Visualisation of macropinosome maturation by the recruitment of sorting nexins

We report that phosphoinositol-binding sorting nexin 5 ( SNX5) associates with newly formed macropinosomes induced by EGF stimulation. We used the recruitment of GFP-SNX5 to macropinosomes to track their maturation. Initially, GFP-SNX5 is sequestered to discrete subdomains of the macropinosome; these subdomains are subsequently incorporated into highly dynamic, often branched, tubular structures. Time-lapse videomicroscopy revealed the highly dynamic extension of SNX5-labelled tubules and their departure from the macropinosome body to follow predefined paths towards the perinuclear region of the cell, before fusing with early endosomal acceptor membranes. The extension and departure of these tubular structures occurs rapidly over 5-10 minutes and is dependent upon intact microtubules. As the tubular structures depart from the macropinosome there is a reduction in the surface area and an increase in tension of the limiting membrane of the macropinosome. In addition to the recruitment of SNX5 to the macropinosome, Rab5, SNX1 and EEA1 are also recruited by newly formed macropinosomes, followed by the accumulation of Rab7. SNX5 forms heterodimers with SNX1 and this interaction is required for endosome association of SNX5. We propose that the departure of SNX5-positive tubules represents a rapid mechanism of recycling components from macropinosomes thereby promoting their maturation into Rab7-positive structures. Collectively these findings provide a detailed real-time characterisation of the maturation process of the macropinocytic endosome.

3D information visualisation: An historical perspective

The use of 3D visualisation of digital information is a recent phenomenon. It relies on users understanding 3D perspectival spaces. Questions about the universal access of such spaces has been debated since its inception in the European Renaissance. Perspective has since become a strong cultural influence in Western visual communication. Perspective imaging assists the process of experimenting by the sketching or modelling of ideas. In particular, the recent 3D modelling of an essentially non-dimensional Cyber-space raises questions of how we think about information in general. While alternate methods clearly exist they are rarely explored within the 3D paradigm (such as Chinese isometry). This paper seeks to generate further discussion on the historical background of perspective and its role in underpinning this emergent field. © 2005 IEEE.