Research Methodology

The scientific aim of LEO is to study the genetic and morphogenetic mechanisms that give rise to form, structure and functional organisation during ontogenic development.

The experimental strategy is based on two main principles: (a) trans-disciplinary approach, and (b) use of in vivo genetic model organisms that allow a bottom-up comparative perspective of ontogeny.  

Multi-disciplinary approach

A unified view of morphogenesis requires expertise and approaches from different scientific disciplines. Consequently, LEO has invested considerable energy to build a trans-disciplinary research environment through the collaborative association with a research group focused on the analysis of biological images. This local partnership has led to: (i) the creation of a unique trans-disciplinary lab atmosphere in which young researchers from the fields of developmental biology, evolution, genetics, mathematics, physics and informatics share space, training and research projects; (ii) the implementation of a confocal microscopy platform that is under continuous development, aimed at visualising events of cellular morphogenesis in vivo, at high temporal and spatial resolution; and (iii) the development and implementation of mathematical algorithms for the analysis and modelling of sub-cellular, cellular and supra-cellular events of morphogenesis.

LEO trans-disciplinary environment is built around four main disciplines [see organic structure] to address the ORIGIN, EXPRESSION, TRANSFORMATION and ALTERATION of biological patterns during ontogenic development (see below).

To study the ORIGIN or genetic control LEO uses reverse genetics to identify novel genes (e.g. candidate gene, subtractive hybridisation, microarray and in silico analyses), mRNA and protein expression assays to determine the temporal and spatial organisation of gene expression (e.g. RT-PCR, in situ hybridisation, immunohistochemistry), and loss and gain of function methodologies to dissect gene function (e.g. mutant analyses, anti-sense gene knockdown, GAL4-UAS transgenesis).

The EXPRESSION of events that generate form, structure, and functional organisation is addressed using in vivo microscopy at high temporal and spatial resolution (e.g. spinning disc / confocal microscopy) combined with image processing and analysis techniques to access the morpho-topology and spatial organisation of morphogenetic components at supra-cellular, cellular and sub-cellular levels. To this aim, LEO has established an open-ended Mutual Enhancement collaborative relationship [see definition] with the Laboratory of Scientific Image Analysis (SCIAN) at the University of Chile.

The TRANSFORMATION occurring throughout evolutionary and developmental time is addressed by comparing the spatial and temporal organisation of development in related model organisms (e.g. zebrafish and medaka), and between distant vertebrate species (e.g. fish, frogs and humans), to dissect events of conservation, loss, acquisition, heterotopy and heterochrony.  

The ALTERATION of ontogenic mechanisms that impact Biomedical research is investigated in the context of experimental models that allow an in vivo systemic phenomenological approach of the biological basis of human disease.

In vivo genetic model organisms for a bottom-up comparative perspective of ontogeny

The scientific approach of LEO uses teleost species such as ZEBRAFISH (Danio rerio), MEDAKA (Oryzias latipes) and ANNUAL KILLIFISH (Austrolebias sp) as model organisms as they offer several advantages for a multi-scale comparative perspective of form, structure and functional organisation during ontogeny.

Zebrafish and medaka are established genetic organisms with open access to genomic data, and available resources and techniques for genetic manipulation.

Embryos and larvae of zebrafish, medaka and Annual killifish develop exo-utero and share optical transparency, which allow the direct visualisation of ontogenic development using in vivo microscopy. Available techniques permit a multi-scale study of ontogeny in these species, from genetics through morphology up to physiology and behaviour, easing the integration of information from different levels of biological organisation, following a bottom-up (gene-to-behaviour) approach (see left).

Zebrafish, medaka and Annual killifish of the genus Austrolebias are related teleosts with variable degree of independent evolution, making them suitable for a systematic comparative approach of ontogeny in the search of conserved and species-specific mechanisms of development (see below).

In addition to these advantages, teleost fish have also recently emerged as valuable experimental models to address the phenomenological basis of several human diseases.