Girardia Dorotocephala Transcriptome Sequence, Assembly, and Validation Through Characterization of Piwi Homologs and Stem Cell Progeny Markers

Overview

Journal Dev Biol

Publisher Elsevier

Specialties Biology
Reproductive Medicine

Date 2017 Aug 5

PMID 28774726

Citations 7

Authors

Eugene Matthew P Almazan

Sydney L Lesko

Michael P Markey

Labib Rouhana

Affiliations

Soon will be listed here.

Abstract

Planarian flatworms are popular models for the study of regeneration and stem cell biology in vivo. Technical advances and increased availability of genetic information have fueled the discovery of molecules responsible for stem cell pluripotency and regeneration in flatworms. Unfortunately, most of the planarian research performed worldwide utilizes species that are not natural habitants of North America, which limits their availability to newcomer laboratories and impedes their distribution for educational activities. In order to circumvent these limitations and increase the genetic information available for comparative studies, we sequenced the transcriptome of Girardia dorotocephala, a planarian species pandemic and commercially available in North America. A total of 254,802,670 paired sequence reads were obtained from RNA extracted from intact individuals, regenerating fragments, as well as freshly excised auricles of a clonal line of G. dorotocephala (MA-C2), and used for de novo assembly of its transcriptome. The resulting transcriptome draft was validated through functional analysis of genetic markers of stem cells and their progeny in G. dorotocephala. Akin to orthologs in other planarian species, G. dorotocephala Piwi1 (GdPiwi1) was found to be a robust marker of the planarian stem cell population and GdPiwi2 an essential component for stem cell-driven regeneration. Identification of G. dorotocephala homologs of the early stem cell descendent marker PROG-1 revealed a family of lysine-rich proteins expressed during epithelial cell differentiation. Sequences from the MA-C2 transcriptome were found to be 98-99% identical to nucleotide sequences from G. dorotocephala populations with different chromosomal number, demonstrating strong conservation regardless of karyotype evolution. Altogether, this work establishes G. dorotocephala as a viable and accessible option for analysis of gene function in North America.

Citing Articles

Somatic piRNA and PIWI-mediated post-transcriptional gene regulation in stem cells and disease.

Patel M, Jiang Y, Kakumani P Front Cell Dev Biol. 2024; 12:1495035.

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RNaseH-based ribodepletion of total planarian RNA improves detection of longer and non-polyadenylated transcripts.

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Molecular characterization of a flatworm Girardia isolate from Guanajuato, Mexico.

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Transcription Factors Active in the Anterior Blastema of .

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Regeneration of Planarian Auricles and Reestablishment of Chemotactic Ability.

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