PMC:1713256 / 19725-22281 JSONTXT

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In Bmp2C/C; Bmp4C/C; Prx1::cre limbs, only the very distal tips of each digit were separated, with the autopod adopting the shape of a notched pallet (Figure 2F). In wild-type development, the individual digits become free from one another through a process of interdigital apoptosis, observable by staining with acridine orange (Figure 2E and 2G, arrow). Interdigital apoptosis is dramatically reduced in the Bmp2C/C; Bmp4C/C; Prx1::cre limbs (Figure 2F and 2H, arrow). (Note that the strong acridine orange staining in Figure 2H is not in the interdigital mesenchyme but rather is in the remnant of the cells of the AER [red asterisk].)\nAnother region of very prominent apoptosis in the wild-type limb bud is within the AER. The AER is a specialized ridge of ectoderm running along the anterior-posterior axis at the distal margin of the early limb bud. One of the phenotypes characterizing the Bmp2C/C; Bmp4C/C; Prx1::cre mutant is expansion of the AER (see below). Moreover, in the Bmp2C/C; Bmp4C/C; Prx1::cre animals, the AER is maintained as late as E15.5, long after it has started to degrade in wild-type limbs. This can be seen morphologically (unpublished data) or by staining for expression of AER-specific marker such as Fgf8 (Figure 2I and 2J). The AER displays intense staining with acridine orange, TUNEL, or other markers for apoptosis at every stage when it is present, reflecting the rapid turnover of cells in this structure [33–35]. The Bmp2C/C; Bmp4C/C; Prx1::cre mutant limbs at E15.5 show strong acridine orange staining in the remnants of AER (Figure 2H, asterisk), in domains of the distal tip where the AER remains the longest (compare staining in Figure 2F and 2J). No significant difference was observed in the extent of apoptosis in the cells of the AER in earlier stage Bmp2C/C; Bmp4C/C; Prx1::cre mutant embryonic limbs (Figure 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