4 resultados para Cre Recombinase

em Biblioteca Digital da Produção Intelectual da Universidade de São Paulo


Relevância:

60.00% 60.00%

Publicador:

Resumo:

The adipocyte-derived hormone leptin is required for normal pubertal maturation in mice and humans and, therefore, leptin has been recognized as a crucial metabolic cue linking energy stores and the onset of puberty. Several lines of evidence have suggested that leptin acts via kisspeptin expressing neurons of the arcuate nucleus to exert its effects. Using conditional knockout mice, we have previously demonstrated that deletion of leptin receptors (LepR) from kisspeptin cells cause no puberty or fertility deficits. However, developmental adaptations and system redundancies may have obscured the physiologic relevance of direct leptin signaling in kisspeptin neurons. To overcome these putative effects, we re-expressed endogenous LepR selectively in kisspeptin cells of mice otherwise null for LepR, using the Cre-loxP system. Kiss1-Cre LepR null mice showed no pubertal development and no improvement of the metabolic phenotype, remaining obese, diabetic and infertile. These mice displayed decreased numbers of neurons expressing Kiss1 gene, similar to prepubertal control mice, and an unexpected lack of re-expression of functional LepR. To further assess the temporal coexpression of Kiss1 and Lepr genes, we generated mice with the human renilla green fluorescent protein (hrGFP) driven by Kiss1 regulatory elements and crossed them with mice that express Cre recombinase from the Lepr locus and the R26-tdTomato reporter gene. No coexpression of Kiss1 and LepR was observed in prepubertal mice. Our findings unequivocally demonstrate that kisspeptin neurons are not the direct target of leptin in the onset of puberty. Leptin signaling in kisspeptin neurons arises only after completion of sexual maturation.

Relevância:

10.00% 10.00%

Publicador:

Resumo:

Are wild bearded capuchin monkeys selective about where they place nuts on anvils, specifically the anvil pits, during nut cracking? In the present study, we examined (1) whether capuchins` preferences for particular pits are influenced by the effectiveness of the pit in cracking the nut and/or by the stability of the nut during striking, (2) how capuchins detect the affordances of novel pits and (3) the influence of social context on their selections. Anvil pits varied in horizontal dimension (small, medium and large) in experiment 1 and in depth (shallow, medium and deep) in experiment 2. In both experiments, three different pits were simultaneously presented, each on one anvil. We coded the capuchins` actions with the nut in each pit, and recorded the outcome of each strike. In both experiments, capuchins preferred the most effective pit, but not the most stabilizing pit, based on the number of first strikes, total strikes and nuts cracked. Their choice also reflected where the preceding individual had last struck. The capuchins explored the pits indirectly, placing nuts in them and striking nuts with a stone. The preference for pits was weaker than the preference for nuts and stones shown previously with the same monkeys. Our findings suggest that detecting affordances of pits through indirect action is less precise than through direct action, and that social context may also influence selection. We show that field experiments can demonstrate embodied cognition in species-typical activities in natural environments. (C) 2010 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved.

Relevância:

10.00% 10.00%

Publicador:

Resumo:

Modern sugarcane cultivars are complex hybrids resulting from crosses among several Saccharum species. Traditional breeding methods have been employed extensively in different countries over the past decades to develop varieties with increased sucrose yield and resistance to pests and diseases. Conventional variety improvement, however, may be limited by the narrow pool of suitable genes. Thus, molecular genetics is seen as a promising tool to assist in the process of developing improved varieties. The SUCEST-FUN Project (http://sucest-fun.org) aims to associate function with sugarcane genes using a variety of tools, in particular those that enable the study of the sugarcane transcriptome. An extensive analysis has been conducted to characterise, phenotypically, sugarcane genotypes with regard to their sucrose content, biomass and drought responses. Through the analysis of different cultivars, genes associated with sucrose content, yield, lignin and drought have been identified. Currently, tools are being developed to determine signalling and regulatory networks in grasses, and to sequence the sugarcane genome, as well as to identify sugarcane promoters. This is being implemented through the SUCEST-FUN (http://sucest-fun.org) and GRASSIUS databases (http://grassius.org), the cloning of sugarcane promoters, the identification of cis-regulatory elements (CRE) using Chromatin Immunoprecipitation-sequencing (ChIP-Seq) and the generation of a comprehensive Signal Transduction and Transcription gene catalogue (SUCAST Catalogue).

Relevância:

10.00% 10.00%

Publicador:

Resumo:

Bacteria activate a regulatory network in response to the challenges imposed by DNA damage to genetic material, known as the SOS response. This system is regulated by the RecA recombinase and by the transcriptional repressor lexA. Leptospira interrogans is a pathogen capable of surviving in the environment for weeks, being exposed to a great variety of stress agents and yet retaining its ability to infect the host. This study aims to investigate the behavior of L. interrogans serovar Copenhageni after the stress induced by DNA damage. We show that L. interrogans serovar Copenhageni genome contains two genes encoding putative LexA proteins (lexA1 and lexA2) one of them being potentially acquired by lateral gene transfer. Both genes are induced after DNA damage, but the steady state levels of both LexA proteins drop, probably due to auto-proteolytic activity triggered in this condition. In addition, seven other genes were up-regulated following UV-C irradiation, recA, recN, dinP, and four genes encoding hypothetical proteins. This set of genes is potentially regulated by LexA1, as it showed binding to their promoter regions. All these regions contain degenerated sequences in relation to the previously described SOS box, TTTGN 5CAAA. On the other hand, LexA2 was able to bind to the palindrome TTGTAN 10TACAA, found in its own promoter region, but not in the others. Therefore, the L. interrogans serovar Copenhageni SOS regulon may be even more complex, as a result of LexA1 and LexA2 binding to divergent motifs. New possibilities for DNA damage response in Leptospira are expected, with potential influence in other biological responses such as virulence