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Fig. 1. Transfection of Nkx2.1-/- slices with
Nkx2.1 cDNA results in rescue of interneuron phenotypes.
(A-D) Schematic showing slice electroporation and transplantation
paradigm. (A) The Nkx2.1 domain is shown in the MGE of an E12.5
wild-type (wt) mouse embryo slice. The MGE*, a region that
expresses a truncated Nkx2.1 transcript
(Sussel et al., 1999), is
shown in the slice from an Nkx2.1 mutant embryo. (B) This
MGE* region is targeted for electroporation, and after 1 day in
vitro (DIV) the region is dissected out, dissociated and transplanted (C,D)
directly into the neocortex of neonatal pups (as in H-N), or plated onto a
high-density culture of neonatal cortical cells [as in Xu et al.
(Xu et al., 2004); see Fig. S2
in the supplementary material]. (E,F) Coronal sections of a
slice from an E12.5 wt embryo that was electroporated with pNkx2.1-GFP,
maintained 1DIV, then fixed and examined for GFP fluorescence (E) and NKX2.1
immunolabeling (F). The right-hand, electroporated side of the slice has
extensive ectopic NKX2.1 expression, whereas only native NKX2.1 expression is
seen on the left-hand side of the slice (arrow in F). (G) A slice from
an Nkx2.1-/- embryo was electroporated with pNkx2.1-GFP.
After 1DIV, cells from the MGE* (outlined in white) were
transplanted into the cortical plate of a neonatal pup and then examined at
postnatal day 30 (P30) in 40 µm coronal sections. (H) Transplanted
GFP-expressing cells scattered through the medial cortex. (I-N)
Examples of co-labeling for GFP and parvalbumin (PV; I,J), somatostatin (SST;
K,L), and neuropeptide Y (NPY; M,N). In control experiments with pGFP vector,
almost no cells expressing any of these markers are detected after
transplantation of Nkx2.1-/- MGE* progenitors
(Table 1). MGE, medial
ganglionic eminence; LGE, lateral ganglionic eminence; Ctx, cerebral cortex.
Scale bars: 100 µm in G,H.
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