RESUMEN
The gastrointestinal tract is in a state of constant motion. These movements are tightly regulated by the presence of food and help digestion by mechanically breaking down and propelling gut content. Mechanical sensing in the gut is thought to be essential for regulating motility; however, the identity of the neuronal populations, the molecules involved, and the functional consequences of this sensation are unknown. Here, we show that humans lacking PIEZO2 exhibit impaired bowel sensation and motility. Piezo2 in mouse dorsal root, but not nodose ganglia is required to sense gut content, and this activity slows down food transit rates in the stomach, small intestine, and colon. Indeed, Piezo2 is directly required to detect colon distension in vivo. Our study unveils the mechanosensory mechanisms that regulate the transit of luminal contents throughout the gut, which is a critical process to ensure proper digestion, nutrient absorption, and waste removal.
Asunto(s)
Tránsito Gastrointestinal , Canales Iónicos , Mecanotransducción Celular , Animales , Humanos , Ratones , Digestión , Canales Iónicos/metabolismo , Neuronas/metabolismoRESUMEN
PIEZOs are mechanosensitive ion channels that convert force into chemoelectric signals1,2 and have essential roles in diverse physiological settings3. In vitro studies have proposed that PIEZO channels transduce mechanical force through the deformation of extensive blades of transmembrane domains emanating from a central ion-conducting pore4-8. However, little is known about how these channels interact with their native environment and which molecular movements underlie activation. Here we directly observe the conformational dynamics of the blades of individual PIEZO1 molecules in a cell using nanoscopic fluorescence imaging. Compared with previous structural models of PIEZO1, we show that the blades are significantly expanded at rest by the bending stress exerted by the plasma membrane. The degree of expansion varies dramatically along the length of the blade, where decreased binding strength between subdomains can explain increased flexibility of the distal blade. Using chemical and mechanical modulators of PIEZO1, we show that blade expansion and channel activation are correlated. Our findings begin to uncover how PIEZO1 is activated in a native environment. More generally, as we reliably detect conformational shifts of single nanometres from populations of channels, we expect that this approach will serve as a framework for the structural analysis of membrane proteins through nanoscopic imaging.
Asunto(s)
Canales Iónicos , Membrana Celular/metabolismo , Fluorescencia , Canales Iónicos/química , Canales Iónicos/metabolismo , Modelos Moleculares , Movimiento , Conformación Proteica , Análisis de la Célula IndividualRESUMEN
Henry Miller stated that "to relieve a full bladder is one of the great human joys". Urination is critically important in health and ailments of the lower urinary tract cause high pathological burden. Although there have been advances in understanding the central circuitry in the brain that facilitates urination1-3, there is a lack of in-depth mechanistic insight into the process. In addition to central control, micturition reflexes that govern urination are all initiated by peripheral mechanical stimuli such as bladder stretch and urethral flow4. The mechanotransduction molecules and cell types that function as the primary stretch and pressure detectors in the urinary tract mostly remain unknown. Here we identify expression of the mechanosensitive ion channel PIEZO2 in lower urinary tract tissues, where it is required for low-threshold bladder-stretch sensing and urethral micturition reflexes. We show that PIEZO2 acts as a sensor in both the bladder urothelium and innervating sensory neurons. Humans and mice lacking functional PIEZO2 have impaired bladder control, and humans lacking functional PIEZO2 report deficient bladder-filling sensation. This study identifies PIEZO2 as a key mechanosensor in urinary function. These findings set the foundation for future work to identify the interactions between urothelial cells and sensory neurons that control urination.
Asunto(s)
Canales Iónicos/metabolismo , Mecanotransducción Celular/fisiología , Células Receptoras Sensoriales/metabolismo , Vejiga Urinaria/inervación , Vejiga Urinaria/fisiología , Micción/fisiología , Urotelio/citología , Animales , Femenino , Humanos , Canales Iónicos/deficiencia , Ratones , Presión , Reflejo/fisiología , Vejiga Urinaria/citología , Vejiga Urinaria/fisiopatología , Sistema Urinario/inervación , Sistema Urinario/metabolismo , Urotelio/metabolismoRESUMEN
Many organs are designed to move: the heart pumps each second, the gastrointestinal tract squeezes and churns to digest food, and we contract and relax skeletal muscles to move our bodies. Sensory neurons of the peripheral nervous system detect signals from bodily tissues, including the forces generated by these movements, to control physiology. The processing of these internal signals is called interoception, but this is a broad term that includes a wide variety of both chemical and mechanical sensory processes. Mechanical senses are understudied, but rapid progress has been made in the last decade, thanks in part to the discovery of the mechanosensory PIEZO ion channels (Coste et al., 2010). The role of these mechanosensors within the interoceptive nervous system is the focus of this review. In defining the transduction molecules that govern mechanical interoception, we will have a better grasp of how these signals drive physiology.
RESUMEN
Touch submodalities, such as flutter and pressure, are mediated by somatosensory afferents whose terminal specializations extract tactile features and encode them as action potential trains with unique activity patterns. Whether non-neuronal cells tune touch receptors through active or passive mechanisms is debated. Terminal specializations are thought to function as passive mechanical filters analogous to the cochlea's basilar membrane, which deconstructs complex sounds into tones that are transduced by mechanosensory hair cells. The model that cutaneous specializations are merely passive has been recently challenged because epidermal cells express sensory ion channels and neurotransmitters; however, direct evidence that epidermal cells excite tactile afferents is lacking. Epidermal Merkel cells display features of sensory receptor cells and make 'synapse-like' contacts with slowly adapting type I (SAI) afferents. These complexes, which encode spatial features such as edges and texture, localize to skin regions with high tactile acuity, including whisker follicles, fingertips and touch domes. Here we show that Merkel cells actively participate in touch reception in mice. Merkel cells display fast, touch-evoked mechanotransduction currents. Optogenetic approaches in intact skin show that Merkel cells are both necessary and sufficient for sustained action-potential firing in tactile afferents. Recordings from touch-dome afferents lacking Merkel cells demonstrate that Merkel cells confer high-frequency responses to dynamic stimuli and enable sustained firing. These data are the first, to our knowledge, to directly demonstrate a functional, excitatory connection between epidermal cells and sensory neurons. Together, these findings indicate that Merkel cells actively tune mechanosensory responses to facilitate high spatio-temporal acuity. Moreover, our results indicate a division of labour in the Merkel cell-neurite complex: Merkel cells signal static stimuli, such as pressure, whereas sensory afferents transduce dynamic stimuli, such as moving gratings. Thus, the Merkel cell-neurite complex is an unique sensory structure composed of two different receptor cell types specialized for distinct elements of discriminative touch.
Asunto(s)
Vías Aferentes , Células Epidérmicas , Epidermis/inervación , Mecanotransducción Celular , Células de Merkel/metabolismo , Tacto/fisiología , Potenciales de Acción , Animales , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Conductividad Eléctrica , Femenino , Canales Iónicos/metabolismo , Masculino , Ratones , Modelos Biológicos , Neuritas/metabolismo , Neuronas Aferentes/metabolismo , Optogenética , PresiónRESUMEN
Bioactive derivatives from the camphor laurel tree, Cinnamomum camphora, are posited to exhibit chemopreventive properties but the efficacy and mechanism of these natural products are not fully understood. We tested an essential-oil derivative, camphor white oil (CWO), for anti-tumor activity in a mouse model of keratinocyte-derived skin cancer. Daily topical treatment with CWO induced dramatic regression of pre-malignant skin tumors and a two-fold reduction in cutaneous squamous cell carcinomas. We next investigated underlying cellular and molecular mechanisms. In cultured keratinocytes, CWO stimulated calcium signaling, resulting in calcineurin-dependent activation of nuclear factor of activated T cells (NFAT). In vivo, CWO induced transcriptional changes in immune-related genes identified by RNA-sequencing, resulting in cytotoxic T cell-dependent tumor regression. Finally, we identified chemical constituents of CWO that recapitulated effects of the admixture. Together, these studies identify T cell-mediated tumor regression as a mechanism through which a plant-derived essential oil diminishes established tumor burden.
Asunto(s)
Alcanfor/administración & dosificación , Carcinoma de Células Escamosas/tratamiento farmacológico , Queratinocitos/efectos de los fármacos , Aceites Volátiles/administración & dosificación , Neoplasias Cutáneas/tratamiento farmacológico , Linfocitos T Citotóxicos/inmunología , Animales , Antracenos/toxicidad , Carcinoma de Células Escamosas/inducido químicamente , Carcinoma de Células Escamosas/inmunología , Proliferación Celular , Células Cultivadas , Femenino , Humanos , Queratinocitos/inmunología , Queratinocitos/patología , Ratones , Factores de Transcripción NFATC/metabolismo , Piperidinas/toxicidad , Neoplasias Cutáneas/inducido químicamente , Neoplasias Cutáneas/inmunología , Linfocitos T Citotóxicos/efectos de los fármacos , Linfocitos T Citotóxicos/metabolismoRESUMEN
Distal arthrogryposis (DA) is a collection of rare disorders that are characterized by congenital joint contractures. Most DA mutations are in muscle- and joint-related genes, and the anatomical defects originate cell-autonomously within the musculoskeletal system. However, gain-of-function mutations in PIEZO2, a principal mechanosensor in somatosensation, cause DA subtype 5 (DA5) through unknown mechanisms. We show that expression of a gain-of-function PIEZO2 mutation in proprioceptive sensory neurons that mainly innervate muscle spindles and tendons is sufficient to induce DA5-like phenotypes in mice. Overactive PIEZO2 causes anatomical defects through increased activity within the peripheral nervous system during postnatal development. Furthermore, botulinum toxin (Botox) and a dietary fatty acid that modulates PIEZO2 activity reduce DA5-like deficits. This reveals a role for somatosensory neurons: Excessive mechanosensation within these neurons disrupts musculoskeletal development.
Asunto(s)
Artrogriposis , Contractura , Canales Iónicos , Mecanotransducción Celular , Células Receptoras Sensoriales , Animales , Ratones , Artrogriposis/genética , Artrogriposis/fisiopatología , Contractura/genética , Contractura/fisiopatología , Mecanotransducción Celular/genética , Mutación , Células Receptoras Sensoriales/fisiología , Canales Iónicos/genéticaRESUMEN
Sensory neurons detect mechanical forces from both the environment and internal organs to regulate physiology. PIEZO2 is a mechanosensory ion channel critical for touch, proprioception, and bladder stretch sensation, yet its broad expression in sensory neurons suggests it has undiscovered physiological roles. To fully understand mechanosensory physiology, we must know where and when PIEZO2-expressing neurons detect force. The fluorescent styryl dye FM 1-43 was previously shown to label sensory neurons. Surprisingly, we find that the vast majority of FM 1-43 somatosensory neuron labeling in mice in vivo is dependent on PIEZO2 activity within the peripheral nerve endings. We illustrate the potential of FM 1-43 by using it to identify novel PIEZO2-expressing urethral neurons that are engaged by urination. These data reveal that FM 1-43 is a functional probe for mechanosensitivity via PIEZO2 activation in vivo and will facilitate the characterization of known and novel mechanosensory processes in multiple organ systems.
Asunto(s)
Canales Iónicos , Mecanotransducción Celular , Ratones , Animales , Mecanotransducción Celular/fisiología , Canales Iónicos/metabolismo , Células Receptoras Sensoriales/fisiología , Compuestos de Piridinio/metabolismoRESUMEN
Multiple senses, including hearing, touch and osmotic regulation, require the ability to convert force into an electrical signal: A process called mechanotransduction. Mechanotransduction occurs through specialized proteins that open an ion channel pore in response to a mechanical stimulus. Many of these proteins remain unidentified in vertebrates, but known mechanotransduction channels in lower organisms provide clues into their identity and mechanism. Bacteria, fruit flies and nematodes have all been used to elucidate the molecules necessary for force transduction. This chapter discusses many different mechanical senses and takes an evolutionary approach to review the proteins responsible for mechanotransduction in various biological kingdoms.
Asunto(s)
Mecanotransducción Celular , Animales , Fenómenos Biomecánicos , Células Eucariotas/citología , Células Eucariotas/metabolismo , Células Procariotas/citología , Células Procariotas/metabolismo , Vertebrados/metabolismo , Vertebrados/fisiologíaRESUMEN
Activation of stretch-sensitive baroreceptor neurons exerts acute control over heart rate and blood pressure. Although this homeostatic baroreflex has been described for more than 80 years, the molecular identity of baroreceptor mechanosensitivity remains unknown. We discovered that mechanically activated ion channels PIEZO1 and PIEZO2 are together required for baroreception. Genetic ablation of both Piezo1 and Piezo2 in the nodose and petrosal sensory ganglia of mice abolished drug-induced baroreflex and aortic depressor nerve activity. Awake, behaving animals that lack Piezos had labile hypertension and increased blood pressure variability, consistent with phenotypes in baroreceptor-denervated animals and humans with baroreflex failure. Optogenetic activation of Piezo2-positive sensory afferents was sufficient to initiate baroreflex in mice. These findings suggest that PIEZO1 and PIEZO2 are the long-sought baroreceptor mechanosensors critical for acute blood pressure control.
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Barorreflejo/fisiología , Presión Sanguínea/fisiología , Canales Iónicos/fisiología , Mecanotransducción Celular/fisiología , Neuronas/fisiología , Presorreceptores/fisiología , Animales , Barorreflejo/genética , Canales Iónicos/genética , Mecanotransducción Celular/genética , Ratones , Ratones Endogámicos C57BL , Ratones Mutantes , Ganglio Nudoso/fisiología , OptogenéticaRESUMEN
The brush of a feather and a pinprick are perceived as distinct sensations because they are detected by discrete cutaneous sensory neurons. Inflammation or nerve injury can disrupt this sensory coding and result in maladaptive pain states, including mechanical allodynia, the development of pain in response to innocuous touch. However, the molecular mechanisms underlying the alteration of mechanical sensitization are poorly understood. In mice and humans, loss of mechanically activated PIEZO2 channels results in the inability to sense discriminative touch. However, the role of Piezo2 in acute and sensitized mechanical pain is not well defined. Here, we showed that optogenetic activation of Piezo2-expressing sensory neurons induced nociception in mice. Mice lacking Piezo2 in caudal sensory neurons had impaired nocifensive responses to mechanical stimuli. Consistently, ex vivo recordings in skin-nerve preparations from these mice showed diminished Aδ-nociceptor and C-fiber firing in response to mechanical stimulation. Punctate and dynamic allodynia in response to capsaicin-induced inflammation and spared nerve injury was absent in Piezo2-deficient mice. These results indicate that Piezo2 mediates inflammation- and nerve injury-induced sensitized mechanical pain, and suggest that targeting PIEZO2 might be an effective strategy for treating mechanical allodynia.
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Hiperalgesia/metabolismo , Canales Iónicos/metabolismo , Mecanotransducción Celular , Dolor/metabolismo , Potenciales de Acción , Animales , Conducta Animal , Capsaicina , Hiperalgesia/complicaciones , Hiperalgesia/patología , Hiperalgesia/fisiopatología , Canales Iónicos/deficiencia , Ratones Noqueados , Neuronas/metabolismo , Nocicepción , Nociceptores/metabolismo , Dolor/complicaciones , Dolor/patología , Dolor/fisiopatologíaRESUMEN
Sensory tissues exposed to the environment, such as skin, olfactory epithelia, and taste buds, continuously renew; therefore, peripheral neurons must have mechanisms to maintain appropriate innervation patterns. Although somatosensory neurons regenerate after injury, little is known about how these neurons cope with normal target organ changes. To elucidate neuronal plasticity in healthy skin, we analyzed the structure of Merkel-cell afferents, which are gentle touch receptors, during skin remodeling that accompanies mouse hair-follicle regeneration. The number of Merkel cells is reduced by 90% and axonal arbors are simplified during active hair growth. These structures rebound within just days. Computational modeling predicts that Merkel-cell changes are probabilistic, but myelinated branch stability depends on Merkel-cell inputs. Electrophysiology and behavior demonstrate that tactile responsiveness is less reliable during active growth than in resting skin. These results reveal that somatosensory neurons display structural plasticity at the cost of impairment in the reliability of encoding gentle touch.
Asunto(s)
Receptores de Superficie Celular/metabolismo , Piel/metabolismo , Tacto/fisiología , Animales , Conducta Animal , Simulación por Computador , Cabello/crecimiento & desarrollo , Células de Merkel/metabolismo , Ratones Endogámicos C57BLRESUMEN
Although the skin's mechanical properties are well characterized in tension, little work has been done in compression. Here, the viscoelastic properties of a population of mouse skin specimens (139 samples from 36 mice, aged 5 to 34 weeks) were characterized upon varying specimen thickness, as well as strain level and rate. Over the population, we observed the skin's viscoelasticity to be quite variable, yet found systematic correlation of residual stress ratio with skin thickness and strain, and of relaxation time constants with strain rates. In particular, as specimen thickness ranged from 211 to 671 µm, we observed significant variation in both quasi-linear viscoelasticity (QLV) parameters, the relaxation time constant (τ1 = 0.19 ± 0.10 s) and steady-state residual stress ratio (G∞ = 0.28 ± 0.13). Moreover, when τ1 was decoupled and fixed, we observed that G∞ positively correlated with skin thickness. Second, as steady-state stretch was increased (λ∞ from 0.22 to 0.81), we observed significant variation in both QLV parameters (τ1 = 0.26 ± 0.14 s, G∞ = 0.47 ± 0.17), and when τ1 was fixed, G∞ positively correlated with stretch level. Third, as strain rate was increased from 0.06 to 22.88 s-1, the median time constant τ1 varied from 1.90 to 0.31 s, and thereby negatively correlated with strain rate. These findings indicate that the natural range of specimen thickness, as well as experimental controls of compression level and rate, significantly influence measurements of skin viscoelasticity.
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Piel/química , Animales , Fuerza Compresiva , Simulación por Computador , Elasticidad , Ratones , Modelos Biológicos , Estrés Mecánico , ViscosidadRESUMEN
Flight maneuvers require rapid sensory integration to generate adaptive motor output. Bats achieve remarkable agility with modified forelimbs that serve as airfoils while retaining capacity for object manipulation. Wing sensory inputs provide behaviorally relevant information to guide flight; however, components of wing sensory-motor circuits have not been analyzed. Here, we elucidate the organization of wing innervation in an insectivore, the big brown bat, Eptesicus fuscus. We demonstrate that wing sensory innervation differs from other vertebrate forelimbs, revealing a peripheral basis for the atypical topographic organization reported for bat somatosensory nuclei. Furthermore, the wing is innervated by an unusual complement of sensory neurons poised to report airflow and touch. Finally, we report that cortical neurons encode tactile and airflow inputs with sparse activity patterns. Together, our findings identify neural substrates of somatosensation in the bat wing and imply that evolutionary pressures giving rise to mammalian flight led to unusual sensorimotor projections.
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Quirópteros/fisiología , Vuelo Animal/fisiología , Corteza Somatosensorial/fisiología , Animales , Técnicas de Trazados de Vías Neuroanatómicas , Tacto/fisiología , Alas de Animales/inervación , Alas de Animales/fisiologíaRESUMEN
Touch is encoded by cutaneous sensory neurons with diverse morphologies and physiological outputs. How neuronal architecture influences response properties is unknown. To elucidate the origin of firing patterns in branched mechanoreceptors, we combined neuroanatomy, electrophysiology and computation to analyze mouse slowly adapting type I (SAI) afferents. These vertebrate touch receptors, which innervate Merkel cells, encode shape and texture. SAI afferents displayed a high degree of variability in touch-evoked firing and peripheral anatomy. The functional consequence of differences in anatomical architecture was tested by constructing network models representing sequential steps of mechanosensory encoding: skin displacement at touch receptors, mechanotransduction and action-potential initiation. A systematic survey of arbor configurations predicted that the arrangement of mechanotransduction sites at heminodes is a key structural feature that accounts in part for an afferent's firing properties. These findings identify an anatomical correlate and plausible mechanism to explain the driver effect first described by Adrian and Zotterman. DOI: http://dx.doi.org/10.7554/eLife.01488.001.
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Simulación por Computador , Mecanorreceptores/ultraestructura , Mecanotransducción Celular , Modelos Neurológicos , Red Nerviosa/ultraestructura , Piel/inervación , Tacto , Potenciales de Acción , Adaptación Fisiológica , Animales , Potenciales Evocados , Femenino , Mecanorreceptores/fisiología , Células de Merkel/fisiología , Células de Merkel/ultraestructura , Ratones Transgénicos , Microscopía Confocal , Red Nerviosa/fisiología , Técnicas de Trazados de Vías Neuroanatómicas , Factores de TiempoRESUMEN
The neural response to touch stimuli is influenced by skin properties as well as the delivery of stimuli. Here, we compare stimuli controlled by displacement and force, and analyze the impact on firing rates of slowly adapting type I afferents as skin thickness and elasticity change. Uniaxial compression tests were used to measure the mechanical properties of mouse hind limb skin (n=5), resulting in a range of skin thickness measurements (211.6-530.6 µm) and hyper- and visco-elastic properties (average coefficient of variation=0.27).Values were integrated to an axisymmetric finite element model using an Ogden strain energy function. This calculated the propagation of surface loads to tactile end-organ locations, where maximum compressive stress and its rate were sampled and linearly regressed to firing rate. For the observed range of skin thickness, firing response was predicted under both force and displacement control of a ramp-and-hold stimulus. Over the ramp phase of stimulation, the variance in predicted firing rate was higher under displacement than under force control (22.2versus 4.9 Hz) with a similar trend in the sustained phase of stimulation (4.6versus1.3Hz). Given that skin thickness varies significantly between specimens, for human skin perhaps seven more so than for mice, the use of force control is predicted to decrease experimental variance in neurophysiological and psychophysical responses.
RESUMEN
The skin is a dynamic organ whose complex material properties are capable of withstanding continuous mechanical stress while accommodating insults and organism growth. Moreover, synchronized hair cycles, comprising waves of hair growth, regression and rest, are accompanied by dramatic fluctuations in skin thickness in mice. Whether such structural changes alter skin mechanics is unknown. Mouse models are extensively used to study skin biology and pathophysiology, including aging, UV-induced skin damage and somatosensory signaling. As the skin serves a pivotal role in the transfer function from sensory stimuli to neuronal signaling, we sought to define the mechanical properties of mouse skin over a range of normal physiological states. Skin thickness, stiffness and modulus were quantitatively surveyed in adult, female mice (Mus musculus). These measures were analyzed under uniaxial compression, which is relevant for touch reception and compression injuries, rather than tension, which is typically used to analyze skin mechanics. Compression tests were performed with 105 full-thickness, freshly isolated specimens from the hairy skin of the hind limb. Physiological variables included body weight, hair-cycle stage, maturity level, skin site and individual animal differences. Skin thickness and stiffness were dominated by hair-cycle stage at young (6-10 weeks) and intermediate (13-19 weeks) adult ages but by body weight in mature mice (26-34 weeks). Interestingly, stiffness varied inversely with thickness so that hyperelastic modulus was consistent across hair-cycle stages and body weights. By contrast, the mechanics of hairy skin differs markedly with anatomical location. In particular, skin containing fascial structures such as nerves and blood vessels showed significantly greater modulus than adjacent sites. Collectively, this systematic survey indicates that, although its structure changes dramatically throughout adult life, mouse skin at a given location maintains a constant elastic modulus to compression throughout normal physiological stages.
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Elasticidad , Fenómenos Fisiológicos de la Piel , Animales , RatonesRESUMEN
Degenerin/epithelial sodium channels (DEG/ENaCs) are luminaries of gentle touch in Caenorhabditis elegans. In this issue of Neuron, Geffeney et al. demonstrate that eponymous DEG-1 channels carry mechanotransduction currents in a polymodal neuron, where they act upstream of transient receptor potential (TRP) channels.
RESUMEN
The sense of touch detects forces that bombard the body's surface. In metazoans, an assortment of morphologically and functionally distinct mechanosensory cell types are tuned to selectively respond to diverse mechanical stimuli, such as vibration, stretch, and pressure. A comparative evolutionary approach across mechanosensory cell types and genetically tractable species is beginning to uncover the cellular logic of touch reception.