Transcriptome-wide selection and validation of a solid set of reference genes for gene expression studies in the cephalopod mollusk Octopus vulgaris

Frontiers in Molecular Neuroscience

Molecular Neuroscience

Pamela Imperadore¹  Francesca Raffini¹  Giovanna Ponte¹  Graziano Fiorito¹  Vittoria Allegretti¹  Caterina Millino²  Stefano Cagnin³ 

[1] Department of Biology and Evolution of Marine Organisms, Stazione Zoologica Anton Dohrn, Napoli, Italy;Department of Biology, University of Padova, Padova, Italy;Department of Biology, University of Padova, Padova, Italy;CIR-Myo Myology Center, University of Padova, Padova, Italy;

关键词: Octopus vulgaris;    reference genes;    qRT-PCR;    cephalopods;    nervous system;    molecular fingerprint;   

DOI  :  10.3389/fnmol.2023.1091305

 received in 2022-11-06, accepted in 2023-02-20,  发布年份 2023

来源: Frontiers

【 摘 要 】

Octopus vulgaris is a cephalopod mollusk and an active marine predator that has been at the center of a number of studies focused on the understanding of neural and biological plasticity. Studies on the machinery involved in e.g., learning and memory, regeneration, and neuromodulation are required to shed light on the conserved and/or unique mechanisms that these animals have evolved. Analysis of gene expression is one of the most essential means to expand our understanding of biological machinery, and the selection of an appropriate set of reference genes is the prerequisite for the quantitative real-time polymerase chain reaction (qRT-PCR). Here we selected 77 candidate reference genes (RGs) from a pool of stable and relatively high-expressed transcripts identified from the full-length transcriptome of O. vulgaris, and we evaluated their expression stabilities in different tissues through geNorm, NormFinder, Bestkeeper, Delta-CT method, and RefFinder. Although various algorithms provided different assemblages of the most stable reference genes for the different kinds of tissues tested here, a comprehensive ranking revealed RGs specific to the nervous system (Ov-RNF7 and Ov-RIOK2) and Ov-EIF2A and Ov-CUL1 across all considered tissues. Furthermore, we validated RGs by assessing the expression profiles of nine target genes (Ov-Naa15, Ov-Ltv1, Ov-CG9286, Ov-EIF3M, Ov-NOB1, Ov-CSDE1, Ov-Abi2, Ov-Homer2, and Ov-Snx20) in different areas of the octopus nervous system (gastric ganglion, as control). Our study allowed us to identify the most extensive set of stable reference genes currently available for the nervous system and appendages of adult O. vulgaris.

【 授权许可】

Unknown   
Copyright © 2023 Imperadore, Cagnin, Allegretti, Millino, Raffini, Fiorito and Ponte.