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Anopheles albimanus lola knockdown immune response differential transcriptome: effect of Anopheles albimanus lola knockdown on the gene expression profile of the midgut in mosquitoes treated with an immune challenge

ABSTRACT: Anopheles mosquitoes are a potential transmitter of viruses and parasites that cause human diseases. An. albimanus is among the main malaria vectors in Mexico and Latin America. Mosquitoes' ability to transmit pathogens (vectorial competence) is directly related to their genetics and immunity. The innate immunity system is the first line of defense against non-self microbial invaders. The mosquito midgut is a crucial entry point for such microbial invaders and for mosquito immune interactions with them. Here, we provide evidence that the lola gene expression is upregulated by immune challenge in three mosquito tissues and cells: the midgut, fat body, and hemocytes. We applied an efficient method for double-stranded RNA (dsRNA)-mediated silencing of this gene expression in vivo as assessed by RT-qPCR. A transcriptome analysis was conducted to investigate the potential involvement of the Lola transcription factor in regulating gene expression. We combined RNAseq with reverse genetics, and our transcriptome analysis revealed that Lola participates in mosquito defense mechanisms. The knockdown of lola showed that it regulates the expression of genes associated with multiple functions, especially those related to detoxification and immunity. Here, we show for the first time a list of mosquito immunity genes that require Lola expression. These lola-dependent genes include antimicrobial peptides, C-lectins, pattern recognition receptors, Clip-domain serine proteases, ML-domain-containing proteins, enzymes that catalyze the generation and detoxification of reactive oxygen species, and components of the JNK immune signaling pathways in the midgut of An. albimanus. Our studies provide direct evidence of a functional link between JNK signaling and Lola factor in the mosquito midgut. Dysregulation of this pathway may contribute to increased pathogen invasion.

ORGANISM(S): Anopheles albimanus

PROVIDER: GSE317204 | GEO | 2026/06/01

REPOSITORIES: GEO

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Project description:The mosquito Anopheles gambiae uses its innate immune system to control bacterial and Plasmodium infection of its midgut tissue. The activation of potent IMD pathway-mediated anti-Plasmodium falciparum defenses is dependent on the presence of the midgut microbiota, which activate this defense system upon parasite infection through a peptidoglycan recognition protein, PGRPLC. We employed transcriptomic and reverse genetic analyses to compare the P. falciparum infection-responsive transcriptomes of septic and aseptic mosquitoes and to determine whether bacteria-independent anti-Plasmodium defenses exist. To examine the impact of P. falciparum infection on the mosquito midgut and carcass transcriptomes in the presence or absence of midgut bacteria, we used A. gambiae whole genome microarrays to compare the mRNA abundance of P. falciparum-infected and -naïve mosquitoes of antibiotic- and non-antibiotic treated cohorts. P. falciparum infection induced changes in the abundance of as many as 2,183 and 2,429 transcripts in whole mosquitoes belonging to a variety of functional groups in aseptic and septic mosquitoes. Ultimately, we were interested in identifying the genes involved in bacteria-independent anti-Plasmodium responses, and therefore we focused on transcripts displaying increased abundance in the parasite-infected aseptic midguts, placing a particular emphasis on those with predicted immune functions. Because of the central role of serine protease cascades in regulating insect immune defenses, we focused the remainder of our analysis on a clip-domain serine protease C2 (CLIPC2, AGAP004317) and a serine protease inhibitor 7 (SRPN7, AGAP007693) that were specifically upregulated in the parasite-infected, aseptic mosquito midgut. We showed that SRPN7 negatively and CLIPC2 positively regulate the anti-Plasmodium defense, independently of the midgut-associated bacteria. Co-silencing assays suggested that these two genes may function together in a signaling cascade. Neither gene was regulated, nor modulated, by infection with the rodent malaria parasite Plasmodium berghei, suggesting that SRPN7 and CLIPC2 are components of a defense system with preferential activity towards P. falciparum. Further analysis using RNA interference determined that these genes do not regulate the anti-Plasmodium defense mediated by the IMD pathway, and both factors act as agonists of the endogenous midgut microbiota, further demonstrating the lack of functional relatedness between these genes and the bacteria-dependent activation of the IMD pathway. This is the first study confirming the existence of a bacteria-independent, anti-P. falciparum defense.

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Anopheles albimanus lola knockdown immune response differential transcriptome: effect of Anopheles albimanus lola knockdown on the gene expression profile of the midgut in mosquitoes treated with an immune challenge

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