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Fig. 4. Abnormal distributions of Islet1+ and Lim1+ cells in Raldh2^L–/– brachial spinal cord. (A) Quantitation of Islet1/2+ motoneurons in the brachial LMC of E11.5 control and Raldh2^L–/– embryos (Y axis: number of motoneurons per hemisection; ct: 96.84±1.77; mut: 86.04±3; mean±s.e.m.; t-test: P=0.009; ten sections spanning the Raldh2 LMC domain were counted in nine embryos of each genotype). (B) Quantitation of Lim1/2/Islet2+ and Islet1+ motoneurons per hemisection in the brachial LMC of E11.5 control and Raldh2^L–/– embryos. The number of Lim1/2/Islet2+ cells was significantly decreased in mutants (Lim1/2/Islet2: ct, 25.78±1.48; mut, 20.50±1.11; mean±s.e.m.; t-test P=0.024; ten sections were counted in five embryos of each genotype), whereas the number of Islet1+ cells was similar in mutants and controls (Islet1: ct, 80.36±2.68, mut, 76.28±2.94; mean±s.e.m.; t-test P=0.336). (C-F) Lim1 (C,D, red) and Islet1 (E,F, red) transcripts on flat-mounted spinal cords combined with Raldh2 immunofluorescence (C,E, green). (C,E) Composite images in which the overlay of the two signals is shown on the left side only. (E,F) The posterior domain of high Islet1 expression, marked by a bracket, is expanded anteriorly in the mutant. Lim1+ and Islet1+ motor columns are indicated by arrowheads. (G-I) Lim1+ (G,H) and Islet1+ (I,J) transcripts on transverse ventral spinal cord sections at levels indicated by horizontal arrows in C-F. The Islet1 and Lim1 subpopulations within the LMC are marked by brackets.

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