organizer regions (NORs) close to the centromeres of the biarmed chromosomes of pairs 1 and 2, the largest chromosome pairs of the complement. In addition to NORs, CMA3 fluorescence revealed positive signals in approximately 40 other chromosomes. DAPI stained mostly centromeric regions of many chromosomes as well as conspicuously massive blocks overlapping NOR sites. C-banding evidenced a large amount of constitutive heterochromatin in somatic chromosomes, with approximately 40 C-positive acrocentric elements completely heterochromatic, corresponding with the 40 CMA3+ chromosomes and positive heterochromatic blocks in pericentromeric regions of chromosome pairs 1 and 2. Polymerase chain reaction (PCR)-based cloning of satellite DNA with primers derived from Petromyzon marinus centromeric sequences was successful for L. zanandreai genomic DNA. The sequence was AT-rich (59%) and characterized by short consensus motifs similar to other centromeric satellite motifs. FISH using satDNA clones as a probe produced a fluorescent signal on a single pair of small chromosomes. This sequence was PCR-amplified also in L. planeri and P. marinus genomic DNA, and the evolution of this repetitive element in the above species was analysed.
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Abbreviations
- CMA3 :
-
Chromomycin A3
- DAPI:
-
4′,6-Diamidino-2 phenyl-indole
- FISH:
-
Fluorescence in situ hybridization
- K2P:
-
Kimura 2-parameters
- NJ:
-
Neighbour-joining
- NOR:
-
Nucleolar organizing region
- rDNA:
-
Major ribosomal sites
- satDNA:
-
Satellite DNA
References
Amemiya CT, Saha NR, Zapata A (2007) Evolution and development of immunological structures in the lamprey. Curr Opin Immunol 19:535–541
Bachmann L, Sperlich D (1993) Gradual evolution of a specific satellite DNA family in Drosophila ambigua. D. tristis and D. obscura. Mol Biol Evol 10:647–659
Bianco PG (1992) Zoogeographical implications of a first record of Lethenteron zanandreai on the Adriatic slope of central Italy (Cyclostomata: Petromyzontidae). Ichthyol Explor Freshwaters 3:183–186
Boán F, Viñas A, Rodríguez JM, Sánchez L, Gómez-Márquez J (1996) A new EcoRI family of satellite DNA in lampreys. FEBS Lett 394:187–190
Canapa A, Nisi Cerioni P, Barucca M, Olmo E, Caputo V (2002) A centromeric satellite DNA may be involved in heterochromatin compactness in gobiid fishes. Chromosome Res 10:297–304
Capriglione T, Cardone A, Olmo E, Odierna G (1991) Evolution of a centromeric satellite and phylogeny of lacertid lizards. Comp Biochem Physiol 100B:641–645
Caputo V, Giovannotti M, Nisi Cerioni P, Splendiani A, Marconi M, Tagliavini J (2009a) Mitochondrial DNA variation of an isolated population of the Adriatic brook lamprey (Lampetra zanandreai Vladykov, 1955): phylogeographic and phylogenetic inferences (Agnatha: Petromyzontidae). J Fish Biol 75:2344–2351
Caputo V, Giovannotti M, Nisi Cerioni P, Splendiani A, Olmo E (2009b) Chromosomal study of native and hatchery trouts from Italy (Salmo trutta complex, Salmonidae): conventional and FISH analysis. Cytogenet Genome Res 124:51–62
Ciobanu DG, Rudykh IA, Grechko VV, Kramerov DA (2001) Molecular mechanisms of satellite DNA evolution in a group of closely related Caucasian rock lizards. In: Piruzian E, Volkova L (eds) Molecular Mechanisms of Genetic Processes and Biotechnology. International Symposium. Thesis: Moskow, Russia, p. 26
Ciobanu DG, Grechko VV, Darevsky IS, Kramerov DA (2004) New satellite DNA in Lacerta s. str. lizards: evolutionary pathways and phylogenetic impact. J Exp Zoolog B Mol Dev Evol 302:505–516
Colomba M, Vitturi R, Libertini A, Gregorini A, Zunino M (2006) Heterochromatin of the scarab beetle, Bubas bison (Coleoptera: Scarabaeidae) II. Evidence for AT-rich compartmentalization and high amount of rDNA copies. Micron 37:47–51
Cremisi F, Vignali R, Batistoni R, Barsacchi G (1988) Heterochromatic DNA in Triturus (Amphibia, Urodela). II. A centromeric satellite DNA. Chromosoma 97:204–211
Dasilva C, Hadji H, Ozouf-Costaz C, Nicaud S, Jaillon O, Weissenbach J, Roest Crollius H (2002) Remarkable compartmentalization of transposable elements and pseudogenes in the heterochromatin of the Tetraodon nigroviridis genome. Proc Natl Acad Sci U S A 99:13636–13641
De la Herrán R, Fontana F, Lanfredi M, Congiu L, Leis M, Rossi R, Ruiz Rejón C, Ruiz Rejón M, Garrido-Ramos MA (2001) Slow rates of evolution and sequence homogenization in an ancient satellite DNA family of sturgeons. Mol Biol Evol 18:432–436
Docker MF, Youson JH, Beamish RJ, Devlin RH (1999) Phylogeny of the lamprey genus Lampetra inferred from mitochondrial cytochrome b and ND3 gene sequences. Can J Fish Aquat Sci 56:2340–2349
Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791
Fontana F, Bruch RM, Binkowski FP, Lanfredi M, Chicca M, Beltrami N, Congiu L (2004) Karyotype characterization of the lake sturgeon, Acipenser fulvescens (Rafinesque 1817) by chromosome banding and fluorescent in situ hybridization. Genome 47:742–746
Fujiwara A, Nishida-Umehara C, Sakamoto T, Okamoto N, Nakayama I, Abe S (2001) Improved fish lymphocyte culture for chromosome preparation. Genetica 111:77–89
Garrido-Ramos MA, Jamilena M, Lozano R, Ruiz Rejón C, Ruiz Rejón M (1995) The EcoRI centromeric satellite DNA of the Sparidae family (Pisces: Perciformes) contains a sequence motive common to other vertebrate centromeric satellite DNAs. Cytogenet Cell Genet 71:345–351
Gess RW, Coates MI, Rubidge BS (2006) A lamprey from Devonian period of South Africa. Nature 443:981–984
Gill HS, Renaud CB, Chapleau F, Mayden RL, Potter IC (2003) Phylogeny of living parasitic lamprey (Petromyzontiformes) based on morphological data. Copeia 2003:687–703
Haaf T, Schmid M, Steinlein C, Galetti PM, Willard H (1993) Organization and molecular cytogenetics of a satellite DNA family from Hoplias malabaricus (Pisces, Erythrinidae). Chromosome Res 1:77–86
Holcik J, Mrakovcic M (1997) First record of Lethenteron zanandreai (Cyclostomata, Petromyzontidae) in the Adriatic basin of the Balkan peninsula and its zoogeographic consequences. Folia Zoologica 46:263–271
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
Howell WM, Denton TE (1969) Chromosomes of ammocoetes of the Ohio brook lamprey Lampetra aepyptera. Copeia 1960:393–395
Howell WM, Duckett CR (1971) Somatic chromosome of the lamprey, Icthyomyzon gagei. Experentia 27:222–223
Kasahara M (2007) The 2R hypothesis: an update. Curr Opin Immunol 19:547–552
Kimura MA (1980) simple method for estimating evolutionary rate of base substitution through comparative studies of nucleotide sequence. J Mol Evol 16:111–120
King LM, Cummings MP (1997) Satellite DNA repeat sequence variation is low in three species of burying beetles in the genus Nicrophorus (Coleoptera: Silphidae). Mol Biol Evol 14:1088–1095
Klinkhardt M, Tesche M, Greven H (1995) Data Base of Fish Chromosomes. Westarp Wissenschaften, Magdeburg
Kojima NF, Kojima KK, Kobayakawa S, Higashide N, Hamanaka C, Nitta A, Koeda I, Yamaguchi T, Shichiri M, Kohno S, Kubota S (2010) Whole chromosome elimination and chromosome terminus elimination both contribute to somatic differentiation in Taiwanese hagfish Paramyxine sheni. Chromosome Res 18:383–400
Kubota S, Takano J, Tsuneishi R, Kobayakawa S, Fujikawa N, Nabeyama M, Kohono S (2001) Highly repetitive DNA families restricted to germ cells in a Japanese hagfish (Eptatretus burgeri): a hierarchical and mosaic structure in eliminated chromosomes. Genetica 111:319–328
Kumar S, Tamura K, Jakobsen IB, Nei M (2001) MEGA2: molecular evolutionary genetics analysis software. Bioinformatics 17:1244–1245
Kuraku S, Kuratani S (2006) Time scale for Cyclostome evolution inferred with a phylogenetic diagnosis of hagfish and lamprey cDNA sequences. Zool Sci 23:1053–1064
Kusakabe R, Kuratani S (2007) Evolutionary perspectives from development of the mesodermal components in the lamprey. Dev Dyn 236:2410–2420
Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) ClustalW and ClustalX version 2.0. Bioinformatics 23:2947–2948
McCauley DW, Kuratani S (2008) Cyclostome studies in the context of vertebrate evolution. Zool Sci 25:953–954
Osório J, Rétaux S (2008) The lamprey in evolutionary studies. Dev Genes Evol 218:221–235
Plohl M, Luchetti A, Meštrović N, Mantovani B (2008) Satellite DNAs between selfishness and functionality: structure, genomics and evolution of tandem repeats in centromeric (hetero)chromatin. Gene 409:72–82
Potter IC, Robinson ES (1981) New developments in vertebrate cytotaxonomy V. Citotaxonomy of lampreys. Genetica 56:149–151
Potter IC, Robinson ES, Walton SM (1968) The mitotic chromosome of the lamprey Mordacia mordax (Agnatha: Petromyzonidae). Experentia 24:966–967
Potter IC, Rothwell B (1970) The Mitotic chromosome of the lamprey, Petromyzon marinus. Experentia 26:429–430
Renaud CB (1997) Conservation status of northern hemisphere lampreys (Petromyzontidae). J Appl Ichthyol 13:143–148
Robinson ES, Potter IC (1981) The chromosome of the southern hemispheric lamprey, Geotria australis Gray. Experentia 37:239–240
Robinson ES, Potter IC, Webb CJ (1974) Homogeneity of holarctic lamprey karyotypes. Caryologia 27:443–454
Robles F, de la Herrán R, Ludwig A, Ruiz Rejón C, Ruiz Rejón M, Garrido-Ramos MA (2004) Evolution of ancient satellite DNAs in sturgeon genomes. Gene 338:133–142
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor
Sauka-Spengler T, Meulemans D, Jones M, Bronner-Fraser M (2007) Ancient evolutionary origin of the neural crest gene regulatory network. Dev Cell 13:405–420
Schmid M, Haaf T, Geile B, Sims S (1983) Chromosome banding in Amphibia. VIII. An unusual XY/XX sex chromosome system in Gastrotheca riobambae (Anura, Hylidae). Chromosoma 88:69–82
Schwarzacher T, Heslop-Harrison P (2000) Practical in situ hybridization. Springer, New York
Schweizer D (1979) Fluorescent chromosome banding in plants: applications, mechanisms and implications for chromosome structure. In: Davies DR, Hopwood RA (eds) The Plant Genome. Proc 4th John Innes Symposium, John Innes Charity, Norwich, pp 61–72
Silver MR, Kawauchi H, Nozaki M, Sower SA (2004) Cloning and analysis of the lamprey GnRH-III cDNA from eigth species representing the three famiglie of Petromyzontiformes. Gen Comp End 139:85–94
Singer MF (1982) Highly repeated sequences in mammalian genomes. Int Rev Cytol 76:67–112
Smith JJ, Antonacci F, Eichler EE, Amemiya CT (2009) Programmed loss of millions of base pairs from a vertebrate genome. Proc Natl Acad Sci U S A 106:11212–11217
Smith JJ, Stuart AB, Sauka-Spengler T, Clifton SW, Amemiya CT (2010) Development and analysis of a germline BAC resource for the sea lamprey, a vertebrate that undergoes substantial chromatin diminution. Chromosoma 119:381–389
Sumner AT (1972) A simple technique for demonstrating centromeric heterochromatin. Expl Cell Res 75:304–306
Suzuki A, Ikeda Y, Nakayama K (1999) Chromosome and Ag-NORs of three species of Lampetra (Petromyzontiformes). Chromosome Sci 3:150
Swarça AC, Fenocchio AS, Cestari MM, Dias AL (2003) Analysis of heterochromatin by combination of C-banding and CMA3 and DAPI staining in two fish species (Pimelodidae, Siluriformes). Genetica 119:87–92
Tutman P, Dulcic J, Glamuzina B (2009) First record of Po brook lamprey, Lethenteron zanandreai (Cephalospidomorpha, Petromyzontiformes, Petromyzontidae) in the Hutoro-Blato wetland, Bosnia and Herzegonina. Acta Ichthyologica Piscatoria 39:55–58
Vissel B, Nagy A, Choo KHA (1992) A satellite III sequence shared by human chromosomes 13, 14 and 21 that is contiguous with alpha satelliteDNA. Cytogenet Cell Genet 61:81–86
Wong AKC, Rattner JB (1988) Sequence organization and cytological localization of the minor satellite of mouse. Nucl Acids Res 16:11645–11661
Zanandrea G, Capanna E (1964) Contributo alla cariologia del genere Lampetra. Boll Zool 31:669–677
Zardoya R, Meyer A (1996) Evolutionary relationships of the coelacanth, lungfishes, and tetrapods based on the 28 S ribosomal RNA gene. Proc Natl Acad Sci U S A 93:5449–5454
Acknowledgements
The authors are very grateful to Petr Ráb (Academy of the Sciences of the Czech Republic, Liběchov, Czech Republic) for valuable comments and suggestions on the manuscript. We wish to thank the provincial Administration of Macerata (Italy) for permitting specimen collection, Mario Marconi (University of Camerino, Italy) for assisting in field activities, and Mrs. Patricia O’Brien for linguistic revision. This research was financed by Università Politecnica delle Marche.
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1
Southern blot hybridisation of the genomic DNA of L. zanandreai (lane 1), P. marinus (lane 2) and L. planeri (lane 3) digested with EcoRI. The hybridization was carried out with a digoxigenin-labelled probe from the centromeric satellite isolated from L. zanandreai by PCR. Arrow indicates the monomer of the EcoRI satDNA. (JPEG 85 kb)
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Caputo, V., Giovannotti, M., Cerioni, P.N. et al. Chromosomal study of a lamprey (Lampetra zanandreai Vladykov, 1955) (Petromyzonida: Petromyzontiformes): conventional and FISH analysis. Chromosome Res 19, 481–491 (2011). https://doi.org/10.1007/s10577-011-9197-4
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DOI: https://doi.org/10.1007/s10577-011-9197-4