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Figure 5 | Molecular Pain

Figure 5

From: BDNF regulates atypical PKC at spinal synapses to initiate and maintain a centralized chronic pain state

Figure 5

BDNF enhances eIF4F complex formation and nascent synthesis of PKCλ and PKMζ in spinal SNSs in an mTOR-dependent fashion. SNSs were prepared from mouse lumbar spinal cord and treated with or without BDNF (100 ng/ml) or the mTORC1 inhibitor temsirolimus (Tem, 100 nM). A) Following 15 min treatment with the indicated conditions, 5′m7-GTP pulldown assays were performed and eIF4A, 4EBP and eIF4E were assessed by Western blot. BDNF treatment led to an increase in eIF4A pulldown and a decrease in 4EBP pulldown in an mTORC1-dependent fashion (N = 6). B) SNSs were assessed for aPKC mRNA expression by qPCR. SNSs contained PKMζ and PKCλ mRNA but not PKCζ (N = 3). C) SNSs were treated with BDNF +/− Tem in the presence of AHA to tag nascently synthesized proteins for 30 min. AHA aPKC was immunoprecipitated and AHA was labeled with biotin using click-chemistry. BDNF treatment led to an increase in nascently synthesized PKCλ and PKMζ in an mTORC1-dependent fashion (N = 6). * p < 0.05, ** p < 0.01, one way ANOVA with Bonferroni post hoc test compared to Veh.

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