Gentle Mechanical Stimulation Research Articles

Spatial Reconstruction of TRPC-Mechanoreceptors of the Ctenophore <i>Mnemiopsis leidyi</i> A. Agassiz, 1865

Ctenophore Mnemiopsis leidyi A. Agassiz, 1865 responds to gentle mechanical stimulation with intense luminescence; however, the mechanism of this phenomenon is unknown. We searched for possible mechanosensitive receptors that initiate signal transduction resulting in photoprotein luminescence. The three ortholog genes of mouse (5z96) and Drosophila (5vkq) TRPC-proteins, such as ML234550a-PA (860 aa), ML03701a-PA (828 aa) and ML038011a-PA (1395 aa), were found in the M. leidyi genome. The latter protein contains a long ankyrin helix consisting of 16 ANK domains. Study of the annotated domains and the network of interactions between the interactome proteins suggests that the ML234550a-PA and ML03701a-PA proteins carry out cytoplasmic, but ML038011a-PA provides intranuclear transduction of mechanical signals. Spatial reconstruction of the studied proteins revealed differences in their structure, which may be related to various functions of these proteins in the cell. The question of which of these proteins is involved in the initiation of luminescence after mechanical stimulation is discussed.

Effects of gentle mechanical skin stimulation on subjective symptoms and joint range of motions in people with chronic neck and shoulder discomfort

This study aimed to examine the efficacy of a 2-week self-administered gentle mechanical skin stimulation on chronic neck and shoulder discomfort. In participants (n = 12) with chronic neck and shoulder discomfort, subjective measures of pain sensation, discomfort, and difficulty in moving using a visual analog scale (VAS, 0–10) and objective measures of 12 different joint range of motions (ROMs) for the cervical and shoulder regions, using a digital goniometer, were collected before and after self-care with contact acupuncture, called microcones. The self-care for 2 weeks significantly (p < 0.001) decreased all VAS scores to 2.2–2.3 from baseline values of 6.0–7.4. Of the 12 ROMs tested, 8 were significantly increased (p < 0.013). This open-label study suggests the use of self-care with microcones in improving subjective symptoms and joint ROMs in people suffering from chronic neck and shoulder discomfort. However, a randomized, double-blind, controlled clinical trial is needed to further investigate the efficacy and safety of microcones.

Gentle Perineal Skin Stimulation for Control of Nocturia.

One of the major causes of nocturia is overactive bladder (OAB). Somatic afferent nerve stimuli are used for treating OAB. However, clinical evidence for the efficacy of this treatment is insufficient due to the lack of appropriate control stimuli. Studies on anesthetized animals, which eliminate emotional factors and placebo effects, have demonstrated an influence of somatic stimuli on urinary bladder functions and elucidated the underlying mechanisms. In general, the effects of somatic stimuli are dependent on the modality, location, and physical characteristics of the stimulus. Recently we showed that gentle stimuli applied to the perineal skin using a soft elastomer roller inhibited micturition contractions to a greater extent than a roller with a hard surface. Studies aiming to elucidate the neural mechanisms of gentle stimulation-induced inhibition reported that 1-10 Hz discharges of low-threshold cutaneous mechanoreceptive Aβ, Aδ, and C fibers evoked during stimulation with an elastomer roller inhibited the micturition reflex by activating the spinal cord opioid system, thereby reducing both ascending and descending transmission between bladder and pontine micturition center. The present review will provide a brief summary of (1) the effect of somatic electrical stimulation on the micturition reflex, (2) the effect of gentle mechanical skin stimulation on the micturition reflex, (3) the afferent, efferent, and central mechanisms underlying the effects of gentle stimulation, and (4) a translational clinical study demonstrating the efficacy of gentle skin stimuli for treating nocturia in the elderly with OAB by using the two roller types inducing distinct effects on rat micturition contractions. Anat Rec, 302:1824-1836, 2019. © 2019 American Association for Anatomy.

Central nervous system mechanisms involved in inhibition of micturition reflex by gentle mechanical skin stimulation

Central nervous system mechanisms involved in inhibition of micturition reflex by gentle mechanical skin stimulation

Age-Related Changes in Neuromodulatory Control of Bladder Micturition Contractions Originating in the Skin.

The brainstem is essential for producing micturition contractions of the urinary bladder. Afferent input from perineal skin evoked by gentle mechanical stimulation inhibits micturition contractions by decreasing both ascending and descending transmissions between the brainstem and spinal cord. Dysfunction of this inhibitory mechanism may be one cause of the increase in the prevalence of overactive bladder in old age. The aim of this study was to examine effect of aging on function of skin afferent fibers that inhibit bladder micturition contractions in rats. We used anesthetized male rats in three different age groups: young adult (4–5 months old), middle aged (6–9 months old), and aged (27–30 months old). The bladder was expanded to produce isovolumetric rhythmic micturition contractions. Skin afferent fibers were activated for 1 min either by electrical stimulation (0.5 ms, 0.2–10 V, 0.1–10 Hz) of the cutaneous branch of the pudendal nerve (CBPN) or by gentle mechanical skin stimulation with an elastomer roller. When skin afferent nerves were activated electrically, micturition contractions were inhibited in a similar manner in all age groups, with long latency inhibition induced by excitation of Aβ fibers and short latency inhibition by additional Aδ and C fiber excitation (at 1–10 Hz). On the contrary, when skin afferent nerves were activated mechanically by rolling, latency of inhibition following rolling stimulation was prolonged in aged rats. Single unitary afferent nerve activity of low-threshold mechanoreceptors (LTMs) from the cutaneous nerve was recorded. The discharge rate during rolling was not significantly reduced in Aβ units but was much lower in Aδ and C units in aged rats (0.4 and 0.5 Hz, respectively) than in young adult rats (3 and 7 Hz). These results suggest that the neural mechanism that inhibits bladder micturition contractions by skin afferent input is well maintained in old age, but the early inhibition by gentle skin stimulation is decreased because of reduced responses of Aδ- and C-LTMs.

Naphthalimide‐Containing Isomeric Urea Derivatives: Mechanoluminescence and Fluoride Recognition

AbstractIsomeric ortho, meta and para naphthalimide–urea derivatives ND‐2U, ND‐3U and ND‐4U were synthesized and characterized. Upon gentle mechanical stimulation, the weakly emissive ortho isomeric sample ND‐2U was transformed into a yellow‐green emissive material (λem=500 nm), with Δλ≈125 nm. The mechanoluminescent emission of this isomer could be “switched off” by exposing the sample to solvent vapour. Moreover, ND‐2U could respond to fluoride anions in solution with a dramatic enhancement of fluorescence at λ=446 nm. The sigmoidal nature of the binding isotherm suggested cooperative binding of fluoride to the ortho isomer. 1H NMR spectroscopic studies provided vital insights into the nature of ND‐2U/F−complexation, highlighting the role of the proximal 1,8‐naphthalimide motif.

Optimal delineation of single C-tactile and C-nociceptive afferents in humans by latency slowing.

C-mechanoreceptors in humans comprise a population of unmyelinated afferents exhibiting a wide range of mechanical sensitivities. C-mechanoreceptors are putatively divided into those signaling gentle touch (C-tactile afferents, CTs) and nociception (C-mechanosensitive nociceptors, CMs), giving rise to positive and negative affect, respectively. We sought to distinguish, compare, and contrast the properties of a population of human C-mechanoreceptors to see how fundamental the divisions between these putative subpopulations are. We used microneurography to record from individual afferents in humans and applied electrical and mechanical stimulation to their receptive fields. We show that C-mechanoreceptors can be distinguished unequivocally into two putative populations, comprising CTs and CMs, by electrically evoked spike latency changes (slowing). After both natural mechanical stimulation and repetitive electrical stimulation there was markedly less latency slowing in CTs compared with CMs. Electrical receptive field stimulation, which bypasses the receptor end organ, was most effective in classifying C-mechanoreceptors, as responses to mechanical receptive field stimulation overlapped somewhat, which may lead to misclassification. Furthermore, we report a subclass of low-threshold CM responding to gentle mechanical stimulation and a potential subclass of CT afferent displaying burst firing. We show that substantial differences exist in the mechanisms governing axonal conduction between CTs and CMs. We provide clear electrophysiological "signatures" (extent of latency slowing) that can be used in unequivocally identifying populations of C-mechanoreceptors in single-unit and multiunit microneurography studies and in translational animal research into affective touch. Additionally, these differential mechanisms may be pharmacologically targetable for separate modulation of positive and negative affective touch information.NEW & NOTEWORTHY Human skin encodes a plethora of touch interactions, and affective tactile information is primarily signaled by slowly conducting C-mechanoreceptive afferents. We show that electrical stimulation of low-threshold C-tactile afferents produces markedly different patterns of activity compared with high-threshold C-mechanoreceptive nociceptors, although the populations overlap in their responses to mechanical stimulation. This fundamental distinction demonstrates a divergence in affective touch signaling from the first stage of sensory processing, having implications for the processing of interpersonal touch.

The Role of PIEZO2 in Human Mechanosensation

BackgroundThe senses of touch and proprioception evoke a range of perceptions and rely on the ability to detect and transduce mechanical force. The molecular and neural mechanisms underlying these sensory functions remain poorly defined. The stretch-gated ion channel PIEZO2 has been shown to be essential for aspects of mechanosensation in model organisms.MethodsWe performed whole-exome sequencing analysis in two patients who had unique neuromuscular and skeletal symptoms, including progressive scoliosis, that did not conform to standard diagnostic classification. In vitro and messenger RNA assays, functional brain imaging, and psychophysical and kinematic tests were used to establish the effect of the genetic variants on protein function and somatosensation.ResultsEach patient carried compound-inactivating variants in PIEZO2, and each had a selective loss of discriminative touch perception but nevertheless responded to specific types of gentle mechanical stimulation on hairy skin. The patients had profoundly decreased proprioception leading to ataxia and dysmetria that were markedly worse in the absence of visual cues. However, they had the ability to perform a range of tasks, such as walking, talking, and writing, that are considered to rely heavily on proprioception.ConclusionsOur results show that PIEZO2 is a determinant of mechanosensation in humans. (Funded by the National Institutes of Health Intramural Research Program.)

Purinergic Signalling: Pathophysiology and Therapeutic Potential

The article begins with a review of the main conceptual steps involved in the development of our understanding of purinergic signalling, including non-adrenergic, non-cholinergic (NANC) neurotransmission; identification of ATP as a NANC transmitter; purinergic cotransmission; recognition of two families of purinoceptors [P1 (adenosine) and P2 (ATP/ADP)]; and, later, cloning and characterisation of P1 (G protein-coupled), P2X (ion channel) and P2Y (G protein-coupled) receptor subtypes. Further studies have established the involvement of ATP in synaptic neurotransmission in both ganglia and in the central nervous system; long-term (trophic) purinergic signalling in cell proliferation, differentiation and death occurring in development and regeneration; and short-term purinergic signalling in neurotransmission, neuromodulation and secretion. ATP is released from most cell types in response to gentle mechanical stimulation and is rapidly degraded to adenosine by ecto-nucleotidases. This review then focuses on the pathophysiology of purinergic signalling in a wide variety of systems, including urinogenital, cardiovascular, airway, musculoskeletal and gastrointestinal. Consideration is also given to the involvement of purinoceptors in pain, cancer and diseases of the central nervous system. Purinergic therapeutic approaches for the treatment of some of these diseases are discussed.

Gentle Mechanical Skin Stimulation Inhibits Micturition Contractions via the Spinal Opioidergic System and by Decreasing Both Ascending and Descending Transmissions of the Micturition Reflex in the Spinal Cord.

Recently, we found that gentle mechanical skin stimulation inhibits the micturition reflex in anesthetized rats. However, the central mechanisms underlying this inhibition have not been determined. This study aimed to clarify the central neural mechanisms underlying this inhibitory effect. In urethane-anesthetized rats, cutaneous stimuli were applied for 1 min to the skin of the perineum using an elastic polymer roller with a smooth, soft surface. Inhibition of rhythmic micturition contractions by perineal stimulation was abolished by naloxone, an antagonist of opioidergic receptors, administered into the intrathecal space of the lumbosacral spinal cord at doses of 2–20 μg but was not affected by the same doses of naloxone administered into the subarachnoid space of the cisterna magna. Next, we examined whether perineal rolling stimulation inhibited the descending and ascending limbs of the micturition reflex. Perineal rolling stimulation inhibited bladder contractions induced by electrical stimulation of the pontine micturition center (PMC) or the descending tract of the micturition reflex pathway. It also inhibited the bladder distension-induced increase in the blood flow of the dorsal cord at L5–S1, reflecting the neural activity of this area, as well as pelvic afferent-evoked field potentials in the dorsal commissure at the lumbosacral level; these areas contain long ascending neurons to the PMC. Neuronal activities in this center were also inhibited by the rolling stimulation. These results suggest that the perineal rolling stimulation activates the spinal opioidergic system and inhibits both ascending and descending transmissions of the micturition reflex pathway in the spinal cord. These inhibitions would lead to the shutting down of positive feedback between the bladder and the PMC, resulting in inhibition of the micturition reflex. Based on the central neural mechanisms we show here, gentle perineal stimulation may be applicable to several different types of overactive bladder.

Stomatin‐domain protein interactions with acid‐sensing ion channels modulate nociceptor mechanosensitivity

Acid-sensing ion channels (ASICs) and their interaction partners of the stomatin family have all been implicated in sensory transduction. Single gene deletion of asic3, asic2, stomatin, or stoml3 all result in deficits in the mechanosensitivity of distinct cutaneous afferents in the mouse. Here, we generated asic3−/−:stomatin−/−, asic3−/−:stoml3−/− and asic2−/−:stomatin−/− double mutant mice to characterize the functional consequences of stomatin–ASIC protein interactions on sensory afferent mechanosensitivity. The absence of ASIC3 led to a clear increase in mechanosensitivity in rapidly adapting mechanoreceptors (RAMs) and a decrease in the mechanosensitivity in both Aδ- and C-fibre nociceptors. The increased mechanosensitivity of RAMs could be accounted for by a loss of adaptation which could be mimicked by local application of APETx2 a toxin that specifically blocks ASIC3. There is a substantial loss of mechanosensitivity in stoml3−/− mice in which ∼35% of the myelinated fibres lack a mechanosensitive receptive field and this phenotype was found to be identical in asic3−/−:stoml3−/− mutant mice. However, Aδ-nociceptors showed much reduced mechanosensitivity in asic3−/−:stoml3−/− mutant mice compared to asic3−/− controls. Interestingly, in asic2−/−:stomatin−/− mutant mice many Aδ-nociceptors completely lost their mechanosensitivity which was not observed in asic2−/− or stomatin−/− mice. Examination of stomatin−/−:stoml3−/− mutant mice indicated that a stomatin/STOML3 interaction is unlikely to account for the greater Aδ-nociceptor deficits in double mutant mice. A key finding from these studies is that the loss of stomatin or STOML3 in asic3−/− or asic2−/− mutant mice markedly exacerbates deficits in the mechanosensitivity of nociceptors without affecting mechanoreceptor function.

Purinergic Signalling: Pathophysiology and Therapeutic Potential

The article begins with a review of the main conceptual steps involved in the development of our understanding of purinergic signalling, including non-adrenergic, non-cholinergic (NANC) neurotransmission; identification of ATP as a NANC transmitter; purinergic cotransmission; recognition of two families of purinoceptors [P1 (adenosine) and P2 (ATP/ADP)]; and, later, cloning and characterisation of P1 (G protein-coupled), P2X (ion channel) and P2Y (G protein-coupled) receptor subtypes. Further studies have established the involvement of ATP in synaptic neurotransmission in both ganglia and in the central nervous system; long-term (trophic) purinergic signalling in cell proliferation, differentiation and death occurring in development and regeneration; and short-term purinergic signalling in neurotransmission, neuromodulation and secretion. ATP is released from most cell types in response to gentle mechanical stimulation and is rapidly degraded to adenosine by ecto-nucleotidases. This review then focuses on the pathophysiology of purinergic signalling in a wide variety of systems, including urinogenital, cardiovascular, airway, musculoskeletal and gastrointestinal. Consideration is also given to the involvement of purinoceptors in pain, cancer and diseases of the central nervous system. Purinergic therapeutic approaches for the treatment of some of these diseases are discussed.

A gentle mechanical skin stimulation technique for inhibition of micturition contractions of the urinary bladder

Effects of gentle skin stimulation of various segmental areas on the micturition contractions of the urinary bladder were examined in anesthetized male rats. The bladder was expanded by infusing saline via urethral cannula until the bladder produced rhythmic micturition contractions as a consequence of rhythmic burst discharges of vesical pelvic efferent nerves. Gentle stimulation was applied for 1min by slowly rolling on top of skin with an elastomer “roller”. Rolling on the perineal area inhibited both micturition contractions and pelvic efferent discharges during and after stimulation. Stimulation of the hindlimb, abdomen and forelimb inhibited micturition contractions after stimulation ended, in this order of effectiveness. During stimulation of the perineal skin, the reflex increase in pelvic efferent discharges in response to bladder distension to a constant pressure was also inhibited up to 45% of its control response. The inhibition of the micturition contractions induced by perineal stimulation was abolished, to a large extent by the opioid receptor antagonist naloxone and completely by severing cutaneous nerves innervating the perineal skin. We recorded unitary afferent activity from cutaneous branches of the pudendal nerve and found that the fibers excited by stimulation were low-threshold mechanoreceptive Aβ, Aδ and C fibers. Discharge rates of afferent C fibers (7.9Hz) were significantly higher than those of Aβ (2.2Hz) and Aδ (2.9Hz) afferents. The results suggest that low frequency excitation of low threshold cutaneous mechanoreceptive myelinated and unmyelinated fibers inhibits a vesico-pelvic parasympathetic reflex, mainly via release of opioids, leading to inhibition of micturition contraction.

Purinergic Signalling: Pathophysiology and Therapeutic Potential

The article begins with a review of the main conceptual steps involved in the development of our understanding of purinergic signalling, including non-adrenergic, non-cholinergic (NANC) neurotransmission; identification of ATP as a NANC transmitter; purinergic cotransmission; recognition of two families of purinoceptors [P1 (adenosine) and P2 (ATP/ADP)]; and, later, cloning and characterisation of P1 (G protein-coupled), P2X (ion channel) and P2Y (G protein-coupled) receptor subtypes. Further studies have established the involvement of ATP in synaptic neurotransmission in both ganglia and in the central nervous system; long-term (trophic) purinergic signalling in cell proliferation, differentiation and death occurring in development and regeneration; and short-term purinergic signalling in neurotransmission, neuromodulation and secretion. ATP is released from most cell types in response to gentle mechanical stimulation and is rapidly degraded to adenosine by ecto-nucleotidases. This review then focuses on the pathophysiology of purinergic signalling in a wide variety of systems, including urinogenital, cardiovascular, airway, musculoskeletal and gastrointestinal. Consideration is also given to the involvement of purinoceptors in pain, cancer and diseases of the central nervous system. Purinergic therapeutic approaches for the treatment of some of these diseases are discussed.

Age-related change in the effect of gentle mechanical cutaneous stimulation on the somato-cardiac sympathetic C-reflex

This study examined whether aging influences the effect of gentle mechanical cutaneous stimulation (touch) on somato-cardiac sympathetic reflexes in anesthetized rats. A single electrical stimulus (15 V, 0.5 ms) applied to the tibial nerve induced somato-cardiac sympathetic A- and C-reflexes, which originated from the myelinated A- and unmyelinated C-fibers of the tibial nerve, respectively. When touch was applied to the inner thigh ipsilateral to the electrical stimulation, the C-reflex was selectively inhibited in adult rats (4-12 months), and this response was abolished by the opioid receptor antagonist, naloxone. Such an inhibitory effect of touch was impaired in aged rats (26-30 months). In contrast, depression of both A- and C-reflexes during a phenylephrine-induced rise in blood pressure was well maintained in the aged group. This study indicates that activation of the opioidergic system by gentle mechanical cutaneous stimulation attenuates with aging.

Gentle mechanical skin stimulation inhibits the somatocardiac sympathetic: C-reflex elicited by excitation of unmyelinated C-afferent fibers

The effects of gentle mechanical skin stimulation on reflex discharges in cardiac sympathetic nerve evoked by somatic afferent stimulation were studied in anesthetized rats. Mass discharges were recorded from cardiac sympathetic efferent nerve while somatocardiac sympathetic A- and C-reflexes were elicited by single electrical stimuli to myelinated A- and unmyelinated C-afferent fibers of the tibial nerve. Continuous touch was applied to inner thigh skin with a force of 0.12 N for 10 min periods by a soft elastomer “brush” (1.1 cm in diameter with 417 microcones). When touch was applied ipsilateral to the stimulated tibial nerve, the C-reflex was inhibited by up to 40% of its pre-touch amplitude, whereas the A-reflex was unaffected. Inhibition of the C-reflex started during the touch period and lasted for 15 min after cessation of touching. Contralateral touch did not inhibit the C-reflex. The opioid receptor antagonist naloxone attenuated the C-reflex inhibition, but did not abolish it. The C-reflex inhibition was abolished after severing cutaneous nerves innervating inner thigh. We recorded unitary afferent activity from thigh branches of the saphenous nerve and found fibers excited by touch were low-threshold mechanoreceptive Aβ, Aδ and C fibers that have rapidly or slowly adapting properties. In all units tested, average discharge rates during touch period were less than 4 Hz. The results suggest that touch-induced excitation of low threshold cutaneous mechanoreceptive fibers inhibits nociceptive transmission conveyed by C-primary-afferents, via the release of both opioid and non-opioid inhibitory mediators.

Loss of the Atp2c1 Secretory Pathway Ca2+-ATPase (SPCA1) in Mice Causes Golgi Stress, Apoptosis, and Midgestational Death in Homozygous Embryos and Squamous Cell Tumors in Adult Heterozygotes

Loss of one copy of the human ATP2C1 gene, encoding SPCA1 (secretory pathway Ca(2+)-ATPase isoform 1), causes Hailey-Hailey disease, a skin disorder. We performed targeted mutagenesis of the Atp2c1 gene in mice to analyze the functions of this Golgi membrane Ca(2+) pump. Breeding of heterozygous mutants yielded a normal Mendelian ratio among embryos on gestation day 9.5; however, null mutant (Spca1(-/-)) embryos exhibited growth retardation and did not survive beyond gestation day 10.5. Spca1(-/-) embryos had an open rostral neural tube, but hematopoiesis and cardiovascular development were ostensibly normal. Golgi membranes of Spca1(-/-) embryos were dilated, had fewer stacked leaflets, and were expanded in amount, consistent with increased Golgi biogenesis. The number of Golgi-associated vesicles was also increased, and rough endoplasmic reticulum had fewer ribosomes. Coated pits, junctional complexes, desmosomes, and basement membranes appeared normal in mutant embryos, indicating that processing and trafficking of proteins in the secretory pathway was not massively impaired. However, apoptosis was increased, possibly the result of secretory pathway stress, and a large increase in cytoplasmic lipid was observed in mutant embryos, consistent with impaired handling of lipid by the Golgi. Adult heterozygous mice appeared normal and exhibited no evidence of Hailey-Hailey disease; however, aged heterozygotes had an increased incidence of squamous cell tumors of keratinized epithelial cells of the skin and esophagus. These data show that loss of the Golgi Ca(2+) pump causes Golgi stress, expansion of the Golgi, increased apoptosis, and embryonic lethality and demonstrates that SPCA1 haploinsufficiency causes a genetic predisposition to cancer.

Stretch‐activated calcium channels relay fast calcium waves propagated by calcium‐induced calcium influx

For nearly 30 years, fast calcium waves have been attributed to a regenerative process propagated by CICR (calcium-induced calcium release) from the endoplasmic reticulum. Here, I propose a model containing a new subclass of fast calcium waves which is propagated by CICI (calcium-induced calcium influx) through the plasma membrane. They are called fast CICI waves. These move at the order of 100 to 1000 microm/s (at 20 degrees C), rather than the order of 3 to 30 microm/s found for CICR. Moreover, in this proposed subclass, the calcium influx which drives calcium waves is relayed by stretch-activated calcium channels. This model is based upon reports from approx. 60 various systems. In seven of these reports, calcium waves were imaged, and, in five of these, evidence was presented that these waves were regenerated by CICI. Much of this model involves waves that move along functioning flagella and cilia. In these systems, waves of local calcium influx are thought to cause waves of local contraction by inducing the sliding of dynein or of kinesin past tubulin microtubules. Other cells which are reported to exhibit waves, which move at speeds in the fast CICI range, include ones from a dozen protozoa, three polychaete worms, three molluscs, a bryozoan, two sea urchins, one arthropod, four insects, Amphioxus, frogs, two fish and a vascular plant (Equisetum), together with numerous healthy, as well as cancerous, mammalian cells, including ones from human. In two of these systems, very gentle local mechanical stimulation is reported to initiate waves. In these non-flagellar systems, the calcium influxes are thought to speed the sliding of actinomyosin filaments past each other. Finally, I propose that this mechanochemical model could be tested by seeing if gentle mechanical stimulation induces waves in more of these systems and, more importantly, by imaging the predicted calcium waves in more of them.

Site- and exercise-related variation in structure and function of cartilage from equine distal metacarpal condyle

Determine (1) the site-associated response of articular cartilage of the equine distal metacarpal condyle to training at a young age as assessed by changes in indentation stiffness and alterations in cartilage structure and composition, and (2) relationships between indentation stiffness and indices of cartilage structure and composition. Experimental animals (n=6) were trained on a track (increasing exercise to 1km/day by 5 months); controls (n=6) were pasture-reared. Animals were euthanized at 18 months and four osteochondral samples were harvested per metacarpal condyle from dorsal-medial, dorsal-lateral, palmar-medial, and palmar-lateral aspects. Cartilage was analyzed for India ink staining (quantified as reflectance score (RS)), short-term indentation stiffness (sphere-ended, 0.4mm diameter), thickness, and biochemical composition. Cartilage structural, biochemical and biomechanical properties varied markedly with site in the joint. Sites just medial and just lateral to the sagittal ridge showed signs of early degeneration, with relatively low RS, indentation stiffness, and collagen content, and relatively high water content. Effects of exercise and side (left vs right) were not detected for any measure. Overall, indentation stiffness correlated positively with RS and collagen content, and inversely with thickness and water content. Gentle exercise-imposed mechanical stimulation did not markedly affect articular cartilage function or structure. However, the marked site-associated variation suggests that biomechanical environment can initiate degenerative changes in immature cartilage during joint growth and maturation.

In Vivo Imaging of C. elegans Mechanosensory Neurons Demonstrates a Specific Role for the MEC-4 Channel in the Process of Gentle Touch Sensation

In the nematode C. elegans, genes encoding components of a putative mechanotransducing channel complex have been identified in screens for light-touch-insensitive mutants. A long-standing question, however, is whether identified MEC proteins act directly in touch transduction or contribute indirectly by maintaining basic mechanoreceptor neuron physiology. In this study, we used the genetically encoded calcium indicator cameleon to record cellular responses of mechanosensory neurons to touch stimuli in intact, behaving nematodes. We defined a gentle touch sensory modality that adapts with a time course of approximately 500 ms and primarily senses motion rather than pressure. The DEG/ENaC channel subunit MEC-4 and channel-associated stomatin MEC-2 are specifically required for neural responses to gentle mechanical stimulation, but do not affect the basic physiology of touch neurons or their in vivo responses to harsh mechanical stimulation. These results distinguish a specific role for the MEC channel proteins in the process of gentle touch mechanosensation.