Differential transcription of ribosomal cistrons denoting nucleolar dominance in hybrids of Drosophila mulleri and Drosophila navojoa (mulleri complex, Repleta group)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Characterization of Mongolian Gerbil Chromatoid Bodies and Their Correlation with Nucleolar Cycle During Spermatogenesis

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

Relationship between the nucleolar cycle and chromatoid body formation in the spermatogenesis of Phrynops geoffroanus (Reptilia Testudines)

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

Nucleolar behaviour during meiosis in four species of phyllostomid bats (Chiroptera, Mammalia)

We analyzed the behavior of the nucleolus, nucleolar structures and nucleolus organizer regions (NORs) during meiotic division in four species of phyllostomid bats that have different numbers and locations of NORs. Nucleoli began disassembly at leptotene, and the subcomponents released from the nucleolus were dispersed in the nucleoplasm, associated with perichromosomal regions, or they remained associated with NORs throughout division. In Phyllostomus discolor, a delay in nucleolus disassembly was observed; it disassembled by the end of pachytene. The RNA complexes identiied by acridine orange staining were observed dispersed in the nucleoplasm and associated with perichromosomal regions. FISH with rDNA probe revealed the number of NORs of the species: one NOR in Carollia per spicillata, one pair in Platyrrhinus lineatus and P. discolor, and three pairs in Artibeus lituratus. During pachytene, there was a temporary dissociation of the homologous NORs, which returned to pairing at diplotene. The variation in the number (from one to three pairs) and location of NORs (in sex or autosomal chromosomes, at terminal or interstitial regions) did not seem to interfere with the nucleolar behavior of the different species because no variation in nucleolar behavior that could be correlated with the variation in the number and chromosomal location of NORs was detected.

NUCLEOLAR CHROMOSOME VARIANTS IN STERNOPYGUS-MACRURUS (PISCES, STERNOPYGIDAE) FROM 3 BRAZILIAN RIVER BASINS

The karyotypes, location of nucleolus organizer regions (NOR) and constitutive heterochromatin pattern of Sternopygus macrurus (Pisces, Gymnotoidei) of natural populations from the Amazon River, Sao Francisco River and Tiete River (the last belonging to the Upper Parana River system) are reported. All specimens had 2n = 46 chromosomes and presented small differences in karyotypic formulae, but populations of each river basin had a different fixed NOR phenotype. The loss of the satellite and a gradual deletion of the heterochromatin block adjacent to the NOR may be the origin of the variants. The possible mechanism of fixation of the NOR phenotypes, and the implications of the occurrence of intraspecific differences in fixed NOR phenotype in this species are discussed.

Patterns of nucleolar activity during spermatogenesis of two triatomines, Panstrongylus megistus and P. herreri

The present work analyzed spermatogenesis in two species of triatomines (genus Panstrongylus) using silver-ion impregnation. The sex chromosomes of P. megistus and P. herreri had nucleolar organizing activity and became strongly impregnated during the phases of meiotic prophase I. Fragmentation of the nucleolus occurred in both species during the meiotic cycle. The nucleolar region could be observed up to diakinesis in meiotic prophase after which only nucleolar bodies and fragments were seen. Postmeiotic reactivation of rRNA synthesis occurred in these two species and was probably related to cell differentiation.

Espermatogênese e comportamento nucleolar em machos de Heteoptera aquáticos

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Análise das possíveis regiões organizadoras nucleolares e da atividade nucleolar em Triatoma melanocephalae T. lenti, importantes vetores da doença de Chagas

Triatoma melanocephalae T. lentisão espécies crípticas de triatomíneos pertencentes ao subcomplexo Brasiliensis. Esses organismos foram agrupados no subcomplexo apenas por caracteres morfológicos e pela disposição geográfica. Sendo assim, estudos citogenéticos são considerados como importantes ferramentas na classificação dos triatomíneos e, com isso, podem auxiliar na criação de um plano de profilaxia da doença. Por meio da técnica citogenética de impregnação por íons prata, foi possível visualizar a atividade nucleolar e as Regiões Organizadoras Nucleolares (RONs) desses insetos. T. melanocephalaapresentou três RONs ativas nos autossomos durante a prófase I. T. lenti apresentou duas RONs ativas nos autossomos durante a prófase I e a metáfase I. Ambas as espécies apresentaram o fenômeno de persistência do material nucleolar encontrado em triatomíneos. Sendo assim, por meio da análise das RONs, foi possível observar que T. lenti, quando comparado com os outros organismos do subcomplexo, apresentou marcações semelhantes à T. tibiamaculata e que T. melanocephalanão apresenta nenhuma relação direta com o subcomplexo. Palavras-chave: Citogenética. Taxonomia. Triatominae. Subcomplexo Brasiliensis. ABSTRACT Analysis of nucleolus organizer regions and nucleolar activity in important vectors of Chagas disease (Triatoma melanocephala and T. lenti) Triatoma melanocephalaand T. lentiare important vectors of Chagas disease. These cryptic species of triatomines are grouped in the subcomplexbrasiliensisdue only to morphological characters and geographical distribution. Cytogenetic studies are important to the classification of insects and can assist in creating a disease prevention plan. The aim of the present study was to determine nucleolar activity and nucleolus organizer regions (NORs) in these insects using the cytogenetic method of silver ion impregnation. T. melanocephalaexhibited three active NORs in autosomes during prophase I. T. lentiexhibited two active NORs in autosomes during prophase I and metaphase I. Both species exhibit the persistent nucleolar material found in triatomines. The analysis of NORs in the present study revealed that T. lenti exhibited labeling similar to that found in T. tibiamaculata, which belongs to the subcomplex, whereas T. melanocephalashows no direct relationship with the subcomplex. Keywords: Cytogenetics. Taxonomy. Triatominae. Brasiliensissubcomplex.

Cooperation between the activin and Wnt pathways in the spatial control of organizer gene expression

The normal expression pattern of the Wnt responsive homeobox gene Siamois is restricted to the dorso-vegetal region of the Xenopus embryo. Because the Wnt signaling pathway (via β-catenin) is active on the entire dorsal side of the early embryo, we have asked why Siamois expression is not seen in the dorsal ectoderm. Only Wnt signaling, via activation of β-catenin, can induce directly Siamois, and signaling via the SMAD1 (BMP2/4) or SMAD2 (activin/Vg-1) pathways cannot. We now directly show that the SMAD2 pathway can cooperate with the Wnt pathway to induce expression of Siamois much more strongly than the Wnt pathway alone, in normal embryos. We demonstrate the significance of this cooperation in normal embryos by blocking the SMAD2 signaling pathway with a dominant negative activin receptor. The activin dominant negative receptor blocks this cooperative effect and reduces the expression of Siamois by threefold in early embryos. Furthermore, we find that this cooperative relationship between the SMAD2 and Wnt pathways is reciprocal. Thus, in normal embryos, the Wnt pathway can enhance induction, by the SMAD 2 pathway, of the organizer genes Gsc and Chd but not the pan-mesodermal marker genes Xbra and Eomes. We conclude that the Wnt and SMAD2 signaling pathways cooperate to induce the expression of Spemann-organizer specific genes and so help to localize their spatial expression.

Identification of a small, very acidic constitutive nucleolar protein (NO29) as a member of the nucleoplasmin family

We report the discovery and molecular characterization of a small and very acidic nucleolar protein of an SDS/PAGE mobility corresponding to Mr 29,000 (NO29). The cDNA-deduced sequence of the Xenopus laevis protein defines a polypeptide of a calculated molecular mass of 20,121 and a pI of 3.75, with an extended acidic region near its C terminus, and is related to the major nucleolar protein, NO38, and the histone-binding protein, nucleoplasmin. This member of the nucleoplasmin family of proteins was immunolocalized to nucleoli in Xenopus oocytes and diverse somatic cells. Protein NO29 is associated with nuclear particles from Xenopus oocytes, partly complexed with protein NO38, and occurs in preribosomes but not in mature ribosomes. The location and the enormously high content of negatively charged amino acids lead to the hypothesis that NO29 might be involved in the nuclear and nucleolar accumulation of ribosomal proteins and the coordinated assembly of pre-ribosomal particles.

Nucleolar Localization Elements of Xenopus laevis U3 Small Nucleolar RNA

The Nucleolar Localization Elements (NoLEs) of Xenopus laevis U3 small nucleolar RNA (snoRNA) have been defined. Fluorescein-labeled wild-type U3 snoRNA injected into Xenopus oocyte nuclei localized specifically to nucleoli as shown by fluorescence microscopy. Injection of mutated U3 snoRNA revealed that the 5′ region containing Boxes A and A′, known to be important for rRNA processing, is not essential for nucleolar localization. Nucleolar localization of U3 snoRNA was independent of the presence and nature of the 5′ cap and the terminal stem. In contrast, Boxes C and D, common to the Box C/D snoRNA family, are critical elements for U3 localization. Mutation of the hinge region, Box B, or Box C′ led to reduced U3 nucleolar localization. Results of competition experiments suggested that Boxes C and D act in a cooperative manner. It is proposed that Box B facilitates U3 snoRNA nucleolar localization by the primary NoLEs (Boxes C and D), with the hinge region of U3 subsequently base pairing to the external transcribed spacer of pre-rRNA, thus positioning U3 snoRNA for its roles in rRNA processing.

Box H and Box ACA Are Nucleolar Localization Elements of U17 Small Nucleolar RNA

The nucleolar localization elements (NoLEs) of U17 small nucleolar RNA (snoRNA), which is essential for rRNA processing and belongs to the box H/ACA snoRNA family, were analyzed by fluorescence microscopy. Injection of mutant U17 transcripts into Xenopus laevis oocyte nuclei revealed that deletion of stems 1, 2, and 4 of U17 snoRNA reduced but did not prevent nucleolar localization. The deletion of stem 3 had no adverse effect. Therefore, the hairpins of the hairpin–hinge–hairpin–tail structure formed by these stems are not absolutely critical for nucleolar localization of U17, nor are sequences within stems 1, 3, and 4, which may tether U17 to the rRNA precursor by base pairing. In contrast, box H and box ACA are major NoLEs; their combined substitution or deletion abolished nucleolar localization of U17 snoRNA. Mutation of just box H or just the box ACA region alone did not fully abolish the nucleolar localization of U17. This indicates that the NoLEs of the box H/ACA snoRNA family function differently from the bipartite NoLEs (conserved boxes C and D) of box C/D snoRNAs, where mutation of either box alone prevents nucleolar localization.

Weather variability, tides, and Barmah Forest Virus Disease in the Gladstone region, Australia

In this study we examined the impact of weather variability and tides on the transmission of Barmah Forest virus (BFV) disease and developed a weather-based forecasting model for BFV disease in the Gladstone region, Australia. We used seasonal autoregressive integrated moving-average (SARIMA) models to determine the contribution of weather variables to BFV transmission after the time-series data of response and explanatory variables were made stationary through seasonal differencing. We obtained data on the monthly counts of BFV cases, weather variables (e.g., mean minimum and maximum temperature, total rainfall, and mean relative humidity), high and low tides, and the population size in the Gladstone region between January 1992 and December 2001 from the Queensland Department of Health, Australian Bureau of Meteorology, Queensland Department of Transport, and Australian Bureau of Statistics, respectively. The SARIMA model shows that the 5-month moving average of minimum temperature (β = 0.15, p-value < 0.001) was statistically significantly and positively associated with BFV disease, whereas high tide in the current month (β = −1.03, p-value = 0.04) was statistically significantly and inversely associated with it. However, no significant association was found for other variables. These results may be applied to forecast the occurrence of BFV disease and to use public health resources in BFV control and prevention.