Controlling Engineered P2X Receptors with Light

This chapter details methods to express and modify ATP-gated P2X receptor channels so that they can be controlled using light. Following expression in cells, a photoswitchable tool compound can be used to covalently modify mutant P2X receptors, as previously demonstrated for homomeric P2X2 and P2X3 receptors, and heteromeric P2X2/3 receptors. Engineered P2X receptors can be rapidly and reversibly opened and closed by different wavelengths of light. Light-activated P2X receptors can be mutated further to impart ATP-insensitivity if required. This method offers control of specific P2X receptor channels with high spatiotemporal precision to study their roles in physiology and pathophysiology

Scanned optogenetic control of mammalian somatosensory input to map input-specific behavioral outputs

Somatosensory stimuli guide and shape behavior, from immediate protective reflexes to longer-term learning and higher-order processes related to pain and touch. However, somatosensory inputs are challenging to control in awake mammals due to the diversity and nature of contact stimuli. Application of cutaneous stimuli is currently limited to relatively imprecise methods as well as subjective behavioral measures. The strategy we present here overcomes these difficulties, achieving ‘remote touch’ with spatiotemporally precise and dynamic optogenetic stimulation by projecting light to a small defined area of skin. We mapped behavioral responses in freely behaving mice with specific nociceptor and low-threshold mechanoreceptor inputs. In nociceptors, sparse recruitment of single action potentials shapes rapid protective pain-related behaviors, including coordinated head orientation and body repositioning that depend on the initial body pose. In contrast, activation of low-threshold mechanoreceptors elicited slow-onset behaviors and more subtle whole-body behaviors. The strategy can be used to define specific behavioral repertoires, examine the timing and nature of reflexes, and dissect sensory, motor, cognitive and motivational processes guiding behavior

Bones' adaptive response to mechanical loading is essentially linear between the low strains associated with disuse and the high strains associated with the lamellar/woven bone transition.

There is a widely held view that the relationship between mechanical loading history and adult bone mass/strength includes an adapted state or "lazy zone" where the bone mass/strength remains constant over a wide range of strain magnitudes. Evidence to support this theory is circumstantial. We investigated the possibility that the "lazy zone" is an artifact and that, across the range of normal strain experience, features of bone architecture associated with strength are linearly related in size to their strain experience. Skeletally mature female C57BL/6 mice were right sciatic neurectomized to minimize natural loading in their right tibiae. From the fifth day, these tibiae were subjected to a single period of external axial loading (40, 10-second rest interrupted cycles) on alternate days for 2 weeks, with a peak dynamic load magnitude ranging from 0 to 14 N (peak strain magnitude: 0-5000 µε) and a constant loading rate of 500 N/s (maximum strain rate: 75,000 µε/s). The left tibiae were used as internal controls. Multilevel regression analyses suggest no evidence of any discontinuity in the progression of the relationships between peak dynamic load and three-dimensional measures of bone mass/strength in both cortical and cancellous regions. These are essentially linear between the low-peak locomotor strains associated with disuse (∼300 µε) and the high-peak strains derived from artificial loading and associated with the lamellar/woven bone transition (∼5000 µε). The strain:response relationship and minimum effective strain are site-specific, probably related to differences in the mismatch in strain distribution between normal and artificial loading at the locations investigated

Radioactive heat production of six geologically important nuclides

Heat production rates for the geologically important nuclides ${}^{26}$Al, ${}^{40}$K, ${}^{60}$Fe, ${}^{232}$Th, ${}^{235}$U, and ${}^{238}$U are calculated on the basis of recent data on atomic and nuclear properties. The revised data differ by several per cent from some older values, but indicate that more recent analyses converge toward values with an accuracy sufficient for all common geoscience applications, although some possibilities for improvement still remain, especially in the case of ${}^{40}$K and with regard to the determination of half-lives. A Python script is provided for calculating heat production (https://github.com/trg818/radheat).Comment: 14 pages, 1 figur

Changes in communication between muscle stem cells and their environment with aging

Aging is associated with both muscle weakness and a loss of muscle mass, contributing towards overall frailty in the elderly. Aging skeletal muscle is also characterised by a decreasing efficiency in repair and regeneration, together with a decline in the number of adult stem cells. Commensurate with this are general changes in whole body endocrine signalling, in local muscle secretory environment, as well as in intrinsic properties of the stem cells themselves. The present review discusses the various mechanisms that may be implicated in these age-associated changes, focusing on aspects of cell-cell communication and long-distance signalling factors, such as levels of circulating growth hormone, IL-6, IGF1, sex hormones, and inflammatory cytokines. Changes in the local environment are also discussed, implicating IL-6, IL-4, FGF-2, as well as other myokines, and processes that lead to thickening of the extra-cellular matrix. These factors, involved primarily in communication, can also modulate the intrinsic properties of muscle stem cells, including reduced DNA accessibility and repression of specific genes by methylation. Finally we discuss the decrease in the stem cell pool, particularly the failure of elderly myoblasts to re-quiesce after activation, and the consequences of all these changes on general muscle homeostasis

Optical cuff for optogenetic control of the peripheral nervous system

OBJECTIVE: Nerves in the peripheral nervous system (PNS) contain axons with specific motor, somatosensory and autonomic functions. Optogenetics offers an efficient approach to selectively activate axons within the nerve. However, the heterogeneous nature of nerves and their tortuous route through the body create a challenging environment to reliably implant a light delivery interface. APPROACH: Here, we propose an optical peripheral nerve interface – an optocuff -, so that optogenetic modulation of peripheral nerves become possible in freely behaving mice. MAIN RESULTS: Using this optocuff, we demonstrate orderly recruitment of motor units with epineural optical stimulation of genetically targeted sciatic nerve axons, both in anaesthetized and in awake, freely behaving animals. Behavioural experiments and histology show the optocuff does not damage the nerve thus is suitable for long-term experiments. SIGNIFICANCE: These results suggest that the soft optocuff might be a straightforward and efficient tool to support more extensive study of the PNS using optogenetics

Two New Low Redshift 21cm Absorbers

As part of a larger program to identify low redshift radio analogues of the damped Lyman-alpha (DLA) absorbers seen in the spectra of high redshift quasars, Westerbork Synthesis Radio Telescope (WSRT) observations have discovered two new HI 21cm absorption lines at z=0.394 and z=0.437 in the spectra of the radio sources B 0248+430 and B 1243-072 respectively. These sightlines and redshifts were selected for study on the basis of the previously known low ionization absorption lines of MgII, and neither has been observed in the Lyman-alpha line. The 21cm line observations provide information on column densities, temperatures and kinematics of the thickest cold neutral clouds in the absorbers.Comment: 6 pages incl. 2 figures, accepted for publication in the Astrophysical Journal Letters, missing reference adde