Journal: Frontiers in Molecular Neuroscience

Article Title: Endoplasmic Reticulum Stress in Spinal Cord Contributes to the Development of Morphine Tolerance

doi: 10.3389/fnmol.2018.00072

Figure Lengend Snippet: Primers for Real-time PCR.

Article Snippet: According to the previous literatures ( , ), recombinant shRNA lentiviral vectors targeting rat ATF6 gene (ATF6 RNAi-LV) and non-specific control lentivirus (NC-LV) were designed by Genechem (Shanghai, China).

Techniques: Sequencing

Journal: Frontiers in Molecular Neuroscience

Article Title: Endoplasmic Reticulum Stress in Spinal Cord Contributes to the Development of Morphine Tolerance

doi: 10.3389/fnmol.2018.00072

Figure Lengend Snippet: Expression of ATF6 in spinal cord. (A,B) The expression of ATF6 were significantly increased in morphine-tolerant rats measured by real-time PCR and western blots, respectively. ( ∗∗ p < 0.01, ∗∗∗ p < 0.001 compared with naive rats, Real-time PCR: n = 4 in each group; western blots: n = 6 in each group). (C) Immunostaining of ATF6 in spinal dorsal horn. ATF6 was extensively expressed in spinal dorsal horn. The immunoreactivity of ATF6 was significantly increased in morphine-tolerant rats. ( ∗∗ p < 0.01 compared with naive rats, n = 3 in each group, scale bar: 200 μm). (D) Double immunostaining of ATF6 and cell-specific markers in morphine-tolerant rats. ATF6 was co-localized with NeuN (indicated by arrows), not with GFAP or Iba1 in spinal dorsal horn. Scale bar: 50 μm. (E) Chronic morphine administration results in ATF6 translocation to the nucleus. ATF6 was only expressed in cytoplasm in naive rats (indicated by arrows), while ATF6 was expressed in cytoplasm and cell nucleus in morphine-tolerant rats (indicated by arrows). Scale bar: 50 μm. ATF6, activating transcription factor 6; NS, normal saline; MT, morphine tolerance.

Article Snippet: According to the previous literatures ( , ), recombinant shRNA lentiviral vectors targeting rat ATF6 gene (ATF6 RNAi-LV) and non-specific control lentivirus (NC-LV) were designed by Genechem (Shanghai, China).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunostaining, Double Immunostaining, Translocation Assay, Saline

Journal: Frontiers in Molecular Neuroscience

Article Title: Endoplasmic Reticulum Stress in Spinal Cord Contributes to the Development of Morphine Tolerance

doi: 10.3389/fnmol.2018.00072

Figure Lengend Snippet: Effect of ATF6 RNAi-Lentivirus on the development of morphine tolerance. (A) Detection of lentivirus transfection in spinal cord. RNAi-LV or NC-LV was injected into lumbar spinal cord 3 days before intrathecal catheterization, respectively. The expression of enhanced green fluorescent protein in lentiviral vectors was detected in spinal dorsal horn n = 3 in each group. Scale bar: 100 μm. (B,C) RNAi-LV could effectively downregulate the expression of ATF6 induced by chronic morphine treatment measured by real-time PCR and western blots, respectively. ( ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 compared with naive rats, # p < 0.05, ## p < 0.01 compared with MT+NC-LV group, Real-time PCR: n = 4 in each group; western blots: n = 6 in each group). (D) Pretreatment with RNAi-LV could attenuate the development of morphine tolerance. ( ∗∗ p < 0.01 compared with MT group; ## p < 0.01 compared with MT+NC-LV group, n = 6–8 in each group). NS, normal saline; MT, morphine tolerance; ATF6, activating transcription factor 6; %MPE, percentage of maximal possible antinociceptive effect.

Article Snippet: According to the previous literatures ( , ), recombinant shRNA lentiviral vectors targeting rat ATF6 gene (ATF6 RNAi-LV) and non-specific control lentivirus (NC-LV) were designed by Genechem (Shanghai, China).

Techniques: Transfection, Injection, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Saline