Biology subjects

Poulet, J. F. A.

Publications and source records attributed to Poulet, J. F. A..

12 recordsLinked to original sources

bioRxiv2025

A specialized cold sensing system in the naked mole-rat

Avoiding cold or seeking warmth are universal animal needs. Homeothermic mammals maintain a constant body temperature despite fluctuating ambient temperatures. One exception is the naked mole-rat which lacks effective thermogenesis. We show that the naked mole-rat has evolved a greatly expanded cold sensing system. Compared to mice, this species has many more cold-sensitive sensory neurons and most express the cold-activated TRPM8 channel. The naked mole-rat TrpM8 gene harbors a unique upstream exon that when translated produces a TRPM8 protein with a 71 amino acid N-terminal extension. When expressed, the N-terminal extension prevents membrane targeting and abolishes TRPM8 function. Splice forms lacking the extension formed functional cold and menthol activated channels in vivo. Additionally, many naked mole-rat sensory neurons use a TRPM8-independent mechanism to detect cold. Thus, we identified molecular changes that confer both sensitivity and flexibility to cold sensing in a species that is critically dependent on following thermal cues.

neuroscience↗

bioRxiv2024

Bidirectionally responsive thermoreceptors encode cool and warm

Thermal sensation is a fundamental sense initiated by the activity of primary afferent thermoreceptors. While considerable attention has been paid to the encoding of noxious temperatures by thermoreceptors, it is far less clear how they encode innocuous cool and warm which are more commonly encountered in the environment. To address this, we sampled the entire thermoreceptor population using in vivo two-photon calcium imaging in the lumbar dorsal root ganglia of awake and anesthetized mice. We found that the vast majority of thermoreceptors respond bidirectionally, with an enhanced response to cool and a suppressed response to warm. Using in vivo pharmacology and computational modelling, we demonstrate that conductance changes in the cool-sensitive TRPM8 channel are sufficient to explain this bidirectional response type. Our comprehensive dataset reveals the fundamental principles of the peripheral encoding of innocuous temperatures and suggests that the same population of thermoreceptors underlie the distinct sensations of cool and warm.

neuroscience↗

bioRxiv2024

The spatial representation of temperature in the thalamus

Although distinct thalamic nuclei encode sensory information for almost all sensory modalities, the existence of a thalamic representation of temperature with a role in thermal perception remains unclear. To address this, we performed high-density electrophysiological recordings across the entire forelimb somatosensory thalamus in awake mice, and identified an anterior and a posterior representation of temperature that spans three thalamic nuclei. We found that these parallel representations show fundamental differences in the cellular encoding of temperature which reflects their cortical output targets. While the anterior representation encodes cool only and the posterior both cool and warm; in both representations cool was more densely represented and showed shorter latency, more transient responses as compared to warm. Moreover, thalamic inactivation showed a major role in thermal perception. Our comprehensive dataset identifies the thalamus as a key structure in thermal processing and highlights a novel posterior pathway in the thalamic representation of warm and cool.

neuroscience↗

bioRxiv2023

Optogenetic determination of dynamic and cell-type-specific chloride equilibrium potentials

Optogenetics has revolutionized neurobiological research by providing tools for modulating neuronal activity. As these tools utilise light-activated ion fluxes, they afford new opportunities to examine the nature of transmembrane ion gradients. Traditional investigation into the equilibrium potential for chloride (ECl) has been limited to studying endogenous chloride-permeable receptors. Here we demonstrate the utility of using a light-activated chloride channel, stGtACR2, to probe somatic ECl in rodent. This agonist-independent optogenetic strategy is validated in vitro and in vivo, captures differences in ECl dynamics following manipulations of endogenous chloride fluxes, and reveals distinct resting ECl across genetically-defined neuronal subpopulations. Using this approach to challenge chloride homeostasis, we uncover cell-specific ECl dynamics that are supported by the differential expression of endogenous handling mechanisms. Our findings establish an optical method for investigating transmembrane chloride gradients and thereby expand the repertoire of optogenetics.

neuroscience↗

bioRxiv2023

Cortical cellular encoding of thermotactile integration

Recent evidence suggests that primary sensory cortical regions play a role in the integration of information from multiple sensory modalities. How primary cortical neurons integrate multisensory information is unclear, partly because multisensory interactions in the cortex are typically weak or modulatory. To address this question, we take advantage of the robust representation of thermal (cooling) and tactile stimuli in mouse forepaw primary somatosensory cortex (fS1). Using a thermotactile detection task, we show that the perception of threshold level cool or tactile information is enhanced when they are presented simultaneously compared to presentation alone. To investigate the cortical correlates of thermotactile integration, we performed in vivo extracellular recordings from fS1 during unimodal and bimodal stimulation of the forepaw. Unimodal stimulation evoked thermal- or tactile- specific excitatory and inhibitory responses of fS1 neurons. The most prominent features of bimodal, thermotactile stimulation are the recruitment of unimodally silent fS1 neurons, non-linear integration features and a change in the response dynamics to favor longer response durations. Together, we identify quantitative and qualitative changes in cortical encoding that may underlie the improvement in perception of multisensory, thermotactile surfaces during haptic exploration.

neuroscience↗

bioRxiv2022

Brain-wide connectivity map of mouse thermosensory cortices

In the thermal system, skin cooling is represented in the primary somatosensory cortex (S1) and the posterior insular cortex (pIC). Whether S1 and pIC are nodes in anatomically separated or overlapping thermal sensorimotor pathways is unclear, as the brain-wide connectivity of the thermal system has not been mapped. We address this using functionally targeted, dual injections of anterograde viruses or retrograde tracers into S1 and pIC. Our data show that inputs to S1 and pIC originate from two non-overlapping populations, supporting the existence of parallel input pathways. While outputs from S1 and pIC were more widespread and share a number of cortical and subcortical regions, within target structures axonal projections were often separable. We observed a high degree of reciprocal connectivity with thalamic and cortical regions from both S1 and pIC, but output to the mid- and hind-brain was unidirectional. Notably, pIC showed exclusive connectivity with regions associated with thermal processing. Together, these data indicate that cutaneous thermal information is routed to the cortex via multiple, parallel streams of information which are forwarded to overlapping downstream regions for the binding of complex somatosensory percepts and integration with ongoing behavior.

neuroscience↗

bioRxiv2022

The cellular coding of temperature in the mammalian cortex

Temperature is a fundamental sensory modality separate from touch, with dedicated receptor channels and primary afferent neurons for cool and warm (Blix, 1882; Filingeri, 2016; Vriens et al., 2014). Unlike other modalities, however, the cortical encoding of temperature remains mysterious, with very few cortical neurons reported that respond to non-painful temperature and the presence of a thermal cortex is debated (Bokiniec et al., 2018; Craig et al., 2000; Hellon et al., 1973; Milenkovic et al., 2014; Tsuboi et al., 1993). Using widefield and two-photon calcium imaging in the mouse forepaw system, here we identify cortical neurons that respond to cooling and/or warming with distinct spatial and temporal response properties. Surprisingly, we observed a representation of cool, but not warm, in the primary somatosensory cortex, but cool and warm in the posterior insular cortex (pIC). The representation of thermal information in pIC is robust, somatotopicallyarranged and reversible manipulations show a profound impact on thermal perception. Intriguingly, despite being positioned along the same one-dimensional sensory axis, the encoding of cool and warm is distinct, both in highly- and broadly- tuned neurons. Together, our results show that pIC contains the primary cortical representation of skin temperature and may help explain how the thermal system generates sensations of cool and warm.

neuroscience↗

bioRxiv2022

Sensory Schwann cells are required for mechanical nociception and touch perception

That sensory neurons alone transduce mechanical stimuli was challenged by the discovery of nociceptive Schwann cells that can initiate pain. Consistent with the existence of inherently mechanosensitive sensory Schwann cells, we found that the mechanosensory function of almost all nociceptors, including those signaling fast pain, were critically dependent on sensory Schwann cells. Furthermore, in polymodal nociceptors, sensory Schwann cells signal mechanical, but not cold or heat pain. Terminal Schwann cells also surround mechanoreceptor nerve-endings within the Meissners corpuscle and in at hair follicle lanceolate endings that both signal vibrotactile touch. Within Meissners corpuscles, two molecularly and functionally distinct sensory Schwann cells positive for Sox10 and Sox2 differentially modulate rapidly adapting mechanoreceptor function. Using optogenetics we show that Meissners corpuscle Schwann cells are necessary for the perception of low threshold vibrotactile stimuli. These results show that sensory Schwann cells within diverse glio-neural mechanosensory end-organs are sensors for mechanical pain as well as necessary for touch perception. Thus, specialized sensory Schwann are central to the transduction of mechanical forces that underpin somatic sensation.

neuroscience↗

bioRxiv2021

Deep tissue scattering compensation with three-photon F-SHARP

Optical imaging techniques are widely used in biological research, but their penetration depth is limited by tissue scattering. Wavefront shaping techniques are able to overcome this problem in principle, but are often slow and their performance depends on the sample. This greatly reduces their practicability for biological applications. Here we present a scattering compensation technique based on three-photon (3P) excitation, which converges faster than comparable two-photon (2P) techniques and works reliably even on densely labeled samples, where 2P approaches fail. To demonstrate its usability and advantages for biomedical imaging we apply it to the imaging of dendritic spines on GFP-labeled layer 5 neurons in an anesthetized mouse.

bioengineering↗

bioRxiv2021

An executive center for the intake of liquids

It has long been known that orofacial movements for feeding can be triggered, coordinated, and often rhythmically organized at the level of the brainstem, without input from higher centers. We uncover two nuclei that can organize the movements for ingesting fluids in mammals. These neuronal groups, defined by unique transcriptional codes and developmental origins, IRtPhox2b and Peri5Atoh1, are located, respectively, in the intermediate reticular formation of the medulla and around the motor nucleus of the trigeminal nerve. They are premotor to all jaw-opening and tongue muscles. Stimulation of either, in awake animals, opens the jaw, while IRtPhox2b alone also protracts the tongue. Moreover, stationary stimulation of IRtPhox2b entrains a rhythmic alternation of tongue protraction and retraction, synchronized with jaw opening and closing, that mimics lapping. Finally, fiber photometric recordings show that IRtPhox2b is active during volitional lapping. Our study identifies one of the long hypothesized subcortical nuclei underpinning a stereotyped feeding behavior.

neuroscience↗

bioRxiv2019

Multiple two-photon targeted whole-cell patch-clamp recordings from monosynaptically connected neurons in vivo

Although we know a great deal about monosynaptic connectivity, transmission and integration in the mammalian nervous system from in vitro studies, very little is known in vivo. This is partly because it is technically difficult to evoke action potentials and simultaneously record small amplitude subthreshold responses in closely (< 150 m) located pairs of neurons. To address this, we have developed in vivo two-photon targeted multiple (2 - 4) whole-cell patch clamp recordings of nearby neurons in superficial layers 1 to 3 of primary somatosensory cortex. Here we describe a step-by-step guide to this approach in the anesthetised mouse, including: the design of the setup, surgery, preparation of pipettes, targeting and acquisition of multiple whole-cell recordings, as well as in vivo and post-hoc histology. The procedure takes ~ 4 hours from start of surgery to end of recording and allows examinations both into the electrophysiological features of unitary excitatory and inhibitory monosynaptic inputs and the synaptic mechanisms of correlated neuronal activity.

neuroscience↗

bioRxiv2018

The sensory coding of warm perception

Humans easily discriminate tiny skin temperature changes that are perceived as warming or cooling. Dedicated thermoreceptors forming distinct thermosensory channels or \"labelled lines\" are thought to underlie thermal perception. We show that mice have similar perceptual thresholds for forepaw warming to humans (~1 {degrees}C change) and do not mistake warming for cooling. Mice perform warm discrimination tasks without dedicated thermoreceptors, but use information carried by unmyelinated polymodal C-fibers. Deletion of the heat-sensitive transduction channels TRPM2 and TRPV1 did not impact warming perception or afferent coding of warm. However, without the cold sensitive TRPM8 channel, afferent coding of cooling was impaired and these mice cannot perceive warming or cooling. Our data is incompatible with the existence of thermospecific labelled lines, but can be reconciled by the existence of central circuits that compare and integrate the input from at least two types of polymodal afferents, hitherto thought to exclusively signal pain.

neuroscience↗