Molecular and Functional Characterization of GABA Receptor Subunits GRD and LCCH3 from Human Louse Pediculus Humanus Humanus

Hashim, Omar; Charvet, Claude; Toubaté, Berthine; Ahmed, Aimun A. E.; Lamassiaude, Nicolas; Neveu, Cédric; Dimier‐Poisson, Isabelle; Debierre‐Grockiego, Françoise; Dupuy, Catherine

doi:10.1124/molpharm.122.000499

Cited by 4 publications

(8 citation statements)

References 42 publications

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“…In the present work, in addition to our previous studies describing three GABA subunits in human body louse, Phh-rdl [21] , Phh-grd and Phh-lcch3 [20] we have now identified another gene encoding for a putative GABA receptor subunit that we named Phh-hocas for Homologous to Cysloop alpha like subunit. Phh-HoCas formed a separate clade distinct from the well-known GRD, LCCH3, RDL and 8916 subunits.…”

Section: Discussionmentioning

confidence: 64%

“…In the genome of P. humanus humanus, our analysis failed to identify the gene encoding for 8916. Assembly of insect genome is challenging due to the large stretch of AT, we cannot exclude defaults in assembly of the human louse genome, as revealed by the missed sequences found in the Phh-GRD genomic annotation and the strikingly very long Phh-GRD gene (> 7 kb) encompassing 3 contigs [20]. Confirmation of the genomic annotation by comparing with the transcriptomics data is a good method to have a complete view of cys-loop LGICs as observed in I. ricinus and I. scapularis [22].…”

Section: Discussionmentioning

confidence: 99%

“…The 5' ends and 3'ends of gamma-aminobutyric-acid receptor alpha-2 subunit precursor (PHUM507160; XM_002430955.1) were characterized by rapid amplification of cDNA ends (RACE-PCR) using Gene Racer ® kit (Invitrogen) as previously described [20] using gene-specific and nested specific primers (Figure 1, Table 1). Cycles for first and second nested PCR was 94°C for 5 min then 35 cycles of 94°C for 30 sec, 58°C for 30 sec, 72°C for 1 min and final extension 72°C for 5 min.…”

Section: Race-pcr and Cloning Of Full Length Transcriptsmentioning

confidence: 99%

“…Quantitative PCR (qPCR) analysis was performed on 100 ng of RNA extracted from adult lice, nits, L1, L2, L3 larvae stages, as well from heads, thoraxes and abdomens of adult louse using the primers listed in (Table 2) and by applying comparative cycle threshold experiment 2 -(ΔCt) with β-actin as endogenous control as described [20]. Data of resulting relative expressions were analyzed by ANOVA test followed by Tukey's multiple comparisons test and plotted as median using GraphPad Prism 7.…”

Section: Spatial and Temporal Expressionmentioning

confidence: 99%

“…Our previous exploration of GABA receptor subunits in human body louse Pediculus humanus humanus revealed that the genome encodes for three GABA receptor subunits Phh-RDL, Phh-GRD, and Phh-LCCH3 that are expressed throughout the developmental stages and in different tissues [20]. Moreover, Phh-RDL was able to reconstitute anion selective functional homomeric receptor while Phh-GRD and Phh-LCCH3 combined to form cation selective hetero-pentameric receptors [20,21].…”

Section: Introductionmentioning

confidence: 99%

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Identification and Spatiotemporal Expression of a Putative New GABA Receptor Subunit in the Human Body Louse Pediculus humanus humanus

HASHIM,

Toubate,

Charvet

et al. 2024

Preprint

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Human louse (Pediculus humanus) is an obligatory blood feeding ectoparasite with two ecotypes: human body louse (Pediculus humanus humanus), a competent vector of several bacterial pathogens and the human head louse (Pediculus humanus capitis) responsible for pediculosis and affecting millions of people around the globe. GABA (γ-aminobutyic acid) receptors, members of the cys-loop ligand gated ion channels superfamily, are among the main pharmacological targets for insecticides. In insects, four subunits of GABA receptors: resistant to dieldrin (RDL), glycin like receptor of drosophila (GRD), ligand gated chloride channel homologue3 (LCCH3), and 8916 are well described and form distinct phylogenetic clades revealing orthologous relationships. Our previous studies in human body louse confirmed that subunits Phh-RDL, Phh-GRD and Phh-LCCH3 are well clustered in their corresponding clades. In the present work, we cloned and characterized a putative new GABA receptor subunit in human body louse that we named HoCas for Homologous to Cys-loop alpha like subunit. Extending our analysis to arthropods, HoCas was found to be conserved and clustered in a new (fifth) phylogenetic clade. Interestingly, the gene encoding this subunit is ancestral and has been lost in some insect orders. Compared to the other studied GABA receptor subunits, HoCas exhibited a relatively higher expression level in all development stages and in different tissues of human body louse. These findings improved our understanding of the complex nature of GABA receptors in Pediculus humanus and more generally in arthropods.

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Section: Discussionmentioning

confidence: 64%

Section: Discussionmentioning

confidence: 99%

Section: Race-pcr and Cloning Of Full Length Transcriptsmentioning

confidence: 99%

Section: Spatial and Temporal Expressionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Identification and Spatiotemporal Expression of a Putative New GABA Receptor Subunit in the Human Body Louse Pediculus humanus humanus

HASHIM,

Toubate,

Charvet

et al. 2024

Preprint

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Molecular cloning, functional characterization and differential expression of two novel GABA_AR‐like subunits from red swamp crayfish Procambarus clarkii

Valladares‐Hernández,

Hernández‐Martínez,

Cuaxospa

et al. 2024

Eur J of Neuroscience

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In this work, we cloned and functionally expressed two novel GABAA receptor subunits from Procambarus clarkii crayfish. These two new subunits, PcGABAA‐α and PcGABAA‐β2, revealed significant sequence homology with the PcGABAA‐β subunit, previously identified in our laboratory. In addition, PcGABAA‐α subunit also shared a significant degree of identity with the Drosophila melanogaster genes DmGRD (GABA and glycine‐like receptor subunits of Drosophila) as well as PcGABAA‐β2 subunit with DmLCCH3 (ligand‐gated chloride channel homolog 3). Electrophysiological recordings showed that the expression in HEK cells of the novel subunits, either alone or in combination, failed to form functional homo‐ or heteromeric receptors. However, the co‐expression of PcGABAA‐α with PcGABAA‐β evoked sodium‐ or chloride‐dependent currents that accurately reproduced the time course of the GABA‐evoked currents in the X‐organ neurons from crayfish, suggesting that these GABA subunits combine to form two types of GABA receptors, one with cationic selectivity filter and the other preferentially permeates anions. On the other hand, PcGABAA‐β2 and PcGABAA‐β co‐expression generated a chloride current that does not show desensitization. Muscimol reproduced the time course of GABA‐evoked currents in all functional receptors, and picrotoxin blocked these currents; bicuculline did not block any of the recorded currents. Reverse transcription polymerae chain reaction (RT‐PCR) amplifications and FISH revealed that PcGABAA‐α and PcGABAA‐β2 are predominantly expressed in the crayfish nervous system. Altogether, these findings provide the first evidence of a neural GABA‐gated cationic channel in the crayfish, increasing our understanding of the role of these new GABAA receptor subunits in native heteromeric receptors.

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Identification and Spatiotemporal Expression of a Putative New GABA Receptor Subunit in the Human Body Louse Pediculus humanus humanus

Hashim,

Toubaté,

Charvet

et al. 2024

Genes

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The human louse (Pediculus humanus) is an obligatory blood feeding ectoparasite with two ecotypes: the human body louse (Pediculus humanus humanus), a competent vector of several bacterial pathogens, and the human head louse (Pediculus humanus capitis), responsible for pediculosis and affecting millions of people around the globe. GABA (γ-aminobutyric acid) receptors, members of the cys-loop ligand gated ion channel superfamily, are among the main pharmacological targets for insecticides. In insects, there are four subunits of GABA receptors: resistant-to-dieldrin (RDL), glycin-like receptor of drosophila (GRD), ligand-gated chloride channel homologue3 (LCCH3), and 8916 are well described and form distinct phylogenetic clades revealing orthologous relationships. Our previous studies in the human body louse confirmed that subunits Phh-RDL, Phh-GRD, and Phh-LCCH3 are well clustered in their corresponding clades. In the present work, we cloned and characterized a putative new GABA receptor subunit in the human body louse that we named HoCas, for Homologous to Cys-loop α like subunit. Extending our analysis to arthropods, HoCas was found to be conserved and clustered in a new (fifth) phylogenetic clade. Interestingly, the gene encoding this subunit is ancestral and has been lost in some insect orders. Compared to the other studied GABA receptor subunits, HoCas exhibited a relatively higher expression level in all development stages and in different tissues of human body louse. These findings improved our understanding of the complex nature of GABA receptors in Pediculus humanus and more generally in arthropods.

show abstract

Molecular and Functional Characterization of GABA Receptor Subunits GRD and LCCH3 from Human Louse Pediculus Humanus Humanus

Cited by 4 publications

References 42 publications

Identification and Spatiotemporal Expression of a Putative New GABA Receptor Subunit in the Human Body Louse <em>Pediculus humanus humanus</em>

Identification and Spatiotemporal Expression of a Putative New GABA Receptor Subunit in the Human Body Louse <em>Pediculus humanus humanus</em>

Molecular cloning, functional characterization and differential expression of two novel GABA_AR‐like subunits from red swamp crayfish Procambarus clarkii

Identification and Spatiotemporal Expression of a Putative New GABA Receptor Subunit in the Human Body Louse Pediculus humanus humanus

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Molecular and Functional Characterization of GABA Receptor Subunits GRD and LCCH3 from Human Louse Pediculus Humanus Humanus

Cited by 4 publications

References 42 publications

Identification and Spatiotemporal Expression of a Putative New GABA Receptor Subunit in the Human Body Louse <em>Pediculus humanus humanus</em>

Identification and Spatiotemporal Expression of a Putative New GABA Receptor Subunit in the Human Body Louse <em>Pediculus humanus humanus</em>

Molecular cloning, functional characterization and differential expression of two novel GABAAR‐like subunits from red swamp crayfish Procambarus clarkii

Identification and Spatiotemporal Expression of a Putative New GABA Receptor Subunit in the Human Body Louse Pediculus humanus humanus

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Molecular cloning, functional characterization and differential expression of two novel GABA_AR‐like subunits from red swamp crayfish Procambarus clarkii