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M Mandrioli

Publications and source records attributed to M Mandrioli.

At least 19 recordsLinked to original sources
PubMed2006

DNA methylation of fly genes and transposons.

The use of anti-5-methylcytosine antibodies in affinity columns allowed the identification of methylated sequences in the genome of Drosophila melanogaster adults. In view of the presence of transposable elements amongst the identified sequences, it has been suggested that DNA methylation is involved in transposon control in the fly genome. On the contrary, a reanalysis of these data furnishes several intriguing elements that could raise new questions about the role that DNA methylation plays in the fly genome. The aim of the present paper is to discuss some features that emerge from the analysis of the identified methylated sequences.

Animals↗
PubMed2004

Epigenetic tinkering and evolution: is there any continuity in the role of cytosine methylation from invertebrates to vertebrates?

The function of DNA methylation has been investigated in depth in vertebrate and plant genomes, establishing that it is involved in gene silencing and transposon control. Data regarding insect methylation, even if still scanty, apparently argue against evolutionary conservation of DNA methylation functions. Cytosine methylation, therefore, proves to be an epigenetic tool repeatedly used to accomplish different functions in different taxa according to a sort of epigenetic tinkering occurring during evolution.

Animals↗
PubMed2004

DNA methylation in insects.

Cytosine DNA methylation has been demonstrated in numerous eukaryotic organisms and has been shown to play an important role in human disease. The function of DNA methylation has been studied extensively in vertebrates, but establishing its primary role has proved difficult and controversial. Analysing methylation in insects has indicated an apparent functional diversity that seems to argue against a strict functional conservation. To investigate this hypothesis, we here assess the data reported in four different insect species in which DNA methylation has been analysed more thoroughly: the fruit fly Drosophila melanogaster, the cabbage moth Mamestra brassicae, the peach-potato aphid Myzus persicae and the mealybug Planococcus citri.

5-Methylcytosine↗
PubMed2002

Cytogenetic characterization of telomeres in the holocentric chromosomes of the lepidopteran Mamestra brassicae.

Telomeres of the Mamestra brassica holocentric chromosomes were studied by Southern blotting, in-situ hybridization and Bal31 assay evidencing the presence of the telomeric (TTAGG)n repeat. Successively, molecular analysis of telomeres showed that TRAS1 transposable elements were present at the subtelomeric regions of autosomes but not in the NOR-bearing telomeres of the Z and W sex chromosomes. TRAS1 appeared to be transcriptionally active and non-methylated, as evaluated by RT-PCR and digestion with MspI and HpaII. Finally, dot-blotting experiments showed that the 2.8 +/- 0.5% of the M. brassicae genome consists of TRAS1.

Animals↗
PubMed2001

Cytogenetic and molecular analysis of the pufferfish Tetraodon fluviatilis (Osteichthyes).

In view of their compact genome, pufferfish (Tetraodontiformes) have been proposed as model animal for the study of the vertebrate genome. Despite such interest, cytogenetic information about puffers is still scanty. To fill this gap, a cytogenetic analysis of T. fluviatilis has been performed using both classical and molecular techniques. C-banding, followed by DAPI staining, evidenced that in T. fluviatilis, like all other puffer species so far examined, heterochromatin is essentially AT-rich and it is located at centromeres, whereas staining with CMA3, silver staining and FISH with a 28S ribosomal RNA gene DNA probe showed 2-4 nucleolar organizing regions (NORs) located in heterochromatic regions in the considered puffer species. FISH with the 5S probe put in evidence both in T. fluviatilis and in T. nigroviridis only a 5S cluster per haploid genome that is physically unlinked with the major ribosomal RNA genes including the 28S rRNA genes. Hybridization with the (TTAGGG)n probe showed in all the puffers brightly fluorescent signals uniform both in size and intensity at the end of all the chromosomes. Finally, mariner-like elements (MLEs) have been identified in T. fluviatilis and they have located into the NOR-associated heterochromatin.

Animals↗
PubMed2001

DNA endoreplication in the clustered supramedullary neurons of the pufferfish Diodon holacanthus L. (osteichthyes).

DNA contents, ranging from 4C to more than 500C, were determined by a quantitative microfluorimetric assay in supramedullary neuron nuclei of the pufferfish Diodon holacanthus. The distribution of C values after ethidium bromide staining indicates an inter- and intra-individual variation in DNA contents which do not correspond to duplications of the total genome, suggesting that DNA replication is specific for particular genes (endoreplication). Moreover, the DNA content appears to be correlated with nuclear size. A relationship between the DNA amounts and the presence of AT- and GC-rich sequences has been shown. This work demonstrates, for the first time, DNA endoreplication in a specific neuronal type in vertebrates.

Animals↗
PubMed2000

Chromosome analysis and molecular characterization of highly repeated DNAs in the aphid Acyrthosiphon pisum (Aphididae, Hemiptera).

Despite the interest in aphid biology, information on chromatin organization of their holocentric chromosomes is still limited to few species. In order to fill this gap, we have performed an extensive survey on pea aphid mitotic chromosomes using both classical and molecular cytogenetic techniques. Our results after silver, CMA3 and DAPI-staining, C-banding and fluorescent in situ hybridization (FISH) using 28S rDNA and 5S rDNA as probes evidenced a tendency of repetitive DNAs to be concentrated on the X chromosomes. FISH experiments with the telomeric probe (TTAGG)n revealed bright hybridization signals on each telomere of all Acyrthosiphon pisum chromosomes. No interstitial signals were seen.

Animals↗
PubMed2000

Chromosomal analysis of repeated DNAs in the rainbow wrasse Coris julis (Pisces, Labridae).

Despite the interest of several authors, the karyotype of the labrid C. julis is still debated and in particular the presence of sex-chromosomes is still contradictory. In order to analyze the karyotype organization of C. julis we have performed an analysis with classical and molecular cytogenetic techniques. Our results after silver-, CMA3- and DAPI-staining, C-banding and fluorescent in situ hybridization (FISH) using 28S rDNA, 5S rDNA, and telomeric repeat (TTAGGG)n, as probes allowed us to characterize the chromosomal location of several repetitive DNAs of C. julis. Finally, regardless of the technique used, no difference in the chromosome complement was found between males and females.

Animals↗
PubMed2000

Cytogenetic analysis of the pufferfish Tetraodon fluviatilis (Osteichthyes).

Because of their compact genome, pufferfish (Tetraodontiformes) have been proposed as a model for the study of the vertebrate genome. The genome of pufferfish is peculiar as it has the structural complexity of the genomes of higher vertebrates, but has small introns and lacks large clusters of highly repetitive sequences. Despite such interest, information about the genetics of pufferfish is still scanty. To fill this gap, we have performed a cytogenetic analysis of the pufferfish, Tetraodon fluviatilis, which can be maintained in an aquarium for a long time and, unlike the pufferfish, Fugu rubripes, it is not difficult to obtain. Karyotype analysis shows that T. fluviatilis has 2n = 42 with two metacentric chromosomes, four submetacentrics, two subtelocentrics and 34 acrocentrics. C-banding, followed by DAPI staining, showed that heterochromatin is essentially AT-rich and is located at centromeres. Staining of the same metaphase plates with CMA3 showed the presence of four heterochromatic regions located on two pairs of submetacentric chromosomes. Silver staining and FISH with a 28S rDNA probe showed that these GC-rich regions are nucleolar organizing regions (NORs). Finally, regardless of the technique used, no difference in the chromosome complement was found between males and females.

Animals↗
PubMed2000

Molecular and cytogenetic analysis of the goby Gobius niger (Teleostei, Gobiidae).

A molecular cytogenetic study of Gobius niger has been conducted by treating its mitotic chromosomes with silver-, CMA3- and DAPI-staining and fluorescent in situ hybridization using four multicopy or repetitive DNAs (the 28S and 5S rDNAs, the TTAGGG telomeric repeat and the mariner-like elements) as probes. In particular, the study proved the presence of NOR heteromorphism and suggested the possible role of the transposable element mariner in its genesis. In situ hybridization with the 5S rDNA probe proved the presence of just one 5S-bringing chromosome pair, whereas hybridization with the telomeric repeat revealed small bright hybridization spots, uniform in size and intensity, on each telomere of all chromosomes but no interstitial signals were noticed.

Animals↗
PubMed2000

18S-28S rDNA is interspersed throughout heterochromatin in the millipede Enologus oxypygum (Diplopoda).

In the present study, somatic metaphase chromosomes of the millipede Enologus oxypygum (Diplopoda: Julida) were hybridized in situ with a sea urchin (Echinodermata) ribosomal probe (prR14) in order to map major RNA genes (rDNA). Chromosomal preparations were also silver stained (Ag-NOR) to evaluate the rDNA transcriptional activity. Our results indicate that RNA genes are throughout heterochromatin in eight chromosomes involving 1/4 of the total heterochromatin which, in this species, is about 67% of the total DNA. Ag-NOR and FISH patterns were not coincident.

Animals↗
PubMed2000

The use of two density gradient centrifugation techniques and the swim-up method to separate spermatozoa with chromatin and nuclear DNA anomalies.

Human semen is heterogeneous in quality, not only between males but also within a single ejaculate. Differences in quality are evident, both when examining the classical parameters of sperm number, motility and morphology and in the integrity of the sperm nucleus. The aim of this study was to determine the efficiency of the PureSperm((R)), Percoll((R)) and swim-up preparation techniques to eliminate spermatozoa with nuclear anomalies. Semen samples were collected, washed and one part of the semen spread on a slide, the remainder was prepared using the swim-up, PureSperm((R)) or Percoll((R)) techniques. Spermatozoa from different fractions were fixed on slides and assessed. Sperm samples (n) from different men were stained using the chromomycin A(3) (CMA(3)) fluorochrome, which indirectly demonstrates a decreased presence of protamine (n = 31 for swim-up; n = 45 for PureSperm((R)); n = 39 for Percoll((R))). Spermatozoa prepared using PureSperm((R)) (n = 35) and Percoll((R)) (n = 37) were also examined for the presence of endogenous DNA nicks. Good quality spermatozoa should not possess DNA nicks and not stain (i.e. fluoresce) with CMA(3). When prepared using the swim-up technique the spermatozoa recovered showed no significant improvement with the CMA(3) staining. When spermatozoa were prepared using the PureSperm((R)) and Percoll((R)) techniques, a significant (P < 0.001) decrease in both CMA(3) positivity and DNA strand breakage was observed. These results indicate that both the PureSperm((R)) and Percoll((R)) techniques can enrich the sperm population by separating out those with nicked DNA and with poorly condensed chromatin.

Centrifugation, Density Gradient↗
PubMed2000

Ceratotoxins: female-specific X-linked genes from the medfly, Ceratitis capitata.

In this paper, we report the chromosomal localization of ceratotoxins, a gene family encoding antibacterial female-specific peptides from the mediterranean fruit fly Ceratitis capitata. The analysis of both polytene and mitotic chromosomes by in situ hybridization shows that ceratotoxins are the first case of female-specific X-linked genes from the medfly C. capitata. Southern blot analysis reveals that the ceratotoxin gene family is not specifically amplified in the female reproductive accessory glands of C. capitata.

Animals↗
PubMed1999

Cytogenetic and molecular characterization of a highly repeated DNA sequence in the peach potato aphid Myzus persicae.

Electrophoresis following digestion of Myzus persicae genomic DNA with HindIII showed the presence of a prominent band of approximately 200 bp whereas a faint electrophoretic band corresponding to DNA fragments of about 3000 bp was observed after digestion with ApaI. In situ digestion with restriction enzymes, followed by in situ nick translation, showed that ApaI targets are localized at the nucleolus organizer-bearing X telomeric region, whereas HindIII restriction sites are clustered in intercalary C-positive areas on the same X chromosome. Fluorescent in situ hybridization (FISH) carried out by using digoxygenin-labeled HindIII repeats as probe fully confirmed overlapping between the hybridization sites of this probe and the AT-rich intercalary heterochromatic bands on the X chromosome. These findings, together with published data, allow us to conclude that the M. persicae genome possesses three classes of C-positive heterochromatin: (i) a GC-rich argentophilic band located on one telomere of the X chromosome that contains ApaI targets; (ii) AT-rich intercalary bands located on the X chromosome containing clustered HindIII fragments; (iii) AT-rich telomeric bands located on autosomes, consisting of HaeIII repeats. Molecular analysis has shown that the length of the HindIII repeat consensus sequence is 189 bp with an AT content of 67%. Southern blotting with HindIII monomers revealed a regular ladder of bands composed of multimers of basic length that are characteristic of satellite DNAs. The HindIII repeat displays other features typical of eukaryotic satellite arrays such as overlapping with heterochromatic bands and a high degree of sequence similarity among monomers (84%-94%). A similarity plot showed that sequences were particularly variable in the 50-100 bp region whereas they proved to be highly conservative in the first 50 bp, thus suggesting that this portion of the repeat might be functionally important.

AT Rich Sequence↗
PubMed1999

NOR heteromorphism within a parthenogenetic lineage of the aphid Megoura viciae.

In parthenogenetic females of a clone of the aphid Megoura viciae (Homoptera, Aphididae), more than 50% of the cells show heteromorphism between homologous NORs which are located on one telomeric region of the two X chromosomes. Using different techniques, such as staining with the CG-specific fluorochrome chromomycin A3, silver staining and in-situ hybridization with an rDNA probe, we have shown that the observed heteromorphism is due to an unequal distribution of ribosomal genes between homologous NOR regions. The total number of rDNA genes per individual aphid remained constant. Moreover, the analysis of cells from single embroys has shown that the observed heteromorphism is not only intraclonal but also intraindividual. These data, together with the finding of X chromosomes connected by chromatin bridges between their NORs, allow us to suggest that mitotic unequal crossing over could be the main cause of NOR heteomorphism in this taxon.

Animals↗
PubMed1999

The role of rDNA genes in X chromosome association in the aphid Acyrthosiphon pisum.

Silver staining of mitotic metaphases of the aphid A. pisum reveals the presence of argentophilic bridges connecting the two X chromosomes. The presence of nucleolar material connecting sex chromosomes seems to be quite a common phenomenon in organisms belonging to very different phyla, and suggests a role of nucleolar proteins in chromosome association and disjunction. In somatic cells of A. pisum, bridges connecting X chromosomes are detectable not only after silver staining but also after CMA3 staining. This finding suggests that GC rich DNA is involved in this type of association. Molecular analysis of rDNA intergenic spacers shows several 247 bp repeats containing short sequences having a high level of homology with the chi sequence of Escherichia coli and with the consensus core region of human hypervariable minisatellites. Moreover, each 247 bp repeat presents a perfect copy of a promoter sequence for polymerase I. These aphid repeats show structural homologies with a 240 bp repeat, which is considered to be responsible for sex chromosome pairing in Drosophila, not only in view of their common presence within rDNA spacers but also for their length and structure. The presence of chi sequences in the IGS of A. pisum, by promoting unequal crossing-over between rDNA genes, could thus give rise to the nucleolar organizing region (NOR) heteromorphism described in different aphid species. Although X pairing at NORs is fundamental in aphid male determination, the presence of heteromorphism of rDNA genes does not inhibit male determination in the A. pisum clone utilized for our experiments.

Animals↗
PubMed1998

Silver staining as a new banding technique to identify aphid chromosomes.

The karyotype of the aphid Aphis sambuci has been studied by means of C-banding, fluorochrome staining and AgNO3 staining. Heterochromatic areas are confined exclusively to the X chromosomes. Interestingly, silver staining produces a clear-cut longitudinal differentiation of all chromosomes, which allows reliable identification of homologues.

Animals↗