Ca²⁺-dependent regulation of Ca²⁺ currents in rat primary afferent neurons: role of CaMKII and the effect of injury. J Neurosci 2012 Aug 22;32(34):11737-49
Date
08/24/2012Pubmed ID
22915116Pubmed Central ID
PMC3723336DOI
10.1523/JNEUROSCI.0983-12.2012Scopus ID
2-s2.0-84865228570 (requires institutional sign-in at Scopus site) 26 CitationsAbstract
Currents through voltage-gated Ca²⁺ channels (I(Ca)) may be regulated by cytoplasmic Ca²⁺ levels ([Ca²⁺](c)), producing Ca²⁺-dependent inactivation (CDI) or facilitation (CDF). Since I(Ca) regulates sensory neuron excitability, altered CDI or CDF could contribute to pain generation after peripheral nerve injury. We explored this by manipulating [Ca²⁺](c) while recording I(Ca) in rat sensory neurons. In uninjured neurons, elevating [Ca²⁺](c) with a conditioning prepulse (-15 mV, 2 s) inactivated I(Ca) measured during subsequent test pulses (-15 mV, 5 ms). This inactivation was Ca²⁺-dependent (CDI), since it was decreased with elimination of Ca²⁺ influx by depolarization to above the I(Ca) reversal potential, with high intracellular Ca²⁺ buffering (EGTA 10 mm or BAPTA 20 mm), and with substitution of Ba²⁺ for extracellular Ca²⁺, revealing a residual voltage-dependent inactivation. At longer latencies after conditioning (>6 s), I(Ca) recovered beyond baseline. This facilitation also proved to be Ca²⁺-dependent (CDF) using the protocols limiting cytoplasmic Ca²⁺ elevation. Ca²⁺/calmodulin-dependent protein kinase II (CaMKII) blockers applied by bath (KN-93, myristoyl-AIP) or expressed selectively in the sensory neurons (AIP) reduced CDF, unlike their inactive analogues. Protein kinase C inhibition (chelerythrine) had no effect. Selective blockade of N-type Ca²⁺ channels eliminated CDF, whereas L-type channel blockade had no effect. Following nerve injury, CDI was unaffected, but CDF was eliminated in axotomized neurons. Excitability of sensory neurons in intact ganglia from control animals was diminished after a similar conditioning pulse, but this regulation was eliminated by injury. These findings indicate that I(Ca) in sensory neurons is subject to both CDI and CDF, and that hyperexcitability following injury-induced loss of CDF may result from diminished CaMKII activity.
Author List
Tang Q, Bangaru ML, Kostic S, Pan B, Wu HE, Koopmeiners AS, Yu H, Fischer GJ, McCallum JB, Kwok WM, Hudmon A, Hogan QHAuthors
Quinn H. Hogan MD Professor in the Anesthesiology department at Medical College of WisconsinWai-Meng Kwok PhD Professor in the Anesthesiology department at Medical College of Wisconsin
Hongwei Yu MD Professor in the Anesthesiology department at Medical College of Wisconsin
MESH terms used to index this publication - Major topics in bold
Analysis of VarianceAnimals
Biophysical Phenomena
Biophysics
Calcium
Calcium Channel Blockers
Calcium Signaling
Calcium-Calmodulin-Dependent Protein Kinase Type 2
Chelating Agents
Dantrolene
Drug Interactions
Egtazic Acid
Electric Stimulation
Enzyme Inhibitors
Ganglia, Spinal
Genetic Vectors
Green Fluorescent Proteins
Hyperalgesia
Laminectomy
Male
Membrane Potentials
Neurons, Afferent
Pain Threshold
Patch-Clamp Techniques
Peripheral Nerve Injuries
Rats
Rats, Sprague-Dawley
Signal Transduction
Time Factors
