Heterochromatin Pattern Research Articles

Chromosomal differentiation of Tribe Cestreae (Solanaceae) by analyses of 18-5.8-26S and 5S rDNA distribution

Tribe Cestreae is monophyletic with three genera: Cestrum, Sessea, and Vestia. Karyotypically, it is outstanding within Solanaceae by several features: (1) basic number x = 8, (2) large chromosome sizes, (3) complex heterochromatin patterns, (4) occurrence of B-chromosomes (Bs) in Cestrum with particular banding patterns and rDNA sites distribution, and (5) absence of Arabidopsis-type telomeres. Seventeen South American Cestreae species from the three genera were studied using fluorescence in situ hybridization (FISH) with ribosomal DNA regions (5S and 18-5.8-26S) as probes, with the aim of recognizing specific or group-specific chromosomal markers and analyzing karyotype diversity in a systematic and evolutionary context. The first chromosome number report for Cestrum euanthes, C. kunthii, C. lorentzianum, and C. tomentosum is included. Variation in number and distribution of rDNA loci was observed among the species, concerning both As and Bs chromosomes. Despite the constancy of the karyotype and numbers of rDNA loci, the mapping of 18-5.8-26S and 5S rDNA loci allowed to differentiate Cestreae genera and species groups within Cestrum, highlighting the importance of these markers as cytotaxonomic character in this tribe.

C-banding patterns and phenotypic characteristics in individuals ofSapajus(Primates: Platyrrhini) and its application to management in captivity

The classification of capuchin monkeys represents one of the greatest taxonomical controversies within primates. The high variability presented by the species, observed mainly in coat coloration, makes the classification of captive animals difficult. The objective of the present study was to analyze the patterns of constitutive heterochromatin in 26 individuals of capuchin monkeys kept in a screening center of wild animals in Rio de Janeiro state, Brazil, and correlate them with phenotypic characteristics, in order to improve the management in captivity. Metaphasic chromosomes were obtained using lymphocyte culture and the constitutive heterochromatic regions were stained by using the C-banding technique. All individuals presented 2n = 54. C-banding was observed on chromosomes 4, 6, 11, 12, 13, 17, and 19. Two types of C-banding staining in chromosome pair 11 were identified: a large terminal heterochromatic block occupying more than 60% of the chromosome and a small intercalar heterochromatic block. In a...

Heterochromatin Dynamics during the Differentiation Process Revealed by the DNA Methylation Reporter Mouse, MethylRO

SummaryIn mammals, DNA is methylated at CpG sites, which play pivotal roles in gene silencing and chromatin organization. Furthermore, DNA methylation undergoes dynamic changes during development, differentiation, and in pathological processes. The conventional methods represent snapshots; therefore, the dynamics of this marker within living organisms remains unclear. To track this dynamics, we made a knockin mouse that expresses a red fluorescent protein (RFP)-fused methyl-CpG-binding domain (MBD) protein from the ROSA26 locus ubiquitously; we named it MethylRO (methylation probe in ROSA26 locus). Using this mouse, we performed RFP-mediated methylated DNA immunoprecipitation sequencing (MeDIP-seq), whole-body section analysis, and live-cell imaging. We discovered that mobility and pattern of heterochromatin as well as DNA methylation signal intensity inside the nuclei can be markers for cellular differentiation status. Thus, the MethylRO mouse represents a powerful bioresource and technique for DNA methylation dynamics studies in developmental biology, stem cell biology, as well as in disease states.

NOR- bearing as a plesiomorphic characteristic in Mimus saturninus (Passeriformes Mimidae)

The order Passeriformes is the largest group of species karyotyped among birds, however little is known about the cytogenetic of the Mimidae family, registering only karyology basic data (giemsa staining). The aim of this study was to analyze the chromosomal complement from the species Mimus saturninus by conventional staining and differential chromosome banding. Diploid number and chromosome morphology were determined, as well as the distribution pattern of constitutive heterochromatin (CBG-banding), GTG-banding andAgNOR staining (NORs). The Chalk-browed Mockingbird has 2n=80. The first and fourth pairs are submetacentric and the second, third and fifth are acrocentric. The remaining chromosomes pairs of the complement have telocentric morphology. The Z chromosome is submetacentric and the W is metacentric. CBG-banding showed positive staining in the pericentromeric region of most macrochromosomes and microchromosomes and also at Z chromosome, differently from W chromosome which appeared totally heterochromatic. The GTG-banding was similar to Gallus gallus and in other species which have already been GTG-banded. The NORs were identified in a pair of microchromosomes characterized by presenting a remarkable secondary constriction. This can be considered as a plesiomorphic characteristic for M. saturninus once baseline groups (Paleognathae) also showed a pair of microchromosomes bearing NORs.

Cytogenetic analysis in three species from genusAstyanax(Pisces; Characiformes) with a new occurrence of B chromosome inAstyanax paranae

Cytogenetic studies were carried out in three species from genus Astyanax: Astyanax paranae (Tagacaba Stream and Taua Stream), Astyanax altiparanae (Iguacu River and Maringa Stream) and Astyanax fasciatus (Maringa Stream). Both populations of A. paranae featured 2n = 50 chromosomes, although with different karyotype formulae and Ag-nucleolar organizer regions systems. Using the FISH technique in the Tagacaba population, it was possible to detect rDNA genes in 14 chromosomes and bitelomeric marking in one chromosome. The constitutive heterochromatin pattern also showed differences between the two populations. Moreover, the Tagacaba population showed macrochromosome B in 50% of analyzed females. Both A. altiparanae populations featured 2n = 50 chromosomes, but also differed in karyotype formula. Both populations featured multiple-NOR systems and quite similar constitutive heterochromatin patterns. For the A. fasciatus population from Maringa Stream, the diploid number was 2n = 46 chromosomes with karyotype ...

Distribution of constitutive heterochromatin in Triatoma melanocephala (Hemiptera, Triatominae).

In principle, Triatoma melanocephala was included in the Brasiliensis subcomplex on the basis of morphological parameters and geographical layout, since there were no other relevant data available in the literature. On the basis of karyotype, it has been proposed to exclude T. melanocephala, as well as of T. vitticeps and T. tibiamaculata, from the subcomplex, which shows fragmentation of the X sex chromosomes, thereby approaching the species of North America. Therefore, the present study aimed to determine the pattern of constitutive heterochromatin of T. melanocephala to provide new data on the cytotaxonomy of this vector of Chagas disease and especially to try to relate this species to some group, complex or subcomplex of triatomine species, aiding in their classification. This species showed no constitutive heterochromatin in the autosomes or X sex chromosome, but only the Y sex chromosome. The number of chromosomes and heterochromatin pattern of T. melanocephala proved to be identical to that described for Panstrongylus lutzi. Thus, the present study demonstrated a tentative relationship between T. melanocephala and P. lutzi. However, we emphasize that other comparative studies should be conducted between these species, such as experimental crosses and molecular, enzymatic, morphological, and morphometric analyses to determine whether these species are actually evolutionarily related or if the number of chromosomes and the heterochromatin pattern emerged as homoplasies in T. melanocephala and P. lutzi.

Distribution of constitutive heterochromatin in species of triatomines with fragmentation of sex chromosomes X.

Cytogenetic analyses of triatomines are considered to be important taxonomic tools. Thus, we analyzed the pattern of constitutive heterochromatin in 7 species of triatomine with fragmentation of the sex chromosome X, focusing on the cytotaxonomy of these triatomines. The species analyzed included Triatoma vitticeps, Triatoma melanocephala, Triatoma tibiamaculata, Triatoma protracta, Meccus pallidipennis, Panstrongylus megistus, and Panstrongylus lignarius. The seminiferous tubules of the adult males were subjected to C-banding. P. megistus and P. lignarius showed differences in chromosome number and disposition of constitutive heterochromatin, as only P. lignarius showed C-blocks in autosomes. C-banding can differentiate these species, since one of the sex chromosome (X) is heterochromatic in T. vitticeps. T. protracta showed C-blocks in both ends of all autosomes, T. tibiamaculata showed terminal C-dots in some autosomal pairs and M. pallidipennis did not show constitutive heterochromatin in autosomes. Thus, we confirmed the heterochromatic pattern of 7 species of insects and emphasized the importance of cytogenetic techniques for C-banding for taxonomy studies of the triatomines, which are important vectors of Chagas disease.

A comparative study of genome organization and inferences for the systematics of two large bushcricket genera of the tribe Barbitistini (Orthoptera: Tettigoniidae: Phaneropterinae)

BackgroundPoecilimon and Isophya are the largest genera of the tribe Barbitistini and among the most systematically complicated and evolutionarily intriguing groups of Palearctic tettigoniids. We examined the genomic organization of 79 taxa with a stable chromosome number using classical (C–banding, silver and fluorochrome staining) and molecular (fluorescence in situ hybridization with 18S rDNA and (TTAGG) n telomeric probes) cytogenetic techniques. These tools were employed to establish genetic organization and differences or similarities between genera or species within the same genus and determine if cytogenetic markers can be used for identifying some taxonomic groups of species.ResultsDifferences between the karyotypes of the studied genera include some general changes in the morphology of the X chromosome in Isophya (in contrast to Poecilimon). The number of major rDNA clusters per haploid genome divided Poecilimon into two main almost equal groups (with either one or two clusters), while two rDNA clusters predominated in Isophya. In both genera, rDNA loci were preferentially located in the paracentromeric region of the autosomes and rarely in the sex chromosomes. Our results demonstrate a coincidence between the location of rDNA loci and active NORs and GC-rich heterochromatin regions. The C/DAPI/CMA3 bands observed in most Poecilimon chromosomes suggest the presence of more families of repetitive DNA sequences as compared to the heterochromatin patterns in Isophya.ConclusionsThe results show both differences and similarities in genome organization among species of the same genus and between genera. Previous views on the systematics and phylogenetic grouping of certain lineages are discussed in light of the present cytogenetic results. In some cases, variation of chromosome markers was observed to correspond with variation in other evolutionary traits, which is related to the processes of ongoing speciation and hybridization in zones of secondary contact. It was concluded that the physical mapping of rDNA sequences and heterochromatin may be used as an additional marker for understanding interspecific relationships in these groups and their routes of speciation.

Evolutionary dynamics of rDNA genes on chromosomes of the Eucinostomus fishes: cytotaxonomic and karyoevolutive implications.

Several chromosomal features of Gerreidae fish have been found to be conserved. In this group, it is unclear whether the high degree of chromosomal stasis is maintained when analyzing more dynamic regions of chromosomes, such as rDNA sites that generally show a higher level of variability. Thus, cytogenetic analyses were performed on 3 Atlantic species of the genus Eucinostomus using conventional banding (C-banding, Ag-NOR), AT- and GC-specific fluorochromes, and fluorescence in situ hybridization mapping of telomeric sequences and 5S and 18S rDNA sites. The results showed that although the karyotypical macrostructure of these species is similar (2n = 48 chromosomes, simple Ag-NORs seemingly located on homeologous chromosomes and centromeric heterochromatin pattern), there are differences in the positions of rDNA subunits 5S and 18S. Thus, the ribosomal sites have demonstrated to be effective cytotaxonomic markers in Eucinostomus, presenting a different evolutionary dynamics in relation to other chromosomal regions and allowing access to important evolutionary changes in this group.

First karyotype analysis, physical rDNA mapping, and genome size assessment in 4 North African Astragalus taxa (Fabaceae)

Four taxa of Algerian Astragalus L. were studied for their chromosome number, karyotype features, and genome size. Fluorochrome banding was done for detection of GC-rich DNA regions, fluorescence in situ hybridization (FISH) for physical mapping of 35S and 5S rRNA genes, and flow cytometry for nuclear DNA content. All the taxa present the same chromosome number (2n = 2x = 16) and single 35S and 5S rDNA loci, but their distributions on chromosomes are different. The GC heterochromatin pattern was different among studied taxa and an unusually high number of chromomycin-positive bands were observed in A. pseudotrigonus Batt. & Trab. The genome size differed between species, ranging from 2C = 1.39 pg in A. cruciatus Link. to 2C = 2.71 pg in A. armatus subsp. tragacanthoides (Desf.) Maire. No difference in nuclear DNA amount was detected between the 2 subspecies of A. armatus. Although Astragalus is a large genus comprising some 3000 species, such morphometric and molecular cytogenetic karyotype analyses, with genome sizes, are particularly scarce therein. Therefore, published genome sizes have also been compiled into one table.

Molecular–cytogenetic studies of ribosomal RNA genes and heterochromatin in three European Fraxinus species

Three European representatives of the genus Fraxinus were studied for the first time for their rDNA and heterochromatin patterns. The physical mapping of two rRNA gene families 5S and 18S–5.8S–26S (45S) and the distributional pattern of GC-rich regions in the chromosomes have been established by means of fluorescence in situ hybridization (FISH) and fluorochrome banding with chromomycin A3. The genome size was assessed by flow cytometry. Heterochromatin and rDNA organization was conserved and almost identical for two species from Fraxinus section (F. angustifolia and F. excelsior). The number and position of rDNA loci in F. ornus (section Ornus) were similar; however, the organization of genes was quite different. In this species the 5S and 45S rRNA genes were colocalized at the level of satellites of two chromosome pairs bearing nucleolar organizing regions (NORs). One 5S locus was also observed under the 45S one of one chromosome pair. In F. angustifolia and F. excelsior, only 45S loci were situated at the level of satellites and secondary constrictions, while 5S was located just under 45S in the distal part of the short arm of one out of two marked pairs. The number and position of GC-rich DNA correspond to those of 45S loci. The genome size (2C value) was of 1.54 and 1.68 pg for F. angustifolia and F. excelsior, respectively. Fraxinus ornus possessed the highest 2C DNA value (1.98 pg). In the light of these cytogenetic features the clear differentiation between two sections (Fraxinus and Ornus) was observed both at the rDNA and genome size levels.

Cytogenetic characterization of the ant Trachymyrmex fuscus Emery, 1934 (Formicidae: Myrmicinae: Attini) with the description of a chromosomal polymorphism

SummaryThe genus Trachymyrmex is a key group in the tribe Attini because of its close phylogenetic relationship to leaf-cutter ants, Acromyrmex and Atta. Cytogenetic data are only available for five taxa of Trachymyrmex, with chromosome numbers of 2n = 12, 18, 20 and 22, and morphology with predominantly metacentric chromosomes. The aim of the present study was to characterize the karyotype of the ant Trachymyrmex fuscus Emery, 1934, by means of the number and morphology of its chromosomes, heterochromatin pattern, CMA3 and DAPI fluorochromes in the population of two nests collected at Paraopeba, state of Minas Gerais, Brazil. Nineteen females presented 2n = 18 chromosomes (16m + 2sm) and a single male presented n = 9 (8m + 1sm). A size chromosomal polymorphism involving the short arm of the submetacentric pair was confirmed by statistical analysis, with three character conditions: heterozygous SB (with a difference in size between the short arms), standard SS (smaller short arms) and homozygote BB (bigger short arms). In the first nest, both SB and SS workers were observed. The other nest contained heterozygous (SB), homozygous (BB), and a male carrying the B chromosome (larger size). The presence of heterochromatin on all centromeric and pericentromeric chromosomes of T. fuscus suggests that the size difference observed in the submetacentric pair in the SB and BB workers is not related to the heterochromatin but to a duplication of euchromatic regions through intra- or inter- chromosomal rearrangements. The fluorochrome CMA3 matched the C-banding markings, indicating that the heterochromatin is rich in GC base pairs. As far as we know, this is the first chromosomal polymorphism reported in the tribe Attini.

Mapping five repetitive DNA classes in sympatric species of Hypostomus (Teleostei: Siluriformes: Loricariidae): analysis of chromosomal variability

Fish belonging to the genus Hypostomus are known for exhibiting a striking diversity in its karyotype structure, however the knowledge concerning the distribution patterns of heterochromatin and location of repetitive DNA sequences in the karyotypes is still limited. Aiming a better understanding of the chromosomal organization in this group, we analyzed three sympatric species of Hypostomus collected in the Hortela stream, a component of the Paranapanema River basin, Botucatu/SP/Brazil. The analyses involved the cytogenetic characterization and chromosomal mapping of repetitive sequences and intra/interspecific comparisons using sequences of the cytochrome C oxidase subunit I. The results revealed that H. ancistroides presents a karyotype with 2n = 68 chromosomes, H. strigaticeps 2n = 72 chromosomes, and H. nigromaculatus 2n = 76 chromosomes. In addition to differences found in the diploid number, it was also observed variations in karyotypic formulae, amount of constitutive heterochromatin, and location of nucleolus organizer regions. The cytogenetic mapping of 5S and 18S rDNA, as well as of the H3 histone gene, disclosed a differential dispersion process among the three species. In some cases the Rex1 transposable element showed to be co-located with 5S rDNA sites. The molecular analyses support the cytogenetic data and represent an additional tool for the characterization of the analyzed species. The results evidenced that chromosomal variations are not restricted to differences in diploid number or karyotypic macrostructure in the genus Hypostomus, indicating that events such as transposition of heterochromatin and rDNA segments may participate in the differentiation process occurred in these species.

Characterization of Brazilian accessions of wild Arachis species of section Arachis (Fabaceae) using heterochromatin detection and fluorescence in situ hybridization (FISH)

The cytogenetic characterization of Arachis species is useful for assessing the genomes present in this genus, for establishing the relationship among their representatives and for understanding the variability in the available germplasm. In this study, we used fluorescence in situ hybridization (FISH) to examine the distribution patterns of heterochromatin and rDNA genes in 12 Brazilian accessions of five species of the taxonomic section Arachis. The heterochromatic pattern varied considerably among the species: complements with centromeric bands in all of the chromosomes (A. hoehnei) and complements completely devoid of heterochromatin (A. gregoryi, A. magna) were observed. The number of 45S rDNA loci ranged from two (A. gregoryi) to eight (A. glandulifera), while the number of 5S rDNA loci was more conserved and varied from two (in most species) to four (A. hoehnei). In some species one pair of 5S rDNA loci was observed adjacent to 45S rDNA loci. The chromosomal markers revealed polymorphism in the three species with more than one accession (A. gregoryi, A. magna and A. valida) that were tested. The previous genome assignment for each of the species studied was confirmed, except for A. hoehnei. The intraspecific variability observed here suggests that an exhaustive cytogenetic and taxonomic analysis is still needed for some Arachis species.

B Chromosomes in the Tree FrogHypsiboas albopunctatus(Anura: Hylidae)

Supernumerary or B chromosomes are one of the main causes for numerical chromosomal variation in higher eukaryotes. These extragenetic elements have been studied for more than a century, with the goal of trying to understand their origin, and how they survive as a polymorphism in natural populations. Hypsiboas albopunctatus is a nocturnal hylid frog distributed in the central-eastern part of South America. Previously, variation in chromosome numbers was described for a population from Rio Claro, Sao Paulo, Brazil, in which a single small-sized, metacentric B chromosome was present in ca. 40% of the analyzed individuals (n ¼ 17). We herein describe the presence of B chromosomes in populations of H. albopunctatus from northeastern Argentina (Corrientes and Misiones provinces), with unusual morphological and structural characteristics. The frequency of B chromosomes varied significantly among analyzed populations. We found four diploid numbers (2N ¼ 22, 22 þ 1B, 22 þ 2B, and 22 þ 3B), and in a few individuals mitotic instability occurred. C banding revealed variations in the heterochromatin (DAPIþ) pattern between Bs with similar morphology, indicating the existence of two new structural variants of these supernumerary elements in H. albopunctatus (B1 and B2). Nucleolar organizer regions marked positively on the eighth pair, coincident with the location of ribosomal DNA as demonstrated with fluorescent in situ hybridization, but Bs did not mark positive with these two techniques. Also, fluorescent in situ hybridization with telomeric probes showed no differences in location and intensity between Bs and autosomal chromosomes. The present communication is the first case of B chromosome polymorphisms in hylid frogs and the sixth reported in Anura.

Distribution of constitutive heterochromatin in two species of triatomines: Triatoma lenti Sherlock and Serafim (1967) and Triatoma sherlocki Papa, Jurberg, Carcavallo, Cerqueira & Barata (2002)

Triatoma lenti and Triatoma sherlocki are hemipterans that belong to the brasiliensis subcomplex. In triatomines, the constitutive heterochromatin pattern is species-specific and allows, in many cases, for the grouping of species. Thus, we cytogenetically analyzed T. sherlocki and T. lenti using C-banding, and we compared the results with previous ones obtained in other species of the brasiliensis subcomplex. Both species were found to have a male diploid chromosome number of 22 chromosomes (2n=20A+XY) with heterochromatic blocks at one or both chromosomal ends of all autosomal pairs. During early meiotic prophase, they showed a large heteropycnotic chromocenter constituted by the association of both sex chromosomes plus two autosomal pairs and many heterochromatic blocks dispersed inside the nucleus. All of these cytogenetic characteristics are similar to those observed in other species of brasiliensis subcomplex, results which confirm the grouping of T. sherlocki and T. lenti within this subcomplex. However, we emphasize the importance of other approaches, such as molecular analysis, to confirm the placement of T. lenti within the brasiliensis subcomplex.

Arrangement of nuclear structures is not transmitted through mitosis but is identical in sister cells

Although it is well known that chromosomes are non-randomly organized during interphase, it is not completely clear whether higher-order chromatin structure is transmitted from mother to daughter cells. Therefore, we addressed the question of how chromatin is rearranged during interphase and whether heterochromatin pattern is transmitted after mitosis. We additionally tested the similarity of chromatin arrangement in sister interphase nuclei. We noticed a very active cell rotation during interphase, especially when histone hyperacetylation was induced or transcription was inhibited. This natural phenomenon can influence the analysis of nuclear arrangement. Using photoconversion of Dendra2-tagged core histone H4 we showed that the distribution of chromatin in daughter interphase nuclei differed from that in mother cells. Similarly, the nuclear distribution of heterochromatin protein 1β (HP1β) was not completely identical in mother and daughter cells. However, identity between mother and daughter cells was in many cases evidenced by nucleolar composition. Moreover, morphology of nucleoli, HP1β protein, Cajal bodies, chromosome territories, and gene transcripts were identical in sister cell nuclei. We conclude that the arrangement of interphase chromatin is not transmitted through mitosis, but the nuclear pattern is identical in naturally synchronized sister cells. It is also necessary to take into account the possibility that cell rotation and the degree of chromatin condensation during functionally specific cell cycle phases might influence our view of nuclear architecture.

Karyomorphology of Caesalpinia Species (Caesalpinioideae: Fabaceae) from Caatinga and Mata Atlantica Biomes of Brazil

Out of 140 Caesalpinia s.l. species, only 20 species have the chromosome numbers presently known, and nine species have the karyomorphological studies available. We determine the karyotype and the chromosome morphometry in five Caesalpinia s.l. spp., and we describe the heterochromatin pattern in four of them. The diploid chromosome number of 24 was reported for the first time in Caesalpinia calycina, Caesalpinia microphylla and Caesalpinia pluviosa var. peltophoroides, and confirmed in Caesalpinia ferrea var. leiostachya and Caesalpinia pulcherrima. Different karyotype formulae were obtained for each of these five species. The chromosome asymmetry index varied from 34.94 % in C. pluviosa to 39.45 % in C. ferrea. The average chromosome length among the five species differed significantly (P<0.010). The heterochromatin blocks were evidenced by C-banding in both terminal and proximal regions of the Caesalpinia spp. chromosomes. This study represents a contribution toward an increased knowledge on cytogenetics and evolution of this genus.

The X chromosome, heterochromatin and speciation in malaria mosquitoes

The available data suggest that the X chromosome of Anopheles had originated independently from the X chromosome of Drosophila. In order to gain insights into a possible role of the X chromosome in speciation of malaria mosquitoes, we studied the pattern of rearrangements, genomic landscapes and heterochromatin variations among closely and distantly related species. • Our cytogenetic analysis of the An. gambiae species complex showed fixed differences in the amount and pattern of the X heterochromatin among sibling species as well as between the M and S forms (Fig. 1). Interestingly, the low-recombination pericentric region displayed the high level of polymorphism in the size of heterochromatin and the rDNA locus (Fig. 2). • The largest region of the high genomic divergence between the incipient species (the M and S forms) co-localizes with the heterochromatin of the An. gambiae X chromosome (Fig. 3). • Our comparative physical mapping of distantly related malaria mosquito species identified the fastest rate of inversion fixation on the X chromosome despite the paucity of inversion polymorphism (Fig. 4). • The analysis of the An. gambiae genome demonstrated the highest density of transposable elements, microsatellites, minisatellites, and satellites in the X chromosome (Fig. 5). • The X chromosome is enriched in genes that may be involved in premating isolation, such as genes encoding for proteins with molecular and signal transduction activity (Fig. 6). Thus, our study points out to the elevated evolutionary plasticity of the euchromatic gene order and heterochromatin structure of the X chromosome suggesting a possible role of the sex chromosome in speciation of malaria mosquitoes. An. gambiae SUA 2La (M) An. gambiae Pimperena (S) An. gambiae Akron (M) An. gambiae Kisumu (S) Pericentric heterochromatin exhibits inter-specific differences An. merus An. quadriannulatus An. arabiensis An. gambiae Zanu (S) Fig. 1. Heterochromatin patterns observed after staining with DAPI. Arrows indicate heterochromatin (darker areas) exhibiting inter and intra-specific differences. While the M forms (Sua, Akron) exhibit single large bands (A, B), S forms (Zanu, Pimperena, Kisumu) exhibit a secondary heterochromatic band in (C-E). Sibling species (FH) also exhibit variation in heterochromatin patterns. An. gambiae Kisumu (S) An. gambiae Akron (M)

Comparative cytogenetics of ten species of cichlid fishes (Teleostei, Cichlidae) from the Araguaia River system, Brazil, by conventional cytogenetic methods

Cichlids represent one of the most species-rich families of fishes and have attracted the attention of evolutionary biologists due to the rapid radiation occurring in some groups and the importance of some species in the world aquaculture. Cytogenetic analysis was conducted in 10 cichlid species from the Araguaia River, Amazon Basin, Brazil. The chromosome number was 2n=48 for all analyzed species except for Laetacara araguaiae Ottoni et Costa, 2009 (2n=44). Chromosomal polymorphism was detected only in Geophagus proximus (Castelnau, 1855), which exhibits an extra large submetacentric and and a dot-like chromosomes. Moreover, the C-banding revealed a general pericentromeric heterochromatic pattern and some additional blocks for some species. The heterochromatic blocks corresponding to AgNOR bearing regions were observed in all species and also corresponded to CMA3 positive blocks, which were observed in terminal regions. Besides the general conserved chromosomal and heterochromatin patterns for South American cichlids, the presence of GC-rich heterochromatin was quite different in the species Biotodoma cupido (Heckel, 1840), Geophagus proximus, Retroculus lapidifer (Castelnau, 1855), Crenicichla strigata Günther, 1862 and Heros efasciatus Heckel, 1840. The results suggest that independent events of heterochromatin modification occurred during chromosome evolution in the group, regardless of the conservation of macro-chromosomal structure.