apetx2  (Alomone Labs)

Journal: Frontiers in Neuroscience

Article Title: Involvement of Acid-Sensing Ion Channel 1b in the Development of Acid-Induced Chronic Muscle Pain

doi: 10.3389/fnins.2019.01247

Figure Lengend Snippet: Effect of amiloride, mambalgin-1, APETx2 and PcTx1 on ASIC1b-expressing muscle afferent DRG neurons. (A) Whole-cell patch clamp recording on an ASIC1b-expressing DRG neuron projecting to gastrocnemius muscle labeled by fluorogold. (B) Mambalgin-1 (MB-1) (1 μM) inhibited acid (pH 5.0)-induced currents in 13 of 14 ASIC1b-expressing muscle afferent DRG neurons. (C) APETx2 (1 μM) inhibited acid (pH 5.0)-induced currents in 6 of 13 ASIC1b-expressing muscle afferent DRG neurons. (D) PcTx1 (100 nM) inhibited acid (pH 5.0)-induced currents in 5 of 11 ASIC1b-expressing muscle afferent DRG neurons.

Article Snippet: Mambalgin-1, APETx2, and psalmotoxin 1 (PcTx1) were purchased from Alomone Labs and prepared by autoclaved water in stock solutions of 200 μM, 1 mM, and 200 μM respectively.

Techniques: Expressing, Patch Clamp, Labeling

Journal: British Journal of Pharmacology

Article Title: Acid‐sensing ion channel 3 blockade inhibits durovascular and nitric oxide‐mediated trigeminal pain. Acid‐sensing ion channel 3 blockade inhibits durovascular and nitric oxide‐mediated trigeminal pain

doi: 10.1111/bph.14990

Figure Lengend Snippet: APETx2 inhibits nitroglycerin‐evoked periorbital mechanosensitivity in mice. There was a significant overall difference in periorbital mechanosensitivity across all groups (a). Acute nitroglycerin (NTG) induced a decreased periorbital mechanosensitivity in mice that was partially reversed by pretreatment with APETx2. Chronic NTG administration in mice (NTG‐primed) produced a basal periorbital mechanosensitivity (b) that reached significance from Day 5 when compared to vehicle control mice. Following withdrawal of NTG periorbital mechanosensitivity returned to that of the non‐sensitised vehicle control group. Subsequent exposure to bright light stress (BLS) resulted in a significant overall difference in periorbital mechanosensitivity (c). NTG‐primed mice demonstrated an increased periorbital mechanosensitivity when compared to vehicle control and APETx2 treated mice. The periorbital mechanosensitivity evoked in response to BLS was blocked by pretreatment with APETx2, returning to that of vehicle control and APETx2 treated mice. Timeline of latent sensitisation to BLS protocol (d). Animals were sensitised with chronic administration of NTG (10 mg·kg −1 , i.p.) every second day for 9 days. Periorbital mechanical sensitivity was assessed with the von Frey assay (black circles) before and 2‐hr post NTG treatment on Days 1, 5, and 9. Animals were recovered for 1 week and then on Day 17 mechanical sensitivity was reassessed prior to 1 hr of BLS. On Day 18, animals were treated with either APETx2 (0.23 mg·kg −1 ) or vehicle control 30 min prior to and again 30 min post BLS. Mechanical sensitivity was then assessed 2‐hr post BLS. * P

Article Snippet: 2.6 Materials APETx2 (Alomone Labs, Israel) was dissolved in water to 500 μg·ml−1 and diluted in 0.9% saline.

Techniques: Mouse Assay, Produced

Journal: British Journal of Pharmacology

Article Title: Acid‐sensing ion channel 3 blockade inhibits durovascular and nitric oxide‐mediated trigeminal pain. Acid‐sensing ion channel 3 blockade inhibits durovascular and nitric oxide‐mediated trigeminal pain

doi: 10.1111/bph.14990

Figure Lengend Snippet: APETx2 reduces durovascular nociceptive‐evoked and spontaneous neuronal activity in the trigeminal nucleus caudalis (TNC). Experimental set‐up in the rat (a). Durovascular nociceptive afferents arising in the trigeminal ganglion (TG) are activated via stimulation of the dura mater surrounding the middle meningeal artery (MMA). Durovascular‐evoked responses are then recorded in the TNC. TNC durovascular nociceptive‐evoked neuronal responses are significantly reduced following APETx2 (b), starting from 45 min post infusion and remain significantly reduced at 1 hr (c). Examples of TNC spontaneous neuronal activity following control (d) and APETx2 infusion (e) that was significantly decreased over the 1‐hr recording window in APETx2 (f), but not vehicle control treated rats compared to baseline. * P

Article Snippet: 2.6 Materials APETx2 (Alomone Labs, Israel) was dissolved in water to 500 μg·ml−1 and diluted in 0.9% saline.

Techniques: Activity Assay

Journal: British Journal of Pharmacology

Article Title: Acid‐sensing ion channel 3 blockade inhibits durovascular and nitric oxide‐mediated trigeminal pain. Acid‐sensing ion channel 3 blockade inhibits durovascular and nitric oxide‐mediated trigeminal pain

doi: 10.1111/bph.14990

Figure Lengend Snippet: APETx2 blocks NO‐induced sensitisation to noxious stimuli. There was a significant overall difference in durovascular‐evoked trigeminal nucleus caudalis neuronal activation across all groups. Sodium nitroprusside (SNP) induced increased durovascular‐evoked responses (a) when compared to vehicle control treated rats. Pretreatment with APETx2 significantly reduced the SNP‐induced increase in durovascular‐evoked responses (a). There was a significant overall difference in noxious pinch‐evoked trigeminal nucleus caudalis neuronal activation (b) across all groups. Sodium nitroprusside induced a non‐significant modest increase in noxious pinch‐evoked responses (b) when compared to vehicle control treated rats. Pretreatment with APETx2 significantly reduced the noxious pinch‐evoked responses when compared to SNP treated rats (b). * P

Article Snippet: 2.6 Materials APETx2 (Alomone Labs, Israel) was dissolved in water to 500 μg·ml−1 and diluted in 0.9% saline.

Techniques: Activation Assay

Journal: Journal of Biomedical Science

Article Title: Local ASIC3 modulates pain and disease progression in a rat model of osteoarthritis

doi: 10.1186/1423-0127-19-77

Figure Lengend Snippet: Fast Blue labeling and immunohistochemistry staining for ASIC3 : (a-b) Naïve- model, (c-d) OA-model, (e-f) APETx2 administration to OA-model in early phase. Photos in each row are the same DRG. In ( b ),( d ),( f ), large arrows indicate Fast Blue labeled, ASIC3 immunoreactive (ASIC3-ir) DRG cells, while ASIC3-ir cells that were not labeled by Fast Blue are indicated by small arrowheads. More than 100 FB-labeled neurons were analyzed from 4 rats in each group. The percentage of ASIC3-ir knee joint afferents was 18 ± 3% (mean ± SD) in naïve models, 46 ± 4% in OA-models ( p = 0.003), and 20 ± 5% in the early-phase APETx2 group ( p = 0.006), respectively. Scale bar: 50 μm.

Article Snippet: In this study, APETx2 was effective only in the early phase because it worked against initial inflammation before the severe cartilage damage.

Techniques: Labeling, Immunohistochemistry, Staining

Journal: Journal of Biomedical Science

Article Title: Local ASIC3 modulates pain and disease progression in a rat model of osteoarthritis

doi: 10.1186/1423-0127-19-77

Figure Lengend Snippet: Histological evaluation of knee joint using Modified Mankin Score. a scale of 0–13 (from 0 (worst) to 13 (best)). Intra-articular injection of APETx2 in early phase prevented OA progression, including breakdown of articular surface and hypocellularity. * p

Article Snippet: In this study, APETx2 was effective only in the early phase because it worked against initial inflammation before the severe cartilage damage.

Techniques: Modification, Injection

Journal: Journal of Biomedical Science

Article Title: Local ASIC3 modulates pain and disease progression in a rat model of osteoarthritis

doi: 10.1186/1423-0127-19-77

Figure Lengend Snippet: Histology of knee joints with Safranin O staining at Day14. Three different magnifications (×1.25, ×10, ×20) were shown in each group. ( a-c ) Naïve model: Full thickness of cartilage. Rich chondrocytes with proteoglycan (red staining by safranin O) ( d-f ) OA model: Severe damage of cartilage surface with loss of chondrocytes in superficial and middle layer (#), hypertrophied chondrocytes in deep zone (white arrow) were observed in ( f ) . Increased thickening of subchondral bone subjacent to the area of severe cartilage lesion was also observed in ( d,e ). ( g-i ) APETx2 administration in early phase: Chondrocytes were well observed in superficial and middle layer (black arrow). Although proteoglycan loss, cartilage surface kept smooth and no apparent thinning (*) in ( i ). ( j-l ) APETx2 in late phase: same findings as OA. Apparent chondroprotective effect was not seen (#) and hypertrophied chondrocytes in deep zone were also observed (white arrow) in ( l ). F: femur, M: meniscus, T: tibia, Scale bar: 1 mm, 100 μm 50 μm.

Article Snippet: In this study, APETx2 was effective only in the early phase because it worked against initial inflammation before the severe cartilage damage.

Techniques: Staining

Journal: Journal of Biomedical Science

Article Title: Local ASIC3 modulates pain and disease progression in a rat model of osteoarthritis

doi: 10.1186/1423-0127-19-77

Figure Lengend Snippet: The effect of intra-articular injection of APETx2 on behavior tests. ( a )weight distribution, ( b ) paw withdrawal reflex (same manner as Figure 1 ). Weight distribution was changed significantly at Day3 in early APETx2 administration group. Frequency of paw withdrawal reflex i.e. secondary hyperalgesia reduced with APETx2 injection. The inhibitory effects on secondary hyperalgesia in both early- and late-phase groups were observed at Day14. * p

Article Snippet: In this study, APETx2 was effective only in the early phase because it worked against initial inflammation before the severe cartilage damage.

Techniques: Injection