摘要：Bats are the only mammals capable of powered flight and their body temperature can reach up to 42 °C during flight. Additionally, bats display robust type I IFN interferon (IFN-I) responses and some species constitutively express IFN-α. Reference genes with stable expression under temperature oscillations and IFN-I release are therefore critical for normalization of quantitative reverse-transcription polymerase chain reaction (qRT-PCR) data in bats. The expression stability of reference genes in
Rousettus aegyptiacus remains elusive, although this species is frequently used in the infection research. We selected
ACTB,
EEF1A1,
GAPDH and
PGK1 as candidate reference genes and evaluated their expression stability in various tissues and cells from this model bat species upon IFN-I treatment at 35 °C, 37 °C and 40 °C by qRT-PCR. We employed two statistical algorithms, BestKeeper and NormFinder, and found that
EEF1A1 exhibited the highest expression stability under all tested conditions.
ACTB and
GAPDH displayed unstable expression upon temperature change and IFN-I treatment, respectively. By normalizing to
EEF1A1, we uncovered that
GAPDH expression was significantly induced by IFN-I in
R. aegyptiacus. Our study identifies
EEF1A1 as the most suitable reference gene for qRT-PCR studies upon temperature changes and IFN-I treatment and unveils the induction of
GAPDH expression by IFN-I in
R. aegyptiacus. These findings are pertinent to other bat species and may be relevant for non-volant mammals that show physiological fluctuations of core body temperature.
