Diploma Thesis from the year 2011 in the subject Medicine - Biomedical
Engineering, grade: 1,0, Technical University of Berlin
(Biotechnologie), language: English, abstract: Pain is an unpleasant
sensory and emotional experience and an essential component of the
body's defense system. Noxious stimuli are detected by special afferent
sensory neurons called nociceptors (or nociceptive neurons), which
connect peripheral tissues with the central nervous system. Noxious
signals travel along the nociceptor to the dorsal horn of the spinal
cord from where they projected to higher brain regions via 2nd order
neurons. After evaluation in brain stem, thalamus, and cortex a complex
sensory perception arises: Pain. Extensive research has been performed
to gain insight into molecular mechanisms of pain sensation and its
modulation. Many of the transduction channels that convert thermal,
mechanical or chemical stimuli into electrical activity are transient
receptor potential (TRP) ion channels, which are widely expressed in
nociceptors and play a prominent role in pain sensation. Still many
transduction molecules that trigger pain have not been discovered. To
identify transcripts enriched in TRPV1-positive neurons I used the
ability to activate these channels with the specific agonists capsaicin
and resiniferatoxin (RTX). The resulting calcium influx changes
structural properties and thus alters the density of TRPV1-positive
neurons. This enables their separation via density gradient
centrifugation. In comparison to a control group (including
TRPV1-positive neurons) this newly developed method allows me to find
predominantly expressed genes in TRPV1-positive neurons. To validate the
depletion I utilized western blot, immunocytochemistry and qPCR. The
results showed effective depletion of TRPV1-postitive neurons for up to
95%. Subsequent gene expression analysis revealed promising candidates
that were highly enriched in TRPV1-positive neurons. The candidate genes
were classified a
