Figure 1. Vegf-c/Vegfr3 Are Expressed by Adult Hippocampal Stem Cells.

(A) Coronal DG section of an adult Vegfr3::YFP mouse. YFP expression is detected both in vessels and SGZ cells. White dotted line indicates SGZ.

(B) isolation of Vegfr3^YFP cells from the DG of adult Vegfr3::YFP mice. After exclusion of CD31⁺ endothelial cells, a subset of Vegfr3^YFP cells were sorted by FACS.

(C) qRT-PCR analysis of FACS-sorted cells from the DG of Vegfr3::YFP adult mice. Vegfr3 transcripts are specifically enriched in Vegfr3^YFP cells compared to total DG neural cells (non-endothelial DG cells) and Vegfr3^YFP-negative cells, n = 3 independent experiments; bars: mean ± SEM; Student’s t test: p < 0.05 (*).

(D and E) Coronal DG section of adult Vegfc^lacZ/+ mouse stained with X-gal and of wild-type mouse labeled with anti-VEGF-C Ab (E). White arrows indicate a higher level of VEGF-C expression in SGZ.

(F) Representative images of coronal DG sections in adult Vegfr3::YFP mice stained with Abs against markers of RGL (GFAP and Nestin), progenitor cells (Ascl1 and Tbr2), neuroblasts (DCX), and neurons (NeuN) (red).

(G) Quantification of Vegfr3^YFP SGZ cells by their morphology. Percentage of Vegfr3^YFP RGL cells expressing GFAP and Nestin.

(H) Quantification of Vegfr3^YFP SGZ cell subtypes according to their antigen expression. y axis: % of YFP⁺ marker⁺ cells/YFP⁺ cells. Total 30–500 cells were counted from three to six sections.

(I) 5% of Vegfr3^YFP SGZ cells has entered the cell cycle (Ki67⁺). n = 100–150 cells/section, counted from seven sections. Bars: mean ± SEM.

(J) Vegfr3^YFP cells represent 60% of BrdU⁺ SGZ cells. Most BrdU⁺Vegfr3^YFP cells are non-radial cells (98%), and only a few of them are RGL cells (2%). n = 30–50 cells counted from five sections.

(K) Vegfr3^YFP SGZ cell morphology and activity. An anti-GFAP Ab labels RGLs (white), but not YFP⁺-non-radial cells. A 3-hr pulse of BrdU (red) labels exclusively non-radial YFP⁺ cells.

(L) Comparison of the expression of Vegfr3^YFP and other stage-specific markers in SGZ cells. Vegfr3^YFP expression is specific to NSCs and progenitor cells.

The scale bars represent 100 µm (A, D, and E) and 16 µm (F and K).

See also Figure S1.