Characterization of a ligand-gated cation channel based on an inositol receptor in the silkworm, Bombyx mori

Kikuta, Shingo; Endo, Haruka; Tomita, Natsuo; Takada, Tomoyuki; Morita, Çhiharu; Asaoka, Kiyoshi; Satō, Ryōichi

doi:10.1016/j.ibmb.2016.04.010

Cited by 36 publications

(54 citation statements)

References 40 publications

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“…The procedures for capped RNA (cRNA) synthesis followed those described in a previous study [9]. Briefly, cRNA was synthesized using an mMESSAGE mMACHINE ® T7 kit (Thermo Fisher Scientific) according to the manufacturer’s instructions and stored at ‒80°C until use.…”

Section: Methodsmentioning

confidence: 99%

“…Negative control oocytes were injected with water and therefore showed only endogenous receptor activities. Whole-cell current was recorded using a two-electrode voltage clamp in a perfusion system with Ringer’s solution [9]. The current was amplified with an OC-725C amplifier (Warner Instruments, Hamden, CT, USA) at a holding potential of ‒70 mV, low-pass filtered at 50 Hz, and digitized at 1 kHz.…”

Section: Methodsmentioning

confidence: 99%

“…Myo -inositol, an indispensable nutrient and a factor that prolongs feeding behavior in B . mori larvae, is recognized via BmGr10 expression in sensory organs [9]. Therefore, Grs in specialist feeders play a key role in detecting specific compounds.…”

Section: Introductionmentioning

confidence: 99%

“…armigera responds to fructose in a Xenopus oocyte expression system [11]. Furthermore, members of the Gr43-like clade in the Gr family, which includes HarmGr4, have been found in various Diptera and Lepidoptera to act as receptors of fructose and myo -inositol [9, 20]. Since Gr43-like genes are also present in T .…”

Section: Introductionmentioning

confidence: 99%

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A mannitol/sorbitol receptor stimulates dietary intake in Tribolium castaneum

2017

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In insects, perception of chemical stimuli is involved in the acceptance or rejection of food. Gustatory receptors (Grs) that regulate external signals in chemosensory organs have been found in many insects. Tribolium castaneum, a major pest of stored products, possesses over 200 Gr genes. An expanded repertoire of Gr genes appears to be required for diet recognition in species that are generalist feeders; however, it remains unclear whether T. castaneum recognizes a suite of chemicals common to many products or whether its feeding is activated by specific chemicals, and whether its Grs are involved in feeding behavior. It is difficult to determine the food preferences of T. castaneum based on dietary intake due to a lack of appropriate methodology. This study established a novel dietary intake estimation method using gypsum, designated the TribUTE (Tribolium Urges To Eat) assay. For this assay, T. castaneum adults were fed a gypsum block without added organic compounds. Sweet preference was determined by adding sweeteners and measuring the amount of gypsum in the excreta. Mannitol was the strongest activator of T. castaneum dietary intake. In a Xenopus oocyte expression, TcGr20 was found to be responsible for mannitol and sorbitol responses, but not for responses to other tested non-volatile compounds. The EC50 values of TcGr20 for mannitol and sorbitol were 72.6 mM and 90.6 mM, respectively, suggesting that TcGr20 is a feasible receptor for the recognition of mannitol at lower concentrations. We used RNAi and the TribUTE assay to examine whether TcGr20 expression was involved in mannitol recognition. The amounts of excreta in TcGr20 dsRNA-injected adults decreased significantly, despite the presence of mannitol, compared to control adults. Taken together, our results indicate that T. castaneum adults recognized mannitol/sorbitol using the TcGr20 receptor, thereby facilitating the dietary intake of these compounds.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A mannitol/sorbitol receptor stimulates dietary intake in Tribolium castaneum

2017

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“…The fragment encoded luciferase was introduced into the pcDNA3.1 vector. The Eluc expression vector was transfected into HEK293T cultured cells in accordance with the procedures in the previous study 43 . Desiccation of cells (4 × 10 7 cells) was examined at 48 h after transfection.…”

Section: Methodsmentioning

confidence: 99%

Towards water-free biobanks: long-term dry-preservation at room temperature of desiccation-sensitive enzyme luciferase in air-dried insect cells

Kikuta

Watanabe

Satō

et al. 2017

Sci Rep

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Desiccation-tolerant cultured cells Pv11 derived from the anhydrobiotic midge embryo endure complete desiccation in an ametabolic state and resume their metabolism after rehydration. These features led us to develop a novel dry preservation technology for enzymes as it was still unclear whether Pv11 cells could preserve an exogenous enzyme in the dry state. This study shows that Pv11 cells protect an exogenous desiccation-sensitive enzyme, luciferase (Luc), preserving the enzymatic activity even after dry storage for 372 days at room temperature. A process including preincubation with trehalose, dehydration, storage, and rehydration allowed Pv11 (Pv11-Luc) cells stably expressing luciferase to survive desiccation and still emit luminescence caused by luciferase after rehydration. Luminescence produced by luciferase in Pv11-Luc cells after rehydration did not significantly decrease in presence of a translation inhibitor, showing that the activity did not derive from de novo enzyme synthesis following the resumption of cell metabolism. These findings indicate that the surviving Pv11 cells almost completely protect luciferase during desiccation. Lacking of the preincubation step resulted in the loss of luciferase activity after rehydration. We showed that preincubation with trehalose associated to induction of desiccation tolerance-related genes in Pv11 cells allowed effective in vivo preservation of enzymes in the dry state.

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Differential expression of a fructose receptor gene in honey bee workers according to age and behavioral role

Takada

Sasaki

Satō

et al. 2017

Arch Insect Biochem Physiol

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Honey bee (Apis mellifera) workers contribute to the maintenance of colonies in various ways. The primary functions of workers are divided into two types depending on age: young workers (nurses) primarily engage in such behaviors as cleaning and food handling within the hive, whereas older workers (foragers) acquire floral nutrients beyond the colony. Concomitant with this age-dependent change in activity, physiological changes occur in the tissues and organs of workers. Nurses supply younger larvae with honey containing high levels of glucose and supply older larvae with honey containing high levels of fructose. Given that nurses must determine both the concentration and type of sugar used in honey, gustatory receptors (Gr) expressed in the chemosensory organs likely play a role in distinguishing between sugars. Glucose is recognized by Gr1 in honey bees (AmGr1); however, it remains unclear which Gr are responsible for fructose recognition. This study aimed to identify fructose receptors in honey bees and reported that AmGr3, when transiently expressed in Xenopus oocytes, responded only to fructose, and to no other sugars. We analyzed expression levels of AmGr3 to identify which tissues and organs of workers are involved in fructose recognition and determined that expression of AmGr3 was particularly high in the antennae and legs of nurses. Our results suggest that nurses use their antennae and legs to recognize fructose, and that AmGr3 functions as an accurate nutrient sensor used to maintain food quality in honey bee hives.

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Characterization of a ligand-gated cation channel based on an inositol receptor in the silkworm, Bombyx mori

Cited by 36 publications

References 40 publications

A mannitol/sorbitol receptor stimulates dietary intake in Tribolium castaneum

A mannitol/sorbitol receptor stimulates dietary intake in Tribolium castaneum

Towards water-free biobanks: long-term dry-preservation at room temperature of desiccation-sensitive enzyme luciferase in air-dried insect cells

Differential expression of a fructose receptor gene in honey bee workers according to age and behavioral role

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