In blowflies and fruit flies, the Malpighian tubules (MTs) plays a key role in regulating Ca2+ homeostasis. However, the renal regulation of Ca2+ balance in hematophagous flies, such as mosquitoes, has not been extensively examined, especially in the context of hematophagy. The objective of our study was to understand how MTs regulates mosquito Ca2+ homeostasis during blood meal digestion. We first examined the chronological changes in Ca2+ content of adult female Ae. aegypti after blood feeding. Adult female mosquitoes were fed blood and dissected at several time points post blood meal; non-fed mosquitoes served as controls. While no significant differences were detected in the Ca2+ level of mosquito whole bodies, we found dynamic changes in the Ca2+ content in MTs post blood feeding. To generate molecular insights into transport mechanisms responsible for these changes in Ca2+ content, we characterized the expressions of plasma membrane Ca2+ ATPase (PMCA) in isolated mosquito MTs before and after blood feeding using RT-QPCR, Western Blot, and immunolocalization. Differential expressions of PMCA on mRNA and protein level were detected in MTs during time periods when the Ca2+ content of MTs was changing, suggesting its potential roles in Ca2+ regulation. The results are expected to enhance our understanding of mosquito physiology.