Figure 3.

Short-term transplantation and in vivo analysis of eGFP-sorted cells. (A-O): naïve mice were transplanted with cells sorted for eGFP expression. Grafts were analyzed 4 weeks post-transplantation. (A): Low-power microphotograph of a graft, showing large numbers of TH+ neurons (the nickel-enhanced 3,3′-diaminobenzidine products appear grayish black; the boxed area is shown enlarged in [B]). Most TH+ neurons appeared to co-express eGFP (C) (co-expression appears yellow; the boxed area is enlarged in [D]); however, some cells expressed only eGFP or only TH. A large proportion of the eGFP+/TH cells also showed overlapping expression with AADC (arrowheads) (co-expression appears purple) (E-H). Many eGFP+/TH+ cells showed overlapping GIRK2 immunoreactivity (arrowheads) (co-expression appears purple), but not all (I-L). Pitx3 was expressed in the nucleus of most eGFP+ cells (M-O). Scale bars = 150 μm (A), 30 μm (B), 100 μm (C), and 50 μm (D, H, L, O) (scale bar in [H] applies to [E-G], [L] applies to [I-K], and [O] applies to [M, N]). Abbreviations: AADC, aromatic L-amino acid decarboxylase; GFP, green fluorescent protein; GIRK2, G protein-activated inwardly rectifying potassium channel 2; Pitx3, paired-like homeodomain transcription factor 3; TH, tyrosine hydroxylase.