Human (Homo sapiens) and chimpanzee (Pan troglodytes) share similar ancestral centromeric alpha satellite DNA sequences but other fractions of heterochromatin differ considerably

The euchromatic regions of chimpanzee (Pan troglodytes) genome share approximately 98% sequence similarity with the human (Homo sapiens), while the heterochromatic regions display considerable divergence. Positive heterochromatic regions revealed by the CBG-technique are confined to pericentromeric areas in humans, while in chimpanzees, these regions are pericentromeric, telomeric, and intercalary. When human chromosomes are digested with restriction endonuclease AluI and stained by Giemsa (AluI/Giemsa), positive heterochromatin is detected only in the pericentromeric regions, while in chimpanzee, telomeric, pericentromeric, and in some chromosomes both telomeric and centromeric, regions are positive. The DA/DAPI technique further revealed extensive cytochemical heterogeneity of heterochromatin in both species. Nevertheless, the fluorescence in situ hybridization technique (FISH) using a centromeric alpha satellite cocktail probe revealed that both primates share similar pericentromeric alpha satellite DNA sequences. Furthermore, cross-hybridization experiments using chromosomes of gorilla (Gorilla gorilla) and orangutan (Pongo pygmaeus) suggest that the alphoid repeats of human and great apes are highly conserved, implying that these repeat families were present in their common ancestor. Nevertheless, the orangutan's chromosome 9 did not cross-hybridize with human probe. The euchromatic regions of chimpanzee (Pan troglodytes) genome share approximately 98% sequence similarity with the human (Homo sapiens), while the heterochromatic regions display considerable divergence. Positive heterochromatic regions revealed by the CBG-technique are confined to pericentromeric areas in humans, while in chimpanzees, these regions are pericentromeric, telomeric, and intercalary. When human chromosomes are digested with restriction endonuclease AluI and stained by Giemsa (AluI/Giemsa), positive heterochromatin is detected only in the pericentromeric regions, while in chimpanzee, telomeric, pericentromeric, and in some chromosomes both telomeric and centromeric, regions are positive. The DA/DAPI technique further revealed extensive cytochemical heterogeneity of heterochromatin in both species. Nevertheless, the fluorescence in situ hybridization technique (FISH) using a centromeric alpha satellite cocktail probe revealed that both primates share similar pericentromeric alpha satellite DNA sequences. Furthermore, cross-hybridization experiments using chromosomes of gorilla (Gorilla gorilla) and orangutan (Pongo pygmaeus) suggest that the alphoid repeats of human and great apes are highly conserved, implying that these repeat families were present in their common ancestor. Nevertheless, the orangutan's chromosome 9 did not cross-hybridize with human probe. © 1995 Wiley-Liss, Inc.     

The AluI-induced bands in great apes and man: implication for heterochromatin characterization and satellite DNA distribution

Restriction endonucleases have recently been proved to be active on fixed chromatin, producing differences in staining of metaphase chromosomes. In this paper we show the results obtained by treating the metaphase chromosomes of Pan troglodytes, Pan paniscus, and Gorilla gorilla with the restriction enzyme AluI. These results demonstrate qualitative differences in the telomeric heterochromatin between Pan and Gorilla despite the fact that these areas appear homogeneous in the two genera by the C-banding method. The results found with individual chromosomes in the different species also appear relevant, in the light of the evolutionary relationships between these nonhuman primates and man. Lastly, the results suggest the presence, in great apes, of some highly repetitive DNA sequences different from the human satellites I-IV.     

Analyse de la structure fine des chromosomes du Gorille (Gorilla gorilla)

Summary Prometaphasic chromosomes of Gorilla, Homo and Pan are compared, using R, Q, T and H-bands techniques in complement of a previous work (Lejeune et al., 1973). Various mechanisms of chromosomal rearrangements are demonstrated with particular reference to heterochromatic segments. Some phylogenic conclusions are proposed.

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Zusammenfassung Prometaphase-Chromosomen von Gorilla, Mensch und Schimpanse werden mit hilfe der R-, Q-, T- und H-Bandentechnik verglichen; frühere Arbeiten (Lejeune et al., 1973) werden dadurch ergänzt. Verschiedene Mechanismen von Chromosomen-Rearrangements werden dargestellt; dabei finden die Heterochromatin-Segmente besondere Beachtung. Einige phylogenetische Folgerungen werden gezogen.

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Travail de l'E.R.A. n^o 47 du C.N.R.S.     

Karyomorphology, heterochromatin patterns and evolution in the genus Ophrys (Orchidaceae)

Karyotype structures and heterochromatin distribution in representative taxa of the genus Ophrys are compared, based on Feulgen-stained and banded somatic metaphase chromosomes. The karyotypes of Ophrys iricolor , O. lupercalis , O. caesiella , O. lutea , O. lunulata , O. x. tardans , O. apifera , O. praecox , O. lacaitae and O. insectifera are described for the first time. The karyological analyses indicate the relationships among the species with respect to asymmetry indices and heterochromatin content. Chromosomal differences have been helpful in clarifying the taxonomic position of Ophrys species that do not have clear affinities. The representative species of Araniferae , Fuciflorae and Ophrys sections exhibited the most asymmetrical karyotypes, while chromosome complements of the O. fusca–O. lutea group, of O. tenthredinifera and of O. bombyliflora proved to be less asymmetrical. Weakly heterochromatic chromosomes, with heterochromatin present mostly in thin centromeric bands, characterize Ophrys C-banded karyotypes. Chromomycin A₃ (CMA) staining revealed that the analysed species exhibit a weak pattern of CMA⁺ bands at centromeric, intercalary or telomeric regions. No DAPI bright blocks were observed. The significance of the karyological data is discussed with regard to the relationships between the analysed species. © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 148 , 87–99.     

Karyotype structure, supernumerary chromosomes and heterochromatin distribution suggest a pathway of karyotype evolution in Dactylorhiza (Orchidaceae)

The relationships between Dactylorhiza romana and D. saccifera from southern Italy were analysed. These two species, both with 2 n = 2 x = 40 chromosomes and belonging to different sections of the genus, were distinguishable on the basis of karyotype structure and heterochromatin amounts and distribution. Their C-banded karyotypes differed considerably. D. saccifera showed most chromosomes with banded regions in the short arms, whereas in D. romana the bands were located mostly at telomeric regions of longer arms. Several individuals of D. romana had one or two very large heterochromatic supernumerary chromosomes. Based on evidence resulting from karyotype structure and heterochromatin distribution in the two species and on the genetic distances derived from the comparison of ITS sequences, it is suggested that D. romana represents a primitive form with respect to D. saccifera and is a possible intermediate step in the evolution of the genus Dactylorhiza from the 42-chromosome Orchis group. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 138 , 85–91.     

Parasitological evidence pertaining to the phylogeny of the hominoid primates

A systematic review of parasitological data pertaining to the phylogeny of hominoid primates revealed considerable internal consistency and congruence with non-parasitological data. Hylobatids are supported as the sister-group of Pongo + Pan + Gorilla , the 'Great Apes'. Within the Great Apes, Pan + Gorilla are sister taxa. Multiple analyses of presence/absence data place Homo with cercopithecids, probably an artefact of humans' widespread occurrence and polymorphic feeding and living habits. Explicit phylogenetic hypotheses are available for only two parasite groups. Hookworms of the genus Oesophagostomum subgenus Conoweberia place Homo as the sister-group of Pan + Gorilla , whereas pinworms of the genus Enterobius place Homo as the sister-group of Pongo + Pan + Gorilla . This disagreement among data sets with regards to the placement of Homo , combined with the complete agreement about the placement of the other hominoids, is consistent with uncertainties in current findings from other sets of data.     

Characterization of constitutive heterochromatin in Cebus apella (Cebidae, Primates) and Pan troglodytes (Hominidae, Primates): comparison to human chromosomes.

Using G bands, some homologies between the chromosomes of Cebus apella (CAP) and human chromosomes are difficult to establish. To solve this problem, we analyzed these homologies by fluorescence in situ hybridization using human whole chromosome probes (ZOO-FISH). The results indicated that 1) the human probe for chromosome 2 partially hybridizes with CAP chromosomes 13 and 5, 2) the human probe for chromosome 3 partially hybridizes with CAP chromosomes 18 and 20, 3) the human probe for chromosome 9 partially hybridizes with CAP chromosome 19, and 4) the human probe for chromosome 14 hybridizes with the p-terminal and q-terminal regions of CAP chromosome 6. However, none of the human probes employed hybridized with the heterochromatic regions of CAP chromosomes. For this reason, we characterized the heterochromatic regions of CAP chromosomes and of the chromosomes of Pan troglodytes (PTR), to allow comparison between CAP, PTR, and human chromosomes using in situ digestion of fixed chromosomes with the restriction enzymes AluI, HaeIII, and RsaI and by fluorescent staining with DA/DAPI. The results show that 1) centromeric heterochromatin is heterogeneous in the three species studied and 2) noncentromeric heterochromatin is homogeneous within each of the three species, but is different for each species. Thus, centromeric heterochromatin undergoes a higher degree of variability than noncentromeric heterochromatin.     

The karyotypes of the thorny catfishes <Emphasis Type="Italic">Wertheimeria maculata</Emphasis> Steindachner, 1877 and <Emphasis Type="Italic">Hassar wilderi</Emphasis> Kindle, 1895 (Siluriformes: Doradidae) and their relevance in doradids chromosomal evolution

We studied the karyotypes of two doradids, the rare and endangered Wertheimeria maculata and a derived Amazonian species, Hassar wilderi. Cytogenetic characterization was assessed using conventional staining (Giemsa), C-banding, and NOR banding. Both species had 2n = 58 chromosomes but differed in their chromosome formulae, 24 m + 14sm + 8st + 12a for W. maculata and 32 m + 16sm + 10st for H. wilderi. In W. maculata heterochromatin was mainly telomeric, and three chromosomes had a fully heterochromatic arm; in H. wilderi heterochromatin was also predominantly telomeric and evident in many more chromosomes. Hassar wilderi also presented one pair of homologues with a fully heterochromatic arm. In both species, nucleolar organizer regions were restricted to one pair of subtelocentric chromosomes. Assuming a basal position for W. maculata, we hypothesized that underlying conserved diploid and NOR-bearing chromosome numbers, chromosomal evolution in doradids has involved pericentric inversions and an increase of heterochromatic blocks.     

Ectocranial suture fusion in primates: pattern and phylogeny

Patterns of ectocranial suture fusion among Primates are subject to species‐specific variation. In this study, we used Guttman Scaling to compare modal progression of ectocranial suture fusion among Hominidae (Homo, Pan, Gorilla, and Pongo), Hylobates, and Cercopithecidae (Macaca and Papio) groups. Our hypothesis is that suture fusion patterns should reflect their evolutionary relationship. For the lateral‐anterior suture sites there appear to be three major patterns of fusion, one shared by Homo‐Pan‐Gorilla, anterior to posterior; one shared by Pongo and Hylobates, superior to inferior; and one shared by Cercopithecidae, posterior to anterior. For the vault suture pattern, the Hominidae groups reflect the known phylogeny. The data for Hylobates and Cercopithecidae groups is less clear. The vault suture site termination pattern of Papio is similar to that reported for Gorilla and Pongo. Thus, it may be that some suture sites are under larger genetic influence for patterns of fusion, while others are influenced by environmental/biomechanic influences. J. Morphol. 275:342–347, 2014. © 2013 Wiley Periodicals, Inc.     

Accentuated polymorphism of heterochromatin and nucleolar organizer regions in Astyanax scabripinnis (Pisces, Characidae): tools for understanding karyotypic evolution

Astyanax scabripinnis has been considered a species complex because it presents high karyotypic and morphological variability among its populations. In this work, individuals of two A. scabripinnis populations from different streams in the same hydrographic basin were analyzed through C‐banding and AgNOR. Although they present distinct diploid numbers, they show meta and submetacentric chromosome groups highly conserved (numerically and morphologically). Other chromosomal characteristics are also shared by both populations, as the pattern of constitutive heterochromatin distribution (large blocks in the telomeric regions of subtelocentric and acrocentric chromosomes) and some nucleolar chromosomes. Inter‐individual variations both in the number and size of heterochromatic blocks, and in the number and localization of NORs were verified in the studied populations, characterizing them as polymorphics for these regions. The mechanisms involved in the dispersion of heterochromatin and NORs through the karyotypes, as well as the possible events related to the generation of polymorphism of those regions are discussed. Furthermore, relationships between these populations and within the context of the scabripinnis complex are also approached. This revised version was published online in July 2006 with corrections to the Cover Date.     

Chromosome banding in Amphibia

The distribution and quantity of constitutive heterochromatin and of the nucleolus organizer regions (NORs) on the chromosomes of 22 species of bufonids and hylids (Amphibia, Anura) was investigated. Three different kinds of constitutive heterochromatin were found and the frequency of brightly fluorescing heterochromatic regions was remarkably high. On almost all chromosomes there is centric and telomeric heterochromatin. Quantitative estimates of heterochromatin demonstrate that large DNA differences among closely related species can not be attributed to differing quantities of constitutive heterochromatin. In all species investigated, only one homologous pair of NORs was found, which lies preferentially in the proximal and interstitial segments of the long chromosome arms. The NORs are always associated with constitutive heterochromatin on both sides. The size variability between homologous NORs is very high. In the euchromatic regions of the metaphase chromosomes, neither Q- nor G-bands can be demonstrated; this can be attributed to an extremely strong contraction of the anuran chromosomes. On the basis of these results various mechanism of the chromosomal evolution in Anura are discussed.     

Comparative cytogenetics and heterochromatic patterns in two species of the genus Acanthostracion (Ostraciidae: Tetraodontiformes)

Some groups of fish, such as those belonging to the Order Tetraodontiformes, may differ significantly in the amount and location of heterochromatin in the chromosomes. There is a marked variation in DNA content of more than seven-fold among the families of this Order. However, the karyoevolutionary mechanisms responsible for this variation are essentially unknown. The largest genomic contents are present in species of the family Ostraciidae (2.20–2.60 pg). The present study cytogenetically characterized two species of the family Ostraciidae, Acanthostracion polygonius and A. quadricornis, using conventional staining, C-bandings, Ag-NOR, CMA₃/DAPI, AluI, PstI, EcoRI, TaqI and HinfI restriction enzymes (REs) and double FISH with 18S and 5S rDNA probes. The karyotypes of both species showed 2n = 52 acrocentric chromosomes (FN = 52; chromosome arms) and pronounced conserved structural characteristics. A significant heterochromatic content was observed equilocally distributed in pericentromeric position in all the chromosome pairs. This condition is unusual in relation to the karyotypes of other families of Tetraodontiformes and probability is the cause of the higher DNA content in Ostraciidae. Given the role played by repetitive sequences in the genomic reorganization of this Order, it is suggested that the conspicuous heterochromatic blocks, present in the same chromosomal position and with apparently similar composition, may have arisen or undergo evolutionary changes in concert providing clues about the chromosomal mechanisms which led to extensive variation in genomic content of different Tetraodontiformes families.     

Heterochromatin and chromosome evolution: a FISH probe of Cebus apella paraguayanus (Primate: Platyrrhini) developed by chromosome microdissection

Neotropical Primate karyotypes are highly variable, particularly in the heterochromatic regions, not only regarding the amount of heterochromatin, but also the composition. G and C banding and FISH techniques provide useful information to characterize interspecific relationships. We used chromosome microdissection to develop a FISH probe of the chromosome 11 heterochromatic block (11qHe+) of Cebus apella paraguayanus (CAPp). Fragments of the 11qHe+ microdissected from fibroblast cell culture were collected in a PCR tube, amplified by degenerate oligonucleotide primer-PCR and subsequently labeled. The specificity of the FISH probe was confirmed in metaphases of some Ceboidea species. Signals were located in the He+ of chromosomes 4, 11, 12, 13, and 19 of CAPp and in the He+ of chromosomes 4, 12 and 13 of C. a. nigritus (CAPn); no signals were observed when other Ceboidea species were analyzed. We propose that the heterochromatin observed in CAPp and CAPn is specific for these species. We consider this C. apella heterochromatin identity as a possible key for the interpretation of chromosomal evolution in these Ceboidea.     

Heterochromatin in the chromosomes of the gorilla: characterization with distamycin A/DAPI, D287/170, chromomycin A3, quinacrine, and 5-azacytidine

The chromosomes of the gorilla were extensively studied with various staining techniques labeling the different classes of heterochromatin. The chromosomal distribution of distamycin A/DAPI-, D287/170-, quinacrine-, and chromomycin A3-positive heterochromatic regions, as well as the nucleolus organizer regions, is described and compared with the karyotypes of other hominoid species. Lymphocyte cultures were treated with low doses of 5-azacytidine during the last hours of culture. This cytidine analog induces distinct undercondensation in 37 heterochromatic regions in the 24 gorilla chromosomes. The 5-azacytidine-induced undercondensations are localized not only in most of the distamycin A/DAPI-bright heterochromatic regions but also in many telomeric C-bands of the chromosomes. Furthermore, 5-azacytidine preserves the somatic pairing between heterochromatic regions from the interphase nuclei into the metaphase stage. The homeologies and differences in the chromosomal localization of the various classes of heterochromatin, 5-azacytidine-sensitive regions, 5-methylcytosine-rich DNA sequences, and satellite DNAs in the gorilla, chimpanzee, orangutan, and man are discussed.     

Comparative mapping of human alphoid satellite DNA repeat sequences in the great apes

Heterochromatic regions of chromosomes contain highly repetitive, tandemly arranged DNA sequences that undergo very rapid variation compared to unique DNA sequences that are predominantly conserved. In this study the chromosomal basis of speciation has been looked at in terms of repeat sequences. We have hybridized twenty-one chromosome-specific human alphoid satellite DNA probes to metaphase spreads of the chimpanzee (Pan troglodytes), gorilla (Gorilla gorilla), and orangutan (Pongo pygmaeus) to investigate the evolutionary relationship of heterochromatic regions among such hominoid species. The majority of the probes did not hybridize to their corresponding equivalent chromosome but presented hybridization signals on non-corresponding chromosomes. Such observations suggest that rapid changes may have occurred in the ancestral alphoid satellite DNA sequence, resulting in divergence among the great ape species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Karyotype analysis of 2 species of gibbons (Hylobates lar and H. concolor) with different banding species]

The mitotic chromosomes of two species of gibbons (Hylobates lar and H. concolor) are examined and compared, using various banding techniques. These two species have very different karyotypes. At the most, seven pairs of chromosomes have a similar banding pattern. The other elements generally differ by complex structrual rearrangements. Thus, it is difficult to propose a scheme for chromosomal evolution at this stage. Comparison with the karyotypes of man and anthropoid apes also shows very important differences; very few chromosomes are common or only slightly modified. Some considerations about the hypothetical origin of particular chromosomal structures are given.     

Evolution of chromosome Y in primates

We have investigated, by fluorescence in situ hybridization (FISH), the cytogenetic evolution of the Y chromosome in primates using 17 yeast artificial chromosomes, representative of the Y-specific euchromatic region of the human chromosome Y. The FISH experiments were performed on great apes (Homo sapiens, Pan troglodytes, Gorilla gorilla and Pongo pygmaeus pygmaeus), and on two Old World monkeys species as an outgroup (Cercopitecidae Macaca fascicularis and Papio anubis). The results showed that this peculiar chromosome has undergone rapid and unconstrained evolution both in sequence content and organization. Received: 16 January 1998; in revised form: 29 May 1998 / Accepted: 24 June 1998     

Identity of euchromatic bands from man to Cercopithecidae

Summary The karyotypes of four species of Cercopithecidae: Cercopithecus aethiops tantalus, C. sabaeus, Erythrocebus patas, and Miopithecus talapoin are analysed with nearly all the banding techniques. They are compared with each other, and with the karyotypes of the Baboon P. papio and with that of man. It can be concluded that the quasi-totality or the totality of the euchromatin is common to all, but has undergone structural rearrangements, generally detectable. The heterochromatin, defined by C-band staining, and late-replicating DNA, in contrast, appears very variable: In particular, E. patas has acquired very large heterochromatic segments. The significance of these modifications is discussed.Technical assistance: A. M. Fosse and M. Lombard     

Karyotypes of the most primitive catfishes (Teleostei: Siluriformas: Diplomystidae)

Karyotypes of Diplomystes composensis and Diplomystes nahuelbutaensis were the same diploid number (n= 56).The chromosome formula for D. composensis was 16 metacentric + 24 submetacentric + 8 subtelocentric + 8 telocentric chromosomes and for D. nahuelbutaensis was 14 metacentric + 26 submetacentric + 8 subtelocentric +8 telocentric chromosomes. In contrast, the differences in the chromosomal C-banding patterns between these species was large. For instance, chromosome pairs 5,6, and 7 of D. nahuelbutaensis showed heterochromatic centromeres and pairs 23, 24, 27, and 28 were entirely heterochromotic. Diplomystes composensis showed conspicuous C-banded blocks in pairs 7, 24, and 25 (chromosome pair 7 had one heterochromatic arm, chromosome pair 24 was entirely heterochromatic, and chromosome pair 25 had heterochromatin close to centromere). Comparison with other ostariophysan karyotypes (e.g. gymnotiforms, characiforms, and cypriniforms), does not allow any conclusions about the ploesiomorphic catfish condition, because the karyotypes of the outgroups are too variable. A synapomorphy shared by characiforms, gymnotiforms, and diplomystid catfishes is the presence of more metacentric to submetacentric than substelocentric to telocentric chromosomes. Cypriniforms are more primitive because they have more subtelocentric to telocentric than metacentric to submetacentric chromosomes.