Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: Cytosolic and mitochondrial calcium concentration of TRPA1- or TRPV1-positive DRG neurons during exposure to 10 µM cisplatin. ( A ) Immunostaining of transient receptor potential ankyrin 1 (TRPA1)- or transient receptor potential vanilloid 1 (TRPV1)-positive DRG neurons and live imaging of Fluo-4 (green) and Rhod-2 (red). ( B ) The cytosolic calcium of TRPV1-positive DRG neurons concentration instantly increased to 1.067 ± 0.018 (* p < 0.05) after 5 min and continuously increased to 1.394 ± 0.068 (** p < 0.01) after 70 min. The mitochondrial calcium concentration declined to 1.095 ± 0.009 (* p < 0.05) after 5 min for TRPV1-positive DRG neurons and steadily declined to 0.577 ± 0.044 after 70 min (** p < 0.01). ( C ) During the exposure of 10 µM cisplatin, the cytosolic calcium concentration of TRPA1-positive neurons increased after 5 min to 1.024 ± 0.011 (** p < 0.01) until 70 min at 1.240 ± 0.074 (*** p < 0.001). The mitochondrial calcium concentration was lower after 10 min at 0.928 ± 0.029 (*** p < 0.001) until it reached 0.790 ± 0.050 at 70 min (*** p < 0.001). ( D ) The cytosolic calcium concentration of TRPV1-positive DRG neurons was increased after 15 min to 65 min compared to TRPA1-positive DRG neurons (* p < 0.05, ** p < 0.01). ( E ) The mitochondrial calcium concentration of TRPA1- or TRPV1-positive DRG neurons was different after 5 min until the end of the experiment (* p < 0.05, ** p < 0.01). TRPV1-positive sensory neurons had a lower mitochondrial calcium concentration during exposure to 10 µM cisplatin after 15 min (* p < 0.05, ** p < 0.01). n = 6 cells per condition. * = significant effect between TRPA1+ and TRPV1+ DRG neurons. Scale = 50 µm.

Article Snippet: The cells were first stained with extracellular TRPA1 or TRPV1 antibodies (Alomone Labs, Jerusalem, Israel).

Techniques: Concentration Assay, Immunostaining, Imaging

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: Cytosolic and mitochondrial calcium concentration of TRPA1- or TRPV1-positive DRG neurons during exposure to 10 µM oxaliplatin. ( A ) Immunostaining of TRPA1- or TRPV1-positive DRG neurons and live imaging of Fluo-4 (green) and Rhod-2 (red) before and after application of 10 µM cisplatin or oxaliplatin. ( B ) TRPA1-positive DRG neurons showed an immediate increase of the cytosolic calcium concentration to 1.037 ± 0.006 (** p < 0.01) after 5 min and continuously increased to 1.372 ± 0.062 after 70 min (*** p < 0.001). The mitochondrial calcium concentration decreased after 5 min to 0.967 ± 0.008 (*** p < 0.001) and declined to 0.712 ± 0.010 after 70 min (*** p < 0.001). ( C ) The TRPV1-positive DRG neurons showed an immediate increase of the cytosolic calcium concentration to 1.049 ± 0.005 (*** p < 0.001) after 5 min and increased to 1.480 ± 0.034 after 70 min (*** p < 0.001). The mitochondrial calcium concentration decreased to 0.959 ± 0.020 (* p < 0.05) after 5 min and to 0.725 ± 0.077 (** p < 0.01) after 70 min (* p < 0.05). ( D ) No difference could be determined in the cytosolic calcium concentration of TRPA1- or TRPV1-positive DRG neurons ( p > 0.05). ( E ) No difference could be determined in the mitochondrial calcium concentration of TRPA1- or TRPV1-positive DRG neurons ( p > 0.05). n = 6 cells per condition. * = significant effect between TRPA1+ and TRPV1+ DRG neurons. Scale = 50 µm.

Article Snippet: The cells were first stained with extracellular TRPA1 or TRPV1 antibodies (Alomone Labs, Jerusalem, Israel).

Techniques: Concentration Assay, Immunostaining, Imaging

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: Relative cytosolic and mitochondrial calcium concentration during exposure to 10 µM cis- or oxaliplatin of TRPA1- or TRPV1-positive DRG neurons. ( A ) No differences in the cytosolic calcium concentration of TRPA1-positive DRG neurons after exposure to 10 µM cis- or oxaliplatin could be determined ( p > 0.05). ( B ) The cytosolic calcium of TRPV1-positive DRG neurons concentration during exposure to cis- or oxaliplatin showed no difference ( p > 0.05). ( C ) During exposure to 10 µM cis- or oxaliplatin, no difference in the mitochondrial calcium concentration of TRPA1-positive DRG neurons could be determined ( p > 0.05). ( D ) While exposing TRPV1-positive DRG neurons, differences between the two chemotherapeutics on the mitochondrial calcium concentration could be determined after 20 min. The mitochondrial calcium concentration of TRPV1-positive DRG neurons exposed to oxaliplatin was higher at 0.856 ± 0.033 compared to 0.727 ± 0.040 (* p < 0.05). The mitochondrial calcium concentration was higher for oxaliplatin-exposed TRPV1-positive DRG neurons until 0.786 ± 0.058 after 40 min (* p < 0.05). n = 6 cells per condition. * = significant effect between cis- and oxaliplatin, ** p < 0.01

Article Snippet: The cells were first stained with extracellular TRPA1 or TRPV1 antibodies (Alomone Labs, Jerusalem, Israel).

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: ROS production of TRPA1- or TRPV1-positive DRG neurons during exposure to 10 µM cisplatin. ( A ) Fluorescence of CellRox (green) in TRPA1- or TRPV1-positive DRG neurons during exposure to 10 µM cisplatin at different time points. ( B ) TRPA1-positive sensory neurons showed an instant increase of the ROS production to 1.431 ± 0.042 (*** p < 0.001) after 5 min and reached its peak at 2.100 ± 0.197 after 30 min (*** p < 0.001). After 50 min, the ROS production dropped below the control level at 50 min to 0.607 ± 0.036 (*** p < 0.001) and further declined to 0.370 ± 0.035 (*** p < 0.001) after 70 min. ( C ) After exposure to 10 µM cisplatin of the TRPV1-positive DRG neurons, the ROS production increased to 1.238 ± 0.042 (*** p < 0.001) after 5 min and dropped below control level at 50 min at 0.873 ± 0.068 (* p < 0.05) and steadily declined to 0.756 ± 0.072 (** p < 0.01) after 70 min. n = 6 cells per condition. * = significant effect compared to control level. Scale = 50 µm.

Article Snippet: The cells were first stained with extracellular TRPA1 or TRPV1 antibodies (Alomone Labs, Jerusalem, Israel).

Techniques: Fluorescence, Control

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: ROS production of TRPA1- or TRPV1-positive DRG neurons during exposure to 10 µM oxaliplatin. ( A ) Fluorescence of CellRox (green) of TRPA1- or TRPV1-positive DRG neurons at different time points. ( B ) The ROS production of TRPA1-positive DRG neurons increased after 5 min to 1.421 ± 0.103 (** p < 0.01) during exposure to 10 µM oxaliplatin and dropped to control level after 40 min at 1.31 ± 0.194 ( p > 0.05). ( C ) The TRPV1-positive sensory neurons had an increased ROS production after 5 min at 1.080 ± 0.020 (** p < 0.01) and dropped to the control level after 10 min at 1.033 ± 0.036 ( p > 0.05). After 30 min, the ROS production declined below the control level at 0.810 ± 0.079 (* p < 0.05) and steadily declined to 0.429 ± 0.050 after 70 min (*** p < 0.001). n = 6 cells per condition. * = significant effect compared to control level. Scale = 20 µm.

Article Snippet: The cells were first stained with extracellular TRPA1 or TRPV1 antibodies (Alomone Labs, Jerusalem, Israel).

Techniques: Fluorescence, Control

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: The ROS production of TRPA1- or TRPV1-positive DRG neurons during exposure to cis- or oxaliplatin. ( A ) A difference could be determined for TRPA1-positive DRG neurons at 30 min with a higher ROS production for cisplatin-exposed DRG neurons at 2.100 ± 0.197 (* p < 0.05). After 45 min (1.194 ± 0.223; * p < 0.05) until 70 min (1.022 ± 0.233; * p < 0.05), the ROS production was higher for oxaliplatin-exposed neurons. ( B ) The ROS production of TRPV1-positive DRG neurons was higher for cisplatin-exposed neurons over the entire experimental period (* p < 0.05, ** p < 0.01). n = 6 cells per condition. * = significant effect compared to control level.

Article Snippet: The cells were first stained with extracellular TRPA1 or TRPV1 antibodies (Alomone Labs, Jerusalem, Israel).

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: Antibodies and dilution used for the TRPA1- and TRPV1-positive staining.

Article Snippet: The cells were first stained with extracellular TRPA1 or TRPV1 antibodies (Alomone Labs, Jerusalem, Israel).

Techniques: Staining

Journal: Molecular Pain

Article Title: HC-030031, a TRPA1 selective antagonist, attenuates inflammatory- and neuropathy-induced mechanical hypersensitivity

doi: 10.1186/1744-8069-4-48

Figure Lengend Snippet: The effect of TRPA1 antagonism on AITC -induced nocifensive behaviors . Both 100 mg/kg (red bar) and 300 mg/kg (blue bar) of HC-030031 significantly reduced lifting behavior relative to vehicle treated rats given plantar injection of 1% AITC (white bar; * p, 0.05).

Article Snippet: Full length human TRPA1 cDNA was purchased from Origene Technologies (Rockville, MD) and subcloned into pcDNA5/FRT/V5-His TOPO TA vector (Invitrogen, Carlsbad, CA).

Techniques: Injection

Journal: Molecular Pain

Article Title: HC-030031, a TRPA1 selective antagonist, attenuates inflammatory- and neuropathy-induced mechanical hypersensitivity

doi: 10.1186/1744-8069-4-48

Figure Lengend Snippet: The effect of TRPA1 antagonism on complete Freund's adjuvant induced mechanical hypersensitivity . A. Paw withdrawal thresholds for the injected hind paw were assessed using the Randall-Selitto device prior to intraplantar CFA injections, prior to dosing with vehicle (white bars), 20 mg/kg naproxen (green bars), 100 mg/kg HC-030031 (red bars) or 300 mg/kg HC-030031 (blue bars), and at one hour post p.o. dosing. Naproxen, 100 mg/kg and 300 mg/kg HC-030031 produced a significant attenuation in mechanical hypersensitivity at one hour post dose (*p < 0.05). B. Paw withdrawal thresholds for the noninjected hind paw assessed using the Randall-Selitto device prior to intraplantar CFA injections, prior to dosing with vehicle (white bars), 20 mg/kg naproxen (green bars), 100 mg/kg HC-030031 (red bars) or 300 mg/kg HC-030031 (blue bars), and at one hour post p.o. dosing. C. Percent reversal seen at one hour post p.o. dosing with vehicle (white bars), 20 mg/kg naproxen (green bars), 100 mg/kg HC-030031 (red bars) or 300 mg/kg HC-030031 (blue bars).

Article Snippet: Full length human TRPA1 cDNA was purchased from Origene Technologies (Rockville, MD) and subcloned into pcDNA5/FRT/V5-His TOPO TA vector (Invitrogen, Carlsbad, CA).

Techniques: Adjuvant, Injection, Randall–Selitto Test, Produced

Journal: Pain Research & Management

Article Title: Exploring the Analgesic Initiation Mechanism of Tuina in the Dorsal Root Ganglion of Minor CCI Rats via the TRPV1/TRPA1-cGMP Pathway

doi: 10.1155/2024/2437396

Figure Lengend Snippet: Primary and secondary antibodies used for Western blotting.

Article Snippet: Primary , Rabbit anti-TRPA1 polyclonal IgG , 1 : 1000 , Proteintech Group, Chicago, USA.

Techniques: Western Blot, Concentration Assay

Journal: PLoS ONE

Article Title: Pharmacogenetic inhibition of lumbosacral sensory neurons alleviates visceral hypersensitivity in a mouse model of chronic pelvic pain

doi: 10.1371/journal.pone.0262769

Figure Lengend Snippet: Primary and secondary antibodies used.

Article Snippet: Rabbit polyclonal anti-TRPA1 , St John’s Laboratory , STJ193121 , 1:100.

Techniques: