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Fig. 1. In vivo labelling of lRL-derived PC neurons by exo utero electroporation
of EYFP. (A) Diagram outlining the method.
Following injection of a plasmid carrying EYFP or Venus
(left panel), electric pulses were applied. Red arrow indicates an
EYFP-transfected region. (B,C) Dorsal (B) and ventral (C) views of the
hindbrain (outlined) that was electroporated with EYFP at E12.5 and
fixed at E14.5. There are two streams of labelled cells, the AEMS and PEMS
(arrowheads in C). The cells in the AEMS take two separate routes (arrows in
the inset of B). Because neurons in the PEMS begin their migration earlier,
most labelled cells therein were located ventrally, leaving few cells behind
(arrowhead in the inset of B). Rostral is towards the left. Asterisks show the
EYFP-transfected side, the broken horizontal lines show the midline.
(D,E) Transverse sections of EYFP-introduced embryos
immunostained for EYFP (green, left panel), after Mbh2 in situ
hybridization (purple, middle panel). The left, middle and right panels in D
and E show the same field of view of metencephalon (D) and myelencephalon (E).
In both, EYFP-labelled migrating cells expressed Mbh2 (arrowheads in
insets of D and E). Broken vertical lines show the midline. Dorsal is upwards
and EYFP-transfected side is towards the left. Cb, cerebellum. Scale
bars: 500 µm in B and C; 250 µm in the inset of B; 100 µm in D; 200
µm in E; 55 µm for right panel insets in D,E.
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