0000000000030819
AUTHOR
Mariastella Colomba
Comparative Cytogenetic Analysis of Three Stylommatophoran Slugs (Mollusca, Pulmonata)
system is still controversial because phylogeny and systematic relationships at the family level are poorly understood. Besides morphological studies, ribosomal RNA genes and the H3/H4 histone gene cluster (Ambruster et al., 2005; Wade et al., 2006) have also been used to resolve the relationships within this order. Recently, by comparison of primary sequence of mitochondrial and nuclear genes, Grande et al. (2004) resolved the Stylommatophora as the early split monophyletic sister group of all the other gastropod taxa. Available data on the cytogenetics of Stylommatophora are extremely poor (reviewed by Patterson, 1969, and Thiriot-Quievreux, 2003) and mostly concern the haploid (n) and/ o…
Organometallic complexes with biological molecules
Novel triorganotin(IV) complexes of two beta-lactamic antibiotics, 6-[D-(-)-beta-amino-p-hydroxyphenyl-acetamido]penicillin (=amoxicillin) and 6-[D-(-)-alpha-aminobenzyl]penicillin (=ampicillin), have been synthesized and investigated both in solid and solution states. The complexes corresponded to the general formula R(3)Sn(IV)antib*H(2)O (R=Me, n-Bu, Ph; antib=amox=amoxicillinate or amp=ampicillinate). Structural investigations about configuration in the solid state have been carried out by interpreting experimental IR and 119Sn Mossbauer data. In particular, IR results suggested polymeric structures both for R(3)Sn(IV)amox.H(2)O and R(3)Sn(IV)amp*H(2)O. Moreover, both antibiotics appear …
Cytogenetics of the land snails Cantareus aspersus and C. mazzullii (Mollusca: Gastropoda: Pulmonata).
A cytogenetic study was carried out on the chromosomes and nuclear DNA contents of the land snails Cantareus aspersus and C. mazzullii (Gastropoda: Pulmonata). Chromosomes were studied using Giemsa staining, banding methods and fluorescent in situ hybridization (FISH) with three repetitive DNA probes [18S rDNA, (GATA)n and (TTAGGG)n]. Results were very similar in the two species both showing (1) 54 bi-armed chromosomes [submetacentrics (SM) C metacentrics (M) C subtelocentrics (ST)]; (2) 10 terminal NORs after sequential application of rDNA FISH and silver staining; (3) uniform DNA fluorescence with CMA3 and DAPI staining and (4) genomic composition considerably enriched both in highly- and…
Cytogenetic characterization ofBrachidontes pharaonis(Fisher P., 1870): Karyotype, banding and fluorescentin situhybridization (fish) (Mollusca: Bivalvia: Mytilidae)
Abstract The mussel Brachidontes pharaonis (Fisher P., 1870) (Bivalvia: Mytilidae) has a diploid chromosomal set of 28 made up of 14 pairs of which eight are mono-armed (ST) and six bi-armed (M+SM). Fourteen bivalents occur in spermatocytes both at pachytene and metaphase-I. The use of combined silver and CMA3 staining reveals that nucleolus organizer regions (NORs) are located terminally on the long arm of a small subtelocentric chromosome pair (pair 14) and are compartmentalized in GC base pairs. A Paracentrotus lividus (Echinodermata) 4.3 kilobase (kb) rDNA probe (prR14) consisting of sequences from the 3′ end of 18S rDNA to the 3′ end of 26S rDNA was used to map the rDNA loci of B. phar…
Chromosomal studies of five tropical scorpaeniform fishes (Teleostei, Scorpaenidae)
Abstract The karyotypes of five species of Scorpaenidae (genera Scorpenopsis, Dendrochirus and Pterois) from the Indian Ocean were analysed using various banding methods and in situ hybridisation with a ribosomal probe. All the species investigated are characterised by a diploid set of 48 chromosomes (mainly acrocentric and/or subtelocentric) and by a NOR location on the small arm of a medium‐sized pair. All the chromosomes stained uniformly with DAPI, whereas C‐banding evidenced a small amount of hete‐rochromatin. Despite the marked morphological differences among these species, the low degree of diversification of the chromosome sets with respect to the ancestral set of teleosts (2n = 48 …
Characterization of mitotic chromosomes of four species of the genus Diplodus: Karyotypes and chromosomal nucleolar organizer region phenotypes
Karyotypes have been described in four Mediterranean species of the genus Diplodus (Teleostei, Sparidae), D. vulgaris, D. puntazzo, D. sargus and D. annularis. Chromosomes were mainly acrocentric in all but D. vulgaris, where certain chromosome pairs were subtelocentric. A remarkable intraspecific heteromorphism in the number of NOR-bearing chromosomes along with a substantial interspecific variability in position of chromosomal Ag signals have been encountered. The presumed origin of multiple NOR-bearing chromosomes in Diplodus species and variation of the NOR location is discussed. ? 1996 The Fisheries Society of the British Isles
Characterization of mitotic and meiotic, chromosomes of the vermetid gastropodDendropoma (Novastoa) Petraeum(Monterosato, 1884) (Mollusca, Caenogastropoda)
Abstract The vermetid gastropod Dendropoma (Novastoa) petraeum from the Northwestern coast of Sicily was investigated karyologically. The count of spermatocyte bivalents at diakinesis gave the haploid number of this species as n=17. The count of mitotic chromosomes in males, females and early developing embryos gave the diploid number as 2n=34. Irrespective of sex, the karyotype of D. petraeum consists of 17 homomorphic chromosome pairs, of which 8 are metacentric, 4 subtelocentric and 5 acrocentric. This species is characterized by the presence of two chromosomes per cell involved in nucleolus organization (NOR) and by a low amount of constitutive heterochromatin.
Organometallic complexes with biological molecules. XVIII. Alkyltin(IV) cephalexinate complexes: synthesis, solid state and solution phase investigations
Abstract Dialkyltin(IV) and trialkyltin(IV) complexes of the deacetoxycephalo-sporin-antibiotic cephalexin [7-( d -2-amino-2-phenylacetamido)-3-methyl-3-cephem-4-carboxylic acid] (Hceph) have been synthesized and investigated both in solid and solution phase. Analytical and thermogravimetric data supported the general formula Alk 2 SnOHceph · H 2 O and Alk 3 Snceph · H 2 O (Alk=Me, n -Bu), while structural information has been gained by FT-IR, 119 Sn Mossbauer and 1 H, 13 C, 119 Sn NMR data. In particular, IR results suggested polymeric structures both for Alk 2 SnOHceph · H 2 O and Alk 3 Snceph · H 2 O. Moreover, cephalexin appears to behave as monoanionic tridentate ligand coordinating th…
Cytogenetics in the sacoglossan Oxynoe olivacea (Mollusca: Opisthobranchia): karyotype, chromosome banding and fluorescent in situ hybridization
Developing embryos and sexually mature follicles of the male portion of ovotestis proved to be a suitable material as a source of cleaving cells for advanced cytological investigations on the sacoglossan species Oxynoe olivacea Rafinesque, 1819 (Mollusca: Opisthobranchia). O. olivacea has a diploid chromosomal number of 30 made up of 15 pairs of which six are metacentric/submetacentric (M/SM), four subtelocentric (ST) and five on the borderline between SM and ST. Correspondingly, 15 bivalents occur in spermatocytes at Metaphase I. Constitutive heterochromatin is scarce and restricted to small C-bands seen in five pachytene bivalents. The use of combined silver staining and fluorescent in si…
Molecular and biometric data on Carabus (Macrothorax) morbillosus Fabricius, 1792 (Coleoptera, Carabidae) from Mid Mediterranean areas
The present study was carried out using molecular and biometric data of Carabus (Macrothorax) morbillosus from mid-Mediterranean areas to determine additional information on basal relationships among its representative subspecies. To this aim, two different kinds of approach were employed, including a morphometric analysis of four morphological parameters (i.e., elytra length, elytra width, pronotum length, pronotum width) of 128 specimens, and a Bayesian genetic analysis of 44 cytochrome oxidase subunit I (COI) partial sequences (i.e., 38 examined for the first time and six retrieved from GenBank database). Representative populations of C. (M.) morbillosus were sampled in four countries, n…
Chromosome analysis using different staining techniques and fluorescent in situ hybridization in Cerithium vulgatum (Gastropoda: Cerithiidae)
In the present paper one population of the “large” subtidal mollusc Cerithium vulgatum Bruguiere, 1792 (Gastropoda: Cerithiidae) from the Northwestern coast of Sicily was investigated from a karyological point of view. The chromosome complement was Giemsa stained, conventionally karyotyped in 18 homomorphic chromosome pairs (10 bi-armed and 8 mono-armed), and subsequently analysed using silver, CMA3 and DAPI staining, and fluorescent in situ hybridization (FISH) with three repetitive DNA probes [ribosomal DNA (rDNA), (TTAGGG)n and (GATA)n]. FISH with the rDNA probe consistently mapped major ribosomal sites (18S-28S rDNA) in the terminal region of the short arms of one small sized mono-armed…
Chromosomes of the Parasitic Isopod Anilocra Physodes
AbstractCounts of mitotic chromosomes have allowed to determine 2n = 12 as the modal diploid number of Anilocra physodes. Application of the C-banding technique reveals that two hetero-chromatic blocks bordering the centromeric region occur in each chromosome. This supports the notion that the actual low chromosome number of A. physodes may have been derived through a process of Robertsonian fusion.
Multiple-chromosome sex systems in the darkling beetles Blaps gigas and Blaps gibba (Coleoptera, Tenebrionidae)
We have studied mitotic and meiotic chromosomes in the males of two species of Blaps: B. gigas and B. gibba. Karyological characteristics such as the occurrence of a multivalent configuration at diakinesis and two types of metaphase-II spreads support the notion that multiple-chromosome sex systems involving five chromosomes in B. gigas and eight chromosomes in B. gibba have developed in these species. Results obtained by means of silver staining and C-banding techniques suggest that the complex sex systems occurring in B. gigas and B. gibba may have originated from exchanges of terminal ribosomal genes among the Y chromosome and some autosomes.
CYTOGENETICS OF THE AMPHIPOD JASSA MARMORATA (COROPHIOIDEA: ISCHYROCERIDAE): KARYOTYPE MORPHOLOGY, CHROMOSOME BANDING, FLUORESCENTIN SITUHYBRIDIZATION, AND NUCLEAR DNA CONTENT
Abstract Developing embryos proved to be a suitable source of cells for advanced cytological investigations on Amphipods. Conventional karyotyping, Ag- and fluorochrome-staining, C-banding, endonuclease digestion, fluorescent in situ hybridization (FISH) and nuclear DNA flow cytometric assay were tested in the Ischyroceridae Jassa marmorata. The karyotype consists of 6 chromosome pairs of which 5 are metacentric and 1 subtelocentric. The rDNA/FISH revealed that major ribosomal cistrons are located on the telomeric regions in the short arm of pair 6. A marked size variation of hybridization signals was observed. Silver and fluorochrome staining enhanced no chromosome regions. Constitutive he…
rDNA (18S-28S and 5S) co-localization and linkage between ribosomal genes and (TTAGGG)n telomeric sequence in the earthworm Octodrilus complanatus (Annelida: Oligochaeta: Lumbricidae) revealed by single- and double-colour FISH
Spermatogonial and metaphase I chromosomes of the lumbricid earthworm Octodrilus complanatus (Annelida: Oligochaeta) were examined using fluorescent in situ hybridization (FISH) with three repetitive DNA probes-5S rDNA, 18S-26S rDNA, and (TTAGGG)(n). Single-color FISH consistently mapped one chromosome pair per spread using either 5S rDNA or 18S-26S rDNA as probes. Simultaneous (18S-26S)-5S and (18S-26S)-(TTAGGG)(n) FISH demonstrated that repeated units of the two ribosomal families were overlapped and closely associated with telomeric sequences.
Repetitive DNAs in the slug Milax nigricans: association of ribosomal (18S–28S and 5S rDNA) and (TTAGGG)n telomeric sequences) in the slug M. nigricans (Mollusca: Gastropoda: Pulmonata)
Spermatocyte chromosomes of the slug Milax nigricans (Mollusca: Gastropoda: Pulmonata) were studied using silver staining (Ag-NOR) and fluorescent in situ hybridization (FISH) with four repetitive DNA probes [18S rDNA, 5S rDNA, (TTAGGG)n and (GATA)n]. Silver impregnation was inadequate to localize the chromosome sites of nucleolus organizer regions (NORs) since no silver dots occurred on the chromosomes at spermatogonial metaphase and a diffuse silver stainability could be observed on the bivalents at metaphase-I. Unlike silver staining, single-colour rDNA FISH consistently mapped major ribosomal sites (18S-28S rDNA) on two small-sized chromosomes in spermatogonial cells and on the correspo…
Brief communication. Karyotype analysis, banding, and fluorescent in situ hybridization in the scarab beetle Gymnopleurus sturmi McLeay (Coleoptera Scarabaeoidea: Scarabaeidae)
Conventional staining, differential banding, and in situ hybridization with both ribosomal and telomeric probes to mitotic chromosomes of Gymnopleurus sturmi (Scarabaeoidea : Scarabaeidae) are described. The karyotype is distinguished by a pericentric inversion polymorphism in chromosome 3, which is either acrocentric or subtelocentric. Silver staining (Ag-NOR) and chromomycin A3 (CMA3), failed to study the detection of nucleolar organizer regions (NORs), due to the extensive silver and CMA3 stainability of all GC-rich heterochromatin. Fluorescent in situ hybridization (FISH) using a Paracentrotus lividus (Echinodermata) rDNA probe mapped the ribosomal RNA genes (rDNA). FISH with the all-hu…
NOR regions of polychaete worms of the genus Ophryotrocha studied by chromosome banding techniques and FISH.
This article reports the results of cytogenetic analyses carried out on 10 species of polychaete worms belonging to the genus Ophryotrocha (Dorvilleidae). Nucleolar organizer regions (NORs) were characterized by Ag staining, C-banding, CMA3 staining, and ribosomal fluorescent in situ hybridization (rDNA FISH). Extensive intraspecific variation in NOR number and distribution were observed in O. costlowi, O. sp. macrovifera, O. notoglandulata, O.l. labronica, O. l. pacifica (2n = 6), O. p. puerilis, O. diadema (2n = 8), O. hartmanni, O. gracilis (2n = 10). In O. sp. robusta (2n = 10), Ag-NORs were always located on a single chromosome pair. CMA3 staining suggests a possible trend toward a GC-…
Chromosomal location polymorphism of major rDNA sites in two Mediterranean populations of the killifih Aphanius fasciatus (Pisces: Cyprinodontidae)
The chromosomes of the Mediterranean killifish, Aphanius fasciatus from two populations, the Lagoon of Venice (LV, 15 specimens) and the Lagoon ‘Stagnone di Marsala’ (Sicily) (SM, 48 specimens), have been investigated using conventional Ag-staining and fluorescent in situ hybridization (FISH) with 18S rDNA probe. The two methods revealed variation in the number of major rDNA sites ranging from 8 to 14 (LV) and from 1 to 4 (SM) per individual. The fact that each individual possessed its own number of sites implies that observed variation was structural. Moreover, overlapping of silver staining and FISH patterns demonstrated that all ribosomal genes were transcriptionally active in each speci…
FISH mapping of 18S-28S and 5S ribosomal DNA, (GATA)n and (TTAGGG)n telomeric repeats in the periwinkle Melarhaphe neritoides (Prosobranchia, Gastropoda, Caenogastropoda)
Spermatocyte chromosomes of Melarhaphe neritoides (Mollusca, Prosobranchia, Caenogastropoda) were studied using fluorescent in situ hybridization (FISH) with four repetitive DNA probes (18S rDNA, 5S rDNA, (TTAGGG)n and (GATA)n). Single-colour FISH consistently mapped one chromosome pair per spread using either 18S or 5S rDNA as probes. The telomeric sequence (TTAGGG)n hybridized with termini of all chromosomes whereas the (GATA)n probe did not label any areas. Simultaneous 18S-5S rDNA and 18S-(TTAGGG)n FISH demonstrated that repeated units of the three multicopy families are closely associated on the same chromosome pair.
"KARYOTYPE, BANDING AND rDNA FISH IN THE SCARAB BEETLE ANOPLOTRUPES STERCOROSUS (COLEOPTERA SCARABAEOIDEA: GEOTRUPIDAE). DESCRIPTION AND COMPARATIVE ANALYSIS
Six specimens of Anoplotrupes stercorosus (Coleoptera Scarabaeoidea: Geotrupidae) were analysed using conventional staining, banding techniques and fluorescent in situ hybridization with a ribosomal probe (rDNA FISH). Detailed karyotype description was also joined to a comparative analysis between present data and those previously reported for Thorectes intermedius [Chromosome Res. 7 (1999) 1]. The two species, both belonging to the tribe Geotrupini, show the same modal number but different autosomal morphology which is in contrast with the high chromosome stability argued for Geotrupinae during the last three decades. Moreover, a detailed comparison reveals the occurrence of a plesiomorphi…
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. p…
Chromosome Analysis and rDNA FISH in The Stag Beetle Dorcus Parallelipipedus L. (Coleoptera: Scarabaeoidea: Lucanidae)
In the present work the chromosome complement (2n = 18; 8AA + XY) of the stag beetle Dorcus parallelipipedus L. (Scarabaeoidea: Lucanidae) is analyzed using conventional Giemsa staining, banding techniques and ribosomal fluorescent in situ hybridization (rDNA FISH). rDNA FISH remains the unique tool for providing a clear-cut identification of Nucleolar Organizer Regions (NORs) when conventional banding methods such as silver- and CMA3-staining proved to be inadequate. The dull, homogeneous CMA3 fluorescence of all chromosomes indicates the absence of markedly GC rich compartmentalized regions in D. parallelipipedus genome. Silver impregnation inadequacy in detecting NOR regions is to be sou…
Karyotypes, Banding Patterns and Nuclear DNA Content inCrepidula unguiformisLamarck, 1822, andNaticarius stercusmuscarum(Gmelin, 1791) (Mollusca, Caenogastropoda)
ABSTRACT The chromosome complement and the nuclear DNA content in two caenogastropod species from the Mediterranean Sea, Crepidula unguiformis (Calyptraeidae) and Naticarius stercusmuscarum (Naticidae), were investigated by the application of both classical and molecular cytogenetic methods. Despite the constancy of haploid chromosome numbers (n = 17 in both species), C. unguiformis and N. stercusmuscarum show genome sizes amounting to 6.36 and 2.63 pg, respectively. Moreover, while N. stercusmuscarum resembles cytogenetically the other neotaenioglossan caenogastropods studied so far, C. unguiformis differs in: (i) number and location of rDNA clusters (ii), composition of telomeric repeats,…
Evidence for male XO sex-chromosome system in Pentodon bidens punctatum (Coleoptera Scarabaeoidea: Scarabaeidae) with X-linked 18S-28S rDNA clusters
In scarab beetle species of the genus Pentodon, the lack of analysis of sex chromosomes in females along with the poor characterization of sex chromosomes in the males, prevented all previous investigations from conclusively stating sex determination system. In this study, somatic chromosomes from females and spermatogonial chromosomes from males of Pentodon bidens punctatum (Coleoptera: Scarabaeoidea: Scarabaeidae) from Sicily have been analyzed using non-differential Giemsa staining. Two modal numbers of chromosomes were obtained: 2n = 20 and 19 in females and males, respectively. This finding along with other karyological characteristics such as the occurrence of one unpaired, heterotypi…
Heterochromatin of the scarab beetle, Bubas bison (Coleoptera: Scarabaeidae) II. Evidence for AT-rich compartmentalization and a high amount of rDNA copies
An unexpected result arising from a previous characterization of the scarab beetle Bubas bison (Coleoptera: Scarabaeidae) heterochromatin was its unusual homogeneous reaction to different staining methods. In particular, silver stainability of heterochromatic ends of all chromosomes prevented identification of the number of rDNA transcriptionally active regions. Data formerly obtained using silver impregnation (Ag-NOR), C- G- and DAPI banding are here improved and completed by application of CMA(3) staining and rDNA FISH with the aim to investigate heterochromatin base composition and locate rDNA regions with respect to NOR-associated heterochromatin. Our results show that B. bison has a hi…
Organometallic complexes with biological molecules: V.In vivo cytotoxicity of diorganotin(IV)-amoxicillin derivatives in mitotic chromosomes ofrutilus rubilio (pisces, cyprinidae)
In order to test in vivo cytotoxicity of diorganotin(IV)-amoxicillin (amox) derivatives, mitotic chromosomes of Rutilus rubilio (Pisces, Cyprinidae) have been analyzed using two different chromosome-staining techniques. Results gathered after exposure of fish to the free amox.3H 2 O, R 2 SnClamox.2H 2 O, and R 2 Snamox 2 .2H 2 O (R = methyl, butyl and phenyl ; amox - = 6-[D(-)-β-amino-p-hydroxyphenylacetamido]penicillinate) suggest that methyl derivatives seem to exert a lower cytotoxicity than butyl and phenyl ones and that R 2 Snamox 2 .2H 2 O derivatives are more toxic than R 2 Snclamox.2H 2 O at both 10 -5 and 10 -7 mol dm -3 concentrations. The following structural lesions have been id…