Expressão das proteínas da matriz na discondroplasia da tíbia

A discondroplasia da tíbia (TD) em aves consiste numa anomalia do esqueleto onde existe uma falha nos processos normais da ossificação endocondral. Esta patologia é caracterizada pela formação de uma cartilagem não vascularizada e não mineralizada que se estende até à metáfise. Uma vez que existem várias anomalias do esqueleto em mamíferos com lesões semelhantes às apresentadas pela TD, este trabalho teve como objectivo a caracterização desta patologia em termos das moléculas que podem estar envolvidas no seu desenvolvimento. Assim, foi estudada a expressão das macromoléculas da matriz extracelular, das enzimas degradadoras da matriz (metaloproteinases da matriz: MMPs), bem como das moléculas envolvidas na proliferação e diferenciação celular, na angiogénese e apoptose. A expressão génica foi realizada, por PCR quantitativo em tempo real, em placas de crescimento normais e discondroplásicas obtidas a partir de frangos de carne (broilers) da estirpe Cobb. Os níveis proteicos de algumas MMPs foram analisados por immunoblotting e zimografia de gelatina. No presente estudo não se verificou alteração na expressão dos genes dos colagénios do tipo II, IX, X e XI, bem como do agrecano, nas lesões discondroplásicas. Observou-se uma redução acentuada nos níveis de mRNA da gelatinase-B (MMP-9), da colagenase-3 (MMP-13) e das estromalisinas -2 (MMP-10) e -3 (MMP-11), bem como nos níveis proteicos da gelatinase-A (MMP-2) e da MMP-13. Por outro lado, a MMP-7 aumentou drasticamente a expressão do seu gene. As moléculas envolvidas na proliferação e diferenciação dos condrócitos, tais como a PTHrP, o Ihh, o Cbfa-1 e o Sox-9, mantiveram a sua expressão génica nas lesões. Por outro lado, o TGF-β reduziu a sua expressão. A caspase-3 também dimimuiu a sua expressão na patologia. Em relação aos factores angiogénicos, o FGF manteve a sua expressão e o VEGF aumentou significativamente nas lesões. Este aumento do VEGF juntamente com o aumento da MMP-7 sugere um aumento da hipoxia nas lesões. Os nossos resultados sugerem que a acumulação da cartilagem observada na discondroplasia é devida a uma diminuição da proteólise da matriz, resultado de uma sub-expressão das MMPs, e não de um aumento da produção das macromoléculas da matriz. Desta forma, os nossos resultados sugerem que a falha na expressão e/ou activação das MMPs poderá estar associada ao desenvolvimento da discondroplasia da tíbia em aves. Finalmente, os nossos resultados vêm suportar os resultados anteriores que sugerem uma ligação entre a expressão das MMPs e anomalias no processo de ossificação endocondral.

Early post-fledgling survival in a fragmented population of a tropical cooperative breeding passerine

During the last years tropical forest has been a target of intense study especially due to its recent big scale destruction. Although a lot still needs to be explored, we start realizing how negative can the impact of our actions be for the ecosystem. Subsequently, the living community have been developing strategies to overcome this problem avoiding bottlenecks or even extinctions. Cooperative breeding (CB) has been recently pointed out as one of those strategies. CB is a breeding system where more than two individuals raise one brood. In most of the cases, extra individuals are offspring that delay their dispersal and independent breeding what allows them to help their parents raising their siblings in the subsequent breeding season. Such behavior is believed to be due, per example, to the lack of mates or breeding territories (ecological constraints hypothesis), a consequence of habitat fragmentation and/or disturbance. From this point, CB is easily promoted by a higher reproductive success of group vs pairs or single individuals. Accordingly, during this thesis I explore the early post-fledging survival of a cooperative breeding passerine, namely the impact of individual/habitat quality in its survival probability during the dependence period of the chicks. Our study species is the Cabanis’s greenbul (Phyllastrephus cabanisi), a medium-sized, brownish passerine, classified within the Pycnonotidae family. It is found over part of Central Africa in countries such as Angola, Democratic Republic of the Congo, Mozambique and Kenya, inhabiting primary and secondary forests, as well as woodland of various types up to 2700m of altitude. Previous studies have concluded that PC is a facultative cooperative breeder. This study was conducted in Taita Hills (TH) at the Eastern Arc Mountains (EAM), a chain of mountains running from Southeast Kenya to the South of Tanzania. TH comprises an area of 430 ha and has been suffering intense deforestation reflecting 98% forest reduction over the last 200 years. Nowadays its forest is divided in fragments and our study was based in 5of those fragments. We access the post-fledging survival through radio-telemetry. The juvenile survey was done through the breeding females in which transmitters were placed with a leg-loop technique. Ptilochronology is consider to be the study of feather growth bars and has been used to study the nutritional state of a bird. This technique considers that the feather growth rate is positively proportional to the individual capability of ingesting food and to the food availability. This technique is therefore used to infer for individual/habitat quality. Survival was lowest during the first 5 days post-fledging representing 53.3%. During the next 15 days, risk of predation decreased with only 14.3% more deceased individuals. This represents a total of only 33% survived individuals in the end of the 50 days. Our results showed yet a significant positive relationship between flock size and post-fledging survival as well as between ptilochronology values and post-fledgling survival. In practice, these imply that on this population, as bigger the flock, as greater the post fledging survival and that good habitat quality or good BF quality, will lead to a higher juvenile survival rate. We believe that CB is therefore an adaptive behaviour to the lack of mates/breeding territory originated from the mass forest destruction and disturbance. Such results confirms the critical importance of habitat quality in the post-fledging survival and, for the first time, demonstrates how flock size influences the living probability of the juveniles and therefore how it impacts the (local) population dynamics of this species. In my opinion, future research should be focus in disentangle individual and habitat quality from each other and verify which relationship exist between them. Such study will allow us to understand which factor has a stronger influence in the post-fledging survival and therefore redirect our studies in that direction. In order to confirm the negative impact of human disturbance and forest fragmentation, it would be of major relevance to compare the reproductive strategies and reproductive success of populations living in intact forests and disturbed patches.