Finding genes that maintain a stable expression level across large developmental windows is a real challenge. In our research, we’ve developed a novel method to normalize gene expression in the sheep heart, covering the important stages from late gestation, through neonatal life, and into adulthood. This is an essential step in understanding how gene regulation changes during heart development and how it impacts the long-term function of the heart.
Normalization to stable reference genes is a key technique in any experimental study. It helps ensure that the changes we see in our genes of interest are real and not just an artifact of technical variations. This gives us confidence in the data we collect. However, during the course of development, changes are so widespread that even genes typically considered stable can show alterations. We found that the global transcription of mRNA itself is not exempt from this phenomenon—there were significant shifts in mRNA levels across the periods we studied.