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Investigating Sex Hormone Influence on Nesfatin‐1 Signaling through qPCR

Nesfatin‐1 is a potent anorectic peptide that participates in the hypothalamo‐pituitary‐adrenal (HPA) axis, and previous research indicates that the expression and function of nesfatin‐1 is influenced by female sex hormones. While nesfatin‐1 signaling in the HPA axis of male animals is well‐studied, nesfatin‐1 signaling in the HPA axis of female animals remains to be investigated. Changes in nesfatin‐1 action and expression in dispersed rat pituitaries as well as hypothalamus, pituitary, and adrenal gland tissues harvested across the estrus cycle was determined through use of qPCR. However, housekeeping genes used for normalization of qPCR data have not been validated for use in studies examining the role of female sex hormones. Therefore, tissues were collected from cycling female rats to perform qPCR experiments measuring expression of the most commonly used housekeeping genes in order to determine which housekeeping genes are stable across the estrus cycle. These studies suggest that nesfatin‐1 expression and signaling in the HPA is influenced by female sex hormones. Furthermore, our data indicate which housekeeping genes are most appropriate for research on sex differences.