Pericentric inversion events in karyotypic distinction of Brazilian lizards of genus Phyllopezus (Squamata, Gekkonidae) detected by chromosomal banding patterns

Cytogenetic investigations based on conventional and differential staining analysis (C-and replication R-banding and Ag-staining) were carried out on eight specimens of Phyllopezus periosus, 17 of P. pollicaris pollicaris, and one of P. pollicaris przewalskii collected from different localities of Brazil. P. periosus and P. p. pollicaris share the same diploid number of 2n = 40 chromosomes, and their karyotypes are very distinctive regarding to the number of biarmed and uniarmed chromosomes. After careful side-by-side comparison of R-banded chromosomes in both taxa, pronounced homology between, at least, eight pairs was revealed. The R-banding patterns allowed us to postulate that karyotype differentiation could be due to pericentric inversion events. P. p. przewalskii (2n = 38) exhibited a very similar karyotype to that found in P. p. pollicaris, except for the presence of one metacentric pair, which probably resulted from a Robertsonian rearrangement. Single and multiple pairs of NOR-bearing chromosomes, showing variation in number and location, were detected among the three forms of Phyllopezus. Similar C-banding patterns were found in P. periosus and P. p. pollicaris. Sex chromosomes were not positively identified.

Cytogenetics of a new species of Paratelmatobius cardosoi group (Anura : Leptodactylidae), with the description of an apparent case of pericentric inversion

The karyotype of a new species of Paratelmatobius from the P cardosoi group is described. As with other Paratelmatobius and Scythrophrys karyotypes, Paratelmatobius sp. (aff. cardosoi) shows a diploid number of 24 chromosomes, in addition to other similarities with the former karyotypes. The Paratelmatobius sp. (aff. cardosoi) karyotype differs from that of P. cardosoi in the morphology of pair 4, the NOR location and the C-bands in pairs 3 and 8 (exclusive to Paratelmatobius sp.) and those of pairs 7 and 9 (exclusive to P. cardosoi). Both karyotypes also differ in the amount of heterochromatin in pair 1. The presence of interstitial heterochromatin in the long arm of pair 1 and the interstitial C-bands in both arms of chromosome 5 are apparently synapomorphic characters of P. cardosoi and Paratelmatobius sp. (aff. cardosoi), since they are absent in the other Paratelmatobius and Scythrophrys karyotypes. In Paratelmatobius sp. (aff. cardosoi), the nucleolus organizer region is on the short arm of a small metacentric chromosome (pair 9), an arrangement similar to the NOR-bearing chromosome pair in the karyotype of P. poecilogaster and in karyotype 11 of Scythrophrys. A conspicuous heteromorphism unrelated to the sex determining mechanism was also observed and probably arose from a pericentric inversion.

Variant Rett Syndrome in a Girl with a Pericentric X-Chromosome Inversion Leading to Epigenetic Changes and Overexpression of the MECP2 Gene

Rett syndrome is a neurodevelopmental disorder caused by mutations in the MECP2 gene. We investigated the genetic basis of disease in a female patient with a Rett-like clinical. Karyotype analysis revealed a pericentric inversion in the X chromosome -46,X,inv(X)(p22.1q28), with breakpoints in the cytobands where the MECP2 and CDKL5 genes are located. FISH analysis revealed that the MECP2 gene is not dislocated by the inversion. However, and in spite of a balanced pattern of X inactivation, this patient displayed hypomethylation and an overexpression of the MECP2 gene at the mRNA level in the lymphocytes (mean fold change: 2.55±0.38) in comparison to a group of control individuals; the expression of the CDKL5 gene was similar to that of controls (mean fold change: 0.98±0.10). No gains or losses were detected in the breakpoint regions encompassing known or suspected transcription regulatory elements. We propose that the de-regulation of MECP2 expression in this patient may be due to alterations in long-range genomic interactions caused by the inversion and hypothesize that this type of epigenetic de-regulation of the MECP2 may be present in other RTT-like patients.

Malformations of Cortical Development in Patients With Mid line Facial Defects and Ocular Hypertelorism

Objectives: We studied the neuroimaging and neurophysiological aspects of 17 patients with midline facial defects with ocular hypertelorism (MFDH). Methods: The investigation protocol included a previous semistructured questionnaire about family history; gestational, neonatal, and postnatal development; and dysmorphologic and neurologic evaluation. Recognized monogenic disorders and individuals with other well-known conditions were excluded. All patients had high resolution magnetic resonance imaging (MRI) with multiplanar reconstruction (MPR) and routine electroencephalograms (EEGs). Results: We detected abnormalities in five patients whose MRIs had been previously reported as normal. MRI showed central nervous system (CNS) structural abnormalities in all patients, which included commissural alterations in 16/17 (94%), malformations of cortical development in 10/17 (58%), disturbances of neural tube closure in 7/17(42%), and posterior fossa anomalies in 6/17 (35%). Some patients had more than one type of malformation occurring at different stages of the embryonary process. EEGs showed epileptiform activity in 4/17 (24%) and background abnormalities in 5/17 (29%) of patients. Conclusion: This study clearly demonstrated the presence of structural and functional neurologic alterations related to MFDH. Therefore, the CNS anomalies cannot be considered incidental findings but an intrinsic part of this condition, which could be related to environmental effects and/or genetic mutations. These findings would provide a basis for future investigations on MFDH and should also be considered when planning rehabilitation.

Novel H3K4me3 marks are enriched at human- and chimpanzee-specific cytogenetic structures.

Human and chimpanzee genomes are 98.8% identical within comparable sequences. However, they differ structurally in nine pericentric inversions, one fusion that originated human chromosome 2, and content and localization of heterochromatin and lineage-specific segmental duplications. The possible functional consequences of these cytogenetic and structural differences are not fully understood and their possible involvement in speciation remains unclear. We show that subtelomeric regions-regions that have a species-specific organization, are more divergent in sequence, and are enriched in genes and recombination hotspots-are significantly enriched for species-specific histone modifications that decorate transcription start sites in different tissues in both human and chimpanzee. The human lineage-specific chromosome 2 fusion point and ancestral centromere locus as well as chromosome 1 and 18 pericentric inversion breakpoints showed enrichment of human-specific H3K4me3 peaks in the prefrontal cortex. Our results reveal an association between plastic regions and potential novel regulatory elements.

Simple method for detection of MYH11 DNA rearrangements in patients with inv(16)(p13q22) and acute myeloid leukemia.

The pericentric inversion on chromosome 16 [inv(16)(p13q22)] and related t(16;16)(p13;q22) are recurrent aberrations associated with acute myeloid leukemia (AML) M4 Eo. Both abberations result in a fusion of the core binding factor beta (CBFB) and smooth muscle myosin heavy chain gene (MYH11). A selected genomic 6.9-kb BamHl probe detects MYH11 DNA rearrangements in 18 of 19 inv(16)/t(16;16) patients tested using HindIII digested DNA. The rearranged fragments were not detectable after remission in two cases tested, while they were present after relapse in one of these two cases tested.

NORs inheritance analysis in crossings including individuals from two stocks of rainbow trout (Oncorhynchus mykiss)

Silver nitrate staining of rainbow trouts (Oncorhynchus mykiss) chromosomes, for the identification of the nucleolar organizing regions (NORs), revealed that in individuals from Nucleo Experimental de Salmonicultura de Campos do Jordao (Brazil) NORs were located in the long arms of a submetacentric pair while in specimens from Mount Shasta (USA) NORs were located in the short arms of a submetacentric pair. Cytogenetic analysis of the offspring, obtained through artificial crosses including individuals from both stocks, allowed the identification of NORs in two submetacentric chromosomes, one in the short arms and the other in the long arms, confirming the effectiveness of the hybridization process. Complementary results obtained using the FISH technique with 18S and 5S rDNA probes showed that NOR-bearing chromosomes exhibited a cluster of 5S genes located in tandem with the 18S gene cluster in both stocks. The results allow us to suggest that the difference in NOR-bearing chromosomes found between the two stocks is likely to be due to a pericentric inversion involving the chromosome segment where 18S and 5S rDNA genes are located. The presence of ribosomal genes in the long arms of a submetacentric chromosome is apparently a particular characteristic of the rainbow trout stock of Campos do Jordao and might be used as a chromosome marker in studies of controlled crosses in this species.

KARYOTYPE and NUCLEOLUS ORGANIZER REGIONS of Ophisternon aenigmaticum (Teleostei: Synbranchiformes: Synbranchidae) FROM VENEZUELA

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mechanisms of karyotype differentiation in Cassidinae sensu lato (Coleoptera, Polyphaga, Chrysomelidae) based on seven species of the Brazilian fauna and an overview of the cytogenetic data

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Chromosome evolution in three Brazilian Leptodactylus species (Anura, Leptodactylidae), with phylogenetic considerations

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Autosomal rearrangement in Gryllus assimilis Fabricius, 1775 (Orthoptera, Gryllidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

POPULATIONAL STUDY OF CHROMOSOME POLYMORPHISM IN ZONOTRICHIA-CAPENSIS (EMBERIZIDAE, PASSERIFORMES, AVES)

The karyotypes of 328 specimens of Zonotrichia capensis from four regions, three in the state of Parana and one in the state of São Paulo (Brazil), were studied. This species presents a chromosomal polymorphism due to pericentric inversion, involving the 3rd and 5th chromosome pairs. Chromosome 3 exists in the 3st and 3sm conditions and chromosome 5, in 5st and 5m conditions. Mitotic metaphase spreads were obtained from bone-marrow material of colchicine-treated animals.At all collection sites the frequency of chromosome 5m was considerably lower than that of the inverted chromosome 3sm.The present work tried to establish a correlation between the frequencies of the 3rd and 5th chromosome pairs with the climatical and geographical parameters at the collecting regions.Data show that the frequency of chromosome 5m is higher in regions with low temperature and high altitude.

Chromosomal studies on five species of the genus Leptodactylus Fitzinger, 1826 (Amphibia, Anura) using differential staining

Cytogenetic studies were carried out on five species of Leptodactylus, namely L. fuscus, L, notoaktites, L. labyrinthicus, L. ocellatus, and L. podicipinus, after standard staining, Ag-NOR and C-banding as well as BrdU incorporation for three of them. The species had 2n = 22 chromosomes and two basic karyotype patterns. Chromosome 8 was a marker bearing a secondary constriction. In all species, this secondary constriction corresponded to the Ag-NOR site. The species had centromeric C-bands in all chromosomes of the complement, but some interstitial or telomeric bands seemed to differentiate some karyotypes, either at the species or the population level. In L. ocellatus, the C-banding pattern confirmed the occurrence of a heteromorphic pericentric inversion in chromosome 8 in specimens from one of the populations. The BrdU incorporation technique showed no detectable difference in the replication patterns of the major bands in the chromosomes of L. noroaktites, L. labyrinthicus, and L. ocellatus.

Chromosome localization of the ribosomal genes 18S and 5S in four stocks of rainbow trout (Oncorhynchus mykiss) from cultivated and naturalized stocks in Brazil

In the present study, fluorescence in situ hybridization (FISH) was employed to determine the chromosomal location of genes 18S rDNA and 5S rDNA in four rainbow trout stocks. In specimens from the stocks of Núcleo Experimental de Salmonicultura de Campos do Jordão and Gavião river, 18S genes were located at a subterminal position in the long arms of two submetacentric chromosomes, whereas in specimens from stocks of Mount Shasta and Teresópolis they were found in the short arms. In all analyzed stocks, 5S genes were located in two chromosome pairs. In a subtelocentric pair, 5S genes were present in the short arms and, in the other submetacentric pair, 5S genes were at an interstitial position. In the latter, 18S and 5S genes were contiguous. Taking into account that both 18S and 5S rDNA genes have been localized in the short arm of a submetacentric chromosome in almost all rainbow trout samples so far studied, the presence of such genes in the long arm, as seen in the samples from Núcleo Experimental de Salmonicultura de Campos do Jordão and Gavião river, supports the hypothesis of a pericentric inversion involving this chromosome segment in the ancestor line of these stocks. The observed polymorphism allowed the identification of a very useful genomic marker, and may therefore constitute an important tool in the genetic management of rainbow trout stocks.