Journal of Mosquito Research 2024, Vol.14, No.2, 61-66 http://emtoscipublisher.com/index.php/jmr 64 Figure 4Gr22+OSNs on the Anopheles gambiae maxillary palps respond to CO2 Figure 5 shows the response of Ir76b+, Ir25a+and orco+ olfactory sensory neurons (OSNs) on the antennae of Anopheles gambiae to specific human-related odor stimuli. Part A presents the response of Ir76b+ neurons to 0.28% trimethylamine and its comparison with a water control; Part B displays the response of Ir25a+neurons to 1% pyridine and its comparison with a water control; Part C shows the response of orco+neurons to 1% hexanal and its comparison with a paraffin oil control. On the left side of each figure are the GCaMP6f activity traces following stimulation, and on the right side is the quantification of the maximum ΔF/F values derived from these traces. Orange and black bars indicate the start and duration of the stimulus, respectively. All charts are plotted with mean ± standard error, and bilateral Wilcoxon signed-rank tests show p-values less than 0.01, indicating that the responses after stimulation are significantly higher than controls. This indicates that these OSNs can specifically respond to odor molecules related to humans, providing important insights into how mosquitoes use olfaction to identify humans. 2 Analysis of Research Findings By delving into the olfactory system of Anopheles gambiae, this study unraveled the complex mechanisms behind this malaria mosquito's response to human odors, demonstrating how scientists can precisely insert the driver-responder-marker (DRM) system into the mosquito genome using CRISPR-Cas9 technology. This enables precise manipulation and tracking of specific chemoreceptor genes in olfactory sensory neurons. Such innovative gene editing methods not only enhance the precision of research but also pave new avenues for experimental design and olfactory studies. Utilizing calcium imaging techniques, the research team further validated the efficiency and accuracy of the created gene expression lines in capturing the mosquito's response to human odor molecules. The combined use of these techniques allows scientists to observe and analyze in real-time the activity of olfactory sensory neurons upon encountering specific odorants, deepening our understanding of how mosquitoes identify human hosts through their olfactory system. These findings not only deepen our knowledge of mosquito olfactory behavior but also provide a valuable scientific basis for developing targeted mosquito control
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