Cobalt uptake enables identification of capsaicin- and bradykinin-sensitive subpopulations of rat dorsal root ganglion cellsin vitro

Nagy, I.; Pabla, Ravinder; Matesz, Clara; Dray, A.; Woolf, Clifford J.; Urbán, László

doi:10.1016/0306-4522(93)90576-2

Cited by 37 publications

(18 citation statements)

References 18 publications

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“…Neurons from different regions of the brain, including primary cultures of neocortical (Turetsky et al, 1994), hippocampal (Burke et al, 1995), retinal (Allcorn et al, 1996), and dorsal root ganglia (Nagy et al, 1993) neurons as well as slices of the hippocampus (Williams et al, 1992) and brainstem (Zhou et al, 1995) have been shown to be stained with cobalt after exposure to AMPA or kainate. These observations indicate that Ca 2ϩ -permeable, non-NMDA receptors exist in the brain, as discussed by Pruss et al (1991).…”

Section: Discussionmentioning

confidence: 99%

Development of calcium-permeable ?-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors in cultured neocortical neurons visualized by cobalt staining

Jensen¹,

Schousboe²,

Pickering³

1998

J. Neurosci. Res.

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The developmental expression of calcium (Ca2+)-permeable alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and kainate receptors in cultured neocortical neurons was evaluated by using cobalt uptake, a histochemical method that identifies cells expressing Ca2+-permeable, non-N-methyl-D-aspartate (non-NMDA) receptors. At a concentration of 500 microM, AMPA was found to stimulate cobalt uptake only late in development, resulting in staining of 2.7%+/-0.3% of the neurons maintained in culture for 12 days in vitro (DIV). When AMPA receptor desensitization was blocked with 50 microM cyclothiazide, the developmental profile of cobalt uptake mediated by 25 microM AMPA changed dramatically. The cobalt staining now appeared in young cultures (5 DIV), and the percentage of stained cells increased from 3.4%+/-0.2% at 5 DIV to 21.7%+/-1.6% at 12 DIV. The effect of 200 microM kainate was similar to that seen with 25 microM AMPA plus 50 microM cyclothiazide, resulting in 17.7%+/-0.3% stained neurons at 12 DIV. The cobalt uptake was specific to AMPA and kainate receptors because NMDA receptors and voltage-gated calcium channels were found not to mediate any cobalt staining. In addition, 10 microM 6-nitro-7-sulphamoylbenzo-[f]-quinoxaline-2,3-dione (NBQX) was able to prevent all staining at 5 and 8 DIV and most of the staining at 12 DIV, indicating that the non-NMDA ionotropic glutamate receptors are involved in cobalt uptake into the neurons. The AMPA receptor-selective antagonist GYKI 53655 was used to differentiate between cobalt influx through AMPA- or kainate-preferring receptors. After pretreatment with concanavalin A (con A), an inhibitor of kainate receptor desensitization, cobalt uptake was assessed after stimulation by 200 microM kainate in the presence of 25 microM GYKI 53655. No cobalt staining was observed under these conditions, indicating that most if not all of the cobalt influx induced by kainate was mediated through AMPA receptor channels.

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Section: Discussionmentioning

confidence: 99%

Development of calcium-permeable ?-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors in cultured neocortical neurons visualized by cobalt staining

Jensen¹,

Schousboe²,

Pickering³

1998

J. Neurosci. Res.

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“…Cobalt taken up by the neurons through activated nonselective cationic channels (Nagy et al, 1993) was precipitated by adding a 2.5% mercaptoethanol solution in buffer for 1 min. Cells were fixed in 70% ethanol.…”

Section: Methodsmentioning

confidence: 99%

Localization of the Endocannabinoid-Degrading Enzyme Fatty Acid Amide Hydrolase in Rat Dorsal Root Ganglion Cells and Its Regulation after Peripheral Nerve Injury

Lever¹,

Robinson²,

Cibelli³

et al. 2009

J. Neurosci.

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Fatty acid amide hydrolase (FAAH) is a degradative enzyme for a group of endogenous signaling lipids that includes anandamide (AEA).AEA acts as an endocannabinoid and an endovanilloid by activating cannabinoid and vanilloid type 1 transient receptor potential (TRPV1) receptors, respectively, on dorsal root ganglion (DRG) sensory neurons. Inhibition of FAAH activity increases AEA concentrations in nervous tissue and reduces sensory hypersensitivity in animal pain models. Using immunohistochemistry, Western blotting, and reverse transcription-PCR, we demonstrate the location of the FAAH in adult rat DRG, sciatic nerve, and spinal cord. In naive rats, FAAH immunoreactivity localized to the soma of 32.7 Ϯ 0.8% of neurons in L4 and L5 DRG. These were small-sized (mean soma area, 395.96 Ϯ 5.6 m 2 ) and predominantly colabeled with peripherin and isolectin B4 markers of unmyelinated C-fiber neurons; 68% colabeled with antibodies to TRPV1 (marker of nociceptive DRG neurons), and Ͻ2% colabeled with NF200 (marker of large myelinated neurons). FAAH-IR was also present in small, NF200-negative cultured rat DRG neurons. Incubation of these cultures with the FAAH inhibitor URB597 increased AEA-evoked cobalt uptake in a capsazepine-sensitive manner. After sciatic nerve axotomy, there was a rightward shift in the cell-size distribution of FAAH-immunoreactive (IR) DRG neurons ipsilateral to injury: FAAH immunoreactivity was detected in larger-sized cells that colabeled with NF200. An ipsilateral versus contralateral increase in both the size and proportion of FAAH-IR DRG occurred after spinal nerve transection injury but not after chronic inflammation of the rat hindpaw 2 d after injection of complete Freund's adjuvant. This study reveals the location of FAAH in neural tissue involved in peripheral nociceptive transmission.

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“…Studying cobalt accumulation, in addition to measuring changes in the intracellular Ca 2+ concentration and membrane currents, is also an established technique to study the expression and activity of various ligand-gated non-selective cationic channels, including TRPV1 (Mahahmud et al 2009;Marin-Burgin et al 2000;Nagy et al 1993;Pruss et al 1991;Sathianathan et al 2003;Singh Tahim et al 2005). Indeed, capsaicin, heat and anandamide produce specific and selective TRPV1-mediated cobalt entry in a concentration and temperature-dependent manner into primary sensory neurons (Mahmud et al 2009;MarinBurgin et al 2000;Sathianathan et al 2003;Singh Tahim et al 2005).…”

Section: Discussionmentioning

confidence: 96%

Effects of cannabinoids on capsaicin receptor activity following exposure of primary sensory neurons to inflammatory mediators

et al. 2010

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Cobalt uptake enables identification of capsaicin- and bradykinin-sensitive subpopulations of rat dorsal root ganglion cellsin vitro

Cited by 37 publications

References 18 publications

Development of calcium-permeable ?-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors in cultured neocortical neurons visualized by cobalt staining

Development of calcium-permeable ?-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors in cultured neocortical neurons visualized by cobalt staining

Localization of the Endocannabinoid-Degrading Enzyme Fatty Acid Amide Hydrolase in Rat Dorsal Root Ganglion Cells and Its Regulation after Peripheral Nerve Injury

Effects of cannabinoids on capsaicin receptor activity following exposure of primary sensory neurons to inflammatory mediators

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