* Wasson JA, Harris G, Keppler-Ross S, Brock TJ, Dar AR, Butcher RA, Fische SEJ, Kagias K, Clardy J, Zhang Y, et al. Neuronal control of maternal provisioning in response to social cues. Science Advances. 2021;7 (34) :8782. Publisher's VersionAbstract

Mothers contribute cytoplasmic components to their progeny in a process called maternal provisioning. Provisioning is influenced by the parental environment, but the molecular pathways that transmit environmental cues between generations are not well understood. Here, we show that, in Caenorhabditis elegans, social cues modulate maternal provisioning to regulate gene silencing in offspring. Intergenerational signal transmission depends on a pheromone-sensing neuron and neuronal FMRFamide (Phe-Met-Arg-Phe)–like peptides. Parental FMRFamide-like peptide signaling dampens oxidative stress resistance and promotes the deposition of mRNAs for translational components in progeny, which, in turn, reduces gene silencing. This study identifies a previously unknown pathway for intergenerational communication that links neuronal responses to maternal provisioning. We suggest that loss of social cues in the parental environment represents an adverse environment that stimulates stress responses across generations.

DOI: 10.1126/sciadv.abf8782

Morud J, Hardege I, Liu H, Wu T, Choi M-K, Basu S, Zhang Y, Schafer WR. Deorphanization of novel biogenic amine-gated ion channels identifies a new serotonin receptor for learning. Current Biology. 2021;2021 (07) :036. Publisher's VersionAbstract

Pentameric ligand-gated ion channels (LGICs) play conserved, critical roles in both excitatory and inhibitory synaptic transmission and can be activated by diverse neurochemical ligands. We have performed a characterization of orphan channels from the nematode C. elegans, identifying five new monoamine-gated LGICs with diverse functional properties and expression postsynaptic to aminergic neurons. These include polymodal anion channels activated by both dopamine and tyramine, which may mediate inhibitory transmission by both molecules in vivo. Intriguingly, we also find that a novel serotonin-gated cation channel, LGC-50, is essential for aversive olfactory learning of pathogenic bacteria, a process known to depend on serotonergic neurotransmission. Remarkably, the redistribution of LGC-50 to neuronal processes is modulated by olfactory conditioning, and lgc-50 point mutations that cause misregulation of receptor membrane expression interfere with olfactory learning. Thus, the intracellular trafficking and localization of these receptors at synapses may represent a molecular cornerstone of the learning mechanism.


DOI: 10.1016/j.cub.2021.07.036

*Yang W, Wu T, Tu S, Choi M-K, Duan F, Zhang Y. Redundant neural circuits regulate olfactory masking. bioRxiv. 2021;2021 (04.19) :440489. Publisher's VersionAbstract

Olfactory masking is a complex olfactory response found in humans. However, the mechanisms whereby the presence of one odorant masks the sensory and behavioral responses elicited by another odorant are poorly understood. Here, we report that Caenorhabditis elegans displays olfactory masking and that the presence of a repulsive odorant, 2-nonanone, that signals threat strongly masks the attraction of other odorants, such as isoamyl alcohol (IAA) or benzaldehyde that signals food. Using a forward genetic screen, we found that several genes, osm-5, osm-1, and dyf-7, known to regulate the structure and function of sensory neurons played a critical role in olfactory masking. Loss of these genes mildly reduces the response to 2-nonanone and disrupts the masking effect of 2-nonanone. Restoring the function of OSM-5 in either AWB or ASH, two sensory neurons known to mediate 2-nonanone-evoked avoidance, is sufficient to rescue olfactory masking. AWB is activated by the removal of 2-nonanone stimulation or the onset of IAA; however, the mixture of 2-nonanone and IAA stimulates AWB similarly as 2-nonanone alone, masking the cellular effect of IAA. The latency of the AWB response is critical for the masking effect. Thus, our results identify redundant neural circuits that regulate the robust masking effect of a repulsive odorant and uncover the neuronal and cellular basis for this complex olfactory task.


* Wasson JA#, Harris G#, Keppler-Ross S, Brock TJ, Dar AR, Butcher RA, Fischer SEJ, Kagias K, Clardy J, Zhang Y&, et al. Neuronal control of maternal provisioning in response to social cues. #: co-first author; &: co-corresponding author. bioRxiv. 2021;10.1101 (2021.02.01) :429208. Publisher's VersionAbstract

Mothers contribute cytoplasmic components to their progeny in a process called maternal provisioning. Provisioning is influenced by the parental environment, but the molecular pathways that transmit environmental cues from mother to progeny are not well understood. Here we show that in C. elegans, social cues modulate maternal provisioning to regulate gene silencing in offspring. Intergenerational signal transmission depends on a pheromone-sensing neuron and neuronal FMRF (Phe-Met-Arg-Phe)-like peptides. Parental FMRF signaling promotes the deposition of mRNAs for translational components in progeny, which in turn reduces gene silencing. Previous studies had implicated FMRF signaling in short-term responses such as modulated feeding behavior in response to the metabolic state1,2, but our data reveal a broader role, to coordinate energetically expensive processes such as translation and maternal provisioning. This study identifies a new pathway for intergenerational communication, distinct from previously discovered pathways involving small RNAs and chromatin, that links sensory perception to maternal provisioning.




Alcedo J, Jin Y, Portman DS, Prahlad V, Raizen D, Rapti G, Xu SXZ, Zhang Y, Wu C-F. Nature's gift to neuroscience. J Neurogenet. 2020;34 ((3-4) :223-224. Publisher's VersionAbstract

PMID: 33446019

DOI: 10.1080/01677063.2020.1841760

*Liu H, Zhang Y. What can a worm learn in a bacteria-rich habitat?. J Neurogenet . 2020;(15) :1-9. Publisher's VersionAbstract

With a nervous system that has only a few hundred neurons, Caenorhabditis elegans was initially not regarded as a model for studies on learning. However, the collective effort of the C. elegans field in the past several decades has shown that the worm displays plasticity in its behavioral response to a wide range of sensory cues in the environment. As a bacteria-feeding worm, C. elegans is highly adaptive to the bacteria enriched in its habitat, especially those that are pathogenic and pose a threat to survival. It uses several common forms of behavioral plasticity that last for different amounts of time, including imprinting and adult-stage associative learning, to modulate its interactions with pathogenic bacteria. Probing the molecular, cellular and circuit mechanisms underlying these forms of experience-dependent plasticity has identified signaling pathways and regulatory insights that are conserved in more complex animals.

PMID: 33054485







  DOI: 10.1080/01677063.2020.1829614

Koterniak B, Pilaka PP, Gracida X, Schneider L-M, Pritišanac I, Zhang Y, Calarco JA. Global regulatory features of alternative splicing across tissues and within the nervous system of C. elegans. Genome Research. 2020;(267328) :120. Publisher's VersionAbstract

Alternative splicing plays a major role in shaping tissue-specific transcriptomes. Among the broad tissue types present in metazoans, the central nervous system contains some of the highest levels of alternative splicing. While many documented examples of splicing differences between broad tissue-types exist, there remains much to be understood about the splicing factors and the cis sequence elements controlling tissue and neuron subtype-specific splicing patterns. Using Translating Ribosome Affinity Purification coupled with deep-sequencing (TRAP-seq) in C. elegans, we have obtained high coverage profiles of ribosome-associated mRNA for three broad tissue classes (nervous system, muscle, and intestine) and two neuronal subtypes (dopaminergic and serotonergic neurons). We have identified hundreds of splice junctions that exhibit distinct splicing patterns between tissue types or within the nervous system. Alternative splicing events differentially regulated between tissues are more often frame-preserving, conserved across Caenorhabditis species and enriched in specific cis regulatory motifs. Utilizing this information, we have identified a likely mechanism of splicing repression by the RNA-binding protein UNC-75/CELF via interactions with cis elements that overlap a 5' splice site. Alternatively spliced exons also overlap more frequently with intrinsically disordered peptide regions than constitutive exons. Moreover, regulated exons are often shorter than constitutive exons but are flanked by longer intron sequences. Among these tissue-regulated exons are several highly conserved microexons less than 27 nucleotides in length. Collectively, our results indicate a rich layer of tissue-specific gene regulation at the level of alternative splicing in C. elegans that parallel the evolutionary forces and constraints observed across metazoa.





PMID: 33127752 DOI: 10.1101/gr.267328.120

* Pereira AG, Gracida X, Kagias K, Zhang Y. C. elegans aversive olfactory learning generates diverse intergenerational effects. J Neurogenet. 2020;10 (1080) :1-11. Publisher's VersionAbstract

Parental experience can modulate the behavior of their progeny. While the molecular mechanisms underlying parental effects or inheritance of behavioral traits have been studied under several environmental conditions, it remains largely unexplored how the nature of parental experience affects the information transferred to the next generation. To address this question, we used C. elegans, a nematode that feeds on bacteria in its habitat. Some of these bacteria are pathogenic and the worm learns to avoid them after a brief exposure. We found, unexpectedly, that a short parental experience increased the preference for the pathogen in the progeny. Furthermore, increasing the duration of parental exposure switched the response of the progeny from attraction to avoidance. To characterize the underlying molecular mechanisms, we found that the RNA-dependent RNA Polymerase (RdRP) RRF-3, required for the biogenesis of 26 G endo-siRNAs, regulated both types of intergenerational effects. Together, we show that different parental experiences with the same environmental stimulus generate different effects on the behavior of the progeny through small RNA-mediated regulation of gene expression.


doi: 10.1080/01677063.2020.1819265
    Morud J, Hardege I, Liu H, Wu T, Basu S, Zhang Y, Schafer WR. Deorphanisation of novel biogenic amine-gated ion channels identifies a new serotonin receptor for learning. bioRxiv . 2020;10 (1101). Publisher's VersionAbstract

    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.

    *Choi, Myung-Kyu, Liu H, Wu T, Yang W, Zhang Y. NMDAR-mediated modulation of gap junction circuit regulates olfactory learning in C. elegans. Nature Communications . 2020;11 (3467). Publisher's VersionAbstract

    Modulation of gap junction-mediated electrical synapses is a common form of neural plasticity. However, the behavioral consequence of the modulation and the underlying molecular cellular mechanisms are not understood. Here, using a C. elegans circuit of interneurons that are connected by gap junctions, we show that modulation of the gap junctions facilitates olfactory learning. Learning experience weakens the gap junctions and induces a repulsive sensory response to the training odorants, which together decouple the responses of the interneurons to the training odorants to generate learned olfactory behavior. The weakening of the gap junctions results from downregulation of the abundance of a gap junction molecule, which is regulated by cell-autonomous function of the worm homologs of a NMDAR subunit and CaMKII. Thus, our findings identify the function of a gap junction modulation in an in vivo model of learning and a conserved regulatory pathway underlying the modulation.

    * Wu T, Duan F, Yang W, Liu H, Caballero A, Fernandes de Abreu DA, Dar AR, Alcedo J, Ch’ng Q, Butcher RA, et al. Pheromones modulate learning by regulating the balanced signals of two insulin-like peptides. Neuron. 2019;2019.09.006. Publisher's VersionAbstract

    Social environment modulates learning through unknown mechanisms. Here, we report that a pheromone mixture that signals overcrowding inhibits C. elegans from learning to avoid pathogenic bacteria. We find that learning depends on the balanced signaling of two insulin-like peptides (ILPs), INS-16 and INS-4, which act respectively in the pheromone-sensing neuron ADL and the bacteria-sensing neuron AWA. Pheromone exposure inhibits learning by disrupting this balance: it activates ADL and increases expression of ins-16, and this cellular effect reduces AWA activity and AWA-expressed ins-4. The activities of the sensory neurons are required for learning and the expression of the ILPs. Interestingly, pheromones also promote the ingestion of pathogenic bacteria while increasing resistance to the pathogen. Thus, the balance of the ILP signals integrates social information into the learning process as part of a coordinated adaptive response that allows consumption of harmful food during times of high population density.



    * Pereira A, Gracida X, Kagias K, Zhang Y. Learning of pathogenic bacteria in adult C. elegans bidirectionally regulates pathogen response in the progeny. BioRxiv. 2019;doi:10.1101/500264. Publisher's VersionAbstract

    Parental experience can generate adaptive changes in the behavioral and physiological traits of the offspring13. However, the biological properties of this intergenerational regulation and the underlying molecular and cellular mechanism are not well understood. Here, we show that the experience of learning to avoid pathogenic bacteria in C. elegans alters the behavioral response to the pathogen in the progeny through the endogenous RNA interference (RNAi) pathway. We previously show that the adult C. elegans learns to avoid the smell of pathogenic bacteria, such as the Pseudomonas aeruginosa strain PA14, after feeding on the pathogen for a few hours4,5. Here, we report that this learning experience can bidirectionally regulate the olfactory response to PA14 in the progeny that are never directly exposed to the pathogen. The olfactory preference for PA14 in these progeny is linearly correlated with the learned avoidance of PA14 in their mothers. If the mothers show strong learning of PA14, their progeny avoid PA14; intriguingly, if the mothers show weak learning of PA14, the progeny prefer PA14, suggesting that the PA14-trained mothers transmit both the negative and positive information of PA14 to their progeny. The intergenerational behavioral effect results from an altered behavioral decision regulated by an olfactory sensorimotor neural circuit. Learning to avoid the pathogen also influences the development of the progeny, which is regulated independently from the behavioral change. Animals mutated for the RRF-3/RNA-directed RNA polymerase, a master regulator for the synthesis of the small interfering RNAs that are maternally inherited or in the soma6,7, display the normal naive and learned response to PA14 but are defective in regulating the olfactory response to PA14 in their progeny. Our results characterize an intergenerational effect that allows the progeny to rapidly adapt to an environmental condition that is critical for survival.

    * Harris G, Wu T, Linfield G, Choi M-K, Liu H, Zhang Y. Molecular and cellular modulators for multisensory integration in C. elegans. PLoS Genetics. 2019;15(3): e1007706. Publisher's VersionAbstract

    In the natural environment, animals often encounter multiple sensory cues that are simultaneously present. The nervous system integrates the relevant sensory information to generate behavioral responses that have adaptive values. However, the neuronal basis and the modulators that regulate integrated behavioral response to multiple sensory cues are not well defined. Here, we address this question using a behavioral decision in C. elegans when the animal is presented with an attractive food source together with a repulsive odorant. We identify specific sensory neurons, interneurons and neuromodulators that orchestrate the decision-making process, suggesting that various states and contexts may modulate the multisensory integration. Among these modulators, we characterize a new function of a conserved TGF-β pathway that regulates the integrated decision by inhibiting the signaling from a set of central neurons. Interestingly, we find that a common set of modulators, including the TGF-β pathway, regulate the integrated response to the pairing of different foods and repellents. Together, our results provide mechanistic insights into the modulatory signals regulating multisensory integration.

    *Donato A, Kagias K, Zhang Y#, Hilliard MA&. Neuronal sub-compartmentalization: a strategy to optimize neuronal function. #: Co-first authors. &:Co-senior authors. Biol Rev Camb Philos Soc. 2018;doi:10.1111/brv.12487. Publisher's VersionAbstract
    Neurons are highly polarized cells that consist of three main structural and functional domains: a cell body or soma, an axon, and dendrites. These domains contain smaller compartments with essential roles for proper neuronal function, such as the axonal presynaptic boutons and the dendritic postsynaptic spines. The structure and function of these compartments have now been characterized in great detail. Intriguingly, however, in the last decade additional levels of compartmentalization within the axon and the dendrites have been identified, revealing that these structures are much more complex than previously thought. Herein we examine several types of structural and functional sub‐compartmentalization found in neurons of both vertebrates and invertebrates. For example, in mammalian neurons the axonal initial segment functions as a sub‐compartment to initiate the action potential, to select molecules passing into the axon, and to maintain neuronal polarization. Moreover, work in Drosophila melanogaster has shown that two distinct axonal guidance receptors are precisely clustered in adjacent segments of the commissural axons both in vivo and in vitro, suggesting a cell‐intrinsic mechanism underlying the compartmentalized receptor localization. In Caenorhabditis elegans, a subset of interneurons exhibits calcium dynamics that are localized to specific sections of the axon and control the gait of navigation, demonstrating a regulatory role of compartmentalized neuronal activity in behaviour. These findings have led to a number of new questions, which are important for our understanding of neuronal development and function. How are these sub‐compartments established and maintained? What molecular machinery and cellular events are involved? What is their functional significance for the neuron? Here, we reflect on these and other key questions that remain to be addressed in this expanding field of biology.
    * Hao Y#, Yang W#, Hall Q, Ren J, Zhang Y&, Kaplan JM&. Thioredoxin shapes C. elegans sensory response to Pseudomonas produced nitric oxide. #: Co-first authors. &: Co-senior authors. eLife. 2018;pii: e36833. Publisher's VersionAbstract

    Nitric oxide (NO) is released into the air by NO-producing organisms; however, it is unclear if animals utilize NO as a sensory cue. We show that C. elegans avoids Pseudomonas aeruginosa (PA14) in part by detecting PA14-produced NO. PA14 mutants deficient for NO production fail to elicit avoidance and NO donors repel worms. PA14 and NO avoidance are mediated by a chemosensory neuron (ASJ) and these responses require receptor guanylate cyclases and cyclic nucleotide gated ion channels. ASJ exhibits calcium increases at both the onset and removal of NO. These NO-evoked ON and OFF calcium transients are affected by a redox sensing protein, TRX-1/thioredoxin. TRX-1’s trans-nitrosylation activity inhibits the ON transient whereas TRX-1’s de-nitrosylation activity promotes the OFF transient. Thus, C. elegans exploits bacterially produced NO as a cue to mediate avoidance and TRX-1 endows ASJ with a bi-phasic response to NO exposure.
    * Liu H, Yang W, Wu T, Duan F, Soucy E, Jin X, Zhang Y. Cholinergic sensorimotor integration regulates olfactory steering. Neuron. 2018;97 (2) :390-405. Publisher's VersionAbstract
    Sensorimotor integration regulates goal-directedmovements. We study the signaling mechanismsunderlying sensorimotor integration inC.elegansduring olfactory steering, when the sinusoidal move-ments of the worm generate an in-phase oscillation inthe concentration of the sampled odorant. We showthat cholinergic neurotransmission encodes theoscillatory sensory response and the motor state ofhead undulations by acting through an acetylcho-line-gated channel and a muscarinic acetylcholinereceptor, respectively. These signals convergeon two axonal domains of an interneuron RIA,where the sensory-evoked signal suppresses themotor-encoding signal to transform the spatial infor-mation of the odorant into the asymmetry betweenthe axonal activities. The asymmetric synaptic out-puts of the RIA axonal domains generate a direc-tional bias in the locomotory trajectory. Experiencealters the sensorimotor integration to generatespecific behavioral changes. Our study reveals howcholinergic neurotransmission, which can representsensory and motor information in the mammalianbrain, regulates sensorimotor integration duringgoal-directed locomotions.
    *Gracida X, Harris G, Zhang Y&, Calarco JA&. An Elongin-Cullin-SOCS-box Complex Regulates Stress-induced Serotonergic Neuromodulation. &: Co-senior authors. Cell Reports. 2017;10.1016 (11) :042. Publisher's VersionAbstract


    Neuromodulatory cells transduce environmental information into long-lasting behavioral responses. However, the mechanisms governing how neuronal cells influence behavioral plasticity are difficult to characterize. Here, we adapted the translating ribosome affinity purification (TRAP) approach in C. elegans to profile ribosome-associated mRNAs from three major tissues and the neuromodulatory dopaminergic and serotonergic cells. We identified elc-2, an Elongin C ortholog, specifically expressed in stress-sensing amphid neuron dual ciliated sensory ending (ADF) serotonergic sensory neurons, and we found that it plays a role in mediating a long-lasting change in serotonin-dependent feeding behavior induced by heat stress. We demonstrate that ELC-2 and the von Hippel-Lindau protein VHL-1, components of an Elongin-Cullin-SOCS box (ECS) E3 ubiquitin ligase, modulate this behavior after experiencing stress. Also, heat stress induces a transient redistribution of ELC-2, becoming more nuclearly enriched. Together, our results demonstrate dynamic regulation of an E3 ligase and a role for an ECS complex in neuromodulation and control of lasting behavioral states.


    *Yu J, Yang W, Liu H, Hao Y, Zhang Y. An Aversive Response to Osmotic Upshift in <i>Caenorhabditis elegans</i>. . eNeuro. 2017;4 (2) :pii: ENEURO.0282-16.2017.Abstract


    Environmental osmolarity presents a common type of sensory stimulus to animals. While behavioral responses to osmotic changes are important for maintaining a stable intracellular osmolarity, the underlying mechanisms are not fully understood. In the natural habitat of Caenorhabditis elegans, changes in environmental osmolarity are commonplace. It is known that the nematode acutely avoids shocks of extremely high osmolarity. Here, we show that C. elegans also generates gradually increased aversion of mild upshifts in environmental osmolarity. Different from an acute avoidance of osmotic shocks that depends on the function of a transient receptor potential vanilloid channel, the slow aversion to osmotic upshifts requires the cGMP-gated sensory channel subunit TAX-2. TAX-2 acts in several sensory neurons that are exposed to body fluid to generate the aversive response through a motor network that underlies navigation. Osmotic upshifts activate the body cavity sensory neuron URX, which is known to induce aversion upon activation. Together, our results characterize the molecular and cellular mechanisms underlying a novel sensorimotor response to osmotic stimuli and reveal that C. elegans engages different behaviors and the underlying mechanisms to regulate responses to extracellular osmolarity.

    eNeuro. 2017 Apr 21;4(2). 

    doi: 10.1523/ENEURO.0282-16.2017. 



    *Ghosh DD, Nitabach MN, Zhang Y, Harris G. Multisensory integration in C. elegans. Curr Opin Neurobiol. 2017;43 (10.1016/j.conb.2017.01.005) :110-118.Abstract


    Multisensory integration is a neural process by which signals from two or more distinct sensory channels are simultaneously processed to form a more coherent representation of the environment. Multisensory integration, especially when combined with a survey of internal states, provides selective advantages for animals navigating complex environments. Despite appreciation of the importance of multisensory integration in behavior, the underlying molecular and cellular mechanisms remain poorly understood. Recent work looking at how Caenorhabditis elegans makes multisensory decisions has yielded mechanistic insights into how a relatively simple and well-defined nervous system employs circuit motifs of defined features, synaptic signals and extrasynaptic neurotransmission, as well as neuromodulators in processing and integrating multiple sensory inputs to generate flexible and adaptive behavioral outputs.

    PMID: 28273525



    * Shen Y#, Wen Q#, Liu H#, Zhong C, Qin Y, Harris G, Kawano T, Wu M, Xu T, Samuel A, et al. An extrasynaptic GABAergic signal modulates a pattern of forward movement in Caenorhabditis elegans. #: Co-first authors. eLife. 2016;5 (e14197).Abstract

    As a common neurotransmitter in the nervous system, γ-aminobutyric acid (GABA) modulates locomotory patterns in both vertebrates and invertebrates. However, the signaling mechanisms underlying the behavioral effects of GABAergic modulation are not completely understood. Here, we demonstrate that a GABAergic signal in C. elegans modulates the amplitude of undulatory head bending through extrasynaptic neurotransmission and conserved metabotropic receptors. We show that the GABAergic RME head motor neurons generate undulatory activity patterns that correlate with head bending and the activity of RME causally links with head bending amplitude. The undulatory activity of RME is regulated by a pair of cholinergic head motor neurons SMD, which facilitate head bending, and inhibits SMD to limit head bending. The extrasynaptic neurotransmission between SMD and RME provides a gain control system to set head bending amplitude to a value correlated with optimal efficiency of forward movement.