Selecting reference genes for RT-qPCR studies involving the presence of B chromosomes in Psalidodon (Characiformes, Characidae)

Author(s)

Vidal, Mateus Rossetto ; Lasmar, Lucas F. ; Nadai, Pamela C. F. ; Oliveira, Claudio ; Silva, Duilio M. Z. A. ; Foresti, Fausto

• Document type: Journal article
• Source: MOLECULAR BIOLOGY REPORTS; v. 51, n. 1, p. 7-pg., 2024-12-01.

Abstract

Background B chromosomes are extra non-essential elements present in several eukaryotes. Unlike A chromosomes which are essential and present in all individuals of a species, B chromosomes are not necessary for normal functioning of an organism. Formerly regarded as genetically inactive, B chromosomes have been discovered to not only express their own genes, but also to exert influence on gene expression in A chromosomes. Recent studies have shown that, in some Psalidodon (Characiformes, Characidae) species, B chromosomes might be associated with phenotypic effects, such as changes in the reproductive cycle and gene expression. Methods and results In this study, we aimed to establish stable reference genes for RT-qPCR experiments conducted on gonads of three fish species within Psalidodon genus, both in the presence and absence of B chromosomes. The stability of five selected reference genes was assessed using NormFinder, geNorm, BestKeeper, and RefFinder algorithms. We determined ppiaa and pgk1 as the most stable genes in P. fasciatus, whereas ppiaa and hmbsa showed the highest stability in P. bockmanni. For P. paranae, tbp and hprt1 were the most stable genes in females, and ppiaa and hprt1 were the most stable in males. Conclusions We determined the most stable reference genes in gonads of three Psalidodon species considering the presence of B chromosomes. This is the first report of reference gene stability in the genus and provides valuable tools to better understand the effects of B chromosomes at gene expression level. (AU)

• FAPESP's process: 17/22447-8 - Comparative Genetic Activity on B Chromosomes of Astyanax species (Teleostei, Characiformes)
• Grantee: Duílio Mazzoni Zerbinato de Andrade Silva
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 20/13433-6 - Phylogeny and molecular identification of fishes of the superorder Ostariophysi (Chordata: Actinopterygii) using genomic approaches
• Grantee: Claudio de Oliveira
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 19/15140-9 - Analysis of the genic content and differential expression of variants of B chromosome in Astyanax (Teleostei, Characiformes)
• Grantee: Mateus Rossetto Vidal
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 20/01775-0 - Functional and evolutionary aspects of the supernumerary chromosomes in fish based on the Astyanax model (Teleostei, Caraciformes), revealed by genomic and structural analysis
• Grantee: Fausto Foresti
• Support Opportunities: Regular Research Grants