Biology subjects

Lee, T.-T.

Publications and source records attributed to Lee, T.-T..

8 recordsLinked to original sources

bioRxiv2026

The sex determination gene doublesex is essential for female-specific flight and hearing properties in Aedes aegypti mosquitoes

The acoustic communication system supporting reproduction in disease-transmitting mosquitoes is sexually dimorphic, from the sounds that mosquitoes make to the properties of their flagellar ears. The molecular mechanisms shaping these dimorphisms, however, remain unclear. Here, we explored the contributions of the female-specific isoform of the sex determination gene doublesex (dsxF) in specifying female-unique properties in the acoustic communication system of Aedes aegypti mosquitoes. CRISPR-Cas9 based generation of a novel dsxF knockout line found that dsxF mutant females were flightless due to lack of expression of a female-specific myosin gene, myo-fem, in the thorax. We found that DsxF directly binds to four different enhancer sites of myo-fem to promote gene expression. We observed that dsxF female ears exhibited intersex neuroanatomical characteristics and contained a more extensive auditory efferent network than control females. At the peripheral hearing function level, dsxF female ears displayed intersex tuning frequencies associated with increased expression of hearing genes in dsxF female ears compared to control females. We identified three hearing genes transcriptionally inhibited by DsxF via direct binding. DsxF is a bifunctional transcription factor, activating female-specific gene expression in thoraces whilst repressing male-biased gene expression in ears to coordinate female-unique features across sensory and motor systems.

neuroscience↗

bioRxiv2026

Pharmacological Inhibition of SLC33A1 Promotes Endoplasmic Reticulum Hyperoxidation and Induces Adaptive IRE1/XBP1s Signaling

The endoplasmic reticulum (ER) transporter solute carrier family 33 member 1 (SLC33A1) has emerged as an attractive therapeutic target in etiologically diverse diseases, ranging from lung cancer to neurodegenerative disorders. Yet, no pharmacologic SLC33A1 modulators have been described. Here, we show that the small molecule IXA4, a highly selective activator of the adaptive IRE1/XBP1s signaling arm of the unfolded protein response (UPR), binds to SLC33A1 and inhibits its activity. Genetic depletion of SLC33A1 phenocopies the selective induction of IRE1/XBP1s signaling brought about by IXA4 treatment. Chemoproteomic analyses and cryo-electron microscopy show that IXA4 binds SLC33A1 within the central channel to inhibit transport of its substrate metabolite(s). Binding of IXA4 to SLC33A1 leads to the accumulation of oxidized glutathione within the ER, hyperoxidizing the ER lumen and inducing activation of adaptive IRE1/XBP1s signaling. Consistent with this function, we find that pharmacologic inhibition of SLC33A1 with IXA4 selectively reduces viability of KEAP1-deficient lung adenocarcinoma cells that have elevated levels of glutathione, mimicking the sensitivity of these cells to genetic deletion of SLC33A1. Our work demonstrates a new physiologic role of SLC33A1 in regulation of ER redox homeostasis and designates IXA4 as a pharmacologic inhibitor of SLC33A1 that can be used to evaluate the biological impact and therapeutic utility of SLC33A1 inhibition in homeostasis and in disease.

cell biology↗

bioRxiv2026

The sex determination gene fruitless is essential for male Aedes aegypti mosquito swarming and attraction to female flight sounds

Aggregating male mosquitoes locate females by showing strong and innate attraction to female flight sounds, with these behaviors forming the basis of mating in the worlds deadliest animal. In contrast, females do not show aggregation or sound attraction behaviors. Despite being documented for almost a century, the molecular underpinnings of these behavioral dimorphisms remain unknown. Here, we found that by disrupting the male-specific isoform of the key sex determination gene fruitless (referred to as fruM) in Aedes aegypti, mutant males behave like females. fruM mutant males show complete abolition of aggregation and sound attraction behaviors, resulting in failure to locate females for mating. We identified strong fruitless expression in control male ears, with the loss of FruM severely disrupting mutant male ability to amplify female flight sounds compared to control males. fruM mutant male ears propagate frequency-specific, sound-evoked electrical signals at about one-sixth the magnitude of control males, significantly reducing the amount of auditory information relayed to the brain. fruitless thus plays a significant role in determining proper male peripheral hearing function, which enables male hearing behaviors essential for mating. Transcriptomic analyses of mosquito ears enabled identification of essential hearing genes under the transcriptional influence of FruM. We propose that FruM fine-tunes the masculinization of mosquito ears through shaping male-specific hearing cellular machineries that support male attraction to female flight sounds. We conclude that FruM is the master regulator of male-specific reproductive behaviors in mosquitoes and propose fruitless, as well as fruitless-dependent genes, as novel targets for control strategies. HighlightsO_LIFruM is the master regulator of male swarming and attraction to female flight sounds C_LIO_LIFruM in male ears is essential for male amplification of female flight sounds C_LIO_LIFruM in male ears modulates sound-evoked electrical signal propagation to the brain C_LIO_LIFruM regulates essential hearing gene expression C_LI

neuroscience↗

bioRxiv2025

cAMP-related second messenger pathways modulate hearing function in Aedes aegypti mosquitoes

The powerful ears of male Aedes aegypti mosquitoes facilitate identification and localization of mating partners via detection of female flight tones. Male hearing function is modulated by the efferent release of neurotransmitters, though the secondary mechanisms underlying this modulation remain unclear. Here, we investigated these mechanisms using octopamine as a model, as octopamine modulates hearing function and the erection status of fibrillar hairs lining male ears. We found that pharmacological interference with octopamine receptors alters hearing function at multiple levels and identified the second messenger cAMP as likely mediating these changes. Furthermore, the erection status of male ear fibrillar hairs could be altered by targeting specific sub-types of octopamine receptors, but these changes were not linked to changes in ear frequency tuning. Finally, we suggest that octopamine 2 receptors linked to fibrillar hair erection may not always produce functional proteins across species, with downstream implications for hearing behaviors. HighlightsO_LIOctopamine and its receptors are found throughout Aedes aegypti mosquito ears C_LIO_LIPharmacological interference with octopamine receptors modulates hearing function C_LIO_LIModulating levels of the second messenger cAMP also alters hearing function C_LIO_LIEar frequency tuning does not correlate with the erection of hairs lining male ears C_LI

neuroscience↗

bioRxiv2024

Sexually dimorphic auditory representation in Aedes aegypti brains

Male attraction to female flight sounds is a vital, reproducible component of courtship in many species of mosquitoes; however, female acoustic behaviours have proven challenging to define. To investigate sexual dimorphisms in acoustic behaviours, previous reports have largely focused on differences in mosquito peripheral ear anatomy and function. Whilst molecular investigations have recently begun on the auditory periphery, sexual dimorphisms in central processing of acoustic information have not yet been explored. Here we used a combination of neurotracing, calcium imaging and molecular analyses to examine sexual dimorphisms in auditory processing in the yellow fever mosquito Aedes aegypti. We identified shared and dimorphic neurons connecting male and female ears to the primary auditory processing centre in the brain, and defined multiple distinct neuronal clusters based on responses to auditory stimulation. We finally used transcriptomic and proteomic analyses to investigate the molecular factors underlying these differences, with motile ciliary-related terms significantly enriched in males.

neuroscience↗

bioRxiv2024

The voltage-gated potassium channel Shal (Kv4) contributes to active hearing in Drosophila

The full complement of ion channels which influence insect auditory mechanotransduction, and the mechanisms by which their influence is exerted, remain unclear. Shal (Kv4), a Shaker family member encoding voltage-gated potassium channels in Drosophila melanogaster, has been shown to localize to dendrites in some neuron types, suggesting a potential role for Shal in Drosophila hearing, including mechanotransduction. A GFP-protein trap was used to visualize the localization of the Shal channel in Johnstons organ neurons responsible for hearing in the antenna. Shal protein was localized to the cell body and the proximal dendrite region of sensory neurons, suggesting its involvement not only in general auditory function, but specifically in mechanotransduction. Electrophysiological recordings conducted to assess neural responses to auditory stimuli in mutant Shal flies revealed significant decreases in auditory responses. Laser Doppler Vibrometer recordings indicated abnormal antennal free fluctuation frequencies in mutant lines, indicating an effect on active antennal tuning, and thus active transduction mechanisms. This suggests that Shal participates in coordinating energy-dependent antennal movements in Drosophila that are essential for tuning the antenna to courtship song frequencies. Significance StatementThe study of fruit fly hearing has revealed mechanosensitive ion channels that participate in mechanotransduction, and as in mammalian hearing, energy-dependent mechanisms actively amplify and tune auditory processes. Identifying distinct roles played by different ion channels is essential to better understand this process. Here, we explore the influence of a specific voltage-gated potassium channel, Shal, on fly hearing, and find that it affects specific parts of the mechanotransduction process. Our research uncovers Shals localization in sensory dendrite regions of auditory neurons, where it contributes to shaping mechanotransduction and active antennal tuning. Understanding Shals involvement in auditory function and mechanotransduction deepens our knowledge of fly hearing and unveils a key player in the coordination of energy-dependent active antennal movements.

neuroscience↗

bioRxiv2023

Proteostasis and lysosomal quality control deficits in Alzheimer's disease neurons

Aging is the most prominent risk factor for Alzheimers disease (AD). However, the cellular mechanisms linking neuronal proteostasis decline to the characteristic aberrant protein deposits in AD brains remain elusive. Here, we develop transdifferentiated neurons (tNeurons) from human dermal fibroblasts as a neuronal model that retains aging hallmarks and exhibits AD-linked vulnerabilities. Remarkably, AD tNeurons accumulate proteotoxic deposits, including phospho-Tau and A{beta}, resembling those in AD patient and APP mouse brains. Quantitative tNeuron proteomics identify aging and AD-linked deficits in proteostasis and organelle homeostasis, most notably in endosome-lysosomal components. Lysosomal deficits in aged tNeurons, including constitutive lysosomal damage and ESCRT-mediated lysosomal repair defects, are exacerbated in AD tNeurons and linked to inflammatory cytokine secretion and cell death. Supporting lysosomal deficits centrality in AD, compounds ameliorating lysosomal function reduce A{beta} deposits and cytokine secretion. Thus, the tNeuron model system reveals impaired lysosomal homeostasis as an early event of aging and AD.

cell biology↗

bioRxiv2022

Ribotoxic collisions on CAG expansions disrupt proteostasis and stress responses in Huntington's Disease

Huntingtons disease (HD) is a devastating neurodegenerative disorder caused by CAG trinucleotide repeat expansions encoding a polyglutamine (polyQ) tract in the Huntingtin (HTT) gene1. Although mutant HTT (mHTT) protein tends to aggregate, the exact causes of neurotoxicity in HD remain unclear2. Here we show that altered elongation kinetics on CAG expansions cause ribosome collisions that trigger ribotoxicity, proteotoxicity and maladaptive stress responses. CAG expansions cause an elongation rate conflict during HTT translation, when ribosomes rapidly decoding the optimal polyQ encounter a flanking slowly-decoded polyproline tract. The ensuing ribosome collisions lead to premature termination and release of aggregation-prone mHTT fragments. Due to the presence of a stress-responsive upstream open reading frame (uORF), HTT translation and aggregation are limited under normal conditions but enhanced under stress, seeding a vicious cycle of dysfunction. mHTT further exacerbates ribotoxicity by progressively sequestering eIF5A, a key regulator of translation elongation, polyamine metabolism and stress responses. eIF5A depletion in HD cells leads to widespread ribosome pausing on eIF5A-dependent sites, impaired cotranslational proteostasis, disrupted polyamine metabolism and maladaptive stress responses. Importantly, drugs that reduce translation initiation attenuate ribosome collisions and mitigate this escalating cascade of ribotoxic stress and dysfunction in HD.

neuroscience↗