Defined Medium Organotypic Cultures of Spinal Cord Put ‘Pain in a Dish’

Biggs, James E.; Lu, Van B.; Kim, Helena J.; Lai, Aaron Y.; Todd, Kathryn G.; Ballanyi, Klaus; Colmers, William F.; Smith, Peter A.

doi:10.1007/978-1-62703-020-5_14

Cited by 16 publications

(28 citation statements)

References 61 publications

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“…DMOTC rat spinal cord slices with attached DRG were prepared as previously described (Biggs et al 2012;Lu et al 2006). Briefly, embryonic day 13 (E13) rat fetuses were delivered by cesarean section from timed-pregnant female Sprague-Dawley rats (Charles River, Saint-Constant, QC, Canada) under 5% isoflurane anesthesia.…”

Section: Methodsmentioning

confidence: 99%

“…Changes in Ca 2ϩ fluorescence intensity were measured in dorsal horn neurons with a confocal microscope equipped with an argon (488 nm) laser and filters (XLUMPlanF1 ϫ20, 0.95 NA objective, Olympus FV300; Olympus, Markham, ON, Canada). Changes were evoked by one of two methods: 1) application of high-K ϩ solution (20 or 35 mM) for 90 s or 2) electrical stimulation of the dorsal root entry zone (DRZ) (Biggs et al 2012). Tungsten bipolar electrodes (WPI) were used with the following stimulating parameters: frequency, 50 Hz; pulse duration, 100 s; number of pulses, 250.…”

Section: Methodsmentioning

confidence: 99%

“…Typical stimulus intensities ranged between 10 and 60 V. A total of 8 stimulus trains (S1-S8) were delivered at 5-min intervals. Since the most stable response amplitudes were seen with the fifth and sixth stimuli (S5 and S6), responses to the fifth set of stimuli were used for all data comparisons (Biggs et al 2012). Full-frame images (512 ϫ 512 pixels) in a fixed xy plane were acquired at a scanning time of 1.08 s per frame and stored as video files for off-line analysis (Ruangkittisakul et al 2006).…”

Section: Methodsmentioning

confidence: 99%

“…We therefore compared the reduction in HVA Ca 2ϩ channel current (I Ca ) seen in DRG neurons following 5-6 days of exposure to gabapentinoids with that produced by a relatively low concentration of Mn 2ϩ (20 M). We then examined the effects of gabapentinoids and 20 M Mn 2ϩ on sEPSCs in neurons in spinal cord slices maintained in defined-medium organotypic culture (DMOTC; Biggs et al 2012;Lu et al 2006).…”

mentioning

confidence: 99%

“…In view of their antiallodynic action, gabapentinoids may exert preferential effects at excitatory synapses and excitatory neurons as opposed to inhibitory synapses and inhibitory neurons. To test these possibilities, we examined long-term actions of gabapentinoids on overall dorsal horn excitability using confocal Ca 2ϩ imaging (Biggs et al 2012) as well as whole cell recordings from putative excitatory and inhibitory substantia gelatinosa neurons in spinal cord slices maintained in DMOTC.…”

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confidence: 99%

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Analysis of the long-term actions of gabapentin and pregabalin in dorsal root ganglia and substantia gelatinosa

Biggs

Boakye

Ganesan

et al. 2014

Journal of Neurophysiology

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of the long-term actions of gabapentin and pregabalin in dorsal root ganglia and substantia gelatinosa. J Neurophysiol 112: 2398 -2412, 2014. First published August 13, 2014; doi:10.1152/jn.00168.2014.-The ␣2␦-ligands pregabalin (PGB) and gabapentin (GBP) are used to treat neuropathic pain. We used whole cell recording to study their long-term effects on substantia gelatinosa and dorsal root ganglion (DRG) neurons. Spinal cord slices were prepared from embryonic day 13 rat embryos and maintained in organotypic culture for Ͼ5 wk (neuronal age equivalent to young adult rats). Exposure of similarly aged DRG neurons (dissociated and cultured from postnatal day 19 rats) to GBP or PGB for 5-6 days attenuated high-voltage-activated calcium channel currents (HVA I Ca ). Strong effects were seen in medium-sized and in small isolectin B 4 -negative (IB 4 Ϫ) DRG neurons, whereas large neurons and small neurons that bound isolectin B 4 (IB 4 ϩ) were hardly affected. GBP (100 M) or PGB (10 M) were less effective than 20 M Mn 2ϩ in suppression of HVA I Ca in small DRG neurons. By contrast, 5-6 days of exposure to these ␣2␦-ligands was more effective than 20 M Mn 2ϩ in reducing spontaneous excitatory postsynaptic currents at synapses in substantia gelatinosa. Spinal actions of gabapentinoids cannot therefore be ascribed to decreased expression of HVA Ca 2ϩ channels in primary afferent nerve terminals. In substantia gelatinosa, 5-6 days of exposure to PGB was more effective in inhibiting excitatory synaptic drive to putative excitatory neurons than to putative inhibitory neurons. Although spontaneous inhibitory postsynaptic currents were also attenuated, the overall long-term effect of ␣2␦-ligands was to decrease network excitability as monitored by confocal Ca 2ϩ imaging. We suggest that selective actions of ␣2␦-ligands on populations of DRG neurons may predict their selective attenuation of excitatory transmission onto excitatory vs. inhibitory neurons in substantia gelatinosa.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%