Francesca Dumas

Phylogenomics of species from four genera of New World monkeys by flow sorting and reciprocal chromosome painting

BackgroundThe taxonomic and phylogenetic relationships of New World monkeys (Platyrrhini) are difficult to distinguish on the basis of morphology and because diagnostic fossils are rare. Recently, molecular data have led to a radical revision of the traditional taxonomy and phylogeny of these primates. Here we examine new hypotheses of platyrrhine evolutionary relationships by reciprocal chromosome painting after chromosome flow sorting of species belonging to four genera of platyrrhines included in the Cebidae family: Callithrix argentata (silvered-marmoset), Cebuella pygmaea (pygmy marmoset), Callimico goeldii (Goeldis marmoset) and Saimiri sciureus (squirrel monkey). This is the first report of reciprocal painting in marmosets.ResultsThe paints made from chromosome flow sorting of the four platyrrhine monkeys provided from 42 to 45 hybridization signals on human metaphases. The reciprocal painting of monkey probes on human chromosomes revealed that 21 breakpoints are common to all four studied species. There are only three additional breakpoints. A breakpoint on human chromosome 13 was found in Callithrix argentata, Cebuella pygmaea and Callimico goeldii, but not in Saimiri sciureus. There are two additional breakpoints on human chromosome 5: one is specific to squirrel monkeys, and the other to Goeldis marmoset.ConclusionThe reciprocal painting results support the molecular genomic assemblage of Cebidae. We demonstrated that the five chromosome associations previously hypothesized to phylogenetically link tamarins and marmosets are homologous and represent derived chromosome rearrangements. Four of these derived homologous associations tightly nest Callimico goeldii with marmosets. One derived association 2/15 may place squirrel monkeys within the Cebidae assemblage. An apparently common breakpoint on chromosome 5q33 found in both Saimiri and Aotus nancymae could be evidence of a phylogenetic link between these species. Comparison with previous reports shows that many syntenic associations found in platyrrhines have the same breakpoints and are homologous, derived rearrangements showing that the New World monkeys are a closely related group of species. Our data support the hypothesis that the ancestral karyotype of the Platyrrhini has a diploid number of 2n = 54 and is almost identical to that found today in capuchin monkeys; congruent with a basal position of the Cebidae among platyrrhine families.

Mapping genomic rearrangements in titi monkeys by chromosome flow sorting and multidirectional in-situ hybridization.

We developed chromosome painting probes for Callicebus pallescens from flow-sorted chromosomes and used multidirectional chromosome painting to investigate the genomic rearrangements in C. cupreus and C. pallescens. Multidirectional painting provides information about chromosomal homologies at the subchromosomal level and rearrangement break points, allowing chromosomes to be used as cladistic markers. Chromosome paints of C. pallescens were hybridized to human metaphases and 43 signals were detected. Then, both human and C. pallescens probes were hybridized to the chromosomes of another titi monkey, C. cupreus. The human chromosome paints detected 45 segments in the haploid karyotype of C. cupreus. We found that all the syntenic associations proposed for the ancestral platyrrhine karyotype are present in C. cupreus and in C. pallescens. The rearrangements differentiating C. pallescens from C. cupreus re one inversion, one fission and three fusions (two tandem and one Robertsonian)that occurred on the C. cupreus lineage. Our results support the hypothesis that karyological evolution in titi monkeys has resulted in reduction in diploid number and that species with higher diploid numbers (with less derived, more ncestral karyotypes)are localized in the centre of the geographic range of the genera, while more derived species appear to occupy the periphery

Chromosome Painting of the Pygmy Tree Shrew Shows that No Derived Cytogenetic Traits Link Primates and Scandentia

We hybridized human chromosome paints on metaphases of the pygmy tree shrew (Tupaia minor, Scandentia). The lack of the ancestral mammalian 4/8 association in both Primates and Scandentia was long considered a cytogenetic landmark that phylogenetically linked these mammalian orders. However, our results show that the association 4/8 is present in Tupaia along with not previously reported associations for 1/18 and 7/10. Altogether there are 11 syntenic associations of human chromosome segments in the pygmy tree shrew karyotype: 1/18, 2/21, 3/21, 4/8, 7/10, 7/16, 11/20, 12/22 (twice), 14/15 and 16/19. Our data remove any cytogenetic evidence that Scandentia has a preferential phylogenetic relationship with Primates.

Neotropical primate evolution and phylogenetic reconstruction using chromosomal data

Abstract Platyrrhini are a group of Neotropical primates living in central and south America, and have been extensively studied through morphological and molecular data in order to shed light on their phylogeny and evolution. Agreement on the main clades of Neotropical primates has been reached using different approaches, but many phylogenetic nodes remain under discussion. Contrasting hypotheses have been proposed, presumably due to different markers and the presence of polymorphisms in the features considered; furthermore, neither Neotropical primate biodiversity nor their taxonomy are entirely known. In our perspective, a cytogenetic approach can help by making an important contribution to the evaluation of the phylogenetic relationships among Platyrrhini. In this work, molecular cytogenetic data regarding the principal nodes of the Neotropical monkey tree have been reviewed; classical cytogenetic data have also been considered, especially when other data have proven elusive, permitting us to discuss highly derived karyotypes characterized by a wide range of diploid numbers of chromosomes and variable chromosomal evolution with different rearrangement and polymorphism rates.

Distribution of Interstitial Telomeric Sequences in Primates and the Pygmy Tree Shrew (Scandentia)

It has been hypothesized that interstitial telomeric sequences (ITSs), i.e., repeated telomeric DNA sequences found at intrachromosomal sites in many vertebrates, could be correlated to chromosomal rearrangements and plasticity. To test this hypothesis, we hybridized a telomeric PNA probe through FISH on representative species of 2 primate infraorders, Strepsirrhini (Lemur catta, Otolemur garnettii, Nycticebus coucang) and Catarrhini (Erythrocebus patas, Cercopithecus petaurista, Chlorocebus aethiops, Colobus guereza), as well as on 1 species of the order Scandentia, Tupaia minor, used as an outgroup for primates in phylogenetic reconstructions. In almost all primate species analyzed, we found a telomeric pattern only. In Tupaia, the hybridization revealed many bright ITSs on at least 11 chromosome pairs, both biarmed and acrocentric. These ITS signals in Tupaia correspond to fusion points of ancestral human syntenic associations, but are also present in other chromosomes showing synteny to only a single human chromosome. This distribution pattern was compared to that of the heterochromatin regions detected through sequential C-banding performed after FISH. Our results in the analyzed species, compared with literature data on ITSs in primates, allowed us to discuss different mechanisms responsible for the origin and distribution of ITSs, supporting the correlation between rearrangements and ITSs.

Exploring Evolution in Ceboidea (Platyrrhini, Primates) by Williams-Beuren Probe (HSA 7q11.23) Chromosome Mapping

The ancestral platyrrhine karyotype was characterised by a syntenic association of human 5 and a small segment of human 7 orthologues. This large syntenic association has undergone numerous rearrangements in various phylogenetic lines. We used a locus-specific molecular cytogenetic approach to study the chromosomal evolution of the human 7q11.23 orthologous sequences (William-Beuren syndrome, WS) in various Ceboidea (Platyrrhini) species. The fluorescent in situ hybridisation of the WS probe revealed a two-way pattern of chromosomal organisation that suggests various evolutionary scenarios. The first pattern (seen in Callimico and Saimiri) includes a fairly simple disruption of the 7/5 syntenic association by a chromosome fission. The second pattern (seen in Atelinae, Alouattinae and in Callicebus) is characterised by an increasing complexity in the 7/5 association as a consequence of a series of inversions and translocations resulting in different syntenic associations. These data support recent proposals for phylogenomic groupings of New World monkeys. The study also illustrates how single-locus probe hybridisations can reveal intrachromosomal rearrangements.

2nd Congress of the European Federation for Primatology

Folia Primatol 2008;79:305–401 DOI: 10.1159/000137690 2nd Congress of the European Federation for Primatology Prague, September 3–7, 2007 Editors: Vaclav Vancata and Marina Vancatova, Prague, Czech Republic Do Capuchin Monkeys (Cebus apella) Deal with Tokens as They Do with Real Food? Elsa Addessi a , Alessandra Mancini a, b , Lara Crescimbene a, b , Elisabetta Visalberghi a a Unit of Cognitive Primatology and Primate Centre, Institute of Cognitive Sciences and Technologies, CNR, Rome, b Università La Sapienza, Rome, Italy E-Mail: elsa.addessi@istc.cnr.it

Evolutionary insight on localization of 18S, 28S rDNA genes on homologous chromosomes in Primates genomes

Abstract We explored the topology of 18S and 28S rDNA units by fluorescence in situ hybridization (FISH) in the karyotypes of thirteen species representatives from major groups of Primates and Tupaia minor (Günther, 1876) (Scandentia), in order to expand our knowledge of Primate genome reshuffling and to identify the possible dispersion mechanisms of rDNA sequences. We documented that rDNA probe signals were identified on one to six pairs of chromosomes, both acrocentric and metacentric ones. In addition, we examined the potential homology of chromosomes bearing rDNA genes across different species and in a wide phylogenetic perspective, based on the DAPI-inverted pattern and their synteny to human. Our analysis revealed an extensive variability in the topology of the rDNA signals across studied species. In some cases, closely related species show signals on homologous chromosomes, thus representing synapomorphies, while in other cases, signal was detected on distinct chromosomes, leading to species specific patterns. These results led us to support the hypothesis that different mechanisms are responsible for the distribution of the ribosomal DNA cluster in Primates.

The evolution of human synteny 4 by mapping sub-chromosomal specific probes in Primates

Comparative cytogenetic data concerning the orthologue to human chromosome 4 in primates shows that this chromosome is conserved between humans and non-human primates. However, the degree of conservation is not as high as previously estimated. In primates it is as a rule a large submetacentric chromosome but many exceptions are known especially in taxa characterized by a high level of chromosomal rearrangements. The rearrangements that have been visualized by chromosome painting so far, which are mostly interchromosomal changes, are in fact only a fraction of the actual chromosomal changes that have occurred during evolution. Intrachromosome changes can be analysed through classical cytogenetic approach or by mapping sub-chromosomal specific probes. In order to study human synteny 4 evolution we mapped diverse subchromosomal specific probes on chromosomes of representative species of the main primate taxa, with the aim to verify markers order conservation along the orthologues to human chromosome 4, allowing us the detection of possible intra-chromosomal rearrangements. The mapping of these probes permitted us to test previous cytogenetic hypothesis on human synteny 4 evolution, and to show marker order conservation between orthologues to human synteny 4 in Catarrhini and Platyrrhini, but with a different position of the centromeres. This data permitted us to hypothesize the occurrence of a new centromere evolution in one of the two lineages. Moreover we analysed literature data regarding HSA4 homologous in primates with particular attention to Platyrrhini, allowing us the reconstruction of the changes that synteny 4 has undergone during evolution. Lastly we highlight the value of the subchromosomal specific probes mapping approach in the detection of intrachromosomal rearrangements that can be crucial for a more refined comparative mapping and for phylogenetic reconstruction.