Journal: Anesthesiology

Article Title: Transcriptional changes in dorsal spinal cord persist after surgical incision despite preemptive analgesia with peripheral resiniferatoxin

doi: 10.1097/ALN.0000000000002006

Figure Lengend Snippet: Gene changes after surgical incision and with preemptive resiniferatoxin based analgesia. Mean gene expression (sFPKM) is expressed for the four experimental groups and fold change is shown for the ratio of expression from control DH tissue to DH tissue from surgically manipulated animals. In order to increase power, experimental groups were grouped as follows for determination of statistical significance: 1) control DH with DH contralateral to surgical incision and 2) DH ipsilateral to surgical incision with and without resiniferatoxin. The threshold defining significant differentially expressed genes for the MAGIC Differential Gene analysis was set to a DEG Score of 65 to maintain the overall false discovery rate below 5% without exceeding an incremental false discovery rate of 20% upon addition of potential differentially expressed genes. See also Supplemental Digital Content 3 .

Article Snippet: The slides were incubated in primary antibody (Iba1, 1:2,000, Abcam, Cambridge, MA) diluted in antibody diluent (1% BSA, 0.05% sodium azide, and 0.1% Tween-20 in PBS) for 1 hour.

Techniques: Expressing

Journal: Anesthesiology

Article Title: Transcriptional changes in dorsal spinal cord persist after surgical incision despite preemptive analgesia with peripheral resiniferatoxin

doi: 10.1097/ALN.0000000000002006

Figure Lengend Snippet: Changes in prodynorphin (Pdyn) and microglial markers. A) Of opioid receptors and peptides, only the transcript for Pdyn, which for prodynorphin, is upregulated. B) Differentially expressed genes were correlated with a public database27 of cell specific gene expression. Tissue-specific expression marker genes were defined as 3-fold increased expression over other cell types. C) Incision mainly upregulated genes that are markers of microglia even after peri-operative resiniferatoxin administration. D) Transcripts for microglial specific marker Itgal are increased after surgical incision with or without resiniferatoxin administration. Microglial activation markers Itgam (E) and Aif1 (F) are increased after surgical incision. G) Dorsal horn immunohistochemical staining for Iba1 confirms microglial activation on POD5 (outline of spinal gray matter shown by dotted line). H) The ipsilateral dorsal spinal cord contained significantly more Iba1+ cells in lamina I and II relative to the contralateral side (p=0.015 for ipsi vs contra, two-way repeated measures ANOVA, n=3 for both treatment groups, data plotted as mean with SD). For (A, D, E, F) * = Score above threshold by MAGIC Differential Gene analysis, # = member gene of the module correlating with surgery from WGCNA analysis.

Article Snippet: The slides were incubated in primary antibody (Iba1, 1:2,000, Abcam, Cambridge, MA) diluted in antibody diluent (1% BSA, 0.05% sodium azide, and 0.1% Tween-20 in PBS) for 1 hour.

Techniques: Expressing, Marker, Activation Assay, Immunohistochemical staining, Staining