Figure 2. Unexpected complexity of protonephridial tubules is revealed by systematic gene expression mapping of slc genes along the protonephridial tubules.
(A) Cartoon shows previous segmentation model of the protonephridial tubule and expression map of slc genes along the protonephridial tubule. (B–G) Representative images show expression domains of selected slc genes in (B) PT1, PT2 and PT3, (C) PT2 and PT3, (D) PT3, (E) DT1, DT2 and CD, (F) DT2 and CD and (G) CD. Fluorescent overlay of the indicated gene (red) with PT marker (AcTub) and distal tubule (DT) marker (CAVII-1). A color-coded scheme of the protonephridial tubule at the end of each panel represents the expression domain of the indicated gene. Images are maximum projections of confocal Z-sections. Scale bars: 50 μm. (H) Longitudinal-section through a worm shows a dorsal-bias expression of slc12a-1. Fluorescent overlay of slc12a-1 with DT marker (CAVII-1) and streptavidin (which labels the basement membrane of several planarian epithelial structures, including the outer epithelium). Inset shows a magnification of CD, visualized by slc12a-1, crossing the basement membrane of the dorsal epithelia. Yellow arrowhead, exterior opening of the CD. Scale bars: 200 μm. (I) TEM image shows CD connected to the dorsal epithelia. Inset shows a magnification of CD connected to the dorsal epithelia. e, epithelia; bm, basement membrane; m, mesenchyme; sj, septate junction; l, lumen; ms, muscle. Scale bars: 5 μm.
Figure 2—figure supplement 1. Solute carrier gene families in the planarian Schmidtea mediterranea.
Figure 2—figure supplement 2. Schematic representation of phylogenetic clusters of γ- (A), δ- (B) groups of slcs and the Tim barrel- (C), IT- (D), Drug/Metabolite (E) transporter clans of slcs.
Planarian homologs are colored in red.
Figure 2—figure supplement 3. Schematic representation of phylogenetic clusters of α-groups of slcs.
Planarian homologs are colored in red.
Figure 2—figure supplement 4. Schematic representation of phylogenetic clusters of β-groups of slcs.
Planarian homologs are colored in red.
Figure 2—figure supplement 5. Schematic representation of phylogenetic clusters of Smed-slc1a (A), Smed-slc5a (B), Smed-slc22a (C), Smed-slc6a (D).
Planarian homologs are colored in red.
Figure 2—figure supplement 6. Schematic representation of phylogenetic clusters of Smed-slc4a (A), Smed-slc7a (B), Smed-slc12 (C), Smed-slc15 (D), Smed-slc20 (E), Smed-slc23 (F), Smed-slc26 (G).
Planarian homologs are colored in red.
Figure 2—figure supplement 7. Schematic representation of phylogenetic clusters of Smed-slc28 (A), Smed-slc30 (B), and Smed-slc42 (C).
Planarian homologs are colored in red.
Figure 2—figure supplement 8. Expression patterns of slc genes that belong to solute carrier families 1–6 in an asexual strain of the planarian S. mediterranea.
Whole-mount expression patterns of slc genes by in situ hybridization (NBT/BCIP development). Scale bars: 500 μm.
Figure 2—figure supplement 9. Expression patterns of slc genes that belong to solute carrier families 7–15 in an asexual strain of the planarian S. mediterranea.
Whole-mount expression patterns of slc genes by in situ hybridization (NBT/BCIP development). Scale bars: 500 μm.
Figure 2—figure supplement 10. Expression patterns of slc genes that belong to solute carrier families 16–23 in an asexual strain of the planarian S. mediterranea.
Whole-mount expression patterns of slc genes by in situ hybridization (NBT/BCIP development). Scale bars: 500 μm.
Figure 2—figure supplement 11. Expression patterns of slc genes that belong to solute carrier families 24–29 in an asexual strain of the planarian S. mediterranea.
Whole-mount expression patterns of slc genes by in situ hybridization (NBT/BCIP development). Scale bars: 500 μm.
Figure 2—figure supplement 12. Expression patterns of slc genes that belong to solute carrier families 30–38 in an asexual strain of the planarian S. mediterranea.
Whole-mount expression patterns of slc genes by in situ hybridization (NBT/BCIP development). Scale bars: 500 μm.
Figure 2—figure supplement 13. Expression patterns of slc genes that belong to solute carrier families 40–47 in an asexual strain of the planarian S. mediterranea.
Whole-mount expression patterns of slc genes by in situ hybridization (NBT/BCIP development). Scale bars: 500 μm.
Figure 2—figure supplement 14. Expression of slc genes in the PT.
Fluorescent overlay of indicated gene (in red) (A-F) with PT2 and PT3 marker (slc6a-13), DT marker (CAVII-1) and AcTub staining. Images are maximum projections of confocal Z-sections. Scale bars: 50 μm. A color-coded scheme of protonephridial tubule at the end of each panel represents the expression domain of the indicated gene.
Figure 2—figure supplement 15. Expression of slc genes in the PT1 segment of the PT.
(A) Fluorescent overlay of indicated gene (in red) with PT1 and PT2 marker (CUBN1), PT2 and PT3 marker (slc6a-13) and AcTub staining. (B–D) Fluorescent overlay of indicated gene (in red) with PT2 and PT3 marker (slc6a-13), DT marker (CAVII-1) and AcTub staining. Images are maximum projections of confocal Z-sections. Scale bars: 50 μm. A color-coded scheme of protonephridial tubule at the end of each panel represents the expression domain of the indicated gene.
Figure 2—figure supplement 16. Expression of slc genes in the PT2 and PT3 segments of the PT.
Fluorescent overlay of indicated gene (in red) with PT2 and PT3 marker (slc6a-13), DT marker (CAVII-1) and AcTub staining. Images are maximum projections of confocal Z-sections. Scale bars: 50 μm. A color-coded scheme of protonephridial tubule at the end of each panel represents the expression domain of the indicated gene.
Figure 2—figure supplement 17. Expression of slc genes in PT3 segment of the PT.
Fluorescent overlay of indicated gene (in red) (A-C) with PT2 and PT3 marker (slc6a-13), DT marker (CAVII-1) and AcTub staining. Images are maximum projections of confocal Z-sections. Scale bars: 50 μm. A color-coded scheme of protonephridial tubule at the end of each panel represents the expression domain of the indicated gene.
Figure 2—figure supplement 18. Expression of slc genes in the DT.
Fluorescent overlay of indicated gene (in red) (A-G) with PT marker (slc6a-13 or CUBN1), DT marker (CAVII-1) or CD marker (slc12a-1 or slc24a-9) together with AcTub staining. Images are maximum projections of confocal Z-sections. Scale bars: 50 μm. A color-coded scheme of protonephridial tubule at the end of each panel represents the expression domain of the indicated gene.
Figure 2—figure supplement 19. Expression of slc genes in the collecting duct.
Fluorescent overlay of indicated gene (in red) (A-P) with PT2 and PT3 marker (slc6a-13), DT marker (CAVII-1) or CD marker (slc24a-9) together with AcTub staining. Images are maximum projections of confocal Z-sections. Scale bars: 50 μm. A color-coded scheme of protonephridial tubule at the end of each panel represents the expression domain of the indicated gene.
Figure 2—figure supplement 20. Expression of slc genes weakly expressed in both proximal and DTs.
Fluorescent overlay of indicated gene (in red) (A-B) with PT2 and PT3 marker (slc6a-13), DT marker (CAVII-1) and AcTub staining. Images are maximum projections of confocal Z-sections. Scale bars: 50 μm. A color-coded scheme of protonephridial tubule at the end of each panel represents the expression domain of the indicated gene.