Canning CA et al. (2011), Developmental expression of the fermitin/kindli...

XB-ART-43549

Dev Dyn 2011 Aug 01;2408:1958-63. doi: 10.1002/dvdy.22683.

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Developmental expression of the fermitin/kindlin gene family in Xenopus laevis embryos.

Canning CA, Chan JS, Common JE, Lane EB, Jones CM.

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Fermitin genes are highly conserved and encode cytocortex proteins that mediate integrin signalling. Fermitin 1 (Kindlin1) is implicated in Kindler syndrome, a human skin blistering disorder. We report the isolation of the three Fermitin orthologs from Xenopus laevis embryos and describe their developmental expression patterns. Fermitin 1 is expressed in the skin, otic and olfactory placodes, pharyngeal arches, pronephric duct, and heart. Fermitin 2 is restricted to the somites and neural crest. Fermitin 3 is expressed in the notochord, central nervous system, cement gland, ventral blood islands, vitelline veins, and myeloid cells. Our findings are consistent with the view that Fermitin 1 is generally expressed in the skin, Fermitin 2 in muscle, and Fermitin 3 in hematopoietic lineages. Moreover, we describe novel sites of Fermitin gene expression that extend our knowledge of this family. Our data provide a basis for further functional analysis of the Fermitin family in Xenopus laevis.

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Species referenced: Xenopus laevis
Genes referenced: fermt1 fermt2 fermt3 odc1

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	Figure 1. Expression analysis of Fermt1, Fermt2, and Fermt3 by RT-PCR. Fermt1 and Fermt2 transcripts are maternally expressed, whereas Fermt3 expression is detectable from stage 15 onwards. Expression of Odc (Ornithine decarboxylase) is analysed as a loading control. Lanes 1, 3, 5, 7, 9, 11, 13, 15, 17, and 19 are samples extracted from stage 2, 6, 9, 10, 13, 15, 18, 20, 23, and 32. Negative controls (minus Reverse Transcriptase) are shown in lanes 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20.
	Figure 2. Expression of Fermt1 and Fermt2 at gastrula stages. A, B: Expression of both Fermt genes is detected throughout the gastrula embryo. Ectoderm, endoderm, and mesoderm were isolated from embryos at stage 10.5 and analysed for Fermt1 and Fermt2 expression by RT-PCR (C). Fermt1 is expressed in the ectoderm (lane 1) and mesoderm (lane 2) and more weakly in the endoderm (lane 3). Fermt2 is expressed in all three germ layers at stage 10.5. ODC serves as a loading control.
	Figure 3. Expression of Fermt1 from stages 12 to 35. A:Fermt1 expression at stage 12, 15, and 18, respectively. Expression in the ectoderm is clearly seen in transverse section. D: Fermt1 expression is detected throughout the ectoderm and weakly in the somites at stage 22. E: At stage 28, Fermt1 becomes restricted to a number of ectodermal derivatives, including otic, olfactory, and pharyngeal placodes, and pronephric anlage. F: By stage 35, Fermt1 transcripts are detected in the otic vesicle (black arrow), olfactory pit (black asterisk), heart (back arrowhead), and in the pronephric duct (white arrowhead). G: Transverse sections are observed through the level of the eye/olfactory pit (G), the otic vesicle (H), and the pronephric duct (I). Inset (F) demarcates the levels at which sections were analysed (G).
	Figure 4. Expression of Fermt2 from stages 7 to 17. A:Fermt2 is ubiquitously expressed at stage 7. By stage 13 (B,C), Fermt2 becomes restricted to the dorso anterior most region of the embryo. D: At stage 14, a clear delineation of Fermt2 is observed in the future head region (white asterisk). E:Fermt2 expression is observed in the neural folds (white asterisk). F,G: Neural crest (white asterisk) and somites (black arrow) express Fermt2 by stage 16. H: Streams of neural crest expressing Fermt2 are evident at stage 17 (white asterisk). In A, animal pole is to the top, and in C, anterior is to the top. In B, D, E, G, and H, anterior is to the left. F represents a frontal view.
	Figure 5. Expression of Fermt2 from stages 22 to 34. A, B: At stage 22, Fermt2 transcripts are detected in the migratory neural crest, clearly revealed by transverse section (B). C: By stage 25, expression persists in the neural crest (arrow) and becomes detectable within the somites (lateral view, C and dorsal view, D). E, F: Transverse sections through mid-axial levels of a stage-29 embryo reveal the somitic expression (F). Fermt2 transcripts are also detected in Rohon Beard neurons (E, boxed area shown in high magnification, inset, black arrows). At stage 30 (G) and 34 (H) expression persists within the neural crest and somites and is clearly seen in a transverse section (I) from a mid axial level. Asterisk in H demarcates cardiac neural crest. White lines in A and H indicate the levels at which sections are shown in B and I, respectively.
	Figure 6. Expression of Fermt3 at stage 28 and 35. A, B:Fermt3 transcripts are detected at stage 28 in the cement gland (asterisk), notochord, anterior ventral blood islands (back arrow), and in early myeloid progenitor cells. White line in A indicates the level at which this section (B) is taken. C: By stage 35, expression of Fermt3 is visible in the cement gland (C, asterisk), central nervous system (C, E, F), ventral blood islands and vitelline veins (5C, black arrow, D, F), notochord (C, F, G), and in myeloid cells (C, D, F, G). Ventral view of a stage-35 embryo is shown is D and transverse sections are presented (E), the levels of which are demarcated by white lines (C).
	fermt1 (FERM domain containing kindlin 1) gene expression in Xenopus laevis embryos, NF stage 35, as assayed by in situ hybridization, lateral view,anterior left, dorsal up.
	fermt2 (FERM domain containing kindlin 2) gene expression in Xenopus laevis embryos, NF stage 28, as assayed by in situ hybridization, lateral view, anterior left, dorsal up.
	fermt3 (FERM domain containing kindlin 3) gene expression in Xenopus laevis embryos, NF stage 35, as assayed by in situ hybridization, lateral view, anterior left, dorsal up.
	fermt1 (gene_name) gene expression in bissected Xenopus laevis embryo, assayed via in situ hybridization, NF stage 10.5, lateral view, anterior left, dorsal up.
	fermt2 (gene_name) gene expression in bissected Xenopus laevis embryo, assayed via in situ hybridization, NF stage 10.5, lateral view, anterior left, dorsal up.
	fermt1 (gene_name) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.
	fermt1 (gene_name) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.
	fermt2 (gene_name) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 7.
	fernt2 st 13
	fermt2 gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 15, dorsal view, anterior left.
	fermt2 () gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 17, lateral view, anterior left, dorsal up.
	fermt2 () gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 25, (C) lateral view, anterior left, dorsal up (D) dorsal view, anterior left.
	fermt2 (fermitin family member 2) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 29, transverse section mid-axial level, dorsal up.
	fermt2 () gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 34, transverse section, dorsal up.
	fermt3 (fermitin family member 3) st 28
	fermt3 (fermitin family member 3) gene expression in Xenopus laevis embryo, NF stage 28, cross-section, anterior trunk region, as assayed by in situ hybridization, lateral view, anterior left, dorsal up
	fermt3 (fermitin family member 3) gene expression in Xenopus laevis embryo, NF stage 35, cross-section, through eye head and heart region, as assayed by in situ hybridization, lateral view, anterior left, dorsal up.
	fermt3 (fermitin family member 3) gene expression in Xenopus laevis embryo, NF stage 35, cross-section, trunk region, as assayed by in situ hybridization, lateral view, anterior left, dorsal up
	fermt3 (fermitin family member 3) gene expression in Xenopus laevis embryo, NF stage 35, cross-section, tail region, as assayed by in situ hybridization, lateral view, anterior left, dorsal up.