Establishment and a comparative transcriptomic analysis of a male-specific cell line from the African malaria mosquito Anopheles gambiae

Abstract

Cell lines allow studying various biological processes that may not be easily tractable in whole organisms. Here, we have established the first male-specific cell line from the African malaria mosquito, Anopheles gambiae. The cells, named AgMM and derived from the sex-sorted neonate larvae, were able to undergo spontaneous contractions for a number of passages following establishment, indicating their myoblast origin. Comparison of their transcriptome to the transcriptome of an A. gambiae-derived Sua5.1 hemocyte cells revealed distinguishing molecular signatures of each cell line, including numerous muscle-related genes that were highly and uniquely expressed in the AgMM cells. Moreover, the AgMM cells express the primary sex determiner gene Yob and support male sex determination and dosage compensation pathways. Therefore, the AgMM cell line represents a valuable tool for molecular and biochemical in vitro studies of these male-specific processes. In a broader context, a rich transcriptomic data set generated in this study contributes to a better understanding of transcribed regions of the A. gambiae genome and sheds light on the biology of both cell types, facilitating their anticipated use for various cell-based assays.

Molecular characterization and genetic authentication assay for Anopheles ‘hemocyte-like’ cell lines 4a-3A and 4a-3B

“…Anopheline cell lines have been used to examine a wide variety of traits, including to determine if the Minos transposable element integrates in a transposase-dependent manner [ 2 ], to characterize the cellular response to Bin toxin, a candidate vector control tool [ 3 , 4 ], to study hemocyte properties and prophenoloxidase (PPO) expression [ 5 ], for studies of phagocytosis and hemocyte gene expression [ 6 ], to characterize the effect of genetic variation enhancer activity [ 7 ], to characterize interaction of proteins with a role in malaria infection [ 8 ] and to perform STARR-seq and establish a genome-wide catalog of transcriptional enhancers [ 9 ]. Establishment of a male-specific anopheline cell line suitable for the study of processes unique to male mosquitoes was also recently reported [ 10 ]. The 4a-3B Anopheles gambiae cell line is the first continuous insect cell line used to study both humoral and cellular immune responses of An.…”