Analysis of Gene Expression During Amphibian Tail Development (2006 JMU Biosymposium)

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Analysis of Gene Expression During Amphibian Tail Development Stephen Turner photo: www.herpsofnc.org JMU Biosymposium 2006

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Tail Development 101 Vertebrates: Formation begins during embryogenesis. But in MOST vertebrates formation of new segments terminates. H. Scutatum adds new tail segments throughout larval, juvenile, and ADULT stages!

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Somites photo: www.ulb.ac.be • Blocks of undifferentiated cells that arise from mesodermal tissue. • Blocks align down the anteroposterior axis flanking neural tube. • Cells in somite fated to become bone, muscle, or skin in trunk and tail segments. Bone Muscle Skin photo: www.animalpicturesarchive.com • First visible segmented structure in developing embryo.

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Visualizing somites Somite 12-101 Antibody staining * * RNA in situ hybridization MyoD gene: expressed in somites Probe RNA mRNA in cell 12-101 ab 2° ab

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12-101 and MyoD Staining in Xenopus Antibody Staining 12-101 RNA in situ MyoD

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12-101 and MyoD Staining in H. scutatum Antibody Staining 12-101 RNA in situ: Using Xenopus MyoD probe Standard protocol used on Xenopus Reduced stringency

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www.herpnet.net Xenopus laevis Hemidactylium scutatum Nieuwkoop and Faber Tail Development in Embryo, Larvae, Metamorphosis, and Adult

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Tailbud genes Images from Linda K. Gont, et al. • Xbra & Xnot important in embryonic mesoderm patterning. • Xbra in particular is implicated in somitogenesis. • Previous research addresses embryonic expression. • When, and where are these genes expressed in post-embryonic tissues? Xbra: Xnot2:

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Xbra expression in Xenopus laevis Stage 25 Stage 27 Stage 31 Stage 37 Stage 47

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Stage 32 Stage 31 Xnot2 expression in Xenopus laevis Stage 27 Stage 47 Stage 40

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Ongoing work & future directions • Examine expression of MyoD, Xbra, and Xnot2 in sections. • Modify 12-101 protocol for use in H. scutatum. • Modify my protocol to examine expression of these genes in late stage Xenopus (larval and metamorphosis stages). • Clone the H. scutatum homolog of these genes for in situ hybridization analysis of tail development in embryonic, larval, juvenile, and adult stages.

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Acknoledgements Carol Hurney Sharon Babcock Also: Monroe Lab Temple Lab Seifert Lab Keller Lab (Xenopus laevis embryos) Grainger Lab (MyoD clone) De Robertis Lab (Xnot2 clone) Smith Lab (XBra clone) Teresa Pelletier (Pretty pictures)