The cytosolic calcium concentration is regulated by calcium-processing proteins such as TRPV5, TRPV6, NCX1, and PMCA1. Those calcium-processing proteins are important for physiological functions in the brain. The effects of steroid hormones on calcium-processing protein expressions in the brains are undescribed. Thus, the effects of steroid hormones on the distribution, localization, and expressions of calcium-processing proteins in the brain were analyzed. Immature female rats were injected for 5 days with estrogen (E2), progesterone (P4), dexamethasone (DEX), and their antagonists (ICI 182,780 and RU486). The localization and expression of calcium-processing proteins in rat brain were observed by immunofluorescence and western blot analyses, respectively. We found that TRPV5 and TRPV6 proteins were highly expressed in the cerebral cortex (CT), hypothalamus (HY), and brain stem (BS) compared to that in the olfactory bulb (OB) and cerebellum (CB). Also, the NCX1 protein was highly expressed in CT and BS compared to that in OB, HY, and CB, and PMCA1 protein was highly expressed in CT compared to that in other brain regions. Furthermore, expression levels of calcium-processing proteins were regulated by E2, P4, and/or DEX in the CT and HY. In summary, calcium-processing proteins are widely expressed in the immature rat brain, and expressions of calcium-processing proteins in CT and HY are regulated by E2, P4, and/or DEX and can be recovered by antagonist treatment. These results indicate that steroid hormone regulation of calcium-processing proteins may serve as a critical regulator of cytosolic calcium absorption and release in the brain.