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Key aspects of accurate normalization of RT-qPCR data in the phylum Poriferaтезисы доклада Электронная публикация Тезисы
- Авторы: Skorentseva K., Melnikov N., Saidova A., Lavrov A.
- Сборник: Беломорская студенческая научная сессия СПбГУ — 2026
- Тезисы
- Год издания: 2026
- Место издания: Свое издательство Санкт-Петербург
- Первая страница: 25
- Последняя страница: 25
- Аннотация: Quantitative real-time PCR (RT-qPCR) is a precise, sensitive, and widely used method to assess gene expression. However, its application requires accurate normalization of the resulting data, especially when it comes to non-model species. Studies on such organisms (for example, invertebrates) can be complicated by intraspecific and seasonal variability, limitations in sample availability, challenges of laboratory maintenance, and other factors. Sponges (phylum Porifera) are the sister group of Eumetazoa. They lack muscle and nervous tissues, yet possess highly labile epithelia (choanoderm and pinacoderm) and represent a promising model in evolutionary developmental biology. Sponges offer unique opportunities to investigate tissue homeostasis, intercellular communication in the absence of neurohumoral regulation, and ultimately the origins of multicellularity. In this study, we analyzed the expression stability of seven commonly used reference genes for RT-qPCR normalization: ACT1, RPL13A, GAPDH, HPRT1, RSP3A, TBP, and LMN1 responsible for key cellular metabolic processes. Stability was assessed during an induced regeneration experiment in the calcareous marine sponge Leucosolenia corallorrhiza and further validated in three additional species: Halisarca dujardinii, Ephydatia fluviatilis, and Lycopodina hypogea. These species were selected to represent broad ecological and phylogenetic diversity spanning calcareous and demosponges, marine and freshwater forms, filter feeders, and a carnivorous species. To evaluate stability, we applied four widely used algorithms: geNorm, NormFinder, BestKeeper, and RefFinder. We also tested how the choice of reference gene affects the quantification of RHOA, a key regulator of the actin cytoskeleton. None of the genes tested showed universal stability in all species examined, underscoring the need for species-specific validation. We further demonstrate that the use of multiple reference genes for normalization not only reduces variability but also helps avoid misinterpretation of biologically meaningful processes. Together, these results provide a robust methodological framework for future molecular studies of sponges, particularly those focused on regeneration, normal development, and interactions with the environment. This work was supported by RSF grants No. 23-74-10005 (research on Leucosolenia corallorrhiza and Halisarca dujardinii) and No. 24-14-00452 (research on Ephydatia fluviatilis and Lycopodina hypogea).
- Добавил в систему: Скоренцева Ксения Витальевна