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Fig. 5. Stromelysin-3 (ST3) expression, but not hedgehog (xhh) or collagenase-3 (Col3) expression is altered by TSA. Albino embryos with- out (control, top of each panel ) or with (bottom of each panel) 100 nM TSA treatment from stage 20/21 were fixed when control embryos reached stage 33-35 (A,B,C), or stage 41 (D,E,F). ST3 (A, D), xhh (B, E) and Col3 (C, F) mRNA expression was examined by whole mount in situ hybridization on fixed animals. ST3 was largely expressed in the head and the dorsal/proctodeal endoderm region (A, D single arrow) of control animals. Following treatment with TSA, the expression of ST3 was largely unchanged with the exception of the reduction in dorsal/ proctodeal endoderm region (A, D arrow). Despite the TSA induced changes in phenotype, the expression patterns of xhh and Col3 were essentially unchanged (compared the top to bottom animal in B, C, E, F). Note that the kink in the tail of TSA-treated embryos was not very obvious compared to that in Fig. 2. This is due to the whole mount in situ procedure. The protease digestions, high temperatures, and dehydration process caused all embryos to be dorsally curved, thereby diminishing the kinked tail phenotype associated with TSA. |
