Abstract
The development of the brain is sex-dimorphic, and as a result so are many neurological disorders. One approach for studying sex-dimorphic brain development is to measure gene expression in biological samples using RT-qPCR. However, the accuracy and consistency of this technique relies on the reference gene(s) selected. We analyzed the expression of ten reference genes in male and female samples over three stages of brain development, using popular algorithms NormFinder, GeNorm and Bestkeeper. The top ranked reference genes at each time point were further used to quantify gene expression of three sex-dimorphic genes (Wnt10b, Xist and CYP7B1). When comparing gene expression between the sexes expression at specific time points the best reference gene combinations are: Sdha/Pgk1 at E11.5, RpL38/Sdha E12.5, and Actb/RpL37 at E15.5. When studying expression across time, the ideal reference gene(s) differs with sex. For XY samples a combination of Actb/Sdha. In contrast, when studying gene expression across developmental stage with XX samples, Sdha/Gapdh were the top reference genes. Our results identify the best combination of two reference genes when studying male and female brain development, and emphasize the importance of selecting the correct reference genes for comparisons between developmental stages.
Highlights
Mammals that reproduce through sexual reproduction have intrinsic differences between male and female sexes
To ensure accurate analysis of gene expression, ten candidate reference genes (Table 1) were investigated to determine how stable each gene is for sex and time-point of development
The candidate reference genes selected included those genes commonly used in Reverse Transcription quantitative Polymerase Chain Reaction (RT-qPCR) experiments and genes were stably expressed in multiple mouse adult tissues, but had not been tested for suitability as reference genes with mouse embryo tissues (Kouadjo et al, 2007)
Summary
Mammals that reproduce through sexual reproduction have intrinsic differences between male and female sexes. The most obvious morphological differences are those of reproductive organs. Many other less apparent organs and tissues are modulated in a sex-dimorphic manner. There exist numerous differences in the human brain between the male and female sexes. The rate of maturation of the female brain reaches its peak maturity at 10 years of age compared to the male brain, which only peaks 4 years later (Giedd et al, 1997). Numerous neurological diseases have sex-specific biases (Fombonne, 2009), including, schizophrenia
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
