FLUOMORPH

Fluorescent biosensors of organelle morphology to study the nuclear envelope dynamics during cell division

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 Obiettivo del progetto (Objective)

'Eukaryotic cells are compartmentalized into membrane-bound organelles, ensuring highly specialized and essential functions such as protection of the genome, protein synthesis and packaging, intracellular calcium signalling... Most organelles harbour a complex morphology, which is an important structural feature for the achievement of their assigned function. Moreover, some organelles are constituted of distinct subcompartments exhibiting specific structures associated to specialized functions. Visualizing the fine structure of organelles is thus essential to understand many aspects of their associated functions. Importantly, organelles architecture is highly dynamic and can be subjected to dramatic rearrangements in response to cellular changes like cell division or differentiation. The methods available so far to render organelle morphology require highly specialized techniques and are most often incompatible with high-rate image acquisition. The aim of this proposal is to develop a method to distinguish organelle substructures in live cells using a simple bi-dimensional confocal imaging setup. Fluorescent probes of organelle morphology will be designed based on the striking membrane sensing properties of a family of amphitpathic helices called ALPS motifs. To establish the usefulness of these tools, we propose to study the dynamics of the nuclear envelope (NE) and endoplasmic reticulum (ER) during mitosis.'

Introduzione (Teaser)

Visualisation of the fine structure of organelles in the cell is essential for understanding their associated functions. The architecture of organelles is highly dynamic and undergoes dramatic rearrangements in response to cellular changes like division or differentiation.

Descrizione progetto (Article)

Eukaryotic cells are compartmentalised. In general there are four main cellular compartments: the nucleus, the intercisternal space between the membranes of the endoplasmic reticulum, organelles and the cytosol. Most organelles have a complex morphology, which is important for their versatile functions. In particular, some organelles are made of distinct sub-compartments exhibiting specific shapes.

Methods currently available to render organelle morphology require highly specialised techniques and are most often incompatible with high-rate image acquisition. The goal of the EU-funded FLUOMORPH proposal is to develop a method to distinguish organelle sub-structures in live cells using a simple bi-dimensional confocal imaging setup.

Fluorescent probes of organelle morphology were designed based on the striking membrane-sensing properties of a family of amphipathic helices called ALPS motifs. To establish the usefulness of these tools, researchers studied the dynamics of the nuclear envelope and endoplasmic reticulum (ER) during mitosis.

Features of the sequence of ALPS motifs that are critical for their specificity have been studied. Scientists found that while the sequence of the backbone surrounding the ALPS motif can be changed, their structural context is important for the sensitivity to curvature. The changes occurring in ER morphology during cell division have been studied using the probe that stains specifically ER tubules. Preliminary results demonstrated the potential of fluorescent probes to sense the shape of organelles.

Visualising the fine structure of organelles is thus essential to understand many aspects of their associated functions. Applications of the technology could reach into many areas including disease diagnostics and therapeutics.