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Changes in Glutaminase mRNA Expression in Rat Dorsal Root Ganglion Post Tail Inicision
Author(s) -
Crosby Heith A,
Spencer Diana,
Miller Kenneth E
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.lb413
Subject(s) - dorsal root ganglion , messenger rna , neurotransmitter , glutamate receptor , glutaminase , ganglion , anatomy , medicine , dorsum , endocrinology , biology , central nervous system , receptor , gene , biochemistry
Aim of Investigation Glutamate is a neurotransmitter used by primary sensory neurons of the dorsal root ganglion(DRG). This excitatory neurotransmitter is released from peripheral nerve terminals and spinal synaptic terminals during nociceptive signaling. In this study, we evaluated the changes in Glutaminase (GLS) mRNA at 1, 2, and 4 hours post tail incision in a surgical incision tail model. Methods Sprague Dawley rats were anesthetized and a 20mm incision made in the proximal one third of the tail. Controls were surgical naïve rats. At 1, 2, and 4 hours, rats were asphyxiated, decapitated and sacral DRG were collected. Samples were stored in RNAlater ™ (Ambion). RNA was isolated using the Vantage Total RNA kit ™ (Origene). Real‐Time Quantitative Polymerase Chain Reaction (RT‐qPCR) was performed on sacral DRG samples using iScript One‐Step RT‐PCR Kit with Sybr Green™ (Biorad). Threshold cycle (Ct) and melt data were collected for GLS and beta‐actin (control mRNA). Results GLS mRNA in Sacral DRG neurons exhibited both depression and elevation in fold change across sacral levels 1–3 at 1, 2 and 4 hours post incision. Conclusions These results provide insight on how GLS mRNA is maintained in sacral dorsal root ganglia and how it responds to tissue injury. From the results of the present study, incisional wounding results in a decrease in GLS mRNA. We hypothesize that GLS mRNA is maintained in a basal pool and that after injury GLS mRNA is rapidly translated thereby decreasing the available reservoir. This, results in an increase in GLS protein observed in our previous immunohistochemisty and Western blot studies. Research Support: NIH Grant AR047410 (KEM) and OSU CHS Intramural Grant