Labeling PIEZO2 activity in the peripheral nervous system.

Sensory neurons detect mechanical forces from both the environment and internal organs to regulate physiology. PIEZO2 is a mechanosensory ion channel critical for touch, proprioception, and bladder stretch sensation, yet its broad expression in sensory neurons suggests it has undiscovered physiological roles. To fully understand mechanosensory physiology, we must know where and when PIEZO2-expressing neurons detect force. The fluorescent styryl dye FM 1-43 was previously shown to label sensory neurons. Surprisingly, we find that the vast majority of FM 1-43 somatosensory neuron labeling in mice in vivo is dependent on PIEZO2 activity within the peripheral nerve endings. We illustrate the potential of FM 1-43 by using it to identify novel PIEZO2-expressing urethral neurons that are engaged by urination. These data reveal that FM 1-43 is a functional probe for mechanosensitivity via PIEZO2 activation in vivo and will facilitate the characterization of known and novel mechanosensory processes in multiple organ systems.

PIEZO1 transduces mechanical itch in mice.

Itch triggers scratching, a behavioural defence mechanism that aids in the removal of harmful irritants and parasites1. Chemical itch is triggered by many endogenous and exogenous cues, such as pro-inflammatory histamine, which is released during an allergic reaction1. Mechanical itch can be triggered by light sensations such as wool fibres or a crawling insect2. In contrast to chemical itch pathways, which have been extensively studied, the mechanisms that underlie the transduction of mechanical itch are largely unknown. Here we show that the mechanically activated ion channel PIEZO1 (ref. 3) is selectively expressed by itch-specific sensory neurons and is required for their mechanically activated currents. Loss of PIEZO1 function in peripheral neurons greatly reduces mechanically evoked scratching behaviours and both acute and chronic itch-evoked sensitization. Finally, mice expressing a gain-of-function Piezo1 allele4 exhibit enhanced mechanical itch behaviours. Our studies reveal the polymodal nature of itch sensory neurons and identify a role for PIEZO1 in the sensation of itch.

The mechanosensitive ion channel Piezo2 mediates sensitivity to mechanical pain in mice.

The brush of a feather and a pinprick are perceived as distinct sensations because they are detected by discrete cutaneous sensory neurons. Inflammation or nerve injury can disrupt this sensory coding and result in maladaptive pain states, including mechanical allodynia, the development of pain in response to innocuous touch. However, the molecular mechanisms underlying the alteration of mechanical sensitization are poorly understood. In mice and humans, loss of mechanically activated PIEZO2 channels results in the inability to sense discriminative touch. However, the role of Piezo2 in acute and sensitized mechanical pain is not well defined. Here, we showed that optogenetic activation of Piezo2-expressing sensory neurons induced nociception in mice. Mice lacking Piezo2 in caudal sensory neurons had impaired nocifensive responses to mechanical stimuli. Consistently, ex vivo recordings in skin-nerve preparations from these mice showed diminished Aδ-nociceptor and C-fiber firing in response to mechanical stimulation. Punctate and dynamic allodynia in response to capsaicin-induced inflammation and spared nerve injury was absent in Piezo2-deficient mice. These results indicate that Piezo2 mediates inflammation- and nerve injury-induced sensitized mechanical pain, and suggest that targeting PIEZO2 might be an effective strategy for treating mechanical allodynia.

Deep anestrous mares under natural photoperiod treated with recombinant equine FSH (reFSH) and LH (reLH) have fertile ovulations and become pregnant.

The most common equine breeding practice to decrease the time to the first ovulation of the year is to use artificial lights starting December 1 in the Northern Hemisphere. It can take 60-90 d for this lighting regimen to induce a fertile ovulation. The success rate for pharmaceutical compounds to carry out the same process has been variable. One compound that did induce an early ovulation was recombinant equine follicle stimulating hormone (reFSH), but neither pregnancy nor cyclicity was established in that study. Starting on December 1, 20 deep-anestrous mares of light horse breeds (4-15 y old) with follicles ≤ 20 mm in diameter and progesterone < 1 ng/mL were maintained under natural photoperiod while 10 control mares were maintained under artificial photoperiod. Starting on February 6, treatment mares were randomly assigned to one of two groups: reFSH (n = 10) or reFSH/reLH (n = 10). Jugular blood samples were collected daily from all mares, and luteinizing hormone (LH), FSH, progesterone (P4), estradiol-17ß (E2) and immunoreactive (ir)-inhibin were analyzed by radioimmunoassay (RIA). When the largest follicle reached ≥32 mm in diameter, reFSH treatment was discontinued in both groups while reLH treatment continued in the reFSH/reLH group until a cohort of follicles reached ≥35 mm in diameter. Human chorionic gonadotropin (hCG) was administered intravenously (iv) to induce ovulation, and mares were bred to a fertile stallion every other day until ovulation. Mares receiving either reFSH or reFSH/reLH developed follicles ≥35 mm within 5-6 d of treatment compared with 15.8 ± 3.4 d in the control group. Both reFSH and reFSH/reLH induced ovulation in 100% of the mares within 10 d after treatment resulting in an 80% conception rate and a 70% pregnancy rate for both groups. Conception and pregnancy were designated as either presence of a 14 d old embryo (n = 16) or a fetal heartbeat at 24 d (n = 14), respectively. Only three mares in the control group ovulated within the same treatment period. Later ovulations in the control group resulted in 100% conception and pregnancy rates. At 25 d post-conception, treated mares that were pregnant (n = 7 per group) were administered prostaglandin (PGF2α) to terminate the pregnancy and later returned to estrus. Treatment with reFSH or reFSH/reLH given to deep-anestrous mares under natural photoperiod induced fertile ovulations that resulted in pregnancy and cyclicity when pregnancies were terminated.