Functional dissection of mosquito humidity sensing reveals distinct dry and moist cell contributions to blood feeding and oviposition

Aedes aegypti (Ae. aegypti) mosquitoes are major vectors of dengue, chikungunya and other arboviral diseases. Ae. aegypti’s capacity to reproduce and to spread disease depends on the female mosquitoes’ ability to obtain blood meals and find water-filled containers in which to lay eggs (oviposit). While humidity sensation (hygrosensation) has been implicated in these behaviors, the specific hygrosensory pathways involved have been unclear. Here we establish the distinct molecular requirements and anatomical locations of Ae. aegypti Dry Cells and Moist Cells, and examine their contributions to behavior. We show that Dry Cell and Moist Cell responses to humidity involve different ionotropic receptor (IR) family sensory receptors, with dry air-activated Dry Cells reliant upon the ionotropic receptor Ir40a, and humid air-activated Moist Cells upon Ir68a. Both classes of hygrosensors innervate multiple antennal sensilla, including sensilla ampullacea near the antennal base as well as two clas..., Calcium imaging:
To monitor intracellular levels of intracellular calcium in Ir40a-positive cells, QUAS-GCaMP7s (marked by 3xP3-CFP) mosquitoes were crossed to the Ir40aRFP and Ir40aEYFP knock-ins (which express QF2 under the control of endogenous Ir40a regulatory sequences). Ir40aRFP/+ ; QUAS-GCaMP7s/+ and Ir40aRFP/Ir40aEFYP; QUAS-GCaMP7s/+ animals were used to assess responses in control animals and Ir40a mutants, respectively. As Ir68a QF2 knock-ins drove weaker GCaMP expression, imaging of Ir68a-positive cells was performed in a yellow mutant background, whose lighter cuticle facilitates imaging. Ir68aRFP/+ ; QUAS-GCaMP7s/+ animals were crossed with a yellow mutant strain for two generations, and yellow/ yellow; Ir68aRFP/+ ; QUAS-GCaMP7s/+ animals were used for control imaging. These mosquitoes were crossed to yellow/ yellow; Ir68aRFP /+ individuals and descendants were molecularly genotyped to create a stable stock of yellow/ yellow; Ir68aRFP/Ir68aRFP; QUAS-GCaMP7s/+ for Ir68a muta..., , # Calcium Imaging Movies

[https://doi.org/10.5061/dryad.c2fqz61jv](https://doi.org/10.5061/dryad.c2fqz61jv)

## Description of the data and file structure

These files contain the raw calcium imaging data presented in Figure 2, Figure S2, Figure S3 and Figure S4 of \"Functional dissection of mosquito humidity sensing reveals distinct Dry and Moist Cell contributions to blood feeding and oviposition\" by Ruocong Tang, Rachel Busby, Willem J. Laursen, Geoffrey Keane and Paul A. Garrity.

### Files and variables

#### File: Figure\_S2.zip

**Description:** Calcium imaging movies presented as .avi movies.

#### File: Figure\_2M.zip

**Description:** Calcium imaging movies presented as a series of .tif files, with metadata provided in accompanying metatdata.txt files.

#### File: Figure\_2H.zip

**Description:** Calcium imaging movies presented as a series of .tif files, with metadata provided in accompanying metatdata.txt files.

#### File: Figure\_S4.zip

**Description:** Calcium imaging...