Developmental waves of mechanosensitivity acquisition in sensory neuron subtypes during embryonic development

Somatic sensation relies on the transduction of physical stimuli into electrical signals by sensory neurons of the dorsal root ganglia. Little is known about how and when during development different types of sensory neurons acquire transduction competence. We directly investigated the emergence of electrical excitability and mechanosensitivity of embryonic and postnatal mouse sensory neurons. We show that sensory neurons acquire mechanotransduction competence coincident with peripheral target innervation. Mechanotransduction competence arises in different sensory lineages in waves, coordinated by distinct developmental mechanisms. Sensory neurons that are mechanoreceptors or proprioceptors acquire mature mechanotransduction indistinguishable from the adult already at E13. This process is independent of neurotrophin-3 and may be driven by a genetic program. In contrast, most nociceptive (pain sensing) sensory neurons acquire mechanosensitive competence as a result of exposure to target-derived nerve growth factor. The highly regulated process of mechanosensory acquisition unveiled here, reveals new strategies to identify molecules required for sensory neuron mechanotransduction

Structure-guided examination of the mechanogating mechanism of PIEZO2

Piezo channels are mechanically activated ion channels that confer mechanosensitivity to a variety of different cell types. Piezos oligomerize as propeller-shaped homotrimers that are thought to locally curve the membrane into spherical domes that project into the cell. While several studies have identified domains and amino acids that control important properties such as ion permeability and selectivity as well as inactivation kinetics and voltage sensitivity, only little is known about intraprotein interactions that govern mechanosensitivity—the most unique feature of PIEZOs. Here we used site-directed mutagenesis and patch-clamp recordings to investigate the mechanogating mechanism of PIEZO2. We demonstrate that charged amino acids at the interface between the beam domain—i.e., a long α-helix that protrudes from the intracellular side of the “propeller” blade toward the inner vestibule of the channel—and the C-terminal domain (CTD) as well as hydrophobic interactions between the highly conserved Y2807 of the CTD and pore-lining helices are required to ensure normal mechanosensitivity of PIEZO2. Moreover, single-channel recordings indicate that a previously unrecognized intrinsically disordered domain located adjacent to the beam acts as a cytosolic plug that limits ion permeation possibly by clogging the inner vestibule of both PIEZO1 and PIEZO2. Thus, we have identified several intraprotein domain interfaces that control the mechanical activation of PIEZO1 and PIEZO2 and which might thus serve as promising targets for drugs that modulate the mechanosensitivity of Piezo channels

Study of solid 4He in two dimensions. The issue of zero-point defects and study of confined crystal

Defects are believed to play a fundamental role in the supersolid state of 4He. We report on studies by exact Quantum Monte Carlo (QMC) simulations at zero temperature of the properties of solid 4He in presence of many vacancies, up to 30 in two dimensions (2D). In all studied cases the crystalline order is stable at least as long as the concentration of vacancies is below 2.5%. In the 2D system for a small number, n_v, of vacancies such defects can be identified in the crystalline lattice and are strongly correlated with an attractive interaction. On the contrary when n_v~10 vacancies in the relaxed system disappear and in their place one finds dislocations and a revival of the Bose-Einstein condensation. Thus, should zero-point motion defects be present in solid 4He, such defects would be dislocations and not vacancies, at least in 2D. In order to avoid using periodic boundary conditions we have studied the exact ground state of solid 4He confined in a circular region by an external potential. We find that defects tend to be localized in an interfacial region of width of about 15 A. Our computation allows to put as upper bound limit to zero--point defects the concentration 0.003 in the 2D system close to melting density.Comment: 17 pages, accepted for publication in J. Low Temp. Phys., Special Issue on Supersolid

Observation of the conversion decay ϕ→π0e+e−\phi\to\pi^0e^+e^- at CMD-2

Using 15.1^{-1} of data collected by CMD-2 in the $\phi$-meson energy range, the branching ratio of the conversion decay $\phi\to\pi^0e^+e^-$ has been measured for the first time: B(\phi\to\pi^0e^+e^-) = (1.22 \pm 0.34 \pm 0.21) \cdot 10 ^{-5}.Comment: 13 pages, 6 PostScript figures, uses refmerge.sty. To be published in Phys. Lett.

The molecular mechanism and physiological role of silent nociceptor activation

Silent nociceptors are sensory afferents that are insensitive to noxious mechanical stimuli under normal conditions but become sensitized to such stimuli during inflammation. Using RNA-sequencing and quantitative RT-PCR we demonstrate that inflammation selectively upregulates the expression of the transmembrane protein TMEM100 in silent nociceptors and electrophysiology revealed that over-expression of TMEM100 is required and sufficient to un-silence silent nociceptors. Moreover, we show that mice lacking TMEM100 do not develop secondary hyperalgesia, i.e. pain hypersensitivity that spreads beyond the site of inflammation, in a mouse model of knee joint inflammation and that AAV-mediated overexpression of TMEM100 in articular afferents in the absence of inflammation is sufficient to induce secondary hyperalgesia in remote skin regions without causing knee joint pain. Thus, our work identifies TMEM100 as a key regulator of silent nociceptor un-silencing and reveals a physiological role for this hitherto enigmatic afferent subclass in triggering spatially remote secondary hyperalgesia during inflammation

Study of conversion decays phi->etae+e-, eta->e+e-g and eta->pi+pi-e+e- at CMD-2

Using 15.1 pb^{-1} of data collected by CMD-2 in the phi-meson energy range, the branching ratios of the following conversion decays have been measured: B(\phi\to\eta e^+e^-) = (1.14\pm0.10\pm0.06)\cdot10^{-4}, B(\eta\to e^+e^-\gamma) = (7.10\pm0.64\pm0.46)\cdot10^{-3}, B(\eta\to\pi^+\pi^-e^+e^-) = (3.7^{+2.5}_{-1.8}\pm0.3)\cdot10^{-4}. The upper limits for the following rare conversion decays have been obtained at the 90% confidence level: B(\phi\to\eta\mu^+\mu^-) < 9.4\cdot10^{-6}, B(\eta\to e^+e^-e^+e^-) < 6.9\cdot10^{-5}.Comment: 16 pages, 7 PostScript figures. To be published in Phys. Lett.

Hairy sensation

The hairs of the skin not only function to prevent heat loss but also have important sensory functions. Recent work has now established that each hair of the skin is innervated by one or more of three types of mechanoreceptor ending. Each of these three mechanoreceptor types possesses distinct molecular features and detects distinctive information about skin touch, which is relayed to specific brain locations in a somatotopic fashion

The transcription factor c-Maf in sensory neuron development

The proto-oncogene c-Maf has been shown to be an important transcriptional regulator in the differentiation of a number of cellular contexts, like the eye and hematopoietic system. Here we discuss the recent progress made in understanding c-Maf function in the nervous system

Differential modulation of voltage-gated sodium channels by nerve growth factor in three major subsets of TrkA-expressing nociceptors

Nerve growth factor is an inflammatory mediator that induces long-lasting hyperalgesia, which can partially be attributed to nerve growth factor-induced sensitization of primary afferent nociceptors. It was shown that nerve growth factor increases the excitability of polymodal C-fibre nociceptors by modulating tetrodotoxin-sensitive and tetrodotoxin-resistant voltage-gated sodium channels, but hitherto only little is known about the effects of nerve growth factor on sodium currents in other nociceptor subtypes that express the nerve growth factor receptor TrkA. We previously characterized two reporter mouse lines that allow the unequivocal identification of two important subclasses of TrkA-expressing nociceptors – i.e. neuropeptide Y receptor type 2 (NPY2R+) Aδ-fibre nociceptors that mediate pinprick pain and nicotinic acetylcholine receptor alpha-3 subunit (CHRNA3+) silent nociceptors, which are the most abundant TrkA+ nociceptors in visceral organs and deep somatic tissues. Here, we utilized these mouse lines to investigate the expression patterns and the possible nerve growth factor-dependent modulation of sodium channels in these neurons using whole-cell patch-clamp recordings and quantitative real-time polymerase chain reaction. We demonstrate that NPY2R+ nociceptors, CHRNA3+ ‘silent’ nociceptors and polymodal C-fibre nociceptors express different combinations of sodium channel α- and β-subunits and accordingly exhibit functionally different sodium currents. Moreover, we demonstrate that nerve growth factor produces robust hyperpolarizing shifts in the half-activation voltage of tetrodotoxin-resistant currents in NPY2R+ nociceptors and polymodal C-fibre nociceptors and also shifts the half-activation of tetrodotoxin-sensitive currents in polymodal C-fibre nociceptors. In silent nociceptors, however, nerve growth factor solely increases the current density of the tetrodotoxin-resistant current but does not alter other sodium channel properties. Considering the different peripheral target tissues and the previously reported roles in different forms of pain of the nociceptor subpopulations that were examined here, our results suggest that nerve growth factor differentially contributes to the development visceral and cutaneous pain hypersensitivity and highlights the importance of developing different therapeutic strategies for different forms of pain