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Transient receptor potential ankyrin 1 that is induced in dorsal root ganglion neurons contributes to acute cold hypersensitivity after oxaliplatin administration.

Yamamoto K, Chiba N, Chiba T, Kambe T, Abe K, Kawakami K, Utsunomiya I, Taguchi K - Mol Pain (2015)

Bottom Line: Oxaliplatin significantly increased co-localization of TRPA1 expression and isolectin B4 binding in DRG neurons.In addition, we found that intrathecal administration of TRPA1 antisense, but not TRPA1 mismatched oligodeoxynucleotides, knocked down TRPA1 expression and decreased oxaliplatin-induced cold hyperalgesia.Intrathecal administration of the p38 MAPK inhibitor, SB203580, significantly decreased oxaliplatin-induced acute cold hypersensitivity.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicinal Pharmacology, Showa Pharmaceutical University, 3-3165 Higashitamagawagakuen, Machida, Tokyo, 194-8543, Japan. k-yamamoto@ac.shoyaku.ac.jp.

ABSTRACT

Background: Peripheral cold neuropathic pain is a serious side effect of oxaliplatin treatment. However, the mechanism of oxaliplatin-induced cold hyperalgesia is unknown. In the present study, we investigated the effects of oxaliplatin on transient receptor potential ankyrin 1 (TRPA1) in dorsal root ganglion (DRG) neurons of rats.

Results: Behavioral assessment using the acetone spray test showed that 3 and 6 mg/kg oxaliplatin (i.p.) induced acute cold hypersensitivity after 1, 2, 4, and 7 days. Real-time PCR showed that oxaliplatin (6 mg/kg) significantly increased TRPA1 mRNA expression in DRGs at days 1, 2, and 4. Western blotting revealed that oxaliplatin significantly increased TRPA1 protein expression in DRGs at days 2, 4, and 7. Moreover, in situ hybridization histochemistry revealed that most TRPA1 mRNA-labeled neurons in the DRGs were small in size. Oxaliplatin significantly increased co-localization of TRPA1 expression and isolectin B4 binding in DRG neurons. Oxaliplatin induced a significant increase in the percent of TRPA1 mRNA-positive small neurons in DRGs at days 1, 2, and 4. In addition, we found that intrathecal administration of TRPA1 antisense, but not TRPA1 mismatched oligodeoxynucleotides, knocked down TRPA1 expression and decreased oxaliplatin-induced cold hyperalgesia. Double labeling showed that p-p38 mitogen-activated protein kinase (MAPK) was co-expressed in TRPA1 mRNA-labeled neurons at day 2 after oxaliplatin administration. Intrathecal administration of the p38 MAPK inhibitor, SB203580, significantly decreased oxaliplatin-induced acute cold hypersensitivity.

Conclusions: Together, these results demonstrate that TRPA1 expression via activation of p38 MAPK in DRG neurons, at least in part, contributes to the development of oxaliplatin-induced acute cold hyperalgesia.

No MeSH data available.


Related in: MedlinePlus

Effect of oxaliplatin on TRPA1 mRNA as seen with in situ hybridization histochemistry in rat DRG (L4–6) neurons. a Oxaliplatin (6 mg/kg, i.p.) increases the co-localization between TRPA1 and p-p38 MAP kinase in rat DRG (L4–6) neurons. The photomicrographs show representative data. Co-localization of TRPA1 mRNA (green) and p-p38 MAPK (red) is shown. Double-labeled neurons (arrowheads) appear yellow in the merged panel. b Histogram shows the percent of TRPA1 mRNA-positive neurons relative to p-p38-positive neurons. TRPA1 mRNA and p-p38 co-localization was significantly higher on day 2 after oxaliplatin administration (6 mg/kg, i.p.) compared to 5 % glucose treatment. Data are the mean ± SEM. n = 4 each for 5 % glucose and oxaliplatin administration. *P < 0.05 versus 5 % glucose. c Size distribution of TRPA1 mRNA- and p-p38 MAPK-co-expressing neurons in DRGs in 5 % glucose- and oxaliplatin-treated rats. TRPA1 mRNA and p-p38 MAPK-co-expressing neurons were distributed among all sizes of neurons, but most co-expression was found in small DRG neurons. Data are the mean ± SEM. n = 4 each for 5 % glucose treatment and oxaliplatin treatment (6 mg/kg, i.p.). d Intrathecal administration of the p38 MAPK inhibitor, SB203580 (0.5 μg μL−1 h−1), significantly prevents the acute cold hypersensitivity induced by oxaliplatin (6 mg/kg, i.p.). The acetone spray test at day 2 after oxaliplatin was measured in SB203580 and 20 % DMSO treatment groups. Data are the mean ± SEM. n = 4 each for 20 % DMSO treatment and oxaliplatin treatment (6 mg/kg, i.p.). **P < 0.01 versus DMSO. ##P < 0.01 versus oxaliplatin
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Fig6: Effect of oxaliplatin on TRPA1 mRNA as seen with in situ hybridization histochemistry in rat DRG (L4–6) neurons. a Oxaliplatin (6 mg/kg, i.p.) increases the co-localization between TRPA1 and p-p38 MAP kinase in rat DRG (L4–6) neurons. The photomicrographs show representative data. Co-localization of TRPA1 mRNA (green) and p-p38 MAPK (red) is shown. Double-labeled neurons (arrowheads) appear yellow in the merged panel. b Histogram shows the percent of TRPA1 mRNA-positive neurons relative to p-p38-positive neurons. TRPA1 mRNA and p-p38 co-localization was significantly higher on day 2 after oxaliplatin administration (6 mg/kg, i.p.) compared to 5 % glucose treatment. Data are the mean ± SEM. n = 4 each for 5 % glucose and oxaliplatin administration. *P < 0.05 versus 5 % glucose. c Size distribution of TRPA1 mRNA- and p-p38 MAPK-co-expressing neurons in DRGs in 5 % glucose- and oxaliplatin-treated rats. TRPA1 mRNA and p-p38 MAPK-co-expressing neurons were distributed among all sizes of neurons, but most co-expression was found in small DRG neurons. Data are the mean ± SEM. n = 4 each for 5 % glucose treatment and oxaliplatin treatment (6 mg/kg, i.p.). d Intrathecal administration of the p38 MAPK inhibitor, SB203580 (0.5 μg μL−1 h−1), significantly prevents the acute cold hypersensitivity induced by oxaliplatin (6 mg/kg, i.p.). The acetone spray test at day 2 after oxaliplatin was measured in SB203580 and 20 % DMSO treatment groups. Data are the mean ± SEM. n = 4 each for 20 % DMSO treatment and oxaliplatin treatment (6 mg/kg, i.p.). **P < 0.01 versus DMSO. ##P < 0.01 versus oxaliplatin

Mentions: We detected TRPA1 mRNA-labeled neurons in DRG (L4–6) neurons at day 2 after oxaliplatin administration using ISHH. We compared the expression of p-p38 (red in Fig. 6a) between oxaliplatin- and 5 % glucose-treated rats. TRPA1 mRNA-labeled neurons (green in Fig. 6a) overlapped with p-p38-positive neurons. Double-labeling experiments revealed a pronounced overlap between DRG neurons expressing TRPA1 mRNA and those expressing p-p38 (yellow in Fig. 6a, merged). The percent of TRPA1 mRNA/p-p38 dual-positive neurons relative to the total p-p38-positive neurons was significantly increased at day 2 after oxaliplatin administration (36.7 ± 5.0 vs. 59.6 ± 3.9 %, P < 0.01, n = 4) (Fig. 6b). Oxaliplatin significantly increased expression of TRPA1 mRNA and p-p38 in DRG neurons. TRPA1 mRNA/p-p38 dual-positive neurons in oxaliplatin-treated rats were mainly small-sized neurons compared to 5 % glucose treatment (Fig. 6c). Thus, oxaliplatin increased the number of small-diameter DRG neurons that co-expressed TRPA1 mRNA and p-p38.Fig. 6


Transient receptor potential ankyrin 1 that is induced in dorsal root ganglion neurons contributes to acute cold hypersensitivity after oxaliplatin administration.

Yamamoto K, Chiba N, Chiba T, Kambe T, Abe K, Kawakami K, Utsunomiya I, Taguchi K - Mol Pain (2015)

Effect of oxaliplatin on TRPA1 mRNA as seen with in situ hybridization histochemistry in rat DRG (L4–6) neurons. a Oxaliplatin (6 mg/kg, i.p.) increases the co-localization between TRPA1 and p-p38 MAP kinase in rat DRG (L4–6) neurons. The photomicrographs show representative data. Co-localization of TRPA1 mRNA (green) and p-p38 MAPK (red) is shown. Double-labeled neurons (arrowheads) appear yellow in the merged panel. b Histogram shows the percent of TRPA1 mRNA-positive neurons relative to p-p38-positive neurons. TRPA1 mRNA and p-p38 co-localization was significantly higher on day 2 after oxaliplatin administration (6 mg/kg, i.p.) compared to 5 % glucose treatment. Data are the mean ± SEM. n = 4 each for 5 % glucose and oxaliplatin administration. *P < 0.05 versus 5 % glucose. c Size distribution of TRPA1 mRNA- and p-p38 MAPK-co-expressing neurons in DRGs in 5 % glucose- and oxaliplatin-treated rats. TRPA1 mRNA and p-p38 MAPK-co-expressing neurons were distributed among all sizes of neurons, but most co-expression was found in small DRG neurons. Data are the mean ± SEM. n = 4 each for 5 % glucose treatment and oxaliplatin treatment (6 mg/kg, i.p.). d Intrathecal administration of the p38 MAPK inhibitor, SB203580 (0.5 μg μL−1 h−1), significantly prevents the acute cold hypersensitivity induced by oxaliplatin (6 mg/kg, i.p.). The acetone spray test at day 2 after oxaliplatin was measured in SB203580 and 20 % DMSO treatment groups. Data are the mean ± SEM. n = 4 each for 20 % DMSO treatment and oxaliplatin treatment (6 mg/kg, i.p.). **P < 0.01 versus DMSO. ##P < 0.01 versus oxaliplatin
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Fig6: Effect of oxaliplatin on TRPA1 mRNA as seen with in situ hybridization histochemistry in rat DRG (L4–6) neurons. a Oxaliplatin (6 mg/kg, i.p.) increases the co-localization between TRPA1 and p-p38 MAP kinase in rat DRG (L4–6) neurons. The photomicrographs show representative data. Co-localization of TRPA1 mRNA (green) and p-p38 MAPK (red) is shown. Double-labeled neurons (arrowheads) appear yellow in the merged panel. b Histogram shows the percent of TRPA1 mRNA-positive neurons relative to p-p38-positive neurons. TRPA1 mRNA and p-p38 co-localization was significantly higher on day 2 after oxaliplatin administration (6 mg/kg, i.p.) compared to 5 % glucose treatment. Data are the mean ± SEM. n = 4 each for 5 % glucose and oxaliplatin administration. *P < 0.05 versus 5 % glucose. c Size distribution of TRPA1 mRNA- and p-p38 MAPK-co-expressing neurons in DRGs in 5 % glucose- and oxaliplatin-treated rats. TRPA1 mRNA and p-p38 MAPK-co-expressing neurons were distributed among all sizes of neurons, but most co-expression was found in small DRG neurons. Data are the mean ± SEM. n = 4 each for 5 % glucose treatment and oxaliplatin treatment (6 mg/kg, i.p.). d Intrathecal administration of the p38 MAPK inhibitor, SB203580 (0.5 μg μL−1 h−1), significantly prevents the acute cold hypersensitivity induced by oxaliplatin (6 mg/kg, i.p.). The acetone spray test at day 2 after oxaliplatin was measured in SB203580 and 20 % DMSO treatment groups. Data are the mean ± SEM. n = 4 each for 20 % DMSO treatment and oxaliplatin treatment (6 mg/kg, i.p.). **P < 0.01 versus DMSO. ##P < 0.01 versus oxaliplatin
Mentions: We detected TRPA1 mRNA-labeled neurons in DRG (L4–6) neurons at day 2 after oxaliplatin administration using ISHH. We compared the expression of p-p38 (red in Fig. 6a) between oxaliplatin- and 5 % glucose-treated rats. TRPA1 mRNA-labeled neurons (green in Fig. 6a) overlapped with p-p38-positive neurons. Double-labeling experiments revealed a pronounced overlap between DRG neurons expressing TRPA1 mRNA and those expressing p-p38 (yellow in Fig. 6a, merged). The percent of TRPA1 mRNA/p-p38 dual-positive neurons relative to the total p-p38-positive neurons was significantly increased at day 2 after oxaliplatin administration (36.7 ± 5.0 vs. 59.6 ± 3.9 %, P < 0.01, n = 4) (Fig. 6b). Oxaliplatin significantly increased expression of TRPA1 mRNA and p-p38 in DRG neurons. TRPA1 mRNA/p-p38 dual-positive neurons in oxaliplatin-treated rats were mainly small-sized neurons compared to 5 % glucose treatment (Fig. 6c). Thus, oxaliplatin increased the number of small-diameter DRG neurons that co-expressed TRPA1 mRNA and p-p38.Fig. 6

Bottom Line: Oxaliplatin significantly increased co-localization of TRPA1 expression and isolectin B4 binding in DRG neurons.In addition, we found that intrathecal administration of TRPA1 antisense, but not TRPA1 mismatched oligodeoxynucleotides, knocked down TRPA1 expression and decreased oxaliplatin-induced cold hyperalgesia.Intrathecal administration of the p38 MAPK inhibitor, SB203580, significantly decreased oxaliplatin-induced acute cold hypersensitivity.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicinal Pharmacology, Showa Pharmaceutical University, 3-3165 Higashitamagawagakuen, Machida, Tokyo, 194-8543, Japan. k-yamamoto@ac.shoyaku.ac.jp.

ABSTRACT

Background: Peripheral cold neuropathic pain is a serious side effect of oxaliplatin treatment. However, the mechanism of oxaliplatin-induced cold hyperalgesia is unknown. In the present study, we investigated the effects of oxaliplatin on transient receptor potential ankyrin 1 (TRPA1) in dorsal root ganglion (DRG) neurons of rats.

Results: Behavioral assessment using the acetone spray test showed that 3 and 6 mg/kg oxaliplatin (i.p.) induced acute cold hypersensitivity after 1, 2, 4, and 7 days. Real-time PCR showed that oxaliplatin (6 mg/kg) significantly increased TRPA1 mRNA expression in DRGs at days 1, 2, and 4. Western blotting revealed that oxaliplatin significantly increased TRPA1 protein expression in DRGs at days 2, 4, and 7. Moreover, in situ hybridization histochemistry revealed that most TRPA1 mRNA-labeled neurons in the DRGs were small in size. Oxaliplatin significantly increased co-localization of TRPA1 expression and isolectin B4 binding in DRG neurons. Oxaliplatin induced a significant increase in the percent of TRPA1 mRNA-positive small neurons in DRGs at days 1, 2, and 4. In addition, we found that intrathecal administration of TRPA1 antisense, but not TRPA1 mismatched oligodeoxynucleotides, knocked down TRPA1 expression and decreased oxaliplatin-induced cold hyperalgesia. Double labeling showed that p-p38 mitogen-activated protein kinase (MAPK) was co-expressed in TRPA1 mRNA-labeled neurons at day 2 after oxaliplatin administration. Intrathecal administration of the p38 MAPK inhibitor, SB203580, significantly decreased oxaliplatin-induced acute cold hypersensitivity.

Conclusions: Together, these results demonstrate that TRPA1 expression via activation of p38 MAPK in DRG neurons, at least in part, contributes to the development of oxaliplatin-induced acute cold hyperalgesia.

No MeSH data available.


Related in: MedlinePlus