jagged 2 loss of function transfates pronephric cells to MCCs. Three markers of MCCs - shippo1 (A-C), flr (D-F) and rfx2 (G-I) - show expanded expression in the pronephros of jagged 2 morphants at 34 hpf. Control invert morpholino (A,D,G) had no effect on MCC-marker expression, whereas ATG blocking (J2atgMO; B,E,H) and exon 20 mis-splicing morpholinos (J2ex20MO; C,F,I) resulted in uniform expression of MCC markers. (J) MCCs exist as single cells (arrowheads; flr-expressing cells) in the wild-type pronephros in cross section. (K,L) Expressions of shippo1 (K) and flr (L) expand to include all cells in tubule cross sections in J2exon20 morphant embryos. (M-O) slc13a1 expression seen in the control (M) is lost in J2atg (N) and J2exon20 (O) morphants. Similarly, the observed expression of trpM7 in control morpholino-injected embryos (P) is lost in J2exon20 morphants (Q). Control jagged 2 morpholino does not alter the single-cell jagged 2 mRNA expression pattern (R), whereas combined injection of both jagged 2 atg and exon 20 antisense morpholinos (U) result in expanded jagged 2 mRNA expression. (S) Na,K-ATPase:GFP transgenic zebrafish express GFP in the pronephros distal segments (arrowheads). (T) GFP expression is lost specifically in the distal segment of jagged 2 exon 20 morphant embryos (arrowheads).
|
