Long-Term Labeling of Hippocampal Neural Stem Cells by a Lentiviral Vector

Abstract

Using a lentivirus-mediated labeling method, we investigated whether the adult hippocampus retains long-lasting, self-renewing neural stem cells (NSCs). We first showed that a single injection of a lentiviral vector expressing a green fluorescent protein (LV PGK-GFP) into the subgranular zone (SGZ) of the adult hippocampus enabled an efficient, robust, and long-term marking of self-renewing NSCs and their progeny. Interestingly, a subset of labeled cells showed the ability to proliferate multiple times and give rise to Sox2+ cells, clearly suggesting the ability of NSCs to self-renew for an extensive period of time (up to 6 months). In addition, using GFP+ cells isolated from the SGZ of mice that received a LV PGK-GFP injection 3 months earlier, we demonstrated that some GFP+ cells displayed the essential properties of NSCs, such as self-renewal and multipotency. Furthermore, we investigated the plasticity of NSCs in a perforant path transection, which has been shown to induce astrocyte formation in the molecular layer of the hippocampus. Our lentivirus (LV)-mediated labeling study revealed that hippocampal NSCs are not responsible for the burst of astrocyte formation, suggesting that signals released from the injured perforant path did not influence NSC fate determination. Therefore, our studies showed that a gene delivery system using LVs is a unique method to be used for understanding the complex nature of NSCs and may have translational impact in gene therapy by efficiently targeting NSCs.

Restoring miR-132 expression rescues adult hippocampal neurogenesis and memory deficits in Alzheimer’s disease

“…First, a lentiviral vector (LV PGK:mCHERRY) expressing an mCHERRY reporter and a miR-132-overexpressing hairpin under the control of the PGK promoter (Suh et al, 2018) was ll used for stereotactic injections into the dentate gyrus (Figure 5B). Considering the possibility that LV PGK:mCHERRY, apart from adult NSCs, may also transduce a proportion of developmentally born granule cells, we additionally used a retroviral approach (RV CAG:GFP) (Zhao et al, 2006) to target miR-132 overexpression to proliferating adult neuronal precursors in a second mouse cohort (Figure 5C).…”

Neural Stem Cells in the Adult Olfactory Bulb Core Generate Mature Neurons in Vivo

“…We analyzed the expression of molecular markers by the cells in the OB core at early time points (Figures 4 and S2) in order to distinguish between cycling cells (MCM2 + ), putative aNSCs (expressing Nestin, GLAST/Slc1a3, Sox2, Prominin-1/CD133, and GFAP), 4,6,7,16,28,29,38,42,[45][46][47][48][49][50][51][52][53][54][55][56] progenitor cells committed to the neuronal lineage (Pax6) 57,58 or to the oligodendroglial lineage (Olig2), 59 neuroblasts and immature neurons (DCX). 60 Although a single marker is not exclusively expressed by NSCs, the combination of Nestin, GLAST, Sox2, Prominin-1, and GFAP is widely used to identify NSCs in the adult brain (see references above).…”

VCAM1 Labels a Subpopulation of Neural Stem Cells in the Adult Hippocampus and Contributes to Spatial Memory

“…After VCAM1+ cells are infected with the VP virus, the Vcam1 promotor initiates Cre transcription, promoting the expression of GFP protein and Cre-mediated recombination through removal of the STOP cassette, which leads to the expression of tdTomato protein in VCAM1 cells in the SGZ. As a result, the progenies of VCAM1-expressing cells should express tdTomato only, and most should migrate out of the SGZ ( Madisen et al., 2010 , Suh et al., 2018 ) ( Figure 4 A). Surprisingly, we observed some GFP+ tdTomato− cells exhibiting typical neuronal morphology and migrating out of the SGZ.…”