Journal: Molecular Pain

Article Title: An LPA species (18:1 LPA) plays key roles in the self-amplification of spinal LPA production in the peripheral neuropathic pain model

doi: 10.1186/1744-8069-9-29

Figure Lengend Snippet: Blockade of nerve injury-induced cPLA 2 and iPLA 2 activations. ( a and b ) Activation of spinal cPLA 2 (panel a ) and iPLA 2 (panel b ) were detected by cPLA 2 and iPLA 2 activity assays at defined time points after nerve injury. The capital letter “C” represents the control group (naive mice). ( c and d ) After pre-treatments of vehicle, MK-801, CP-99994, AACOCF3, BEL (each 10 nmol, i.t.) and minocycline (30 mg/ml, i.p.) before nerve injury, the activities of spinal cPLA2 (panel c ) and iPLA2 (panel d ) at 1 h after injury were evaluated. The “veh”, “MK”, “CP”, “AA” and “mino” represent vehicle, MK-801, CP-99994, AACOCF3 and minocycline, respectively. ( e and f ) Activities of cPLA 2 (panel e ) and iPLA 2 (panel f ) were measured at 1 h after injury using the Lpar1 - and Lpar3 -deficient mice. “WT” represents the wide-type mice. ( g ) PLA 1 activity in the spinal dorsal horn was measured by PLA 1 activity assay at time-course points after nerve injury. Data represent means ± SEM from experiments using 3-6 mice. * p < 0.05, versus with the control group; # p < 0.05, versus with the vehicle/WT-injury group.

Article Snippet: The protein concentration of the supernatant was determined by the Lowry method, and the assays were performed using a cPLA 2 assay kit (Cayman Chemicals, Ann Arbor, MI, USA) to evaluate the cPLA 2 activity or a modified cPLA 2 assay kit (Cayman Chemicals, Ann Arbor, MI, USA) to evaluate the iPLA 2 activity, as described previously [ ].

Techniques: Activation Assay, Activity Assay

Journal: Molecular Pain

Article Title: An LPA species (18:1 LPA) plays key roles in the self-amplification of spinal LPA production in the peripheral neuropathic pain model

doi: 10.1186/1744-8069-9-29

Figure Lengend Snippet: Cell-type identification of activated cPLA 2 in spinal cord. ( a-f ) Immunohistochemical double labeling of p-cPLA 2 (green) and NeuN (red) as well as p-cPLA 2 (green) and Iba1 (red) in the spinal cord at 1 h after nerve injury. White arrows represent NeuN- or Iba1-co-localized p-cPLA 2 signals. ( g ) Low magnification of double immunostaining with p-cPLA 2 (green) and NeuN (red) in the spinal cord at 1 h after injury. ( h ) Diagram of Figure g. Red dots represent NeuN co-localized p-cPLA 2 signals. Scale bar (including high and low magnifications), 50 μm.

Article Snippet: The protein concentration of the supernatant was determined by the Lowry method, and the assays were performed using a cPLA 2 assay kit (Cayman Chemicals, Ann Arbor, MI, USA) to evaluate the cPLA 2 activity or a modified cPLA 2 assay kit (Cayman Chemicals, Ann Arbor, MI, USA) to evaluate the iPLA 2 activity, as described previously [ ].

Techniques: Immunohistochemical staining, Labeling, Double Immunostaining

Journal: Molecular Pain

Article Title: An LPA species (18:1 LPA) plays key roles in the self-amplification of spinal LPA production in the peripheral neuropathic pain model

doi: 10.1186/1744-8069-9-29

Figure Lengend Snippet: Blockade of p-cPLA 2 signals in spinal neurons after nerve injury. ( a-i ) Vehicle, MK-801, CP-99994, AACOCF3, BEL (each 10 nmol, i.t.) and minocycline (30 mg/ml, i.p.) were pre-treated before nerve injury, and the spinal cord were collected at 1 h after injury, which were used for the double immunohistochemistry with p-cPLA 2 (green) and NeuN (red). Control as well as Lpar1 - and Lpar3 -deficient mice were also used in this experiment. White arrows represent NeuN-co-localized p-cPLA 2 signals. The capital letter “C” and “I” represent control and nerve injury, respectively. The “veh”, “MK”, “CP”, “AA” and “mino” represent vehicle, MK-801, CP-99994, AACOCF3 and minocycline, respectively. Scale bar, 50 μm.

Article Snippet: The protein concentration of the supernatant was determined by the Lowry method, and the assays were performed using a cPLA 2 assay kit (Cayman Chemicals, Ann Arbor, MI, USA) to evaluate the cPLA 2 activity or a modified cPLA 2 assay kit (Cayman Chemicals, Ann Arbor, MI, USA) to evaluate the iPLA 2 activity, as described previously [ ].

Techniques: Immunohistochemistry