Tazaki A et al. (2005), Macroarray-based analysis of tail regeneration ...

XB-ART-1708

Dev Dyn 2005 Aug 01;2334:1394-404. doi: 10.1002/dvdy.20472.

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Macroarray-based analysis of tail regeneration in Xenopus laevis larvae.

Tazaki A , Kitayama A , Terasaka C , Watanabe K , Ueno N , Mochii M .

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Xenopus larvae possess a remarkable ability to regenerate their tails after they have been severed. To gain an understanding of the molecular mechanisms underlying tail regeneration, we performed a cDNA macroarray-based analysis of gene expression. A Xenopus cDNA macroarray representing 42,240 independent clones was differentially hybridized with probes synthesized from the total RNA of normal and regenerating tails. Temporal expression analysis revealed that the up-regulated genes could be grouped into early or late responding genes. A comparative expression analysis revealed that most genes showed similar expression patterns between tail development and regeneration. However, some genes showed regeneration-specific expression. Finally, we identified 48 up-regulated genes that fell into several categories based on their putative functions. These categories reflect the various processes that take place during regeneration, such as inflammation response, wound healing, cell proliferation, cell differentiation, and control of cell structure. Thus, we have identified a panel of genes that appear to be involved in the process of regeneration.

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Species referenced: Xenopus laevis
Genes referenced: anxa2 atf5 atf5.2 c3 csrp2 ctsb h3-5 hmgb2 hmgn2 ifitm1 krt18 krt8 myh9 plg rps17 serpine1 sfrp2 snrnp27 tf wdr54

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	Figure 3. Whole mount in situ hybridization of up-regulated genes, grouped according to function as wound healing, inflammation response, and cell proliferation genes. Tailbud stage embryo (A, D, G, J, L, N), regenerating tail on day 2 (B, E, H), and regenerating tail on day 5 (C, F, I, K, M, O) were hybridized with DIG-labeled RNA probes. Lateral view, dorsal side up, anterior to the left. White arrowheads, notochord; white arrows, blood cells; red arrowhead, uncharacterized cells; black arrowheads, wound epidermis; broken line, the plane of amputation. A–C: Expression of annexin II (XL073e04). D–F: Expression of PAI1 (XL074b06). G–I: Expression of cathepsin B (XL040b13). J, K: Expression of complement component C3 (XL028a02). L, M: Expression of histone H3.3B (XL092g11). N, O: Expression of serotransferrin precursor (XL040d11).
	Figure 4. Whole mount in situ hybridization of up-regulated genes categorized according to function as cell signaling, cell differentiation, and cell structure genes. Tailbud stage embryo (A, C, E, G, I, K, M, O) and regenerating tail on day 5 (B, D, F, H, J, L, N, P). Lateral view, dorsal side up, anterior to the left. Red arrowheads, the proximal region of the regenerating tail; white arrowheads, notochord; black arrows, spinal cord; white arrows, fin epidermis; broken line, the plane of amputation. A, B: Expression of SFRP2 (XL040a14). C, D: Expression of KLG/PTK7 (XL005m14). E, F: Expression of ATF5 (XL049c20). G, H: Expression of cysteine-rich protein (XL014c20). I, J: Expression of HMG-17 (XL034a15). K, L: Expression of HMG-X (XL069l01). M, N: Expression of endo B keratin (XL024e24). O, P: Expression of keratin 8 (XL080o09).
	Figure 5. Whole mount in situ hybridization of unclassified and novel up-regulated genes. Tailbud stage embryo (A, C, E, G, I, K) and regenerating tail on day 5 (B, D, F, H, J, L). Lateral view, dorsal side up, anterior to the left. White arrowhead, regenerating muscles; black arrowhead, the terminal vesicle of the regenerating spinal cord; black arrows, spinal cord; white arrows, blood cells; broken line, the plane of amputation. A, B: Expression of 14kDa transmembrane protein (XL070a09). C, D: Expression of glutamine synthetase (XL106n21). E, F: Expression of XL064e17. G, H: Expression of XL025c16. I, J: Expression of XL051a01. K, L: Expression of XL063b01.
	anxa2 (annexin A2) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.
	serpine1 (serpin peptidase inhibitor, clade E (nexin, plasminogen activator inhibitor type 1), member 1) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.
	ctsb (cathepsin B) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.
	h3-5 (H3.5 histone) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.
	tf (transferin) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.
	atf5.2 (activating transcription factor 5, gene 2) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.
	ifitm1 (interferon induced transmembrane protein 1) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.
	wdr54 (WD repeat domain 54) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.
	snrnp27 (small nuclear ribonucleoprotein 27kDa (U4/U6.U5)) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.
	rps17 (ribosomal protein S17) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.