Abstract

Benjamin Klapholz (Gurdon Institute)

Talin mediates integrin function: multiple molecular mechanisms for one general function

We used genetic and advanced imaging methods to show that talin can adopt distinct, tissue-specific, mechanisms of action to mediate integrin function. Such diverse configurations may allow talin to sense a variety of forces that affect cell-matrix adhesions.

Julian Ng and Ben Sutcliffe

Genetically encoded probes for chemical labelling in Drosophila

Fluorescent proteins (FP) and antibodies are powerful tools for analysing the structure of molecules and cells embedded within tissues. However, each method has drawbacks. FPs are readily detected when expressed in cells, but they have weaker photon yields and limited spectral flexibility compared with organic dyes. Antibodies, conjugated to small dyes, can highlight any endogenous protein. However their bulkiness limits the labelling efficiency in thick tissue blocks during the immunohistochemistry process, which can last for hours – days. Furthermore, weakly interacting antibodies can lead to off-target effects and high background staining. This talk will highlight recently published [1] and unpublished work on chemical based tagging in Drosophila. We find that SNAP and Halo tag based technologies can overcome some of the problems associated with FPs and immunostaining, as fluorescent chemical tagging leads to very fast, even, high-signal and low-background labelling of thick tissues. We have generated and validated in the brain a variety of UAS constructs along with two knockin lines (Brp, Dlg) that should have applications in most fly tissues.

[1] see http://www.pnas.org/content/111/36/E3805.full and jefferislab.org/si/tagging