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Development of a sequential multicolor-FISH approach with 13 chromosome-specific painting probes for the rapid identification of river buffalo (Bubalus bubalis, 2n = 50) chromosomes.
No full textThe development of new molecular techniques (array CGH, M-FISH, SKY-FISH, etc.) has led to great advancements in the entire field of molecular cytogenetics. However, the application of these methods is still very limited in farm animals. In the present study, we report, for the first time, the production of 13 river buffalo (Bubalus bubalis, 2n = 50) chromosome-specific painting probes, generated via chromosome microdissection and the DOP-PCR procedure. A sequential multicolor-FISH approach is also proposed on the same slide for the rapid identification of river buffalo chromosome/arms, namely, 1p-1q, 2p-2q, 3p-3q, 4p-4q, 5p-5q, 18, X, and Y, using both conventional and late-replicating banded chromosome preparations counterstained by DAPI. The provided 'bank' of chromosome-specific painting probes is useful for any further cytogenetic investigation not only for the buffalo breeds, but also for other species of the family Bovidae, such as cattle, sheep, and goats, for chromosome abnormality diagnosis, and, more generally, for evolutionary studies
CYTOGENETIC INVESTIGATIONS IN SHEEP REARED IN SOUTHERN-ITALY BY USING BOTH CHROMOSOME BANDING AND FISH-MAPPING TECHNIQUES
No full textAdvanced comparative cytogenetic analysis of X chromosomes in river buffalo, cattle, sheep, and human
No full textBased on a recently generated comprehensive
gene map for Ovis aries chromosome X (OARX)
with an approximately even locus distribution, we
assigned selected bacterial artificial chromosome
(BAC) probes corresponding to these OARX loci to
Bubalus bubalis (BBU) and Bos taurus (BTA) by
comparative fluorescence in-situ hybridization (FISH)
to improve cytogenetically the X chromosome maps in these species. Twenty-five added loci in BBUX and
BTAX, respectively, contribute to a more detailed
description of the cytogenetic organization of these
chromosomes. Further seven loci were identified in
OARX and two DNA probes were assigned to X and
Y chromosomes in river buffalo, cattle, and sheep,
respectively, and thus identified loci in the pseudoautosomal
region. The additional assignments double the
number of cytogenetic loci in BBUX and increase
their number in BTAX and OARX. The larger quantity
of cytogenetic anchors allows a more precise morphological
comparison of bovid X chromosomes among
each other and with the Homo sapiens (HSA) X chromosome.
The anchor loci confirm and refine syntenic
fragments in HSAX and identify several evolutionary
breakpoints between the compared chromosomes. The
cytogenetic assignments in BBUX, BTAX, and
OARX represent useable anchors for the ongoing genome
sequence assembly in Bovidae
Cytogenetic elaboration of a novel reciprocal translocation in sheep
No full textReciprocal translocations represent one of the most common structural chromosomal rearrangements observed in both humans and domestic animals. In these translocations, the balanced forms are most frequent but may remain undetected because the carriers show a normal phenotype. For this reason, routine cytogenetic analysis of domestic animals should necessarily rely on banded karyotypes. In fact, during a screening analysis, carried out on phenotypically normal young sheep (Ovis aries, OAR, 2n = 54) from Laticauda-Comisana hybrids, a new structural rearrangement was detected. Two abnormal acrocentric chromosomes (the smallest and the largest one) were found in all metaphases of this carrier animal, suggesting the presence of a reciprocal translocation (rcp). CBA and RBA banding were performed in order to characterize the translocation, and FISH with chromosome-specific BAC probes and telomere probes was applied to confirm the cytogenetic data. The translocation was classified as rcp(4q;12q)(q13;q25)
A new translocation t(1p;18) in an Italian Mediterranean river buffalo (Bubalus bubalis, 2n = 50) bull: cytogenetic, fertility and inheritance studies.
No full textCytogenetic investigations in sheep reared in the southern-Italy by using both chromosome banding and fish-mapping techniques
Full text linkProtective Effects of Anthocyanin and α-Tocopherol Against Titanium Dioxide Nanoparticle-Induced DNA Damage in Human Sperm Cells
No full textThe male reproductive system is known for its high complexity and sensitivity, particularly concerning environmental stressors. Titanium dioxide nanoparticles (TiO2-NPs) could alter male reproductive function by increasing sperm DNA damage caused by an overproduction of ROS. The ability of anthocyanin and α-tocopherol to counteract the effects induced by TiO2-NPs was assessed in human sperm cells after 30, 45, and 90 min of in vitro exposure. Antigenotoxicity was evaluated by comet and TUNEL assays, the RAPD‒PCR technique, and relative genomic template stability analyses. The intracellular reactive oxygen species (ROS) concentration was detected by the probe dichlorofluorescein. Anthocyanin and α-tocopherol attenuated sperm DNA damage induced by TiO2-NPs in vitro, likely by activating the natural defence and DNA repair systems in sperm cells. Interestingly, compared with α-tocopherol, anthocyanin exhibited a more pronounced antigenotoxic effect, potentially due to the inherent ability of anthocyanin molecules to stabilise DNA. These results provide valuable insights into the beneficial and protective effects of anthocyanin and α-tocopherol on human germ cells treated with NPs, representing a potential therapeutic strategy to prevent nanomaterial-induced toxicity
TiO2-NPs and cadmium co-exposure: in vitro assessment of genetic and genomic DNA damage on Dicentrarchus labrax embryonic cells
No full textThe increased titanium dioxide nanoparticles (TiO2-NPs) spread and their interaction with organic and inorganic pollutants arouses concern for the potential hazards for organisms and environment. This study tested in vitro the genotoxic effects of TiO2-NPs (1 μg/mL) and cadmium (Cd) (0.1 μg/mL) co-exposure using Dicentrarchus labrax embryonic cells (DLEC) as experimental model. The genotoxicity tests (Comet assay, Diffusion Assay and Random Amplification of Polymorphic DNA (RAPD-PCR) were conducted after 3, 24 and 48 hours of exposure to TiO2-NPs and Cd alone and in combination. The results showed that the percentage of DNA damage and apoptotic cells increases following 48 hours TiO2-NPs exposure, while DNA instability was detected for all the times tested. Cd induced genotoxic effects starting from 3 hour-exposure and for all the treatment times. Cd + TiO2-NPs co-exposure did not cause any genomic damage or apoptosis for all the exposure times. The possibility that Cd and TiO2-NPs form aggregates no longer able of penetrating the nucleus and damaging the genetic material is discussed. Graphical abstract: [Figure not available: see fulltext.]
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