Genotoxic effects of the fungicide thiophanate-methyl on Podarcis sicula assessed by micronucleus test, comet assay and chromosome analysis.

The increasing use of pesticides in modern agriculture has raised the need to evaluate their potential threat to animal and human health. In the present study, the genotoxic effects of environmentally relevant exposure to the fungicide thiophanate-methyl (TM) were assessed in the lizard Podarcis sicula (Reptilia, Lacertidae) using micronucleus test, chromosome aberration analysis and single-cell gel electrophoresis (comet) assay. The number of micronuclei increased significantly with exposure time in lizard specimens exposed to 1.5% TM for 30-40 days. In situ hybridization with the specific HindIII centromeric satellite was positive in 18.7% of the micronuclei observed, suggesting an aneugenic effect of TM during mitosis. DNA damage, evaluated by the comet assay, documented a significant gain in comet length in relation to exposure time that was paralleled by a reduction in head size. Finally, cytogenetic analysis showed a significant increase in chromosome aberrations in exposed animals compared with controls. Our data suggest that long-term TM exposure induces a genomic damage that is positively correlated to exposure time. If such genotoxic effects arise so clearly in an ectothermal vertebrate, such as P. sicula, prolonged exposure TM must be considered as a cytogenetic hazard.

Cytogenetic analysis shows extensive genomic rearrangements between red howler (Alouatta seniculus, Linnaeus) subspecies.

A comparison of the G-banded karyotypes of two red howler subspecies, Alouatta seniculus arctoidea and A. s. sara, showed that they differed by at least 14 chromosomal rearrangements. Genomic reshuffling is so great that homologs between subspecies could not be found for some chromosome, while the assignment of homology for other chromosomes remains uncertain. The two red howlers, however, share an unusual X1X2Y1Y2/ X1X1X2X2 sex-chromosome system that resulted from a Y-autosome translocation, probably in a common ancestor. The great chromosomal variability resulting from rapid chromosomal evolution in howlers indicates that cytogenetic data could make an important contribution to resolving phylogenetic and conservation problems in this group of highly conspicuous New World Monkeys. © 1995 Wiley-Liss, Inc.

Differential DMRT1 expression in the gonads of Podarcis sicula (Reptilia: Lacertidae).

DMRT genes encode a large family of transcription factors which share an unusual cysteine-rich DNA-binding motif, the DM domain. DM family members have been studied in the context of sexual development; in particular, the DMRT1 gene appeared to be the one most directly involved in sex determination, but its activity is largely unexplored and possible downstream targets of this factor have yet to be identified. DMRT1 of the lacertid lizard Podarcis sicula (PsDMRT1) was isolated as a model to study differential gene expression during the seasonal reproductive cycle of an ectothermal species. The adult testis of P. sicula exhibits full activity in spring, complete regression in summer and a slow autumnal recrudescence without spermiation. We cloned a 591-bp partial ORF of the PsDMRT1 fragment, whose putative amino acid sequence contains the conserved DM domain. Northern blot analysis of mRNA from different tissues of P. sicula individuals captured in spring demonstrated DMRT1 transcripts only in testis. Semi-quantitative RT-PCR and in situ hybridization experiments showed peak PsDMRT1 expression in spring, lower expression in autumn and no expression during the period of gonad regression. A possible correlation between androgen level variations and PsDMRT1 transcripts is hypothesized and discussed. Finally, data showed that PsDMRT1 is expressed only in spermatogenic cells, before the second meiotic division, suggesting that its role is confined to the proliferation and maintenance of spermatogonia and spermatocytes.

Mapping chromosomal homology between humans and the black-handed spider monkey by fluorescence in situ hybridization.

We hybridized human chromosome-specific DNA probes to metaphases of the New World monkey Ateles geoffroyi to map the chromosomal homology between these two species. In the haploid Ateles geoffroyi karyotype the total number of signals was 51 for the 22 human autosomal probes used. Compared with Old World monkeys, the number of translocations found in the black-handed spider monkey karyotype was quite striking. The majority of these translocations are apparently Robertsonian and no reciprocal translocations were revealed. Nine autosomal human chromosome probes (11, 13, 14, 17, 18, 19, 20, 21, 22) provided only two signals each per metaphase, but six of these were translocated to subregions of different spider monkey chromosomes. The other 13 autosomal human chromosome paints (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 16) provided fragmented signals. Three human probes (5, 8, 10) provided signals located on two pairs of spider monkey chromosomes. Four human paints (2, 3, 4, 12) provided hybridization signals on three pairs of chromosomes. Probes 6, 7, 15 provided six signals each on two pairs of chromosomes; probe 16 gave eight signals on two pairs of spider monkey chromosomes and probe 1 gave 12 signals on four pairs of chromosomes. The synteny between segments to human 18/8 appears to be an apomorphic ancestral condition for all New World monkeys. A synteny between regions homologous to human 16/10, 5/7 and 2/16 HSA is probably an apomorphic ancestral condition for all Cebidae. The syntenic association 3/15 and 4/1 is an apomorphic condition for the Atelinae.