Biology subjects

Hardege, I.

Publications and source records attributed to Hardege, I..

7 recordsLinked to original sources

bioRxiv2025

Mechanistic dissection of a dopamine-gated cation channel from Daphnia reveals key determinants of ligand selectivity and sensitivity

The pentameric ligand-gated ion channels (pLGICs) are a deeply conserved superfamily that includes nicotinic acetylcholine (nAChR), GABAARs, glycine, and serotonin receptors and mediates fast neurotransmission. While vertebrate pLGICs have been extensively characterized, recent discoveries in invertebrates have revealed atypical members with novel ligand specificities, including dopamine-gated ion channels (dop-LGICs) that enable direct ionotropic dopaminergic signaling. Here, we investigate the molecular determinants of catecholamine sensitivity and selectivity in a cation-permeable dopamine-gated receptor from Daphnia magna (Dm-DopC1), which shares high structural homology with vertebrate nAChRs. We find that mutations in loop B (D188N) near-abolish catecholamine responses, while substitutions in loops A (S123P) and C (S227G, E230P) significantly alter sensitivity and ligand selectivity, highlighting the cooperative role of these loops in shaping receptor pharmacology. Complementary face mutations in loops D and E (N92T, A158G) further refined dopamine versus norepinephrine sensitivity. Finally, we also show that this dopamine receptor can be antagonized by both nicotinic and dopaminergic compounds, underscoring its hybrid pharmacological profile. Together, these results reveal how subtle changes in conserved binding motifs can shift receptor specificity from cholinergic to dopaminergic signaling, providing new insights into the evolutionary flexibility of pLGICs and the molecular basis of ionotropic catecholamine transmission in invertebrates.

biochemistry↗

bioRxiv2025

Novel dopaminergic neurotransmission in the Octopus visual system

Octopus vulgaris, the common octopus, has a complex brain that evolved independently from that of vertebrates, raising the possibility that it may be built using novel circuit motifs or non-canonical neurochemistry. Through systematic characterization of octopus ligand-gated ion channel genes, we identified novel ionotropic receptors for two neurotransmitters, dopamine and acetylcholine, that play key roles in visual circuits. One of these, DopC1, encodes a dopamine-gated cation channel expressed in deeper layers of the optic lobe (inner granular layer and medulla). A second, AChRB1, encodes an acetylcholine-gated anion channel expressed in photoreceptors and dopaminergic outer granular layer neurons. Dopamine drives excitation, particularly in deeper, DopC1 enriched layers, and thereby may play an important role in feed-forward excitation, whereas acetylcholine evokes inhibition, possibly mediating negative feedback from deeper layers. The octopus visual system thus shows fundamental differences in both neurochemistry and wiring compared to mammals, implying distinct mechanisms of visual information processing.

neuroscience↗

bioRxiv2021

Neuropeptide signalling shapes feeding and reproductive behaviours in male C. elegans

Sexual dimorphism occurs where different sexes of the same species display differences in characteristics not limited to reproduction. For the nematode Caenorhabditis elegans, in which the complete neuroanatomy has been solved for both hermaphrodites and males, sexually dimorphic features have been observed both in terms of the number of neurons and in synaptic connectivity. In addition, male behaviours, such as food-leaving to prioritise searching for mates, have been attributed to neuropeptides released from sex-shared or sex-specific neurons. In this study, we show that the lury-1 neuropeptide gene shows a sexually dimorphic expression pattern; being expressed in pharyngeal neurons in both sexes but displaying additional expression in tail neurons only in the male. We also show that lury-1 mutant animals show sex differences in feeding behaviours, with pharyngeal pumping elevated in hermaphrodites but reduced in males. LURY-1 also modulates male mating efficiency, influencing motor events during contact with a hermaphrodite. Our findings indicate sex-specific roles of this peptide in feeding and reproduction in C. elegans, providing further insight into neuromodulatory control of sexually dimorphic behaviours.

neuroscience↗

bioRxiv2021

Distinct roles for two Caenorhabditis elegans acid-sensing ion channels in an ultradian clock

Biological clocks are fundamental to an organisms health, controlling periodicity of behavior and metabolism. Here, we identify two acid-sensing ion channels, with very different proton sensing properties, and describe their role in an ultradian clock, the defecation motor program (DMP) of the nematode Caenorhabditis elegans. An ACD-5-containing channel, on the apical membrane of the intestinal epithelium, is essential for maintenance of luminal acidity, and thus the rhythmic oscillations in lumen pH. In contrast, the second channel, composed of FLR-1, ACD-3 and/or DEL-5, located on the basolateral membrane, controls the intracellular Ca2+ wave and forms a core component of the master oscillator that controls timing and rhythmicity of the DMP. flr-1 and acd-3/del-5 mutants show severe developmental and metabolic defects. We thus directly link the proton-sensing properties of these channels to their physiological roles in pH regulation and Ca2+ signaling, the generation of an ultradian oscillator, and its metabolic consequences. One-Sentence SummaryTwo acid-sensing DEG/ENaC channels play distinct roles in controlling different aspects of rhythmic proton and Ca2+ oscillations in the C. elegans intestine.

physiology↗

bioRxiv2021

Neuronally Produced Betaine Acts via a Novel Ligand Gated Ion Channel to Control Behavioural States

Trimethyl glycine, or betaine, is an amino acid derivative found in diverse organisms, from bacteria to plants and animals. It can function as an osmolyte to protect cells against osmotic stress, and building evidence suggests betaine may also play important functional roles in the nervous system. However, despite growing interest in betaines roles in the nervous system, few molecular mechanisms have been elucidated. Here we identify the expression of betaine synthesis pathway genes in the nervous system of the nematode worm, C. elegans. We show that betaine, produced in a single pair of interneurons, the RIMs, can control complex behavioural states. Moreover, we also identify and characterise a new betaine-gated inhibitory ligand gated ion channel, LGC-41, which is required for betaine related behavioural changes. Intriguingly we observed expression of LGC-41 in punctate structures across several sensory and interneurons, including those synaptically connected to the RIMs. Our data presents a neuronal molecular mechanism for the action of betaine, via a specific receptor, in the control of complex behaviour within the nervous system of C. elegans. This may suggest a much broader role for betaine in the regulation of animal nervous systems than previously recognised.

neuroscience↗

bioRxiv2021

A Novel and Functionally Diverse Class of Acetylcholine-gated Ion Channels

Fast cholinergic neurotransmission is mediated by acetylcholine-gated ion channels; in particular, excitatory nicotinic acetylcholine receptors play well established roles in virtually all nervous systems. Acetylcholine-gated inhibitory channels have also been identified in some invertebrate phyla, yet their roles in the nervous system are less well understood. We report the existence of multiple new inhibitory acetylcholine-gated ion channels with diverse ligand binding properties in C. elegans. We identify three channels, LGC-40, LGC-57 and LGC-58, whose primary ligand is choline rather than acetylcholine, as well as the first evidence of a truly polymodal channel, LGC-39, which is activated by both cholinergic and aminergic ligands. Using our newly deorphanised channel evidence we uncover the surprising extent to which a single neuron expresses both excitatory and inhibitory channels, not only for acetylcholine but also the other major neurotransmitters. The results presented in this study offer a new insight into the potential evolutionary benefit of a vast and diverse repertoire of LGICs to generate complexity in an anatomically compact nervous system.

neuroscience↗

bioRxiv2020

Deorphanisation of novel biogenic amine-gated ion channels identifies a new serotonin receptor for learning

Pentameric ligand-gated ion channels (LGCs) play conserved, critical roles in fast synaptic transmission, and changes in LGC expression and localisation are thought to underlie many forms of learning and memory. The C. elegans genome encodes a large number of LGCs without a known ligand or function. Here, we deorphanize five members of a family of Cys-loop LGCs by characterizing their diverse functional properties that are activated by biogenic amine neurotransmitters. To analyse the neuronal function of these LGCs, we show that a novel serotonin-gated cation channel, LGC-50, is essential for aversive olfactory learning. lgc-50 mutants show a specific defect in learned olfactory avoidance of pathogenic bacteria, a process known to depend on serotonergic neurotransmission. Remarkably, the expression of LGC-50 in neuronal processes is enhanced by olfactory conditioning; thus, the regulated expression of these receptors at synapses appears to represent a molecular cornerstone of the learning mechanism.

neuroscience↗