Identification of differentially expressed genes from multipotent epithelia at the onset of an asexual development

• Published in: Scientific reports Vol. 6; no. 1; p. 27357
• Main Authors: Ricci, Lorenzo, Chaurasia, Ankita, Lapébie, Pascal, Dru, Philippe, Helm, Rebecca R., Copley, Richard R., Tiozzo, Stefano
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 06-06-2016; Nature Publishing Group
• Subjects: 38/90; 38/91; 631/114; 631/136; 631/532/489; Animals; Epithelium - embryology; Gene expression; Gene Expression Profiling - methods; Gene Expression Regulation, Developmental; Genetics; Genomes; Humanities and Social Sciences; Life Sciences; Morphology; multidisciplinary; Nucleic Acid Amplification Techniques; Nucleic Acid Hybridization; Reproduction, Asexual; Reproductive Biology; Science; Sequence Analysis, RNA; Transcriptome; Urochordata - embryology
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep27357

Organisms that have evolved alternative modes of reproduction, complementary to the sexual mode, are found across metazoans. The chordate
Botryllus schlosseri
is an emerging model for asexual development studies.
Botryllus
can rebuild its entire body from a portion of adult epithelia in a continuous and stereotyped process called blastogenesis. Anatomy and ontogenies of blastogenesis are well described, however molecular signatures triggering this developmental process are entirely unknown. We isolated tissues at the site of blastogenesis onset and from the same epithelia where this process is never triggered. We linearly amplified an ultra-low amount of mRNA (<10ng) and generated three transcriptome datasets. To provide a conservative landscape of transcripts differentially expressed between blastogenic
vs.
non-blastogenic epithelia we compared three different mapping and analysis strategies with a
de novo
assembled transcriptome and partially assembled genome as references, additionally a self-mapping strategy on the dataset. A subset of differentially expressed genes were analyzed and validated by
in situ
hybridization. The comparison of different analyses allowed us to isolate stringent sets of target genes, including transcripts with potential involvement in the onset of a non-embryonic developmental pathway. The results provide a good entry point to approach regenerative event in a basal chordate.