title: Alteration of the mu opioid receptor: Ca2+ channel signaling pathway in a subset of rat sensory neurons following chronic femoral artery occlusion.
abstract: The exercise pressor reflex, a crucial component of the cardiovascular response under physiological and pathophysiological states, is activated via metabolic and mechanical mediators that originate from contracting muscles and stimulate group III and IV afferents. We reported previously that stimulation of mu opioid receptors (MOR), expressed in both afferents, led to a significant attenuation of the reflex in rats whose femoral arteries had been occluded for 72 h. The present study examined the effect of arterial occlusion on the signaling components involved in the opioid-mediated modulation of Ca(2+) channels in rat dorsal root ganglion neurons innervating the triceps surae muscles. We focused on neurons that were transfected with cDNA coding for enhanced green fluorescent protein whose expression is driven by the voltage-gated Na(+) channel 1.8 (Na(V)1.8) promoter region, a channel expressed primarily in nociceptive neurons. With the use of a small interference RNA approach, our results show that the pertussis toxin-sensitive Gα(i3) subunit couples MOR with Ca(2+) channels. We observed a significant leftward shift of the MOR agonist [D-Ala2-N-Me-Phe4-Glycol5]-enkephalin concentration-response relationship in neurons isolated from rats with occluded arteries compared with those that were perfused freely. Femoral occlusion did not affect Ca(2+) channel density or the fraction of the main Ca(2+) channel subtype. Furthermore, Western blotting analysis indicated that the leftward shift did not result from either increased Gα(i3) or MOR expression. Finally, all neurons from both groups exhibited an inward current following exposure of the transient potential receptor vanilloid 1 (TRPV1) agonist, 8-methyl-N-vanillyl-6-nonenamide. These findings suggest that sensory neurons mediating the exercise pressor reflex express Na(V)1.8 and TRPV1 channels, and femoral occlusion alters the MOR pharmacological profile.
authors.affiliation: Department of Anesthesiology, Penn State College of Medicine, Hershey, Pennsylvania; and.
authors.forename: Bassil
authors.initials: B
authors.lastname: Hassan
authors.affiliation: Heart and Vascular Institute, Penn State College of Medicine, Hershey, Pennsylvania.
authors.forename: Joyce S
authors.initials: JS
authors.lastname: Kim
authors.affiliation: Department of Anesthesiology, Penn State College of Medicine, Hershey, Pennsylvania; and.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Farrag
authors.affiliation: Heart and Vascular Institute, Penn State College of Medicine, Hershey, Pennsylvania.
authors.forename: Marc P
authors.initials: MP
authors.lastname: Kaufman
authors.affiliation: Department of Anesthesiology, Penn State College of Medicine, Hershey, Pennsylvania; and vruizvelasco@psu.edu.
authors.forename: Victor
authors.initials: V
authors.lastname: Ruiz-Velasco
mesh_terms: D000818:Animals; D015220:Calcium Channels / Q000378:metabolism*; D020013:Calcium Signaling; D005263:Femoral Artery / Q000293:injuries; D044385:GTP-Binding Protein alpha Subunits / Q000378:metabolism; D005727:Ganglia, Spinal / Q000166:cytology / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000032:analysis; D008297:Male; D018482:Muscle, Skeletal / Q000294:innervation; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D017450:Receptors, Opioid, mu / Q000378:metabolism*; D011984:Sensory Receptor Cells / Q000378:metabolism*
chemical_list: D015220:Calcium Channels; D044385:GTP-Binding Protein alpha Subunits; D012333:RNA, Messenger; D017450:Receptors, Opioid, mu; D049452:Green Fluorescent Proteins
keywords: Nav1.8 promoter region; exercise pressor reflex; whole-cell patch clamp