The Impact of Thrombin and Phospholipase C Inhibitor on Motor Axon Development in Caenorhabiditis elegans

PAC Gym

Thrombin accumulation following spinal cord injuries is associated with decreased repair and survival of neurons. Using Caenorhabiditis elegans, past studies in our lab have shown that thrombin prevents the development of axons which are essential for normal functioning of the nervous system. Although C. elegans neurons possess the thrombin receptor, PAR-1, the specific mechanism following PAR-1 activation has not been reported. In other systems, PAR-1 has been shown to use one of two pathways mediated by phospholipase C (PLC) or Rho. Since PLC is a regulator of axonal growth cones, we hypothesize that axonal interruption involves the PLC pathway. To address this question, eggs collected from C. eleganscontaining the insertion of a green fluorescent protein (GFP) promoter in their GABAergic motor neurons were exposed to thrombin, a PLC inhibitor, or in combination and allowed to develop into adults. This particular strain of C. elegansallows for direct visualization of neurons including their axons. Using fluorescence microscopy, axons were categorized as complete or incomplete. If PLC inhibition produces complete axons, then we can determine that thrombin induces its effects by increasing PLC levels. Understanding how thrombin interferes with axon development is important to understanding and developing treatments for spinal cord injury.