A neurogenetic toolkit to decodeAnopheles gambiaeolfaction DOI Open Access
Diego Giraldo, Andrew Hammond,

Wu Jinling

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Aug. 17, 2023

Summary The African malaria mosquito Anopheles gambiae exhibits a strong innate sensory drive to seek out human scent. To detect odorants, An. uses olfactory neurons (OSNs) that can be divided into different classes by unique repertoires of chemoreceptor gene expression. We applied CRISPR-Cas9-mediated T2A-In Frame Fusions and the QF2/QUAS system gain genetic access specific OSN subsets in expressing genes Ir25a , Ir76b Gr22 orco . first optimized methods generate cell-type QF2 driver QUAS responder lines map expression patterns these chemoreceptors across appendages. next transcuticular calcium imaging record neurophysiological responses select human-related odorants for each class. This neurogenetic toolkit tiling including those responsive CO 2 stands support systematic efforts decode olfaction this prolific disease vector at high-resolution combat malaria.

Language: Английский

Mosquito Cell Atlas: A single-nucleus transcriptomic atlas of the adultAedes aegyptimosquito DOI Creative Commons
Olivia V. Goldman,

Alexandra E. DeFoe,

Yanyan Qi

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 25, 2025

The female mosquito's remarkable ability to hunt humans and transmit pathogens relies on her unique biology. Here, we present the Mosquito Cell Atlas (MCA), a comprehensive single-nucleus RNA sequencing dataset of more than 367,000 nuclei from 19 dissected tissues adult male Aedes aegypti , providing cellular-level resolution mosquito We identify novel cell types expand our understanding sensory neuron organization chemoreceptors all tissues. Our analysis uncovers male-specific cells sexually dimorphic gene expression in antenna brain. In mosquitoes, find that glial brain, rather neurons, undergo most extensive transcriptional changes following blood feeding. findings provide insights into cellular basis behavior sexual dimorphism. MCA aims serve as resource for vector biology community, enabling systematic investigation cell-type specific across

Language: Английский

Citations

1
Neurogenetic identification of mosquito sensory neurons DOI Creative Commons
Joanna K. Konopka, Darya Task, Danny Poinapen

et al.

iScience, Journal Year: 2023, Volume and Issue: 26(5), P. 106690 - 106690

Published: April 19, 2023

Anopheles mosquitoes, as vectors for the malaria parasite, are a global threat to human health. To find and bite human, they utilize neurons within their sensory appendages. However, identity quantification of appendage lacking. Here we use neurogenetic approach label all in coluzzii mosquitoes. We homology assisted CRISPR knock-in (HACK) generate T2A-QF2w synaptic gene bruchpilot. membrane-targeted GFP reporter visualize brain quantify major chemosensory appendages (antenna, maxillary palp, labella, tarsi, ovipositor). By comparing labeling brp>GFP Orco>GFP predict extent expressing ionotropic receptors (IRs) or other receptors. This work introduces valuable genetic tool functional analysis mosquito neurobiology initiates characterization that guide behavior.

Language: Английский

Citations

13
Adaptive genomic signatures of globally invasive populations of the yellow fever mosquito Aedes aegypti DOI Creative Commons
Alejandro Nabor Lozada‐Chávez, Irma Lozada-Chávez, Niccolò Alfano

et al.

Nature Ecology & Evolution, Journal Year: 2025, Volume and Issue: unknown

Published: March 28, 2025

Language: Английский

Citations

0
Impact of elevated CO2 level and egg quiescence duration on gene expression in the peripheral olfactory system of Aedes aegypti DOI Creative Commons

Sukritha Nalikkaramal,

Sharon R. Hill, Rickard Ignell

et al.

Scientific Reports, Journal Year: 2025, Volume and Issue: 15(1)

Published: April 24, 2025

Abstract Elevation in CO 2 can significantly impact the biology of various organisms, affecting life-history traits both aquatic and terrestrial forms, including disease-vectoring mosquitoes. For mosquitoes, this effect is accentuated by egg quiescence duration, resulting a change foraging adult females. Female mosquitoes rely on their olfactory system for locating resources, such as nectar blood. This study employs transcriptomic approach to investigate how projected elevation level, under worst-case scenario, interacts with extended duration modulate molecular machinery peripheral system, antennae maxillary palps, yellow fever mosquito, Aedes aegypti . The transcriptome analysis demonstrates significant changes abundance genes related metabolism, xenobiotics degradation chemosensory function, most pronounced effects observed sensing tissue, palp. provides novel insights into anthropogenic climate sensory disease vectors, which may have cascading resource-seeking behaviour.

Language: Английский

Citations

0
Optimized genetic tools for neuroanatomical and functional mapping of the Aedes aegypti olfactory system DOI Creative Commons
Shruti Shankar, Diego Giraldo,

Genevieve M. Tauxe

et al.

G3 Genes Genomes Genetics, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 24, 2025

Abstract The mosquito Aedes aegypti is an emerging model insect for invertebrate neurobiology. We detail the application of a dual transgenesis marker system that reports nature transgene integration with circular donor template CRISPR–Cas9-mediated homology-directed repair at target chemoreceptor genes. Employing this approach, we demonstrate establishment cell-type-specific T2A-QF2 driver lines A. olfactory co-receptor genes Ir8a and orco via canonical CO2 receptor complex gene Gr1 noncanonical involving duplication intended cassette separated by intervening plasmid sequence. Using Gr1+ sensory neurons as example, show introgression such QUAS responder transgenes into yellow cuticular pigmentation mutant strain facilitates transcuticular calcium imaging CO2-evoked neural activity on maxillary palps enhanced sensitivity relative to wild-type mosquitoes enveloped dark melanized cuticle. further apply Cre-loxP excision derive marker-free in-frame fusions clearly map axonal projection patterns from expressing these 3 chemoreceptors antennal lobe devoid background interference 3xP3-based fluorescent markers. clear recording responses in central brain region using genetically encoded indicators GCaMP6s CaMPARI2. Systematic optimized methods different stands enable mapping circuits peripheral levels coding high resolution.

Language: Английский

Citations

0
The conserved IR75 subfamily mediates carboxylic acid detection in insects of public health and agricultural importance DOI Creative Commons

Matthew Cooke,

Michael Solomon Chembars,

R. Jason Pitts

et al.

Journal of Insect Science, Journal Year: 2025, Volume and Issue: 25(1)

Published: Jan. 1, 2025

Insects perceive and respond to carboxylic acids (CAs), amines, aldehydes primarily via conserved ionotropic receptors (IRs). These form the basis for a second olfactory system distinct from well-characterized odorant receptors. Neurons expressing IRs are housed in dedicated sensilla innervate glomeruli, separate those innervated by receptor neurons. The IR8a co-receptor is highly across insect orders and, together with tuning receptors, detects CAs. conservation of genes anatomical separation neural pathways underscore importance these compound classes CAs, specifically chemical ecology. We provide summary acid detection insects, focusing on dipteran lepidopteran species significance public health agriculture. An overview behavior toward CAs provided, as well comprehensive update function insects. Phylogenetic analysis publicly available genome databases reveals several that encode express homologs previously deorphanized highlighting avenues future research.

Language: Английский

Citations

0
An expanded neurogenetic toolkit to decode olfaction in the African malaria mosquito Anopheles gambiae DOI Creative Commons
Diego Giraldo, Andrew Hammond,

Wu Jinling

et al.

Cell Reports Methods, Journal Year: 2024, Volume and Issue: 4(2), P. 100714 - 100714

Published: Feb. 1, 2024

Anopheles gambiae uses its sense of smell to hunt humans. We report a two-step method yielding cell-type-specific driver lines for enhanced neuroanatomical and functional studies olfactory system. first integrated driver-responder-marker (DRM) system cassette consisting linked T2A-QF2 driver, QUAS-GFP responder, gut-specific transgenesis marker into four chemoreceptor genes (Ir25a, Ir76b, Gr22, orco) using CRISPR-Cas9-mediated homology-directed repair. The DRM facilitated rapid selection in-frame integrations via screening GFP+ sensory neurons (OSNs) in G

Language: Английский

Citations

2
Transcriptome‐wide analysis uncovers regulatory elements of the antennal transcriptome repertoire of bumblebee at different life stages DOI Creative Commons
Fatih Dikmen, Tunç Dabak, Burcu Daşer Özgişi

et al.

Insect Molecular Biology, Journal Year: 2024, Volume and Issue: 33(6), P. 571 - 588

Published: April 27, 2024

Abstract Bumblebees are crucial pollinators, providing essential ecosystem services and global food production. The success of pollination relies on the interaction between sensory organs environment. antenna functions as a versatile multi‐sensory organ, pivotal in mediating chemosensory/olfactory information, governs adaptive responses to environmental changes. Despite an increasing number RNA‐sequencing studies insect antenna, comprehensive antennal transcriptome at different life stages were not elucidated systematically. Here, we quantified expression profile dynamics coding/microRNA genes larval head tissues from early‐ late‐stage pupa adult Bombus terrestris suitable model organism among pollinators. We further performed Pearson correlation analyses gene profiles tissue stages, exploring both positive negative trends. positively correlated coding primarily enriched perception chemical stimuli, ion transport, transmembrane transport processes olfactory receptor activity. Negatively mainly organic substance biosynthesis regulatory mechanisms underlying body patterning formation juvenile structures. As post‐transcriptional regulators, miR‐1000‐5p, miR‐13b‐3p, miR‐263‐5p miR‐252‐5p showed correlations, whereas miR‐315‐5p, miR‐92b‐3p, miR‐137‐3p, miR‐11‐3p miR‐10‐3p exhibited correlations tissue. Notably, based inverse relationship, negatively microRNA (miRNA)–mRNA target pairs revealed that differentially expressed miRNAs predictively targeted involved development, shaping structures regulating antenna‐specific functions. Our data serve foundation for understanding stage‐specific transcriptomes large‐scale comparative analysis insects.

Language: Английский

Citations

1
Prospects on non-canonical olfaction in the mosquito and other organisms: why co-express? DOI Creative Commons
Florencia Fernández-Chiappe, Gabriel Koch Ocker,

Meg A. Younger

et al.

Current Opinion in Insect Science, Journal Year: 2024, Volume and Issue: unknown, P. 101291 - 101291

Published: Oct. 1, 2024

Language: Английский

Citations

1
Hybridization Chain Reaction RNA Whole-Mount Fluorescence <em>In situ</em> Hybridization of Chemosensory Genes in Mosquito Olfactory Appendages DOI
Joshua I. Raji, Christopher J. Potter

Journal of Visualized Experiments, Journal Year: 2023, Volume and Issue: 201

Published: Nov. 17, 2023

Mosquitoes are effective vectors of deadly diseases and can navigate their chemical environment using chemosensory receptors expressed in olfactory appendages. Understanding how spatially organized the peripheral appendages offer insights into odor is encoded mosquito system inform new ways to combat spread mosquito-borne diseases. The emergence third-generation hybridization chain reaction RNA whole-mount fluorescence situ (HCR WM-FISH) allows for spatial mapping simultaneous expression profiling multiple genes. Here, we describe a stepwise approach performing HCR WM-FISH on Anopheles antenna maxillary palp. We investigated sensitivity this technique by examining profile ionotropic receptors. asked if described was suitable multiplexed studies tethering probes three spectrally distinct fluorophores. Results provided evidence that robustly sensitive simultaneously detect genes palp Further investigations attest suitability co-expression double triple targets. This technique, when applied with modifications, could be adaptable localize interest tissues other insect species or

Language: Английский

Citations

1