With neither protocol did the MGE cell transplantation cells alter baseline mechanical thresholds in the absence of nerve injury. Several studies demonstrated that a peripheral nerve injury reduces GABAergic signaling, PD0332991 clinical trial including a reduction of the expression of GAD, the biosynthetic enzyme for GABA (Moore et al., 2002). Here, we used quantitative PCR to determine whether MGE transplants alter GAD mRNA levels, relative to those in nerve-injured animals, where a decrease was predicted. For this analysis, we transplanted MGE cells 1 week

after SNI and assayed both GAD65 and 67 levels 1 week later. We chose this time point as Moore et al. (2002) reported that GAD protein levels were significantly reduced 2 weeks after nerve injury. Figure 8B demonstrates that compared to uninjured animals, nerve injury induced a small but significant decrease in the spinal cord levels of GAD65 mRNA, ipsilateral to the injury; GAD67 levels did not differ. By contrast, in the MGE-transplanted group, we recorded significantly greater GAD65

MDV3100 manufacturer (6.7%) and GAD67 (7.1%) levels compared to levels in the medium-injected group. However, GAD mRNA levels in the MGE-transplanted group did not differ from those in the naive group. Taken together, these results indicate that MGE transplantation does not significantly raise basal GAD mRNA expression (in uninjured animals) but that the transplant can overcome the decrease of GAD mRNA (and presumptive inhibitory tone and consequent mechanical hypersensitivity) triggered by peripheral nerve injury. Finally, we asked whether transplantation of MGE GABAergic precursor neurons also has antinociceptive effects in a model of the other major class of chronic pain, namely, a chemical nociception pain model associated with inflammation. Hindpaw injection of formalin produces significant inflammation of the paw

and two distinct phases of pain behavior, characterized by flinching and/or Pregnenolone licking of the paw. In this condition, GABA transmission is not compromised and, in fact, GABAergic tone may be increased in the dorsal horn in the setting of tissue injury (Hossaini et al., 2010). In our studies, we injected 1% formalin into the hindpaw 4 weeks after the mice had received a dorsal horn transplant (ipsilateral to the transplant). Figure 8C illustrates that the transplant did not reduce pain behaviors in either the first or the second phases of this test. There was a slight decrease at all time points during phase II but the reduction was not statistically significant. We conclude that mechanical allodynia in a standard model of neuropathic pain, but not the pain behavior evoked in a common chemical nociception model associated with inflammation, can be reversed by spinal cord transplantation of GABAergic precursor cells.