Emergence of functional sensory subtypes as defined by transient receptor potential channel expression

The existence of heterogeneous populations of dorsal root ganglion (DRG) neurons conveying different somatosensory information is the basis for the perception of touch, temperature, and pain. A differential expression of transient receptor potential (TRP) cation channels contributes to this functional heterogeneity. However, little is known about the development of functionally diverse neuronal subpopulations. Here, we use calcium imaging of acutely dissociated mouse sensory neurons and quantitative reverse transcription PCR to show that TRP cation channels emerge in waves, with the diversification of functional groups starting at embryonic day 12.5 (E12.5) and extending well into the postnatal life. Functional responses of voltage-gated calcium channels were present in DRG neurons at E11.5 and reached adult levels by E14.5. Responses to capsaicin, menthol, and cinnamaldehyde were first seen at E12.5, E16.5, and postnatal day 0 (P0), when the mRNA for TRP cation channel, subfamily V, member 1 (TRPV1), TRP cation channel, subfamily M, member 8 (TRPM8), and TRP cation channel, subfamily A, member 1 (TRPA1), respectively, was first detected. Cold-sensitive neurons were present before the expression or functional responses of TRPM8 or TRPA1. Our data support a lineage relationship in which TRPM8- and TRPA1-expressing sensory neurons derive from the population of TRPV1-expressing neurons. The TRPA1 subpopulation of neurons emerges independently in two distinct classes of nociceptors: around birth in the peptidergic population and after P14 in the nonpeptidergic class. This indicates that neurons with similar receptive properties can be generated in different sublineages at different developmental stages. This study describes for the first time the emergence of functional subtypes of sensory neurons, providing new insight into the development of nociception and thermoreception

Behavioral and other phenotypes in a cytoplasmic Dynein light intermediate chain 1 mutant mouse

The cytoplasmic dynein complex is fundamentally important to all eukaryotic cells for transporting a variety of essential cargoes along microtubules within the cell. This complex also plays more specialized roles in neurons. The complex consists of 11 types of protein that interact with each other and with external adaptors, regulators and cargoes. Despite the importance of the cytoplasmic dynein complex, we know comparatively little of the roles of each component protein, and in mammals few mutants exist that allow us to explore the effects of defects in dynein-controlled processes in the context of the whole organism. Here we have taken a genotype-driven approach in mouse (Mus musculus) to analyze the role of one subunit, the dynein light intermediate chain 1 (Dync1li1). We find that, surprisingly, an N235Y point mutation in this protein results in altered neuronal development, as shown from in vivo studies in the developing cortex, and analyses of electrophysiological function. Moreover, mutant mice display increased anxiety, thus linking dynein functions to a behavioral phenotype in mammals for the first time. These results demonstrate the important role that dynein-controlled processes play in the correct development and function of the mammalian nervous system

Functional imaging in microfluidic chambers reveal sensory neuron sensitivity is differentially regulated between neuronal regions

Primary afferent sensory neurons are incredibly long cells, often traversing distances of over one metre in humans. Cutaneous sensory stimuli are transduced in the periphery by specialised endorgans or free nerve endings which code the stimulus into electrical action potentials that propagate towards the central nervous system. Despite significant advances in our knowledge of sensory neuron physiology and ion channel expression, many commonly used techniques fail to accurately model the primary afferent neuron in its entirety. In vitro experiments often focus on the cell somata and neglect the fundamental processes of peripheral stimulus transduction and action potential propagation. Despite this, these experiments are commonly used as a model for cellular investigations of the receptive terminals. We demonstrate that ratiometric calcium imaging performed in compartmentalised sensory neuron cultures can be used to directly and accurately compare the sensitivity and functional protein expression of isolated neuronal regions in vitro. Using microfluidic chambers, we demonstrate that the nerve terminals of cultured DRG neurons can be depolarised to induce action potential propagation, which has both TTX-resistant and TTXsensitive components. Furthermore, we show that there is a differential regulation of proton sensitivity between the sensory terminals and somata in cultured sensory neurons. We also demonstrate that capsaicin sensitivity is highly dependent on embryonic dissection age. This approach enables a comprehensive method to study the excitability and regional sensitivity of cultured sensory neurons on a single cell level. Examination of the sensory terminals is crucial to further understand the properties and diversity of DRG sensory neurons

Acoustic full-waveform to elastic pre-stack seismic inversion of the Yakutat Terrane, Gulf of Alaska

The Yakutat-North American collision in the Gulf of Alaska has developed a complex subduction zone followed by major deformations such as the Chugach-St Elias mountain range creation, intensified exhumation, fold and thrust-fault formation. I generate a compressional velocity model of the Yakutat microplate using two-dimensional acoustic and isotropic time-domain full waveform inversion (FWI) of marine seismic reflection and refraction data from the STEEP project (ST. Elias Erosion/tectonics Project). FWI is a non-linear data-fitting algorithm that aims to recover subsurface parameters from the recorded seismic wavefield. Seismic wave propagation along the Yakutat terrane is simulated using a staggered-grid finite difference modeling scheme. Drawbacks associated with FWI is cycle skipping during the minimization process, which results in converging to the wrong velocity model. Starting with a good initial model that contains the low-frequency information can help mitigate this issue. The starting velocity model input to FWI in this case is generated by a traveltime tomographic inversion of ocean-bottom seismometer and streamer seismic data. Data preconditioning includes muting, filtering, noise removal and amplitude rescaling of the field seismic data to match the corresponding amplitudes of the synthetic traces. The forward model is able to produce a good match between the observed and the modeled wavefield within half the propagated wavelength. I use the FWI result, which shows good correlation with the industry well, as an input to two additional seismic inversion methods: acoustic post-stack and elastic pre-stack seismic inversion in order to recover shear impedance and density models along the seismic line. Extending the problem to the elastic medium is important to support more advanced seismic interpretation. Both techniques were able to produce higher-resolution images of the Yakutat terrane that are well correlated with the well response. Structural complexities identified in the generated models include the northwest-dipping Pamplona fault system, the offshore folding zone, thickening of the Yakutat basement and, lastly, significantly lower velocities in the Poul Creek formation compared to the younger Yakataga formation, which may be attributed to high-fluid pressure within that formation.Geological Science

Reduced-dimension model for the Rayleigh-Taylor instability in a Hele-Shaw cell

Thesis: S.M., Massachusetts Institute of Technology, Computation for Design and Optimization Program, 2019Cataloged from PDF version of thesis.Includes bibliographical references (pages 93-94).In this thesis we present a reduced-dimension model for the density-driven hydrodynamic Rayleigh-Taylor instability. We motivate the project with experimental findings of a little-understood stabilizing effect of geometry and deviations of measured instability wavelength from theoretical predictions. We present novel methods of data analysis for the experimental data. We then present a reduced-dimension model for the governing equations of the system, Stoke's equations and Fick's law, using polynomial trial functions. We discuss the results and conduct a linear stability analysis of the reduced system. We compare the model to a finite element simulation of the full governing equations using COMSOL, and propose an optimization framework for the basis functions of the reduced model. The reduced model helps in developing physical intuition for the behavior of the instability in this confined geometry, and understanding the effects of certain parameters that are difficult to study experimentally or by simulating the full equations.by Samar Alqatari.S.M.S.M. Massachusetts Institute of Technology, Computation for Design and Optimization Progra