Karyotype differentiation between two species of carangid fishes,genusTrachurus (Perciformes: Carangidae)

A karyological study ofTrachurus trachurus andT. mediterraneus (Perciformes: Carangidae) was conducted by standard, fluorochrome staining (CMA₃, mithramycin, quinacrine mustard, DAPI), C-, Ag-NOR, and Alu-I banding methods. The karyotypes of both species consisted of 2n = 48 chromosomes, but of different FN:T. trachurus possessed a chromosome complement of 2 metacentric and 46 acrocentric elements, fundamental number (FN) = 50 andT. mediterraneus, a chromosome complement of 4 metacentric, 4 submetacentric, 14 subtelocentric and 26 acrocentric chromosomes, FN = 70. In neither of the two taxa investigated were heteromorphic sex chromosomes observed. The nucleolar organizer region was interstitially located on the long arm of the Ist pair of chromosomes in both species, intermediate inT. mediterraneus and subterminal inT. trachurus. Constitutive heterochromatin was found in nearly all centromeric and telomeric regions inT. trachurus; inT. mediterraneus it formed less intense telomeric and centromeric bands and thin interstitial bands on eight chromosome pairs. In addition, the C-positive material reacted differently to the digestion with endonuclease Alu-l in the two species. The results are discussed and compared with karyological data known for other species of Carangidae.     

Karyotype analysis of the sea urchinParacentrotus lividus (Echinodermata): evidence for a heteromorphic chromosome sex mechanism

A consistent diploid number of 2n = 36 was determined for the sea urchinParacentrotus lividus from the Gulf of Palermo by analysis of mitotic chromosomes of both early developing embryos and male gonads. The haploid numbern = 18 was determined by counts of spermatocyte bivalents at diakinesis. A heteromorphic chromosome sex mechanism of the XY type is likely present in this species. This is indicated by the occurrence of a chromosomal pair, pair No. 2, which is heteromorphic in both morphology and size in about 50% of the mitotic figures (metaphases and anaphases) of einbryos. In addition, heteromorphism of the same pair of chromosomes occurred during spermatogonial metaphases in the five male specimens investigated here. The detection of a low chromosome number (2n = 36) compared to other echinoids (2n = 42 to 44), a heteromorphic chromosome sex system and the involvement of three chromosome pairs in nucleolar organization (NORs) provide evidence of the specialization of theP. lividus karyotype.     

Le caryotype d'Aulacomya ater regia (Mollusca: Bivalvia: Mytilidae) des Iles Kerguelen

In 1983, specimens of the antarctic mussel Aulacomya ater regia Powell, 1957 were collected from the Kerguelen Islands and transported alive to the Zoological Station in Villefranche-sur-Mer, France. The chromosome complement of the mussel, as determined from branchial tissue, was found to consist of 13 pairs of chromosomes with terminal or subterminal centromeres. This species differs from the other species of Mytilidae studied up to the present in the number and the morphology of the chromosomes. A. ater regia is also unique among the known bivalves in displaying only terminal or subterminal centromeres.     

Cytogenetic analysis of Oedalechilus labeo (Pisces: Mugilidae), with a report of NOR variability

 A cytogenetic analysis by several staining techniques was carried out on Oedalechilus labeo specimens from the west coast of Italy, in order to extend the karyological knowledge on Mediterranean mullets. The karyotype of the species is composed of 46 acrocentric and 2 subtelocentric chromosomes. Constitutive heterochromatin was found to be restricted to the centromeres. Fluorochrome staining revealed a uniform base composition along the chromosome arms. Nucleolus organizer regions (NORs) are located on the short arms of chromosome pair 9 and can be either homomorphic or quite heteromorphic in size. In some specimens chromomycin A₃-staining and fluorescence in situ hybridization with 18S rDNA revealed a third additional and inactive NOR on the short arms of a medium-sized chromosome. This is the first report of NOR variability in Mugilidae, a family generally characterized by a quite conservative karyotype. Heterochromatin composition and NOR location differentiate O. labeo from a cytotaxonomical point of view from the Liza species studied, Liza being considered the genus from which the genus Oedalechilus derived. Received: 10 April 1999 / Accepted: 21 October 1999     

Karotypic characterization of turbot (Scophthalmus maximus) with conventional,fluorochrome and restriction endonuclease-banding techniques

A karyotypic analysis was carried out in turbot (Scophthalmus maximus) using conventional banding techniques, fluorochromes and restriction endonucleasebanding. The standard karyotype of turbot is 2n=44 chromosomes, with 48 chromosome arms. Nucleolar organizer regions, which were localized in the short arm of Chromosome Pair No. 3, showed C-and chromomycin A₃-positive staining. C-bands were mainly located at the centromeres, but were also present in some interstitial and telomeric locations. No differences were revealed among constitutive heterochromatin bands after treatment of fixed chromosomes with fluorochrome staining and restriction endonucleases. Digestion with DdeI restriction enzyme produced a pattern of interstitial banding which suggested some degree of differentiation along the chromosome arms in turbot. Neither variation in chromosome number of arm number nor polymorphism in nucleolar organizer regions was revealed in the turbot analyzed.     

Cytogenetic analysis of Epinephelus marginatus (Pisces: Serranidae), with the chromosome localization of the 18S and 5S rRNA genes and of the (TTAGGG)n telomeric sequence

 A cytogenetic analysis was carried out on specimens of Epinephelus marginatus (Lowe, 1834) from three localities in the Mediterranean Sea, to deepen knowledge of the chromosome complement of the species and identify any possible population-specific cytogenetic markers. All specimens had a 2n = 48 acrocentric karyotype with two Ag-, chromomycin A₃- and C-positive NORs (nucleolar organizer regions) in the subcentromeric region of the smallest chromosome pair. The constitutive heterochromatin was distributed centromerically. Except for NORs, neither eu- nor heterochromatin show fluorescence after fluorochrome staining, i.e. there is no localized increase of AT- or GC-rich DNA. In all populations, the (TTAGGG) _n telomeric sequences are restricted to the telomeres, and the 18S and the 5S rDNA clusters are located on different chromosome pairs. In specimens from Sardinia, additional signals after C-banding, Ag-staining and FISH (fluorescence in situ hybridization) with 18S rDNA can be observed in the telomeric region of one or two large-sized chromosomes, classified as No. 2. This suggests that additional and variable NORs could be detected in the species in addition to those on the genus-specific, NOR-bearing Chromosome Pair 24. Received: 20 October 1999 / Accepted: 14 April 2000     

Cytogenetic studies in green mussel, Perna viridis (Mytiloida: Pteriomorphia), from West Coast of India

Chromosomes of Perna viridis were characterized by karyotype analysis, C-banding and nucleolus organizer regions (NORs). The diploid chromosome number was confirmed as 30 and the karyotype is composed of ten metacentric and five submetacentric chromosome pairs. Constitutive heterochromatin blocks were found on all chromosomal pairs except chromosomal pair 15, which showed uniform staining throughout the entire chromosomes. Silver staining revealed nucleolus organizer regions on telomeric region of four chromosomal pairs, viz. 1, 3, 7 and 11. This is the first comprehensive study undertaken on chromosomes of Perna viridis.     

Karyotypic characterization of the great sturgeon, Huso huso, by multiple staining techniques and fluorescent in situ hybridization

A karyotype analysis by several staining techniques was carried out on the great sturgeon, Huso huso (Linnaeus, 1758). The karyotype (2n = 118 ± 2) was composed of 42 pairs of meta-/submetacentric chromosomes and 17 pairs of acrocentrics/microchromosomes. Constitutive heterochromatin was mainly located at the centromeric regions of the acrocentric chromosomes. The biarmed chromosomes showed weak C-bands. Fluorescent staining with GC-specific chromomycin A₃ showed clearly recognizable fluorescent regions, whereas a more uniform base composition was revealed by the AT-specific 4,6-diamidino-2 phenylindole. After Ag-staining, nucleolar organizer regions could be observed on the short arms of two medium-sized submetacentrics and on two acrocentrics. Digoxigenated 28S and 5S rDNA probes, prepared from Acipenser naccarii DNA and hybridized to metaphase chromosomes, showed signals on six and two chromosomes, respectively. The telomeric sequence (TTAGGG) _n detected by FISH was located at both ends of each chromosome. Results are discussed in relation to karyotype organization and evolution in sturgeons. Received: 1 April 1998 / Accepted: 21 July 1998     

Structural chromosomal polymorphism in the dog-whelk Nucella lapillus (Mollusca: Neogastropoda)

On the English and French Channel coasts, the dog-whelk Nucella lapilus (L.) exhibits variation in chromosome number which appears to correlate with the degree of wave action on the shore. The more common, 2n=26 morph is typically found on exposed shores subjected to a high degree of wave action, whereas those with higher chromosome numbers, up to the recorded maximum of 2n=36, are restricted to more sheltered environments. The polymorphism is thought to be Robertsonian in nature, involving centric (centromere) fission or fusion, but detailed analysis of the polymorphism has been restricted by lack of success in labelling individual chromosomes. Using a silver-staining technique for the nucleolar organiser regions (NORs), three pairs of chromosomes, in the basic 2n=26 karyotype, have been positively identified. A series of structural chromosomal rearrangements (pericentric and paracentric inversions) affecting one pair of chromosomes involved in the numerical polymorphism is described. Significant differences exist between populations with respect to this character. These chromosomal rearrangements have the potential to reduce the level of interbreeding between the different types, and may act as isolating mechamisms between breeding groups. Structural chromosomal polymorphism is likely, therefore, to have greater significance in relation to adaptation than simple numerical variation. This finding raises important questions concerning the (cyto)taxonomic status of N. lapillus in different parts of its range.     

Karyotype analyses reveal inter-individual polymorphism and association of nucleolus-organizer-carrying chromosomes in Capros aper (Pisces: Zeiformes)

Three different karyomorphs with 2n=46, 2n=44 and 2n=42 for Capros aper (L., 1758) (Zeiformes) collected from the Gulf of Lion, near Banyuls-sur-Mer, France, in July 1990 were determined. Karyomorphs were characterized by the same arm number [Fundamental number (FN)=50], suggesting that chromosome variations are due to Robertsonian fusions. In somatic metaphase spreads stained with silver nitrate, nucleolus organizer regions (NORs) consistently occupied a terminal position on the short arms of two small submetacentric chromosomes. Ca. 30% of silver-stained metaphases in each specimen showed NOR chromosomes associated in pairs by their nucleolus organizer regions.     

The chromosomes of Trypetesa lampas (Cirripedia,Acrothoracica)

Observations were made of the chromosomes of the burrowing barnacle Trypetesa lampas (Hancock). A method of squash preparation was used, which incorporated staining the material in a solution of 2% orcein in 45% acetic acid. The diploid number of chromosomes of T. lampas was found to be 12 and the haploid number 6, in both males and females. No obvious chromosomal mechanism of sex determination was found. There was, therefore, no cytogenetic confirmation of Kühnert's view (1934) that the sex of the larva is predetermined genetically. During mitosis in the females and embryos of Trypetesa lateralis Tomlinson and Kochlorine floridana Wells and Tomlinson, the diploid number of chromosomes was found to be about 14. The numbers of chromosomes in the 3 species of Acrothoracica studied were approximately half those observed in all 12 species of Thoracica and all 5 species of Rhizocephala previously investigated.     

氧化石墨烯遗传毒性的实验研究

研究氧化石墨烯(GO)的遗传毒性,考察其致突变作用,为GO在生物领域的安全应用提供依据。采用Ames试验、体外CHL细胞染色体畸变试验和小鼠体内染色体畸变试验,分别在细菌水平、细胞水平及整体动物水平研究GO的遗传毒性。GO各剂量组的Ames试验结果为阴性。CHL试验中,CHL细胞染色体畸变率随GO浓度的增加而升高,其中1.000 mg·mL~(-1)剂量组(+S_9)和0.500 mg·mL~(-1)剂量组(-S_9)畸变率显著升高(P 0.05)。小鼠骨髓细胞染色体畸变试验中,骨髓细胞染色体畸变率同样随GO浓度的增加而升高,1.000和0.500 mg·kg~(-1)剂量组的畸变率显著提高(P 0.05)。虽然Ames试验结果没有反映出GO的遗传毒性,但在体外及体内染色体畸变试验中,GO均表现出对哺乳动物细胞染色体有潜在的遗传毒性。     

Karyology of the toadfish Porichthys plectrodon (Jordan and Gilbert, 1882) (Batrachoididae) from Margarita Island,Venezuela

This paper reports the results of cytogenetic analyses carried out on Porichthys plectrodon using conventional Giemsa staining, C-banding and silver staining techniques. A diploid chromosome count of 2n=44 was observed, consisting of 8 metacentric, 10 submetacentric, 6 subtelocentric and 20 acrocentric chromosomes. Differences in length made it possible to identify homologous chromosomes within the metacentric group. Constitutive heterochromatin was distributed as large pericentromeric blocks in pairs 1 and 2, while the rest of the chromosomes were marked in centromeric regions, some more conspicuously than others. One pair of small-sized acrocentric NOR-bearing chromosomes (21) was identified by the nucleolar regions located terminally on their short arms.Communicated by P.W. Sammarco, Chauvin     

Les caryotypes de quelques espèces de bivalves et de gastéropodes marins

The karyotypes of the bivalves Ostrea edulis (2 n=20), Crassostrea gigas (2 n=20), Mytilus galloprovincialis (2 n=28) and M. edulis (2 n=28) were obtained from mitotic metaphases of branchial tissue by means of a cellular suspension technique. The occurrence of metacentric, submetacentric and telocentric chromosomes in O. edulis as well as differences in size between Pair 1 and Pair 10 emphasize the cytological interest of this species compared to C. gigas which possesses only metacentric and submetacentric chromosomes. The variation in Pair 2, which is telocentric in M. galloprovincialis and metacencentric in M. edulis, has been observed for the first time and may support taxological separation of these two species. The karyotypes of the mesogastropods Rissoa ventricosa (2 n=32) and Littorina neritoides (2 n=34) were obtained from mitotic metaphases of gonadal tissue by the same technique. Sexual chromosomes were identified in the karyotypes of R. ventricosa. At the beginning of spermatogenesis, the gonad of L. neritoides shows both haploïd and polyploïd meïotic metaphases which correspond to typical (germinal cells) and atypical (nurse cells) forms, respectively.     

Cytogenetic characterization of the greater amberjack, Seriola dumerili (Pisces: Carangidae), by different staining techniques and fluorescence in situ hybridization

A cytogenetic analysis by several staining techniques was carried out on specimens of Seriola dumerili from the Tyrrhenian coast of Sicily. The karyotype was found to be composed of 48 chromosomes in all the specimens examined. All chromosomes are subtelo- and acrocentrics, except for one pair of small-sized submetacentric chromosomes. After Ag-staining, nucleolus organizer regions (NORs) can be observed on the short arms of a medium-sized subtelocentric chromosome pair, and after chromomycin A₃-staining a bright fluorescent signal is evident at the same sites. The 18S rDNA mapping by fluorescence in situ hybridization confirms this unique NOR location. The constitutive heterochromatin is mainly restricted to centromeres and the fluorochrome-staining by chromomycin A₃ and 4′,6-diamidino-2-phenylindole revealed a uniform base composition along the chromosomal arms; therefore stock/population-specific marker chromosomes cannot be identified. Such cytogenetic features, however, rule out the presence of morphologically differentiated sex-chromosomes in the greater amberjack. Received: 16 January 1997 / Accepted: 31 January 1997     

Chromosomal complements of two forms of Neanthes japonica (Polychaeta,Nereididae) with evidence of male-heterogametic sex chromosomes

Karyotypes of the brackish-water polychaeteNeanthes japonica (Izuka) collected from five rivers in Japan from 1984 to 1988 were examined with air-drying and flame-drying methods using materials consisting of regenerating tails, clumps of spermatogonia and youngN. japonica specimens (embryos, larvae or juveniles). A diploid number of 28 was determined in well-spread metaphase chromosomes of mitotic cells. The presence of an XX-XY (male heterogametic) sex chromosome system was established for the first time in polychaetes. The Y chromosome was larger than the X chromosome. Slight differences in karyotype were found between two forms (the small- and large-egg forms), which are very similar in adult morphology but can be distinguished by reproductive and developmental characteristics.     

Number and morphology of chromosomes in three geographical populations of Ascidiella aspersa (Ascidiacea)

In contrast with some previous reports, we have found the same number of chromosomes (n=9, 2n=18) for Ascidiella aspersa Müller from the Gulf of Naples, the west coast of Scotland and the Lagoon of Venice. It follows that the hypothesis, that different geographical populations of A. aspersa might possess different chromosome numbers, is no longer tenable. The karyogram of A. aspersa shows 9 pairs of metacentric autosomes. The oocyte bivalents show chiasmata and are characterized by the faintness with which they are stained.This investigation was supported by a grant from C.N.R. for Marine Biology, n. 69.00117 14.43.3.     

Multiple sex-chromosome system and other karyological characterizations of Pterotrachea hippocampus (Mollusca: Mesogastropoda)

Two modal diploid numbers of chromosomes were found for Pterotraches hippocampus Philippi (Mollusca: Mesogastropoda) collected from the Gulf of Palermo in 1990: 2n=31 and 32 for males and females, respectively. This, along with other karyological characteristics such as the occurrence of a trivalent configuration at diakinesis and two types of metaphase-II spreads in spermatocytes, supports the notion that a X₁X₂Y/X₁X₁X₂X₂ sex mechanism operates in the species investigated here. Silver nitrate procedure revealed an intraindividual variation in the Ag-staining pattern occurring in this species. The majority of the chromosome pair displayed terminal and/or interstitial heterochromatic blocks after C-banding.     

Spermatocyte chromosome banding studies inBuccinulum corneum (Prosobranchia: Neogastropoda): Variation in silver-NOR banding pattern

Diploid number (2n=72), and haploid number (n=36) forBuccinulum corneum (L. 1758) collected from the Gulf of Palermo in December 1987 were determined. A simple method to obtain nucleolar organizer regions (NOR), and constitutive heterochromatin regions (C-bands) of chromosomes ofB. corneum is described. Analyses of silver-stained chromosome preparations ofB. corneum suggest that a within-individual variability in NOR-banding pattern is present in each of the five specimens analysed.     

Chromosomal nucleolar organizer region (NOR) phenotypes in nine species of the genus Ophryotrocha (Polychaeta: Dorvilleidae)

Chromosomal nucleolar organizer region (NOR) phenotypes have been characterized in nine species of the genus Ophryotrocha (Polychaeta: Dorvilleidae), namely O. notoglandulata, O. sp. macrovifera, O. sp. labronica pacifica, O. labronica labronica, O. puerilis puerilis, O. diadema, O. sp. robusta, O. gracills and O. hartmanni. Irrespective of chromosome number and morphology, Ag positive regions were terminally located in all but one species, O. diadema, where the NORs were pericentromerical in a metacentric pair. The presence of a single chromosome pair bearing NOR in invertebrates is considered an ancestral trait. According to this assumption, O. sp. robusta, O. dialema, and perhaps O. p. puerilis appear to be more ancestral than the other species. On the contrary, O. notoglandulaia, O. sp. macrovifera, O. sp. labronica oacifica, with two chromosomal pairs bearing NOR sites, seem to represent examples of further evolution within the genus Ophryotrocha.