PMC:7463108 / 87586-92237 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/7463108","sourcedb":"PMC","sourceid":"7463108","source_url":"https://www.ncbi.nlm.nih.gov/pmc/7463108","text":"Mature BDNF (mBDNF) activates tyrosine receptor kinase B (TrkB) and is neuroprotective, while the precursor to BDNF, pro-BDNF, binds p75NTR and can activate cell death pathways. Based on findings of significant, reversible reductions in glially produced BDNF after exposure to HIV-infected cell supernatant ± morphine (Masvekar et al. 2014), altered BDNF processing in lymphocytes from PWH (Avdoshina et al. 2011), and following exposure to HIV-1 gp120 (Bachis et al. 2012), the pro-BDNF:mBDNF ratio was analyzed in supernatants from wild-type vs. CCR5-null striatal glial cultures exposed to Tat ± morphine for 6 or 24 h (Kim et al. 2018) (Fig. 6). CCR5-deficiency reduced this ratio by over 2-fold in cells treated with Tat and morphine after 6 h (Fig. 6), indicating a relative increase in mBDNF that may partially protect neurons in the CCR5-deficient glial environment.\nFig. 6 Role of CCR5 and BDNF in mediating HIV-1 Tat and morphine-induced interactive cytotoxicity in striatal medium spiny neurons (MSNs). A proportion of glial fibrillary acidic protein (GFAP)-immunolabeled striatal astrocytes display punctate patterns of μ-opioid receptor (MOR) (a) and CCR5 (b) (43.8 ± 2.4%) immunofluorescence—with some faint immunoreactivity extending into the cell processes; scale bars = 10 μm (a-b). HIV-1 Tat and morphine are no longer toxic to MSNs when CCR5 is deleted from glia (c-f). In C57BL/J wild-type mixed glia-MSN co-cultures, Tat is neurotoxic (*p = 0.001 vs. controls), and co-exposure to morphine enhanced Tat-induced toxicity over a 72-h period (**p \u003c 0.001 vs. controls, p \u003c 0.05 vs. Tat) and antagonized by naloxone (c). Naloxone or morphine by themselves had no effect on neuronal survival (c). In co-cultures with CCR5-deficit glia and wild-type neurons, exposure to Tat by itself is significantly toxic (*p \u003c 0.001 vs. controls); however, the enhanced toxicity seen with combined morphine exposure was eliminated (d). Unexpectedly, morphine co-treatment entirely abolished the toxic effects of Tat, restoring MSN survival to control levels. Pre-treatment with naloxone re-established Tat toxicity, suggesting that the paradoxical protective effects of morphine are mediated by MOR (or perhaps another opioid receptor type) (d) (see Kim et al. 2018). The neurotoxic patterns seen in CCR5-deficient MSNs and wild-type glial co-cultures are similar to wild-type co-cultures (e). Co-cultures in which MSNs and glia are both deficient in CCR5 are similar to those in which CCR5 is only deficient in glia (f). CCR5 deletion alters the expression and processing of BDNF precursor (pro-BDNF) to mature (mBDNF) by mixed-glial cultures (g). BDNF is expressed by both astroglia and microglia; mBDNF is neuroprotective, while pro-BDNF can promote programmed cell death. mBDNF and pro-BDNF levels were analyzed in conditioned media from wild-type or CCR5-deficient mixed glia treated with Tat ± morphine after 6 h or 24 h to assess pro-BDNF and mBDNF levels. The proportion of pro-BDNF/mBDNF levels was significantly higher in wild-type compared to CCR5-null glia at 24 h (lower row; g), suggesting reduced neuronal support. Although morphine significantly decreased pro-BDNF in CCR5-deficient glia at both 6 h and 24 h compared to control levels (not shown), the pro-BDNF/mBDNF ratios were unaltered (upper and lower rows; g). By contrast, combined Tat and morphine significantly decreased the pro-BDNF/mBDNF ratio at 6 h, suggesting transient protection with CCR5 deficiency that was not fully sustained at 24 h (p = 0.17) (*p \u003c 0.05, wild-type vs. CCR5-null) (g). Exogenous mBDNF is neuroprotective against combined Tat and morphine treatment (h). Wild-type, mixed glial-MSN co-cultures were treated with mBDNF and Tat, or combined Tat and morphine (represented by dotted survival curves). Tat alone was neurotoxic (*p \u003c 0.05), and Tat was significantly worsened by co-exposing MSNs to morphine (**p \u003c 0.0001). The addition of mBDNF (50 ng/ml; 72 h) fully protected MSNs against combined Tat and morphine toxicity, but only tended to protect (albeit not significantly) MSNs treated with Tat alone (#)(h). Overall, the results in c-h suggest (1) an important role for glial CCR5 in mediating HIV-1 and opiate neurotoxic interactions, (2) that CCR5 deficiency influences signaling through MOR, and (3) that CCR5 (and perhaps MOR) act via a BDNF intermediary to promote or obstruct neuronal survival (Kim et al. 2018). (a-b) Modified and reprinted with permission from Podhaizer et al. (2012). (c-h) Modified and reprinted from Kim et al. (2018), which is an open access article distributed under the terms of the Creative Commons CC BY 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