Adult male C57Bl6 mice were utilized in all experiments (20–25 g, Taconic Farms, Derwood, MD). Mice were group housed and received food and water ad libitum with a 12:12 h light cycle. A total of 111 male mice were used for this study; 13 mice were removed from the study due to post-surgical complications. All experiments complied fully with the principles set forth in the “Guide for the Care and Use of Laboratory Animals” and were approved by the Uniformed Services University IACUC.
All subjects undergoing surgery received isoflurane (Primal Healthcare, Andhra Pradesh, India). Mice received a laminectomy, followed by a contusion simulating moderate SCI using the Infinite Horizons Impactor (50 kdyn; Precision Systems and Instrumentation, Fairfax Station, VA). Mice were immediately given an intrathecal injection of either gp91ds-tat or scrambled ds-tat (AnaSpec, Inc., Fremont, CA) diluted to 50 μM in saline in a 5 μl volume at the lesion epicenter. After expelling the liquid, the needle was held under the dura for 30 s prior to removal. The incision was then closed, and animals were maintained on heating pads until mice regained movement. Acetaminophen (Children’s Tylenol, 200 mg/kg) was added to drinking water for 72 h post-injury. Manual bladder expression was performed daily until normal bladder expression returned. Naïve mice did not undergo surgery or receive isoflurane.
All subjects undergoing surgery received isoflurane (Primal Healthcare, Andhra Pradesh, India). Mice received a laminectomy, followed by a contusion simulating moderate SCI using the Infinite Horizons Impactor (50 kdyn; Precision Systems and Instrumentation, Fairfax Station, VA). Mice were immediately given an intrathecal injection of either gp91ds-tat or scrambled ds-tat (AnaSpec, Inc., Fremont, CA) diluted to 50 μM in saline in a 5 μl volume at the lesion epicenter. After expelling the liquid, the needle was held under the dura for 30 s prior to removal. The incision was then closed, and animals were maintained on heating pads until mice regained movement. Acetaminophen (Children’s Tylenol, 200 mg/kg) was added to drinking water for 72 h post-injury. Manual bladder expression was performed daily until normal bladder expression returned. Naïve mice did not undergo surgery or receive isoflurane.