Studying the Activity of Neuropeptides and Other Regulators of the Excretory System in the Adult Mosquito

Lajevardi, Aryan; Sajadi, Farwa; Donini, Andrew; Paluzzi, Jean‐Paul

doi:10.3791/61849

Cited by 3 publications

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“…Before investigating the effects on transepithelial transport across MTs isolated from DTKR knockdown flies, the activity of DTK-1 was examined in MTs from wild-type flies. Using ion-selective microelectrodes (ISME) to measure select ion concentrations in the primary urine droplets from MTs [46], we first quantified Na + concentration in the secreted fluid and its transport rate following treatment with DTK-1, which revealed a significant decrease in Na + concentration in the secreted fluid ( Fig 4A ) but a significant increase in Na + transport rates ( Fig 4B ) compared to unstimulated MTs. Similarly, there was a significant decrease in the K + concentration ( Fig 4C ) in the secreted fluid and a significant increase in K + transport rate ( Fig 4D ) when wild-type MTs were treated with DTK-1.…”

Section: Resultsmentioning

confidence: 99%

“…Glass capillaries for microelectrodes (TW150-4, World Precision Instruments, Sarasota, FL, USA) and reference electrodes (1B200F-4, World Precision Instruments, Sarasota, FL, USA) were prepared using a Sutter P-97 Flaming Brown pipette puller (Sutter Instruments, San Raffael, CA, USA). The ion-selective microelectrodes were silanized using N, N-dimethyltrimethylsilylamine (Sigma-Aldrich, Oakville, ON) as previously described [46]. The reference electrode was backfilled with 500 mmol l -1 KCl.…”

Section: Methodsmentioning

confidence: 99%

“…The tip of the ionselective microelectrodes were forward-filled with an ion-specific ionophore: Na + ionophore (sodium ionophore II cocktail A, Fluka), K + ionophore (potassium ionophore I cocktail B, Fluka), and Cl - ionophore (Chloride ionophore I - Cocktail A, Sigma-Aldrich). The tips of the electrodes were coated with polyvinyl chloride dissolved in tetrahydrofuran (Sigma-Aldrich) to prevent the movement of the ionophore when submerged in the mineral oil as described previously [46].…”

Section: Methodsmentioning

confidence: 99%

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Another fly diuretic hormone: tachykinins increase fluid and ion transport by adultDrosophila melanogasterMalpighian ‘renal’ tubules

Agard,

Zandawala,

Paluzzi

2024

Preprint

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Due to dynamic environmental conditions including temperature, water and food availability, insects like the model organism Drosophila melanogaster, must modulate their internal physiology to withstand these challenges. Regulation of the excretory system by peptidergic hormones is one mechanism by which insects maintain their internal homeostasis. Tachykinins are a family of neuropeptides that have been shown to stimulate fluid secretion from the Malpighian ‘renal’ tubules (MTs) in some insect species, but it is unclear if that is the case in the fruit fly, D. melanogaster. A central objective of the current study was to examine the physiological role of tachykinin signaling in the MTs of adult D. melanogaster. Using the genetic toolbox available in this model organism along with in vitro and whole animal bioassays, our results indicate that Drosophila tachykinins (DTKs) function as diuretic hormones by binding to the DTK receptor (DTKR) expressed in stellate cells of the MTs. Specifically, DTK activates cation and anion transport across the stimulated MTs, which negatively impacts their survival in response to desiccation due to their inability to conserve water. Thus, besides their previously described roles in neuromodulation of pathways controlling locomotion and food search, olfactory processing, aggression, lipid metabolism and metabolic stress, processing of noxious stimuli and hormone release, DTKs also appear to function as bona fide endocrine factors regulating the excretory system and appear essential for the maintenance of hydromineral balance.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Another fly diuretic hormone: tachykinins increase fluid and ion transport by adultDrosophila melanogasterMalpighian ‘renal’ tubules

Agard,

Zandawala,

Paluzzi

2024

Preprint

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“…In order to determine fluid secretion rates, modified Ramsay assays were performed as described previously (4, 80). Female adults (3-6 day old) were dissected under physiological Aedes saline prepared as previously described (60), and MTs were removed and placed in a Sylgard-lined Petri dish containing 20 μL bathing droplets (1:1 mixture of Schneider’s Insect Medium (Sigma-Aldrich): Aedes saline, immersed in hydrated mineral oil to prevent evaporation.…”

Section: Methodsmentioning

confidence: 99%

The V-type H⁺-ATPase is targeted in anti-diuretic hormone control of the Malpighian ‘renal’ tubules

Sajadi

Paluzzi

2022

Preprint

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Like other insects, secretion by mosquito MTs is driven by the V-type H+-ATPase (VA), which is densely packed in the apical membrane of principal cells. The anti-diuretic neurohormone CAPA inhibits tubule secretion stimulated by select diuretic hormones; however, the cellular effectors of this inhibitory signaling cascade remain unclear. Herein, we show that the VA inhibitor bafilomycin selectively inhibits serotonin (5HT)- and calcitonin-related diuretic hormone (DH31)-stimulated secretion. VA activity increases in DH31-treated MTs, whereas CAPA abolishes this increase. A critical feature of VA activation involves the reversible association of the cytosolic (V1) and membrane (Vo) complexes and results revealed higher V1 protein abundance in membrane fractions of DH31-treated MTs whereas CAPA significantly decreased V1 protein abundance in membrane fractions while increasing it in cytosolic fractions. Immunolocalization of V1 was observed strictly in the apical membrane of MTs treated with DH31 alone whereas immunoreactivity was dispersed following CAPA treatment. Elevated levels of cGMP in CAPA-treated MTs suggests activation of a phosphodiesterase (PDE), resulting in decreased levels of DH31-stimulated cAMP. Therefore, CAPA inhibition of MTs involves reducing VA activity and promotes complex dissociation hindering secretion. Collectively, these findings reveal a key target necessary for hormone-mediated inhibition of MTs required in countering diuresis providing a deeper understanding of this critical physiological process.Significance StatementThe V-type H+-ATPase (VA) proton pump, provides the driving force for transepithelial ion and fluid secretion in insect Malpighian tubules (MTs). While studies have shown that diuretic stimulation activates various signaling pathways that culminate with increased VA activity, our understanding of anti-diuretic signaling and its potential regulation involving the VA remains rudimentary. Here we show that a CAPA neuropeptide acts through the NOS/cGMP/PKG pathway that inhibits VA activity, supporting the notion that anti-diuretic regulation is mediated through reversible dissociation of the VA complexes. Our results demonstrate a critical role of VA inhibition and trafficking necessary for anti-diuretic signaling, advancing our understanding of the complex neuroendocrine regulatory mechanisms of the MTs in an important human disease vector mosquito.

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“…Thus, the potential for evaporative water loss at a given relative humidity is inherently sensitive to changes in temperature (Figure 1). of adaptations to conserve water, like physiological changes in skin or cuticular permeability (Rajpurohit et al, 2008;Wu & Wright, 2015), differential regulation of urine and faeces production (Durant et al, 2021;Durant & Donini, 2019;Lajevardi et al, 2021;Weihrauch et al, 2012), and behavioural changes in activity (Kühnholz & Seeley, 1997;Ostwald et al, 2016). Insects also can mitigate water loss by regulating water intake via changes in water utilization, food sources and selection of specific habitats (Benoit, 2010;Bezerra Da Silva et al, 2019;Hagan et al, 2018)).…”

Section: T H E E F F Ect S Of H U M I Di T Y On Mo Squ I To F I T N E...mentioning

confidence: 99%

Humidity – The overlooked variable in the thermal biology of mosquito‐borne disease

et al. 2023

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Vector-borne parasites are common, important biological enemies of humans, animals and plants, transmitted by one living organism to another. Despite the recent gains in reducing the overall global burden for parasites like malaria (Ashepet et al., 2021;Bhatt et al., 2015;Gething et al., 2010), vector-borne diseases still account for 17% of all infectious diseases and cause 700,000 deaths in humans annually (WHO, 2020). Livestock and crop systems are also plagued by vector-borne diseases, which place serious constraints on agricultural production globally (Döring, 2017;Garros et al., 2017), and vector-borne diseases can be devastating in wildlife populations, particularly when introduced to new areas. Collectively, tens of billions of dollars are spent every year on control, medical interventions and mitigating loss of productivity (

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Studying the Activity of Neuropeptides and Other Regulators of the Excretory System in the Adult Mosquito

Cited by 3 publications

References 0 publications

Another fly diuretic hormone: tachykinins increase fluid and ion transport by adultDrosophila melanogasterMalpighian ‘renal’ tubules

Another fly diuretic hormone: tachykinins increase fluid and ion transport by adultDrosophila melanogasterMalpighian ‘renal’ tubules

The V-type H⁺-ATPase is targeted in anti-diuretic hormone control of the Malpighian ‘renal’ tubules

Humidity – The overlooked variable in the thermal biology of mosquito‐borne disease

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Studying the Activity of Neuropeptides and Other Regulators of the Excretory System in the Adult Mosquito

Cited by 3 publications

References 0 publications

Another fly diuretic hormone: tachykinins increase fluid and ion transport by adultDrosophila melanogasterMalpighian ‘renal’ tubules

Another fly diuretic hormone: tachykinins increase fluid and ion transport by adultDrosophila melanogasterMalpighian ‘renal’ tubules

The V-type H+-ATPase is targeted in anti-diuretic hormone control of the Malpighian ‘renal’ tubules

Humidity – The overlooked variable in the thermal biology of mosquito‐borne disease

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The V-type H⁺-ATPase is targeted in anti-diuretic hormone control of the Malpighian ‘renal’ tubules