Critical Role of Beclin1 in HIV Tat and Morphine-Induced Inflammation and Calcium Release in Glial Cells from Autophagy Deficient Mouse
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We previously showed that autophagy is an important component in human immunodeficiency virus (HIV) replication and in the combined morphine-induced neuroinflammation in human astrocytes and microglia. Here we further studied the consequences of autophagy using glial cells of mice partially lacking the essential autophagy gene Atg6 (Beclin1) exposed to HIV Tat and morphine. Tat is known to cause an inflammatory response, increase calcium release, and possibly interact with autophagy pathway proteins. Following Tat exposure, autophagy-deficient (Becn1+/−) glial cells had significantly and consistently reduced levels in the pro-inflammatory cytokine IL-6 and the chemokines RANTES and MCP-1 when compared to Tat-treated cells from control (C57BL/6J) mice, suggesting an association between the inflammatory effects of Tat and Beclin1. Further, differences in RANTES and MCP-1 secretion between C57BL/6J and Becn1+/− glia treated with Tat and morphine also suggest a role of Beclin1 in the morphine-induced enhancement. Analysis of autophagy maturation by immunoblot suggests that Beclin1 may be necessary for Tat, and to a lesser extent morphine-induced arrest of the pathway as demonstrated by accumulation of the adaptor protein p62/SQSTM1 in C57BL/6J glia. Calcium release induced by Tat alone or in combination with morphine in C57BL/6J glia was significantly reduced in Becn1+/− glia while minimal interactive effect of Tat with morphine in the production of reactive oxygen or nitrogen species was detected in glia derived from Becn1+/− or C57BL/6J. Overall, the data establish a role of Beclin1 in Tat and morphine-mediated inflammatory responses and calcium release in glial cells and support the notion that autophagy mediates Tat alone and combined morphine-induced neuropathology.
KeywordsNeuroinflammation HIV Tat Opioid Beclin1-deficient mouse model
We gratefully acknowledge the support of the National Institutes of Health (NIH)-National Institute on Drug Abuse (NIDA) grants R01 DA036154 awarded to NEH for funding the study. We also acknowledge the financial support of NIH/NIDA R01 DA036154-S1 Diversity Supplement awarded to NEH to support JL and Presidential Fellowship provided to CRO by the Florida International University Graduate School.
JL performed and analyzed the experiments shown in Figs. 1, 2, 3, 4, 5, 6 and 7, Fig. S1–3, and wrote the manuscript. MR performed and analyzed the experiment shown in Fig. 1e and Fig. 5a-b. CRO assisted in the editing of the manuscript. NEH designed, analyzed, and coordinated the study and edited the manuscript. All authors reviewed the results and approved the final version of the manuscript.
Compliance with Ethical Standards
Conflict of Interest
Author Jessica Lapierre declares that she has no conflict of interest. Author Myosotys Rodriguez declares that she has no conflict of interest. Author Chet Raj Ojha declares that he has no conflict of interest. Author Nazira El-Hage declares that she has no conflict of interest.
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