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. 2015 Nov 19;5(6):988–995. doi: 10.1016/j.stemcr.2015.10.014

Figure 1.

Figure 1

Metformin Enhances the Self-Renewal and Neuronal Differentiation of Adult Neural Precursors

(A–C) Primary neurospheres cultured from the adult SVZ were grown in the presence or absence of 500 nM metformin (Met) and quantified 6 days later, as pictured in (A). Scale bar, 200 μm. (B) Quantification of primary neurosphere (NS) number following metformin or PBS exposure. ∗∗∗p ≤ 0.001 (n = 5 for each group). (C) Quantification of the neurosphere diameter following metformin or PBS exposure, as in (A). ∗∗∗p ≤ 0.001 (pooled data from five independent experiments).

(D and E) Primary neurospheres were passaged either once (secondary) or twice (tertiary) into untreated media, and the number (D) and diameter (E) of spheres was quantified 4 days later. ∗∗∗p ≤ 0.001 (n = 4 for each group).

(F) Quantitative analysis of number of secondary neurospheres in the absence and presence of metformin (1 μM). ∗p < 0.05 (pooled data from four independent experiments).

(G) Immunofluorescence images of βIII-tubulin (βIII tub)-positive neurons generated from SVZ adult neurospheres in the absence and presence of metformin. Arrows denote βIII-tubulin-positive cells. Scale bar, 50 μm.

(H) Quantitative analysis of the percentage of βIII-tubulin-positive (β III tubulin+ive) neurons generated in the absence and presence of metformin (200 nM), as shown in (G). ∗∗∗p < 0.001 (pooled data from four independent experiments).

(I–L) In (I) and (K), fluorescence photomicrographs are shown of coronal SVZ (I) and SGZ (K) sections from 3-month-old WT mice that received PBS or metformin daily by injection (i.p., 200 mg/kg) for 7 days, followed by a single BrdU (100 mg/kg) injection 24 hr before. Sections were stained for BrdU (green) and counterstained with Hoechst 33258 (blue). Scale bar, 100 μm. (J and L) Quantitative analysis of total number of BrdU-positive (BrdU+ve) precursor cells within the SVZ (J) and SGZ (L) regions, determined from sections as shown in (I) and (K). ∗∗∗p < 0.001 (pooled data from three independent experiments). Arrows denote BrdU-positive cells.

Error bars indicate SEM.

See also Figure S1.