Skin Sensory Neurons Research Articles

Biomimetic Tactile Sensors Based on Nanomaterials.

Tactile sensor technology has been researched extensively in response to the increasing demand for robotic and wearable healthcare systems. Studies on tactile sensory systems have primarily focused on achieving two goals: (1) developing technologies with high sensing abilities that mimic the biological functions and characteristics of the sensory systems of human skin and (2) satisfying the requirements of wearable device applications by fabricating mechanically flexible devices with low-power data analysis and processing abilities. In this Perspective, we present recent advances in artificial tactile sensory systems, which are based on biomimetic technologies that exhibit functional features of biological systems including mechanoreceptors and human skin sensory neurons for human-machine interfaces. We also discuss the opportunities, current challenges, potential solutions, and future investigative directions pertaining to this field.

Impairment of proprioceptive movement and mechanical nociception in Drosophila melanogaster larvae lacking Ppk30, a Drosophila member of the Degenerin/Epithelial Sodium Channel family.

The mechanosensory neurons of Drosophila larvae are demonstrably activated by diverse mechanical stimuli, but the mechanisms underlying this function are not completely understood. Here we report a genetic, immunohistochemical, and electrophysiological analysis of the Ppk30 ion channel, a member of the Drosophila pickpocket (ppk) family, counterpart of the mammalian Degenerin/Epithelial Na+ Channel family. Ppk30 mutant larvae displayed deficits in proprioceptive movement and mechanical nociception, which are detected by class IV sensory (mdIV) neurons. The same neurons also detect heat nociception, which was not impaired in ppk30 mutant larvae. Similarly, Ppk30 mutation did not alter gentle touch mechanosensation, a distinct mechanosensation detected by other neurons, suggesting that Ppk30 has a functional role in mechanosensation in mdIV neurons. Consistently, Ppk30 was expressed in class IV neurons, but was not detectable in other larval skin sensory neurons. Mutant phenotypes were rescued by expressing Ppk30 in mdIV neurons. Electrophysiological analysis of heterologous cells expressing Ppk30 did not detect mechanosensitive channel activities, but did detect acid-induced currents. These data show that Ppk30 has a role in mechanosensation, but not in thermosensation, in class IV neurons, and possibly has other functions related to acid response.

TRPA1 Channel as a Regulator of Neurogenic Inflammation and Pain: Structure, Function, Role in Pathophysiology, and Therapeutic Potential of Ligands.

TRPA1 is a cation channel located on the plasma membrane of many types of human and animal cells, including skin sensory neurons and epithelial cells of the intestine, lungs, urinary bladder, etc. TRPA1 is the major chemosensor that also responds to thermal and mechanical stimuli. Substances that activate TRPA1, e.g., allyl isothiocyanates (pungent components of mustard, horseradish, and wasabi), cinnamaldehyde from cinnamon, organosulfur compounds from garlic and onion, tear gas, acrolein and crotonaldehyde from cigarette smoke, etc., cause burning, mechanical and thermal hypersensitivity, cough, eye irritation, sneezing, mucus secretion, and neurogenic inflammation. An increased activity of TRPA1 leads to the emergence of chronic pruritus and allergic dermatitis and is associated with episodic pain syndrome, a hereditary disease characterized by episodes of debilitating pain triggered by stress. TRPA1 is now considered as one of the targets for developing new anti-inflammatory and analgesic drugs. This review summarizes information on the structure, function, and physiological role of this channel, as well as describes known TRPA1 ligands and their significance as therapeutic agents in the treatment of inflammation-associated pain.

A TR(I)P to Pruritus Research: Role of TRPV3 in Inflammation and Itch

Pruritus in eczema and other inflammatory skin diseases is an unsolved problem. Recent findings strongly indicate that histamine is not the only mediator of pruritus. Subtypes of the transient receptor potential (TRP) ion channel superfamily are expressed by sensory nerves, keratinocytes, and certain leukocytes. Temperature, pH changes, and certain toxins activate TRPs. New evidence indicates that the vanilloid type 3 (TRPV3) channel is crucially involved in pruritic dermatitis, making it a good candidate for future therapy in skin inflammation and pruritus.

ASIC3, a sensor of acidic and primary inflammatory pain

Acid-sensing ion channels (ASICs) are cationic channels activated by extracellular acidosis that are expressed in both central and peripheral nervous systems. Although peripheral ASICs seem to be natural sensors of acidic pain (e.g., in inflammation, ischaemia, lesions or tumours), a direct demonstration is still lacking. We show that approximately 60% of rat cutaneous sensory neurons express ASIC3-like currents. Native as well as recombinant ASIC3 respond synergistically to three different inflammatory signals that are slight acidifications (approximately pH 7.0), hypertonicity and arachidonic acid (AA). Moderate pH, alone or in combination with hypertonicity and AA, increases nociceptors excitability and produces pain suppressed by the toxin APETx2, a specific blocker of ASIC3. Both APETx2 and the in vivo knockdown of ASIC3 with a specific siRNA also have potent analgesic effects against primary inflammation-induced hyperalgesia in rat. Peripheral ASIC3 channels are thus essential sensors of acidic pain and integrators of molecular signals produced during inflammation where they contribute to primary hyperalgesia.

Myelinated Skin Sensory Neurons Project Extensively throughout Adult Mouse Substantia Gelatinosa

The substantia gelatinosa (SG) of the dorsal horn of the spinal cord is a recipient zone for unmyelinated sensory neurons in adults. Recent studies of the central anatomy of physiologically identified skin sensory neurons in neonatal mice have shown that this region also receives substantial inputs from a variety of myelinated afferents. The present experiments were performed to determine whether these neonatal inputs represent a transient phenotype that retracts from the SG. Studies were conducted in an in vivo spinal cord preparation from adult mice; thoracic levels were targeted to facilitate comparisons with previous in vitro findings. We show that the SG continues to receive substantial projections from myelinated skin sensory neurons throughout life. A large population of myelinated nociceptors conducting in the upper A delta and low A beta range maintained extensive projections throughout all areas of the SG well into adulthood; the latter gave rise to dorsally recurving "flame"-shaped arbors extending into the marginal layer that were identical to afferents described in neonates and after nerve injury in adult rats. Furthermore, exquisitely sensitive down hair follicle afferents projected throughout the inner half of the SG (i.e., lamina IIi) and sent dense clusters of terminals well into the outer SG (IIo), where they intermingled with those of unmyelinated nociceptors. Arguments are presented that the SG likely plays a predominant role in tactile processing under normal conditions, but that this role switches rapidly to nociceptive-only during environmental exigencies imposed by temperature extremes.

Physiological Properties of Mouse Skin Sensory Neurons Recorded Intracellularly In Vivo: Temperature Effects on Somal Membrane Properties

Recent combined analyses of the structural, functional, and molecular attributes of individual skin sensory neurons using semi-intact in vitro preparations from mice have provided a wealth of novel insights into nociceptor biology. How these findings translate to more natural conditions nevertheless remains unresolved. Toward this end, intracellular recordings were obtained from 362 physiologically identified dorsal root ganglion (DRG) neurons in a new in vivo mouse preparation developed for combined structure/function analyses of individual skin sensory neurons. Recordings were conducted at thoracic levels in adult decorticate mice for comparison with in vitro findings from the same trunk region. In all, 270 neurons were recorded at DRG temperatures tightly regulated at normal core values to establish a baseline and 137 skin sensory neurons were included in detailed analyses of somal properties for comparisons with similar data obtained under reduced temperatures mirroring in vitro conditions. Recovery of Neurobiotin-labeled central projections was crucial for verifying perceived afferent identity of certain neurons. Further, profound temperature dependency was seen across diverse physiological properties. Indeed, the broad, inflected somal spikes normally viewed as diagnostic of myelinated nociceptors were found to be a product of reduced temperatures and were not present at normal core values. Moreover, greater complexity was observed peripherally in the mechanical and thermal sensitivity profile of nociceptive and nonnociceptive populations than that seen under in vitro conditions. This novel in vivo preparation therefore holds considerable promise for future analyses of nociceptor function and plasticity in normal and transgenic models of pain mechanisms.

The Spinal Interneurons and Properties of Glutamatergic Synapses in a Primitive Vertebrate Cutaneous Flexion Reflex

Unlike the monosynaptic "stretch" reflex, the exact neuronal pathway for a simple cutaneous reflex has not yet been defined in any vertebrate. In young frog tadpoles, we made whole-cell recordings from pairs of spinal neurons. We found direct, excitatory, glutamatergic synapses from touch-sensitive skin-sensory neurons to sensory pathway interneurons, and then from these sensory interneurons to motoneurons and premotor interneurons on the other side of the body. We conclude that the minimal pathway for this primitive reflex, in which stroking the skin on one side leads to flexion on the other side, is disynaptic. This detailed circuit information has allowed us to ask whether the properties of glutamatergic synapses during the first day of CNS development are tuned to their function in the tadpole's responses. Stroking the skin excites a few sensory neurons. These activate primarily AMPA receptors producing short, strong excitation that activates many sensory pathway interneurons but only allows temporal summation of closely synchronous inputs. In contrast, the excitation produced in contralateral neurons by the sensory pathway interneurons is weak and primarily mediated by NMDA receptors. As a result, considerable summation is required for this excitation to lead to postsynaptic neuron firing and a contralateral flexion. We conclude that from their early functioning, synapses from sensory neurons are strong and those from sensory pathway interneurons are weak. The distribution of glutamate receptors at synapses in this developing circuit is tuned so that synapses have properties suited to their roles in the whole animal's reflex responses.

Widespread Projections from Myelinated Nociceptors throughout the Substantia Gelatinosa Provide Novel Insights into Neonatal Hypersensitivity

Skin sensory neurons have long been thought to undergo major changes in anatomy and physiology over the first few weeks of postnatal life. Low-threshold mechanoreceptors (LTMRs) are believed to project extensively throughout superficial dorsal horn laminas initially and provide the afferent limb for hyperactive nocifensive reflexes. However, our recent studies revealed that neonatal LTMRs do not project into "pain-specific" regions; instead, they exhibit adult-like anatomy shortly after birth. We sought to determine whether the same might be true for myelinated high-threshold mechanoreceptors (HTMRs). We used an intact, ex vivo somatosensory system preparation from neonatal mice to allow intrasomal recording and neurobiotin labeling of individual sensory neurons characterized via natural skin stimuli. Neonatal HTMRs displayed a number of key hallmarks of their adult counterparts; relative to LTMRs, they exhibited broader, inflected somal spikes and higher mechanical thresholds and/or responded in an increasingly vigorous manner to incrementally graded forces in a manner capable of encoding stimulus intensity. Two types were discerned on the basis of central anatomy: one subset projected to superficial laminas (I/II); the other gave rise to diffuse, dorsally recurving collateral arbors extending throughout the entire dorsal horn (I-V). The latter represent a novel cutaneous afferent morphology that persists in older animals. These studies reveal that inputs from myelinated afferents to superficial pain-specific laminas in neonates arise from HTMRs and not LTMRs as commonly thought. This frequently overlooked population is in a position, therefore, to contribute substantially to paradoxical nocifensive behaviors in neonates and various pain states in adults.

Sensory Activation and Role of Inhibitory Reticulospinal Neurons that Stop Swimming in Hatchling Frog Tadpoles

Activity in neuronal networks underlying locomotion and other rhythmic actions can start and stop in response to specific sensory stimuli. In vertebrate locomotion, some reticulospinal neurons such as Mauthner neurons can initiate activity, but the neurons controlling stopping are not defined. We have studied swimming in tadpoles of the frog,Xenopus, which is started by touching the skin and stops when the head contacts a solid surface. Using an immobilized tadpole preparation, the same stimuli control fictive swimming. When head contact is imitated by pressure to the head skin sensory neurons in the trigeminal ganglion are active, spinal neurons receive GABAergic inhibition, and swimming stops. Here we record intracellularly from neurons in the hindbrain that are excited by pressure or electrical stimulation to the head skin. By intracellular filling with neurobiotin, we identify these anatomically as mid-hindbrain reticulospinal neurons (MHRs). These have contralateral descending projections to the spinal cord and GABA-like immunoreactivity. They are rhythmically inhibited during fictive swimming. Individual MHRs reliably stopped ongoing swimming when brief firing was induced by intracellular current injection. The ability of individual MHRs to stop swimming was blocked by the GABAAantagonist bicuculline. Our evidence indicates that MHRs receive direct excitation from trigeminal sensory neurons and in turn release GABA to directly inhibit spinal neurons and turn off the swimming central pattern generator.

Sensory activation and role of inhibitory reticulospinal neurons that stop swimming in hatchling frog tadpoles.

Activity in neuronal networks underlying locomotion and other rhythmic actions can start and stop in response to specific sensory stimuli. In vertebrate locomotion, some reticulospinal neurons such as Mauthner neurons can initiate activity, but the neurons controlling stopping are not defined. We have studied swimming in tadpoles of the frog, Xenopus, which is started by touching the skin and stops when the head contacts a solid surface. Using an immobilized tadpole preparation, the same stimuli control fictive swimming. When head contact is imitated by pressure to the head skin sensory neurons in the trigeminal ganglion are active, spinal neurons receive GABAergic inhibition, and swimming stops. Here we record intracellularly from neurons in the hindbrain that are excited by pressure or electrical stimulation to the head skin. By intracellular filling with neurobiotin, we identify these anatomically as mid-hindbrain reticulospinal neurons (MHRs). These have contralateral descending projections to the spinal cord and GABA-like immunoreactivity. They are rhythmically inhibited during fictive swimming. Individual MHRs reliably stopped ongoing swimming when brief firing was induced by intracellular current injection. The ability of individual MHRs to stop swimming was blocked by the GABA(A) antagonist bicuculline. Our evidence indicates that MHRs receive direct excitation from trigeminal sensory neurons and in turn release GABA to directly inhibit spinal neurons and turn off the swimming central pattern generator.

Central anatomy of individual rapidly adapting low‐threshold mechanoreceptors innervating the “hairy” skin of newborn mice: Early maturation of hair follicle afferents

Adult skin sensory neurons exhibit characteristic projection patterns in the dorsal horn of the spinal gray matter that are tightly correlated with modality. However, little is known about how these patterns come about during the ontogeny of the distinct subclasses of skin sensory neurons. To this end, we have developed an intact ex vivo somatosensory system preparation in neonatal mice, allowing single, physiologically identified cutaneous afferents to be iontophoretically injected with Neurobiotin for subsequent histological analyses. The present report, centered on rapidly adapting mechanoreceptors, represents the first study of the central projections of identified skin sensory neurons in neonatal animals. Cutaneous afferents exhibiting rapidly adapting responses to sustained natural stimuli were encountered as early as recordings were made. Well-stained representatives of coarse (tylotrich and guard) and fine-diameter (down) hair follicle afferents, along with a putative Pacinian corpuscle afferent, were recovered from 2–7-day-old neonates. All were characterized by narrow, uninflected somal action potentials and generally low mechanical thresholds, and many could be activated via deflection of recently erupted hairs. The central collaterals of hair follicle afferents formed recurrent, flame-shaped arbors that were essentially miniaturized replicas of their adult counterparts, with identical laminar terminations. The terminal arbors of down hair afferents, previously undescribed in rodents, were distinct and consistently occupied a more superficial position than tylotrich and guard hair afferents. Nevertheless, the former extended no higher than the middle of the incipient substantia gelatinosa, leaving a clear gap more dorsally. In all major respects, therefore, hair follicle afferents display the same laminar specificity in neonates as they do in adults. The widely held misperception that their collaterals extend exuberant projections into pain-specific regions of the dorsal horn during early postnatal life is shown to have multiple, deep-rooted underpinnings. J. Comp. Neurol. 436:304–323, 2001. © 2001 Wiley-Liss, Inc.

Central anatomy of individual rapidly adapting low‐threshold mechanoreceptors innervating the “hairy” skin of newborn mice: Early maturation of hair follicle afferents

Adult skin sensory neurons exhibit characteristic projection patterns in the dorsal horn of the spinal gray matter that are tightly correlated with modality. However, little is known about how these patterns come about during the ontogeny of the distinct subclasses of skin sensory neurons. To this end, we have developed an intact ex vivo somatosensory system preparation in neonatal mice, allowing single, physiologically identified cutaneous afferents to be iontophoretically injected with Neurobiotin for subsequent histological analyses. The present report, centered on rapidly adapting mechanoreceptors, represents the first study of the central projections of identified skin sensory neurons in neonatal animals. Cutaneous afferents exhibiting rapidly adapting responses to sustained natural stimuli were encountered as early as recordings were made. Well-stained representatives of coarse (tylotrich and guard) and fine-diameter (down) hair follicle afferents, along with a putative Pacinian corpuscle afferent, were recovered from 2-7-day-old neonates. All were characterized by narrow, uninflected somal action potentials and generally low mechanical thresholds, and many could be activated via deflection of recently erupted hairs. The central collaterals of hair follicle afferents formed recurrent, flame-shaped arbors that were essentially miniaturized replicas of their adult counterparts, with identical laminar terminations. The terminal arbors of down hair afferents, previously undescribed in rodents, were distinct and consistently occupied a more superficial position than tylotrich and guard hair afferents. Nevertheless, the former extended no higher than the middle of the incipient substantia gelatinosa, leaving a clear gap more dorsally. In all major respects, therefore, hair follicle afferents display the same laminar specificity in neonates as they do in adults. The widely held misperception that their collaterals extend exuberant projections into pain-specific regions of the dorsal horn during early postnatal life is shown to have multiple, deep-rooted underpinnings.

The pattern of sensory discharge can determine the motor response in young Xenopus tadpoles.

Young Xenopus tadpoles were used to test whether the pattern of discharge in specific sensory neurons can determine the motor response of a whole animal. Young Xenopus tadpoles show two main rhythmic behaviours: swimming and struggling. Touch-sensitive skin sensory neurons in the spinal cord of immobilised tadpoles were penetrated singly or in pairs using microelectrodes to allow precise control of their firing patterns. A single impulse in one Rohon-Beard neuron (= light touch) could sometimes trigger "fictive" swimming. Two to six impulses at 30-50 Hz (= a light stroke) reliably triggered fictive swimming. Neither stimulus evoked fictive struggling. Twenty-five or more impulses at 30-50 Hz (= pressure) could evoke a pattern of rhythmic bursts, distinct from swimming and suitable to drive slower, stronger movements. This pattern showed some or all the characteristics of "fictive" struggling. These results demonstrate clearly that sensory neurons can determine the pattern of motor output simply by their pattern of discharge. This provides a simple form of behavioural selection according to stimulus.

Skin sensory innervation patterns in embryonic chick hindlimbs deprived of motoneurons

During normal development and following a variety of experimental manipulations (e.g., neural tube rotations, limb shifts), sensory neurons in the chick grow to their correct targets. L. Landmesser and M. G. Honig (1986, Dev. Biol. 118, 511–531) have suggested that sensory innervation may be precise, not because sensory neurons respond to limb-derived guidance cues, but because sensory neurons interact with motoneurons, which do respond to such cues. To test this hypothesis for skin sensory neurons, the ventral neural tube, including the motoneuron precursors, was removed from chick embryos prior to sensory axon outgrowth and the resulting patterns of dermatomes and axonal projections were mapped physiologically and anatomically. As reported previously, dorsal root ganglia (DRGs) and cutaneous nerves formed in their usual locations following the early removal of motoneurons, while most muscle nerves and the plexus region were reduced substantially (A. C. Taylor, 1944, J. Exp. Zool. 96, 159–185 ; L. Landmesser and M. G. Honig, 1986, Dev. Biol. 118, 511–531 ; G. J. Swanson and J. Lewis, 1986, J. Embryol. Exp. Morphol. 95, 37–52) . The patterns of axonal projections and dermatomes were surprisingly, although not entirely, normal. In particular, cutaneous nerves in motoneuron-depleted embryos were derived from the same DRGs in approximately the same proportions as normal. Thus, while motoneurons may play a facilitative role in the development of the segmental pattern of skin sensory innervation, they do not appear to be essential.

The development of skin sensory innervation patterns

During embryonic development, skin sensory innervation in the chick hindlimb is established in a precise and orderly fashion. From their earliest outgrowth, axons of each dorsal root ganglion (DRG) project through the limb via the correct pathways to establish their dermatome in the appropriate location. By making experimental manipulations in young embryos and mapping the resulting innervation patterns, we have begun to reveal some of the mechanisms involved. For example, when several DRG are eliminated, the axonal projections of intact DRG are shifted and the dermatomes are enlarged, suggesting that competition among axons is important in determining both the contribution of DRG to axonal pathways and the borders between dermatomes. Further, when the rostrocaudal order of DRG is reversed, skin sensory innervation patterns tend to mimic the motor projection patterns, implying that motor axons may guide the growth of sensory axons. However, neither of these mechanisms can accountfully for the precision with which skin sensory innervation patterns are established. Thus, it is likely that skin sensory neurons use multiple, redundant developmental cues in selecting their appropriate pathways and targets.

Skin sensory innervation patterns in embryonic chick hindlimb following dorsal root ganglion reversals.

During embryonic development skin sensory neurons in lumbosacral dorsal root ganglia (DRGs) establish their dermatomes and axonal projections in a precise, orderly fashion in the chick. To investigate mechanisms responsible for this specific outgrowth, the rostrocaudal order of DRGs T7-LS3 was reversed by rotating the corresponding segments of neural crest, either alone or together with the underlying neural tube in St.15-16 embryos. The resulting skin sensory innervation patterns, mapped physiologically or anatomically at St.29-40, differed between the two experimental groups. Following neural tube rotations DRGs tended to establish innervation patterns that were consonant with their original position in the embryo. Axons from these rotated DRGs generally projected into the appropriate pathways and innervated the appropriate region of skin. Neural crest rotations left the ventral neural tube (including the motor neuron precursors) largely intact. In this case rotated DRGs tended to establish innervation patterns in accordance with their new position in the embryo, almost as if no rotation had been made. These results cannot be explained solely by the inherent specificity of sensory neurons. Instead, the results are largely consistent with the suggestion (Honig et al., 1986; Landmesser and Honig, 1986) that motor axons can direct the outgrowth of sensory axons and thereby influence the establishment of sensory innervation patterns. Other mechanisms that may also affect the development of sensory innervation patterns are discussed.