XXII. BIOCHEMICKÝ ZJAZD - Jesseniova lekárska fakulta

Lectures
UTILITY OF SINGLE CELL RT-PCR (SCrT-PCR) FOR THE STUDY OF
PRIMarY affERENT NEURONS - PRELIMINarY vaLIDATION
Lenka Surdeníková 1 , Fei Ru 2 and Marian Kollárik 1,2#
1
Pathophysiology, Jessenius Medical School, 2 Medicine, Johns Hopkins School
of Medicine, # correspondence: kollarik@jhmi.edu
We addressed the hypothesis that scRT-PCR detection of selected receptors in primary
sensory neurons provides information predictive of their functional response mediated
by these receptors. scRT-PCR was performed on individual mouse or guinea pig primary
sensory neurons. Functional response of the neurons or their putative peripheral nerve
terminals was evaluated by calcium imaging, whole cell patch clamp or single nerve fiber
recordings. scRT-PCR detection of MrgPrA 3
receptor in neurons correlated with their
responsiveness to the MrgPrA 3
selective agonist chloroquine (1mM) in calcium imaging
studies. MrgPrA 3
mRNA was detected in 8 of 9 chloroquine-responsive neurons but not
in 11 chloroquine -unresponsive neurons. Similarly, detection of TRPV1 receptor correlated
with the responsiveness to the TRPV1 agonist capsaicin (1microM) in patch clamp
studies (n=9). Detection of the purinergic receptors P2X 2
/P2X 3
in the nodose nociceptive
neurons correlated with the P2X 2
/P2X 3
current signature in these neurons in patch
clamp studies. scRT-PCR detection of the adenosine A 1
and A 2A
receptors, and the TRPA1
receptor in the vagal nodose nociceptive neurons correlated with the responsiveness of
their putative nerve terminals to the selective adenosine A 1
and A 2A
receptor agonists
and TRPA1 agonists, respectively, in single fiber recording studies. We conclude that
scRT-PCR detection of all receptors evaluated thus far in our studies in primary sensory
neurons correlated with the functional response mediated by these receptors. Our data
indicate that scRT-PCR on the individual primary sensory neurons provides information
predictive of their functional response and is a suitable complementary tool for the
study of these neurons.
XXII. Biochemistry Congress, Martin
99