Differential crosstalk between estrogen receptor (ER) alpha and ER beta and the thyroid hormone receptor isoforms results in flexible regulation of the consensus ERE

Crosstalk between nuclear receptors is important for conversion of external and internal stimuli to a physiologically meaningful response by cells. Previous studies from this laboratory have demonstrated crosstalk between the estrogen (ER) and thyroid hormone receptors (TR) on two estrogen responsive physiological promoters, the preproenkephalin and oxytocin receptor gene promoter. Since ERa and ERb are isoforms possessing overlapping and distinct transactivation properties, we hypothesized that the interaction of ERa and b with the various TR isoforms would not be equivalent. To explore this hypothesis, the consensus estrogen response element (ERE)derived from the Xenopus vitellogenin gene is used to investigate the differences in interaction between ERa and b isoforms and the different TR isoforms in fibroblast cells. Both the ER isoforms transactivate from the consensus ERE, though ERa transactivates to a greater extent than ERb. Although neither of the TRb isoforms have an effect on ERa transactivation from the consensus ERE, the liganded TRa1 inhibits the ERa transactivation from the consensus ERE. In contrast, the liganded TRa1 facilitates ERb-mediated transactivation. The crosstalk between the TRb isoforms with the ERa isoform, on the consensus ERE, is different from that with the ERb isoform. The use of a TRa1 mutant, which is unable to bind DNA, abolishes the ability of the TRa1 isoform to interact with either of the ER isoforms. These differences in nuclear receptor crosstalk reveal an important functional difference between isoforms, which provides a novel mechanism for neuroendocrine integration.

Functional analysis of a novel missense NBC1 mutation and of other mutations causing proximal renal tubular acidosis

Mutations in the Na+-HCO3- cotransporter NBC1 cause severe proximal tubular acidosis (pRTA) associated with ocular abnormalities. Recent studies have suggested that at least some NBC1 mutants show abnormal trafficking in the polarized cells. This study identified a new homozygous NBC1 mutation (G486R) in a patient with severe pRTA. Functional analysis in Xenopus oocytes failed to detect the G486R activity due to poor surface expression. In ECV304 cells, however, G486R showed the efficient membrane expression, and its transport activity corresponded to approximately 50% of wild-type (WT) activity. In Madin-Darby canine kidney (MDCK) cells, G486R was predominantly expressed in the basolateral membrane domain as observed for WT. Among the previously identified NBC1 mutants that showed poor surface expression in oocytes, T485S showed the predominant basolateral expression in MDCK cells. On the other hand, L522P was exclusively retained in the cytoplasm in ECV304 and MDCK cells, and functional analysis in ECV304 cells failed to detect its transport activity. These results indicate that G486R, like T485S, is a partial loss of function mutation without major trafficking abnormalities, while L522P causes the clinical phenotypes mainly through its inability to reach the plasma membranes. Multiple experimental approaches would be required to elucidate potential disease mechanism by NBC1 mutations.

WNT5A Mutations in Patients With Autosomal Dominant Robinow Syndrome

Robinow syndrome is a skeletal dysplasia with both autosomal dominant and autosomal recessive inheritance patterns. It is characterized by short stature, limb shortening, genital hypoplasia, and craniofacial abnormalities. The etiology of dominant Robinow syndrome is unknown; however, the phenotypically more severe autosomal recessive form of Robinow syndrome has been associated with mutations in the orphan tyrosine kinase receptor, ROR2, which has recently been identified as a putative WNT5A receptor. Here, we show that two different missense mutations in WNT5A, which result in amino acid substitutions of highly conserved cysteines, are associated with autosomal dominant Robinow syndrome. One mutation has been found in all living affected members of the original family described by Meinhard Robinow and another in a second unrelated patient. These missense mutations result in decreased WNT5A activity in functional assays of zebrafish and Xenopus development. This work suggests that a WNT5A/ROR2 signal transduction pathway is important in human craniofacial and skeletal development and that proper formation and growth of these structures is sensitive to variations in WNT5A function. Developmental Dynamics 239:327-337, 2010. (C) 2009 Wiley-Liss, Inc.

Cloning and characterization of a zebrafish homologue of human AQP1: a bifunctional water and gas channel

Chen LM, Zhao J, Musa-Aziz R, Pelletier MF, Drummond IA, Boron WF. Cloning and characterization of a zebrafish homologue of human AQP1: a bifunctional water and gas channel. Am J Physiol Regul Integr Comp Physiol 299: R1163-R1174, 2010. First published August 25, 2010; doi:10.1152/ajpregu.00319.2010.-The mammalian aquaporins AQP1, AQP4, and AQP5 have been shown to function not only as water channels but also as gas channels. Zebrafish have two genes encoding an AQP1 homologue, aqp1a and aqp1b. In the present study, we cloned the cDNA that encodes the zebrafish protein Aqp1a from the 72-h postfertilization (hpf) embryo of Danio rerio, as well as from the swim bladder of the adult. The deduced amino-acid sequence of aqp1a consists of 260 amino acids and is 59% identical to human AQP1. By analyzing the genomic DNA sequence, we identified four exons in the aqp1a gene. By in situ hybridization, aqp1a is expressed transiently in the developing vasculature and in erythrocytes from 16 to 48 h of development. Later, at 72 hpf, aqp1a is expressed in dermal ionocytes and in the swim bladder. Western blot analysis of adult tissues reveals that Aqp1a is most highly expressed in the eye and swim bladder. Xenopus oocytes expressing aqp1a have a channel-dependent (*) osmotic water permeability (P(f)*) that is indistinguishable from that of human AQP1. On the basis of the magnitude of the transient change in surface pH (Delta pHS) that were recorded as the oocytes were exposed to either CO(2) or NH(3), we conclude that zebrafish Aqp1a is permeable to both CO(2) and NH(3). The ratio (Delta pHS*)CO2/P(f)* is about half that of human AQP1, and the ratio (Delta pHS*)NH3/P(f)* is about one-quarter that of human AQP1. Thus, compared with human AQP1, zebrafish Aqp1a has about twice the selectivity for CO(2) over NH(3).

A metodologia de análise de dados espaciais aplicada a plantas forrageiras

Dois experimentos e um levantamento por amostragem foram analisados no contexto de dados espaciais. Os experimentos foram delineados em blocos completos casualizados sendo que no experimento um (EXP 1) foram avaliados oito cultivares de trevo branco, sendo estudadas as variáveis Matéria Seca Total (MST) e Matéria Seca de Gramíneas (MSGRAM) e no experimento dois (EXP 2) 20 cultivares de espécies forrageiras, onde foi estudada a variável Percentagem de Implantação (%IMPL). As variáveis foram analisadas no contexto de modelos mistos, sendo modelada a variabilidade espacial através de semivariogramas exponencias, esféricos e gaussianos. Verificou-se uma diminuição em média de 19% e 14% do Coeficiente de Variação (CV) das medias dos cultivares, e uma diminuição em média de 24,6% e 33,3% nos erros padrões dos contrastes ortogonais propostos em MST e MSGRAM. No levantamento por amostragem, estudou-se a associação espacial em Aristida laevis (Nees) Kunth , Paspalum notatum Fl e Demodium incanum DC, amostrados em uma transecção fixa de quadros contiguos, a quatro tamanhos de unidades amostrais (0,1x0,1m; 0,1x0,3m; 0,1x0,5m; e 0,1x1,0m). Nas espécies Aristida laevis (Nees) Kunth e Paspalum notatum Fl, existiu um bom ajuste dos semivariogramas a tamanhos menores das unidades amostrais, diminuíndo quando a unidade amostral foi maior. Desmodium incanum DC apresentou comportamento contrario, ajustando melhor os semivariogramas a tamanhos maiores das unidades amostrais.

Combinação dos efeitos da carga animal com métodos físicos e químicos para o controle de plantas indesejáveis em pastagem nativa da Serra do Sudeste do RS

O experimento foi conduzido no município de Cachoeira do Sul de out/2001 a out/2002, objetivando avaliar a dinâmica vegetacional e o desempenho animal (31/05/02 a 16/09/02), em uma pastagem nativa submetida a diferentes métodos de controle de plantas indesejáveis. O local corresponde à região fisiográfica denominada Serra do Sudeste, com predominância das espécies Paspalum notatum (grama forquilha), Andropogon lateralis (capim caninha), além de Arachis burkartii e Desmodium incanum. Como plantas classificadas como indesejáveis encontram-se Baccharis trimera (carqueja), Vernonia nudiflora (alecrim) e Psidium luridum (araçá do campo). O delineamento experimental utilizado foi um fatorial em blocos (4 x 2), com duas repetições de campo. Os tratamentos foram os seguintes: testemunha (T); roçada de primavera (P); roçada de primavera + outono (O) e roçada de primavera + controle químico (Q), todos em duas ofertas de forragem, média (8%) e alta (14%). Os animais testes foram três novilhas por repetição, mantidas em pastejo contínuo com lotação variável. A massa de forragem foi obtida através de avaliações visuais usando padrões de referência, o acúmulo de forragem através do uso de gaiolas de exclusão e a composição botânica pelo método do ponto. Avaliaram-se os parâmetros de desempenho individual, carga animal, ganho de peso vivo por área e a dinâmica vegetacional. Os dados foram submetidos a análise de variância através do pacote estatístico SAS. O nível alto de oferta permitiu uma menor perda de peso vivo por área. Os tratamentos P e Q proporcionaram um melhor desempenho individual e menor perda de peso vivo por área, quando comparados a T. Os tratamentos P, O e Q foram eficientes no controle de Baccharis trimera. A. burkartii e Eriyngium horridum foram incrementados pela média intensidade de pastejo. Aristida laevis teve sua participação aumentada pela interação tratamento Q e baixa intensidade de pastejo.

Processos de degradação e regeneração da vegetação campestre do entorno de areais do sudoeste do Rio Grande do Sul

No Sudoeste do Rio Grande do Sul ocorrem manchas de substrato arenoso sem cobertura vegetal, conhecidos regionalmente como areais. A tese reexamina o problema e suas causas, enfatizando o papel da vegetação natural e propondo estratégias para a revegetação e prevenção à arenização em sistemas pastoris. Evidências foram obtidas em levantamentos e experimento avaliando processos de degradação e regeneração da vegetação campestre do entorno de areais. Levantamento da vegetação, realizado em 41 parcelas de 4,5 x 9,0 m na borda de 11 areais indicou a existência de dois tipos de comunidades. Areais de Manoel Viana e aqueles usados pelo gado em São Francisco de Assis, com alto percentual de substrato exposto, são caracterizados principalmente por Elyonurus sp., Axonopus pressus e Butia paraguayensis. Areais de Alegrete e aqueles excluídos de pastejo em São Francisco de Assis, com menor percentual de substrato exposto, são caracterizados principalmente por Andropogon lateralis e Aristida laevis. Foi também avaliada a dinâmica da vegetação em gradientes de arenização, usando quadros (0,25 m2) contíguos em 16 transecções de 10 m localizadas em cinco areais. A dinâmica da vegetação foi associada ao uso das áreas pelo gado, pois houve aumento, após 14 meses, de substrato exposto em comunidades de areais sob pastoreio, enquanto que aquelas sem gado apresentaram uma dinâmica espacial-temporal de maior estabilidade da cobertura vegetal. Um experimento foi realizado para avaliar, durante 8 meses, o efeito de níveis controlados de soterramento por areia (0, 5, 10 e 20 cm) em comunidades do entorno de dois areais sob pastoreio. Comunidades caracterizadas por Elyonurus sp. e Axonopus pressus foram mais tolerantes ao soterramento. As evidências indicam que a exclusão do gado de areais pode ser uma alternativa eficaz para a revegetação de areais por espécies das comunidades naturais do entorno. Ademais, a arenização pode ser prevenida pelo uso adequado dos campos que mantenha a cobertura vegetal natural protegendo o solo dos processos erosivos hídrico e eólico.

Caracterização e composição florística de uma comunidade savânica no Rio Grande do Norte, Brasil : subsídios para a conservação

Studies on Brazilian biodiversity are still very few and can observe the difference of knowledge between the different regions of the country. This affirmation can be verified in the investigation to identify the rare species in Brazil and the key biodiversity areas (ACBs). In that study were identified for Brasil 2.256 rare species and 752 ACBs. The Rio Grande do Norte (RN) was the only Brazilian state that has not been identified any rare species and no ACBs, possibly due to the lack of floristic studies in this state. A particular area was selected for this study: an area of ecological tension with savanna physiognomy in Rio do Fogo, RN. This savanna community is represented in RN in a fragment and immersed in restinga and caatinga and was identified and described only through of radar imagens there are no studies to date in loco . We have prepared the following questions about this community savanna: 1) The region delimited and described by through of radar images by the RADAMBRASIL, 1976, can be associated of the Cerrado, in terms of floristic ?; 2) What is the floristic composition of this area? This area includes rare species, endemic or endangered? 3) What is the geographical and phytogeographical distribution of plant species registered in this area? 4) Those plant species registered are endemic or have affinity with other areas phytoecological Brazilian? To answer these questions we performed a floristic inventory of the August 2007 to September/2009. The results are presented in two chapters (manuscripts). The first chapter, titled "The Savannah Rio Grande do Norte: floristic links with other plant formations in the Northeast and Center-West Brazil" was submitted to the Revista Brasileira de Botância. Chapter 1 discusses the phytogeographical distribution of the species, by comparing floristic studies conducted in the Cerrado, Caatinga and Restinga in the Northeast and Cerrado of the Central Brazil. The analysis of data of this study and compilation with other studies indicated that: i) the record of 94 plant species; ii) of total species, about 64% are associated with the Cerrado, the second specialized bibliography, and about 78% as the List of Species of Flora of Brazil. However, about 73% of total species (94) are also distributed in the Caatinga, the Atlantic forest 64%, the Amazon forest 64%, the Pantanal 15% and the Pampa 12%. Floristically the data show that the community studied is influenced by other floras, has a structure where grasses dominate and also because of his appearance the same savanna then be classified as a Savana gramíneo-lenhosa do tabuleiro . Chapter 2, titled "Considerations on the flora of a savanna community in Rio Grande do Norte, northeastern Brazil: Subsidy key area for conservation" was submitted to the Revista Natureza e Conservação. This has the objective of improve the knowledge of flora of Rio Grande do Norte and to identify possible rare species and consequently increase the key areas of biodiversity in Brazil. The data indicated that: i) of 94 species registered in the study area, 40 were new records for the Rio Grande do Norte state; ii) These citations to unpublished state, Stylosanthes montevidensis Vogel (Fabaceae) and Aristida laevis (Nees) Kunth (Poaceae) are indicated for the first time to the Northeast of Brazil; iii) are registered in the area 24 species endemic to Brazil and 63 non-endemic; iv) Aspilia procumbens Baker (Asteraceae) registered in the area is considered a restricted species and micro endemic Rio Grande do Norte, ie rare species; v) Aspilia procumbens is also cited in the category of critically endangered species and Stilpnopappus cearensis Hubber (Asteraceae) a species vulnerable to extinction. This study shows a new area phytoecological in Rio Grande do Norte and indicates the area's potential to contribute with the sites of global significance for biodiversity conservation, either locally, regionally and nationally. This will certainly contribute to respond some targets set by the Global Strategy for Plant Conservation and the Convention on Biological Diversity such as the inventory of vegetal diversity in a region with little collection, which will provide data that contributes to questions and themes related to biodiversity.

Contrasting responses to induction cues by ants inhabiting Maieta guianensis (Melastomataceae)

As duas formigas que mais comumente se associam com a mirmecofita de sub-bosque Maieta guianensis (Melastomataceac) respondem de maneiras diferentes a simulac (o) over tilde es experimentais de herbivoria. Enquanto Pheidole minutula & induzida tanto por danos fisicos e extratos de tecido foliar, Crematogaster laevis 6 induzida apenas pelos danos fisicos. Sugerimos que tal variac (a) over tildeo interespecifica em respostas induzidas e comum e pode influenciar a qualidade das defesas contra herbivoros que as formigas oferecem as plantas que elas ocupam.

Interspecific variation in the defensive responses of obligate plant-ants: experimental tests and consequences for herbivory

The aggressive behavior of ants that protect plants from herbivores in exchange for rewards such as shelter or food is thought to be an important form of biotic defense against herbivory, particularly in tropical systems. To date, however, no one has compared the defensive responses of different ant taxa associated with the same plant species, and attempted to relate these differences to longer-term efficacy of ant defense. We used experimental cues associated with herbivory-physical damage and extracts of chemical volatiles from leaf tissue-to compare the aggressive responses of two ant species obligately associated with the Amazonian myrmecophyte Tococa bullifera (Melastomataceae). We also conducted a colony removal experiment to quantify the level of resistance from herbivores provided to plants by each ant species. Our experiments demonstrate that some cues eliciting a strong response from one ant species elicited no response by the other. For cues that do elicit responses, the magnitude of these responses can vary interspecifically. These patterns were consistent with the level of resistance provided from herbivores to plants. The colony removal experiment showed that both ant species defend plants from herbivores: however, herbivory was higher on plants colonized by the less aggressive ant species. Our results add to the growing body of literature indicating defensive ant responses are stimulated by cues associated with herbivory. However, they also suggest the local and regional variation in the composition of potential partner taxa could influence the ecology and evolution of defensive mutualisms in ways that have previously remained unexplored.

Pigmentação testicular em Physalaemus nattereri (Steindachner) (Amphibia, Anura) com observações anatômicas sobre o sistema pigmentar extracutâneo

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

The neuranatomical basis of central control of cardiorespiratory interactions in vertebrates

It seems that a dual location for vagal preganglionic neurones (VPNs) has important functional correlates in all vertebrates. This may be particularly the case with the central control exerted over the heart by cardiac VPNs (CVPNs). About 30 % of VPNs but up to 70 % of CVPNs are in the nucleus ambiguus (NA) of mammals. There is a similar proportional representation of VPNs between the major vagal nuclei in amphibians and turtles but in fish and crocodilians; the proportion of VPNs in the NA is closer to 10% and in some lizards and birds it is about 5%. However, the CVPNs are distributed unequally between these nuclei so that 45 % of the CVPNs are located in the NA of the dogfish, and about 30% in the NA of Xenopus and the duck. This topographical separation of CVPNs seems to be of importance in the central control of the heart. Cells in one location may show respiration-related activity (e.g those in the dorsal vagal nucleus (DVN) of dogfish and in the NA of mammals) while cells in the other locations do not. Their different activities and separate functions will be determined by their different afferent inputs from the periphery or from elsewhere in the CNS, which in turn will relate to their central topography. Thus, CVPNs in the NA of mammals receive inhibitory inputs from neighbouring inspiratory neurones, causing respiratory sinus arrythmia (RSA), and the CVPNs in the DVN of the dogfish may generate cardiorespiratory synchrony (CRS).

Ant-tended hemiptera in Amazonian myrmecophytes: Patterns of abundance and implications for mutualism function (Hymenoptera: Formicidae)

We assessed how the abundance of ant-tended Hemiptera associated with two Amazonian myrmecophytes, Tococa bullifera and Maieta guianensis, varied as a function of resident ant species. We collected five species or morpho-species of adult hemiptera in the domatia of M. guianensis, with four of these species also found in Tococa bullifera. Maieta guianensis plants inhabited by Crematogaster laevis had over four-fold more hemiptera in them than plants inhabited by Pheidole minutula. In contrast, the density of hemiptera in Tococa bullifera domatia was independent of the species of ant resident. For each of the two ant species inhabiting Maieta guianensis, there was a positive and significant relationship between the abundance of hemiptera and workers inhabiting a plant. This relationship was also significant and positive for the Tococa bullifera plants inhabited by C. laevis. However, there was no relationship between Azteca worker and hemipteran density, although there was a trend towards a positive relationship. Our results indicate that hemipteran abundance can vary significantly between different myrmecophyte species, but that the nature of this relationship is mediated by the identity of the ant associate. Because hemipterans are herbivores, the costs and benefits of different ant partners to the host plant may vary in ways that are often overlooked.

Ontogenia do trato digestório e desenvolvimento gonadal de Betta splendens: aspectos morfológicos

Pós-graduação em Aquicultura - FCAV

Big, bad, and beautiful: phylogenetic relationships of the horned frogs (Anura: Ceratophryidae)

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