Merkel cells transduce and encode tactile stimuli to drive Aß-afferent impulses.

Sensory systems for detecting tactile stimuli have evolved from touch-sensing nerves in invertebrates to complicated tactile end organs in mammals. Merkel discs are tactile end organs consisting of Merkel cells and Aß-afferent nerve endings and are localized in fingertips, whisker hair follicles, and other touch-sensitive spots. Merkel discs transduce touch into slowly adapting impulses to enable tactile discrimination, but their transduction and encoding mechanisms remain unknown. Using rat whisker hair follicles, we show that Merkel cells rather than Aß-afferent nerve endings are primary sites of tactile transduction and identify the Piezo2 ion channel as the Merkel cell mechanical transducer. Piezo2 transduces tactile stimuli into Ca(2+)-action potentials in Merkel cells, which drive Aß-afferent nerve endings to fire slowly adapting impulses. We further demonstrate that Piezo2 and Ca(2+)-action potentials in Merkel cells are required for behavioral tactile responses. Our findings provide insights into how tactile end-organs function and have clinical implications for tactile dysfunctions.

Phosphorylation of phase-separated p62 bodies by ULK1 activates a redox-independent stress response.

NRF2 is a transcription factor responsible for antioxidant stress responses that is usually regulated in a redox-dependent manner. p62 bodies formed by liquid-liquid phase separation contain Ser349-phosphorylated p62, which participates in the redox-independent activation of NRF2. However, the regulatory mechanism and physiological significance of p62 phosphorylation remain unclear. Here, we identify ULK1 as a kinase responsible for the phosphorylation of p62. ULK1 colocalizes with p62 bodies, directly interacting with p62. ULK1-dependent phosphorylation of p62 allows KEAP1 to be retained within p62 bodies, thus activating NRF2. p62S351E/+ mice are phosphomimetic knock-in mice in which Ser351, corresponding to human Ser349, is replaced by Glu. These mice, but not their phosphodefective p62S351A/S351A counterparts, exhibit NRF2 hyperactivation and growth retardation. This retardation is caused by malnutrition and dehydration due to obstruction of the esophagus and forestomach secondary to hyperkeratosis, a phenotype also observed in systemic Keap1-knockout mice. Our results expand our understanding of the physiological importance of the redox-independent NRF2 activation pathway and provide new insights into the role of phase separation in this process.

Plus moist HS-W®: a new nasal packing material for the middle meatus in endoscopic sinus surgery.

PURPOSE: Removal of the current calcium alginate packing materials to the middle meatus in endoscopic sinus surgery (ESS) is usually accompanied by discomfort or pain owing to the hard and brittle nature of these materials. Plus moist HS-W® is a new calcium alginate packing material released in 2022 developed to overcome this issue by changing the uronic acid component. We aimed to compare the discomfort/pain during the removal of Plus moist HS-W® with Kaltostat®, as well as their suitability as packing materials in ESS. METHODS: Kaltostat® and Plus moist HS-W® were used as packing materials in 22 and 21 patients who underwent ESS in 2021 and 2022, respectively. Patients were asked to rate the pain during the packing removal 10 days after ESS using the Numerical Rating Scale (NRS). The ratio of residual packing materials, number of suctions (insertions/extractions of the suction cannula), and time required to remove packing materials were measured. Postoperative complications such as hemorrhage, local infection, lateralization of the middle turbinate, and synechia of the middle meatus were also evaluated. RESULTS: The Plus moist HS-W® group exhibited significantly lower NRS pain scores, a lower ratio of residual packing materials, a reduced number of suctions, and a shorter time required to remove the packing. No obvious postoperative complications occurred in both groups except for one suspicious case of a slight infection in the Kaltostat® group. CONCLUSION: Compared with Kaltostat®, Plus moist HS-W®, characterized by better gelatinization than Kaltostat®, benefits patients by minimizing discomfort/pain during removal. LEVEL OF EVIDENCE: Level 3.

[Introduction and Activities of Nurse Practitioner in Japan in a University Hospital].

Nurse practitioner (NP) is widely known to be an essential position of medical team in the United States, but has not yet been established as an official qualification in Japan. NP in Japan (NP-J) is accepted instead of NP, but they are not the same. We summarized the actual activities of NP-J at our hospital and had an insight into the roles of NP-J in a university hospital and the problems of introduction of NP in the future. The benefits of working as a NP-J at a university hospital are the safe acquisition of procedures at an educational institution and the involvement of various departments. In the future, the education of NP-J in a university hospital may lead to the training of NP-J working in public and private hospitals. The problem of introduction of NP in the future is the legislation. The importance of task shifting and education of NP-J in a university hospital may lead to the spread of NP in the future in Japan.

Expression of Piezo mRNA is unaffected in a rat model of knee osteoarthritis.

Osteoarthritis of the knee impairs activities of daily living of those affected. Its irreversible degenerative changes to the knee joint induce functional disturbance and unpleasant arthralgia. The pain has inflammatory components and often is manifested with mechanical allodynia and hyperalgesia. Sustained weight bearing and joint movements increase pain sensitivity in knee osteoarthritis. Understanding the mechanisms underlying the mechanical allodynia and hyperalgesia might provide a therapeutical target for pain relief in patients with such symptoms. Piezo channel is a mechanically activated ion channel that may be involved in mechanical transduction in the articular cartilage. Although it has been shown that inflammation potentiates Piezo channel current induced by mechanical stimulation, whether Piezo expression levels are influenced by knee osteoarthritis has remained unknown. We measured Piezo mRNA in knee joints and dorsal root ganglia after establishing a model of knee osteoarthritis in rats using monosodium iodoacetate and found Piezo mRNA level is not upregulated. This finding raises a question as whether and how Piezo channels may be involved in mechanically induced pain in osteoarthritis.

Morphological Fabrication of Rubber Cutaneous Receptors Embedded in a Stretchable Skin-Mimicking Human Tissue by the Utilization of Hybrid Fluid.

Sensors are essential in the haptic technology of soft robotics, which includes the technology of humanoids. Haptic sensors can be simulated by the mimetic organ of perceptual cells in the human body. However, there has been little research on the morphological fabrication of cutaneous receptors embedded in a human skin tissue utilizing artificial materials. In the present study, we fabricated artificial, cell-like cutaneous receptors embedded in skin tissue mimicking human skin structure by utilizing rubber. We addressed the fabrication of five cutaneous receptors (free nerve endings, Krause and bulbs, Meissner corpuscles, Pacinian corpuscles and Ruffini endings). In addition, we investigated the effectiveness of the fabricated tissue for mechanical and thermal sensing. At first, in the production of integrated artificial skin tissue, we proposed a novel magnetic, responsive, intelligent, hybrid fluid (HF), which is suitable for developing the hybrid rubber skin. Secondly, we presented the fabrication by utilizing not only the HF rubber but our previously proposed rubber vulcanization and adhesion techniques with electrolytic polymerization. Thirdly, we conducted a mechanical and thermal sensing touch experiment with the finger. As a result, it demonstrated that intelligence as a mechanoreceptor or thermoreceptor depends on its fabric: the HF rubber sensor mimicked Krause and bulbs has the thermal and pressing sensibility, and the one mimicked Ruffini endings the shearing sensibility.

Impact of posterior femoral condylar cartilage and posterior intercondylar distance on rotation of femoral component in total knee arthroplasty.

BACKGROUND: Greater accuracy is needed when determining the final femoral component (FC) rotation during total knee arthroplasty (TKA), because this parameter affects soft tissue balance during flexion and patellar tracking. Anatomical markers, such as the epicondylar axis, are typically used to determine the final FC rotation, although intraoperative confirmation may be challenging. Therefore, rotational position is frequently determined with the posterior condylar axis (PCA) as a landmark. However, the thickness of the posterior condylar cartilage has not been considered and may not be represented on preoperative images. We used plain X-rays to measure the thickness of the medial and lateral posterior condylar cartilage fragments postoperatively, and investigated the effects of differences in cartilage thickness on final FC rotation. METHODS: Fifty knees (19 men, 31 women) underwent primary TKA to treat medial knee osteoarthritis at our hospital between August 2015 and May 2017. All knees were treated using an Attune PS (DePuy Synthes, Inc., Warsaw, IN). We first measured the distance between the posterior femoral condyles, resected the posterior condyle, and measured the thickness of the resected cartilage fragments. We then took X-ray images from a direction tangential to the osteotomy surface, secured the cartilage fragments with digital calipers, and measured the thickness of the cartilage. We investigated the effects of differences in cartilage thickness on final FC rotation of the residual medial and lateral cartilage with a trigonometric function. RESULTS: Medial condylar cartilage thickness averaged 0.6 ± 0.5 mm and the lateral condylar thickness averaged 1.8 ± 0.6 mm; posterior intercondylar distance averaged 46.1 ± 3.3 mm and average impact on rotation of the cartilage remnant was 1.5 ± 0.9° (- 0.1-3.9°). There may be measurement error of up to 4° in the maximum values compared with the preoperative plan in cases with short intercondylar distance. CONCLUSIONS: In cases where the FC external rotation angle is determined using the posterior condyles as landmarks, this angle can be affected by the intercondylar distance, especially in Japanese women who have small physical stature. This angle can potentially be much larger, so caution is advised. Our results suggest that several anatomical landmarks should be referenced to achieve accurate FC rotation.

Pre-operative hemoglobin level and use of sedative-hypnotics are independent risk factors for post-operative delirium following total knee arthroplasty.

BACKGROUND: Delirium is a well-known complication following surgery, especially with the increasing age of patients undergoing surgery. The increasing demands resulting from a prolonged healthy life expectancy has resulted in more arthroplasties despite their age and existing comorbidities. The purpose of this study is to explore the various risk factors that may contribute to delirium in unilateral and bilateral total knee arthroplasties in the elderly population. METHODS: 170 patients who underwent unilateral or bilateral total knee arthroplasties were analyzed retrospectively for delirium. Age, sex, comorbidities, use of sedative-hypnotics, peri-operative blood loss, pre- and post-operative laboratory blood test results were investigated and analyzed. RESULTS: The incidence of post-operative delirium was 6.5% (11 out of 170 patients) with a mean age of 79.5 (± 6.9) years, compared to 73.0 (± 9.0) years in the non-delirium group. Higher age, use of sedative-hypnotics, low pre-operative Hb and Ht, low post-operative Hb, Ht and BUN were observed in the delirium group. Multivariate logistic regression analysis identified that the use of sedative-hypnotics and pre-operative Hb level were independent risk factors for post-operative delirium after TKA. The odds ratios for the use of sedative-hypnotics and pre-operative Hb level were 4.6 and 0.53, respectively. Receiver operating characteristic curve analysis showed that pre-operative Hb of less than 11.1 g/dL was a predictor for the development of delirium, with a sensitivity of 54.6% and a specificity of 91.6%. CONCLUSION: Patients with a pre-operative Hb level of < 11.1 g/dL or those using sedative-hypnotics are associated with post-operative delirium. Peri-operative management and preventative measures are therefore needed to reduce the risks of post-operative delirium in such patients.

Enhancement of Diversity in Production and Applications Utilizing Electrolytically Polymerized Rubber Sensors with MCF: The Second Report on Various Engineering Applications.

We investigated the proposed hybrid skin (H-Skin) for the requirement of haptic sensibility in rubber using our proposed consummate fabrication process together with a multi-layered magnetic compound fluid (MCF) rubber and stocking-like porous rubber permeated by liquids, which was demonstrated in our previous report. The objective was to assess its applicability to sensing normal force and temperature, as well as fields dominated by shear force. For normal force, we investigated the piezo-electricity and electric current induced voltage, as well as the piezo-resistivity of the MCF rubber sensor under pressure. Additionally, we clarified the viability of measuring the softness and texture of materials using the MCF rubber sensor. For the shear motion, we clarified the characteristics of the friction coefficient using the MCF rubber sensor. The MCF rubber sensor can capture the reactions of paper, cloth, convex- and concave-shaped objects such as plant leaves and metal, and the skin of the human finger. Therefore, it is useful to investigate its texture and biological surfaces. Our obtained outstanding results indicated the feasibility of sensing the surface texture for any material in fields such as paper, fashion, apparel manufacturing, and cosmetic industries, which was impossible until now.

Enhancement of Diversity in Production and Application Utilizing Electrolytically Polymerized Rubber Sensors with MCF: 1st Report on Consummate Fabrication Combining Varied Kinds of Constituents with Porous Permeant Stocking-Like Rubber.

To satisfy the requirement of haptic sensibility in rubber such as in the proposed hybrid skin (H-Skin), the authors have demonstrated a new method for solidifying rubber using electrolytic polymerization together with configured magnetic clusters of magnetic compound fluid (MCF) incorporated into the rubber by the application of a magnetic field. However, the rubber and magnetic fluid (MF) involved in the MCF rubber were water-soluble. In addition, the authors have demonstrated the practicability of using electrolytic polymerization with an emulsifier, polyvinyl alcohol (PVA), in which natural rubber (NR) or chloroprene rubber (CR) and silicone rubber (Q) can be mixed as water-soluble and water-insoluble rubbers, respectively. In this study, to enhance production, the feasibility of solidifying rubber by electrolytic polymerization is verified using varied water-insoluble rubber, varied water-insoluble MF, and varied surfactants to aid emulsion polymerization, except in the case of other kinds of rubber and MF which have been demonstrated until recent by the authors. Based on these diverse constituents, the authors propose a consummate fabrication process for multi-layered MCF rubber, which involves porous stocking-like rubber that can be permeated by any liquid. The investigation of this application is presented in the sequential second report.

Merkel disc is a serotonergic synapse in the epidermis for transmitting tactile signals in mammals.

The evolution of sensory systems has let mammals develop complicated tactile end organs to enable sophisticated sensory tasks, including social interaction, environmental exploration, and tactile discrimination. The Merkel disc, a main type of tactile end organ consisting of Merkel cells (MCs) and Aß-afferent endings, are highly abundant in fingertips, touch domes, and whisker hair follicles of mammals. The Merkel disc has high tactile acuity for an object's physical features, such as texture, shape, and edges. Mechanisms underlying the tactile function of Merkel discs are obscured as to how MCs transmit tactile signals to Aß-afferent endings leading to tactile sensations. Using mouse whisker hair follicles, we show herein that tactile stimuli are transduced by MCs into excitatory signals that trigger vesicular serotonin release from MCs. We identify that both ionotropic and metabotropic 5-hydroxytryptamine (5-HT) receptors are expressed on whisker Aß-afferent endings and that their activation by serotonin released from MCs initiates Aß-afferent impulses. Moreover, we demonstrate that these ionotropic and metabotropic 5-HT receptors have a synergistic effect that is critical to both electrophysiological and behavioral tactile responses. These findings elucidate that the Merkel disc is a unique serotonergic synapse located in the epidermis and plays a key role in tactile transmission. The epidermal serotonergic synapse may have important clinical implications in sensory dysfunctions, such as the loss of tactile sensitivity and tactile allodynia seen in patients who have diabetes, inflammatory diseases, and undergo chemotherapy. It may also have implications in the exaggerated tactile sensations induced by recreational drugs that act on serotoninergic synapses.

Development of a Magnetic Compound Fluid Rubber Stability Sensor and a Novel Production Technique via Combination of Natural, Chloroprene and Silicone Rubbers.

Expanding on our previous report, we investigate the stability of a magnetic compound fluid (MCF) rubber sensor that was developed for a variety of engineering applications. To stabilize this sensor, we proposed a novel combination technique that facilitates the addition of dimethylpolysiloxane (PDMS) to natural rubber (NR)-latex or chloroprene rubber (CR)-latex using polyvinyl alcohol (PVA) by experimentally and theoretically investigating issues related to instability. This technique is one of several other novel combinations of diene and non-diene rubbers. Silicone oil or rubber with PDMS can be combined with NR-latex and CR-latex because of PVA's emulsion polymerization behavior. In addition, owing to electrolytic polymerization based on the combination of PDMS and PVA, MCF rubber is highly porous and can be infiltrated in any liquid. Hence, the fabrication of novel intelligent rubbers using any intelligent fluid is feasible. By assembling infiltrated MCF rubber sheets and by conducting electrolytic polymerization of MCF rubber liquid with a hydrate using the adhesive technique as presented in a previous paper, it is possible to stabilize the MCF rubber sensor. This sensor is resistant to cold or hot water as well as γ-irradiation as shown in the previous report.

Novel Adhesion Technique Using Metallic or Non-Metallic Hydrous Oxide of Metal Complexes Involving Magnetic Compound Fluid Rubber under Electrolytic Polymerization and Magnetic Field for Producing Sensors.

As per sequential studies on new types of soft rubber for the artificial skin of robots, smart sensors, etc., we have proposed and investigated hybrid skin (H-Skin) and haptic sensors by using magnetic compound fluid (MCF), compounding natural rubber latex (NR-latex), and applying electric and magnetic fields. Through electrolytic polymerization, the MCF rubber is solidified. The MCF rubber has hybrid sensing functions and photovoltaic effects, and electric charge as battery. In case of the production of soft rubber sensors, however, the problem of adhesion between metal electrodes and rubber is very important. In the present study, we propose a novel adhesive technique for bonding the metal electrodes and MCF rubber by using metallic or non-metallic hydrous oxide, which is a metal complex, via electrolytic polymerization. The anionic radical hydrate reacts with the isoprene molecules of NR-latex or chloroprene rubber latex (CR-latex) such that they are cross-linked and the MCF rubber with the hydrate is solidified, which can be represented via a chemical reaction equation. By means of this adhesive technique, we presented five cases of sensors fabricated using metal electrodes and rubbers. This technique is applicable for novel cohesion between rubber and metal.

Regulation of Piezo2 Mechanotransduction by Static Plasma Membrane Tension in Primary Afferent Neurons.

The Piezo2 channel is a newly identified mammalian mechanical transducer that confers rapidly adapting mechanically activated (RA-MA) currents in primary afferent neurons. The Piezo2 channels sense rapid membrane displacement, but it is not clear whether they are sensitive to osmotic swelling, which slowly increases static plasma membrane tension (SPMT). Here, we show that SPMT exerts a profound impact on the mechanical sensitivity of RA-MA channels in primary afferent neurons. RA-MA currents are greatly enhanced, and the mechanical threshold was reduced in both primary afferent neurons of rat dorsal root ganglia (DRG) and HEK293 cells heterologously expressing Piezo2 when these cells undergo osmotic swelling to increase SPMT. Osmotic swelling switches the kinetics of RA-MA currents to the slowly adapting type in both cultured DRG neurons and HEK293 cells heterologously expressing Piezo2. The potentiation of RA-MA currents is abolished when cultured DRG neurons are treated with cytochalasin D, an actin filament disruptor that prevents SPMT of cultured DRG neurons from an increase by osmotic swelling. Osmotic swelling significantly increases DRG neuron mechano-excitability such that a subthreshold mechanical stimulus can result in action potential firing. Behaviorally, the mechanical hind paw withdrawal threshold in rats is reduced following the injection of a hypotonic solution, but this osmotic effect is abolished when cytochalasin D or Gd(3+) is co-administered with the hypo-osmotic solution. Taken together, our findings suggest that Piezo2-mediated mechanotransduction is regulated by SPMT in primary afferent neurons. Because SPMT can be changed by multiple biological factors, our findings may have broad implications in mechanical sensitivity under physiological and pathological conditions.

Serotonergic transmission at Merkel discs: modulation by exogenously applied chemical messengers and involvement of Ih currents.

The Merkel disc is a main type of tactile end organ consisting of Merkel cells and Aß-afferent endings that responds to tactile stimulation with slowly adapting type 1 (SA1) afferent impulses. Our recent study has shown that Merkel discs in whisker hair follicles are serotonergic synapses using endogenous serotonin to transmit tactile signals from Merkel cells to Aß-afferent endings. In this study, we hypothesize that tactile sensitivity of Merkel discs can be modulated by chemical messengers. We tested this hypothesis by determining whether and how SA1 responses of mouse whisker hair follicles may be affected by exogenously applied chemical messengers. We found that SA1 responses were potentiated by serotonin at low concentration (10 µM) but almost completely occluded by serotonin at high concentration (2 mM). In contrast, SA1 responses were not significantly affected by ATP and its metabolically stable analog α,ß-methylene-ATP, glutamate, γ-aminobutyric acid (GABA), and histamine. SA1 responses were also not affected by antagonists for P2X receptors, ionotropic glutamate receptors, and ionotropic GABA and glycine receptors. Whole-cell patch-clamp recordings reconfirm the presence of both ionotropic and metabotropic 5-HT receptors on afferent neurons and their terminals innervating whisker hair follicles. All whisker afferent neurons expressed hyperpolarization-activated inward currents (Ih ), which are potentiated by serotonin through the activation of metabotropic 5-HT receptors. Taken together, the findings substantiate the serotonergic mechanism of tactile transmission at Merkel discs and identify the involvement of Ih currents in postsynaptic excitatory actions of serotonin. In addition, the findings do not favor any significant involvement of ATP, glutamate, histamine, GABA, or glycine in tactile transmission at the Merkel discs of whisker hair follicles.

Potentiation of NMDA receptor-mediated synaptic transmission at the parabrachial-central amygdala synapses by CGRP in mice.

The capsular part of the central amygdala (CeC) is called the "nociceptive amygdala," as it receives nociceptive information from various pathways, including monosynaptic input from the lateral part of the parabrachial nucleus (LPB), a major target of ascending neurons in the spinal and medullary dorsal horn. LPB-CeC synaptic transmission is mediated by glutamate but the fibers from the LPB also contain calcitonin gene-related peptide (CGRP) and the CeC is rich in CGRP-binding sites. CGRP might be released in response to strong nociception and activate these CGRP receptors. Though it has been shown that CGRP affects the excitatory postsynaptic current (EPSC) amplitude at this synapse in a manner sensitive to NMDA receptor (NMDA-R) blockers, the effect of CGRP on postsynaptic NMDA-R-mediated current recorded in isolation has never been directly examined. Thus, we evaluated the effects of CGRP on NMDA-R-mediated EPSCs that were pharmacologically isolated in brain slices from naïve mice. CGRP significantly increased the amplitude of EPSCs mediated by NMDA-Rs in a manner dependent on protein kinase A activation, but not that mediated by alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors, in concentration-dependent and antagonist-sensitive manners. This CGRP-induced potentiation of synaptic NMDA-R function would have a potent impact on the strengthening of the nociception-emotion link in persistent pain.

Accuracy of Computed Tomography-Based Navigation-Assisted Total Knee Arthroplasty: Outlier Analysis.

BACKGROUND: Achieving neutral limb alignment during total knee arthroplasty (TKA) has been identified as a potential factor in long-term prosthesis survival. This study aimed to analyze the accuracy of component orientation and postoperative alignment of the leg after computed tomography (CT)-based navigation-assisted TKA, compare these parameters with those of a conventional technique, and analyze differences in the data of outliers. METHODS: We retrospectively compared the alignment of 130 TKAs performed with a CT-based navigation system with that of 67 arthroplasties done with a conventional system. The knee joints were evaluated using radiographs. RESULTS: Mean hip-knee-ankle (HKA) angle, frontal femoral component angle, and frontal tibial component angle were 180.7°, 88.8°, and 90.6°, respectively, for the navigation-assisted arthroplasties and 181.1°, 88.7°, and 90.2°, respectively, for the conventional arthroplasties. All preoperative leg axes of 10 outliers in the navigation group were >193°, whereas the data of 17 outliers in the conventional group were scattered. CONCLUSION: This study demonstrates significant improvements in component positioning with the CT-based navigation system. Furthermore, when analyzing cases with preoperative HKA angles ≤192°, no outliers were found in the navigation group, indicating high alignment accuracy. However, in cases with preoperative HKA angles ≥193°, outliers were found in both groups, and no significant difference between the groups was observed (P = .08). Detailed analysis of the outlier cases in the navigation group revealed that the femoral component was placed in the varus position. These findings indicate that the varus knee is an important factor influencing accurate positioning of the femoral component and the postoperative leg axis.

Electrophysiological property and chemical sensitivity of primary afferent neurons that innervate rat whisker hair follicles.

Whisker hair follicles are sensory organs that sense touch and perform tactile discrimination in animals, and they are sites where sensory impulses are initiated when whisker hairs touch an object. The sensory signals are then conveyed by whisker afferent fibers to the brain for sensory perception. Electrophysiological property and chemical sensitivity of whisker afferent fibers, important factors affecting whisker sensory processing, are largely not known. In the present study, we performed patch-clamp recordings from pre-identified whisker afferent neurons in whole-mount trigeminal ganglion preparations and characterized their electrophysiological property and sensitivity to ATP, serotonin and glutamate. Of 97 whisker afferent neurons examined, 67% of them are found to be large-sized (diameter ≥45 µm) cells and 33% of them are medium- to small-sized (diameter <45 µm) cells. Almost every large-sized whisker afferent neuron fires a single action potential but many (40%) small/medium-sized whisker afferent neurons fire multiple action potentials in response to prolonged stepwise depolarization. Other electrophysiological properties including resting membrane potential, action potential threshold, and membrane input resistance are also significantly different between large-sized and small/medium-sized whisker afferent neurons. Most large-sized and many small/medium-sized whisker afferent neurons are sensitive to ATP and/or serotonin, and ATP and/or serotonin could evoke strong inward currents in these cells. In contrast, few whisker afferent neurons are sensitive to glutamate. Our results raise a possibility that ATP and/or serotonin may be chemical messengers involving sensory signaling for different types of rat whisker afferent fibers.

In situ patch-clamp recordings from Merkel cells in rat whisker hair follicles, an experimental protocol for studying tactile transduction in tactile-end organs.

Mammals use tactile end-organs to perform sensory tasks such as environmental exploration, social interaction, and tactile discrimination. However, cellular and molecular mechanisms underlying tactile transduction in tactile end-organs remain poorly understood. The patch-clamp recording technique may be the most valuable approach for detecting and studying tactile transduction in tactile end-organs, but it is technically challenging because tactile transduction elements in an end-organ are normally inaccessible by patch-clamp recording electrodes. Here we describe an in situ patch-clamp recording protocol for the study of tactile transduction in Merkel cells of rat whisker hair follicles, one of the most sensitive tactile end-organs in mammals. This technique offers an opportunity to explore the identities and properties of ion channels that are involved in tactile transduction in whisker hair follicles, and it may also lend a useful tool for researchers to study other tactile end-organs. The experimental protocol describes procedures for 1) tissue dissection and whisker hair follicle preparation, 2) device setup and steps for performing patch-clamp recordings from Merkel cells in a whisker hair follicle, 3) methods of delivering mechanical stimuli, and 4) intra-follicle microinjection for receptor knockdown in whisker hair follicles. The main procedures in this protocol, from tissue preparation to whole-cell patch-clamp recordings, can be completed in a few hours.