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Fig. 4. RA induces intermediate character in early dorsal telencephalic cells. (A) A stage 8 chick embryo (dorsal view, rostral is upwards). Black square indicates prospective dorsal telencephalic neuroectoderm explant region. (B) Expression of transcription factors in stage 8 dorsal (D, st 8) explants cultured alone or with all-trans retinoic acid (RA) or a combination of all-trans retinoic acid and FGF8 for 48-52 hours. Stage 8 D explants cultured alone (n=20) generated PAX6⁺ cells (78±4%) but no MEIS2⁺, ISL1⁺, NKX2.1⁺or EMX1⁺ cells were detected. Stage 8 D explants (n=20) exposed to all-trans retinoic acid generated MEIS2⁺ cells (82±4%) distributed throughout the explants, while ISL1⁺ cells (17±3%) and PAX6⁺ cells (65±5%) were expressed in complementary regions in the explant. No NKX2.1⁺ or EMX1⁺ cells were detected. Stage 8 D explants (n=15) exposed to a combination of all-trans retinoic acid and FGF8 generated PAX6⁺ cells (63±5%), but no MEIS2⁺, NKX2.1 ISL1⁺ or EMX1⁺ cells were detected. Scale bar: 75 µm. (C) Expression of transcription factors in the ventral-most domain of the prospective intermediate telencephalon in a stage 22 chick embryo. ISL1⁺ cells were located in the periphery of the proliferating neuroepithelium, while MEIS2⁺ cells were situated throughout the neuroepithelium overlapping with the ISL1⁺ domain. No PAX6⁺ cells were detected in this ventral subdomain of the intermediate telencephalon.

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