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Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type Xenopus tropicalis Tail and Notochord During Metamorphosis

ABSTRACT: Our experiment revealed the notochord specific metamorphosis-induced genes to clarify the molecular mechanisms of tail regression.

ORGANISM(S): Xenopus tropicalis

PROVIDER: GSE133220 | GEO | 2022/06/14

REPOSITORIES: GEO

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Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type Xenopus tropicalis Tail and Notochord During Metamorphosis

Project description:Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type Xenopus tropicalis Tail and Notochord During Metamorphosis

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Project description:Tail regression is one of the most prominent transformations observed during anuran metamorphosis. A tadpole tail that is twice as long as the tadpole trunk nearly disappears within 3 days in Xenopus tropicalis. Several years ago, it was proposed that this phenomenon is driven by an immunological rejection of larval-skin-specific antigens, Ouro proteins. We generated ouro-knockout tadpoles using the TALEN method to reexamine this immunological rejection model. Both the ouro1- and ouro2-knockout tadpoles expressed a very low level of mRNA transcribed from a targeted ouro gene, an undetectable level of Ouro protein encoded by a target gene and a scarcely detectable level of the other Ouro protein from the untargeted ouro gene in tail skin. Furthermore, congenital athymic frogs were produced by Foxn1 gene modification. Flow cytometry analysis showed that mutant frogs lacked splenic CD8(+) T cells, which play a major role in cytotoxic reaction. Furthermore, T-cell-dependent skin allograft rejection was dramatically impaired in mutant frogs. None of the knockout tadpoles showed any significant delay in the process of tail shortening during the climax of metamorphosis, which shows that Ouro proteins are not essential to tail regression at least in Xenopus tropicalis and argues against the immunological rejection model.

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Expression data from stage 54 Xenopus laevis larvae treated with 20 nM triiodothyronine (T3)

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Expression data from stage 54 Xenopus laevis larvae treated with 20 nM triiodothyronine (T3)

Project description:Induction of Xenopus laevis larvae metamorphosis is dependent on exposure to TH. Metamorphosis involves the regression, growth or remodeling of almost all the tissues in the animals body. Metamorphosis in frogs is induced by thyroid hormone. Each organ system has a unique morphological and genetic program that it follows while undergoing metamorphosis involving both the upregulation and downregulation of genes. In this array we examined the change in gene expression in the tail of larvae undergoing precocious metamorphosis following induction with thyroid hormone. Whole stage 54 xenopus larvae were exposed to either vehicle for 48 hours or 20 nM T3 for 6 or 48 hours. Total RNA was then purified from tail tissue and the samples were examined by hybridization to the Affymetrix Xenopus array. Developmental stage 51-54 Xenopus larvae were selected because these stages do not express high levels of endogenous T3 but are still capable of morphologically responding to exposure.

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