AAV ablates neurogenesis in the adult murine hippocampus.

Stephen Johnston,Cooper Bloyd,Sarah L Parylak,Matthew Shtrahman,Christian D Saavedra,Nolan Mac,Alexander Newberry,Iryna Gallina,Stacy Kim,Fred H Gage,Ondrej Novak,J Tiago Gonçalves,Christina Lim

doi:10.7554/elife.59291

Abstract

Recombinant adeno-associated virus (rAAV) has been widely used as a viral vector across mammalian biology and has been shown to be safe and effective in human gene therapy. We demonstrate that neural progenitor cells (NPCs) and immature dentate granule cells (DGCs) within the adult murine hippocampus are particularly sensitive to rAAV-induced cell death. Cell loss is dose dependent and nearly complete at experimentally relevant viral titers. rAAV-induced cell death is rapid and persistent, with loss of BrdU-labeled cells within 18 hr post-injection and no evidence of recovery of adult neurogenesis at 3 months post-injection. The remaining mature DGCs appear hyperactive 4 weeks post-injection based on immediate early gene expression, consistent with previous studies investigating the effects of attenuating adult neurogenesis. In vitro application of AAV or electroporation of AAV2 inverted terminal repeats (ITRs) is sufficient to induce cell death. Efficient transduction of the dentategyrus (DG)- without ablating adult neurogenesis- can be achieved by injection of rAAV2-retro serotyped virus into CA3. rAAV2-retro results in efficient retrograde labeling of mature DGCs and permits in vivo two-photon calcium imaging of dentate activity while leaving adult neurogenesis intact. These findings expand on recent reports implicating rAAV-linked toxicity in stem cells and other cell types and suggest that future work using rAAV as an experimental tool in the DG and as a gene therapy for diseases of the central nervous system should be carefully evaluated.

Highlights

The subgranular zone (SGZ) of the hippocampal dentate gyrus (DG) is one of only a few regions of the mammalian brain that continues to exhibit neurogenesis into adulthood
Motivated by our own efforts to study the role of adult neurogenesis and the DG in learning and memory, we discovered that neural progenitor cells (NPCs) and immature neurons in the DG are highly susceptible to Recombinant adeno-associated virus (rAAV)-induced death at a range of experimentally relevant titers (3 E11 gc/mL and above)
We found that the delivery of calcium indicators using rAAV at doses equivalent to or below previously reported doses resulted in ablation of adult neurogenesis (Fig. S1A)

Summary

Introduction

The subgranular zone (SGZ) of the hippocampal dentate gyrus (DG) is one of only a few regions of the mammalian brain that continues to exhibit neurogenesis into adulthood. Adult-born dentate granule cells (abDGCs) are continuously generated from a pool of largely quiescent neural stem cells that undergo proliferation, differentiation, and fate specification before maturing into neurons that are indistinguishable from developmentally derived dentate granule cells (DGCs) While the specific role that immature DGCs play in hippocampal function, including the formation of memories, is not fully established, progress has been achieved through recent work focused on precisely modulating and measuring the activity of immature and mature DGCs within the DG of animals during behavior (Anacker et al, 2018; Danielson et al, 2016, 2017; GoodSmith et al, 2017; Hainmueller and Bartos, 2018; Hayashi et al, 2017; Kirschen et al, 2017; Leutgeb et al, 2007; Nakazawa, 2017; Pilz et al, 2016; Senzai and Buzsáki, 2017)

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Conclusion