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102 records · Page 6
All sourcesNASA NTRSDOE OSTI
NASA NTRS2011

Infrared-Bolometer Arrays with Reflective Backshorts

Integrated circuits that incorporate square arrays of superconducting-transition- edge bolometers with optically reflective backshorts are being developed for use in image sensors in the spectral range from far infrared to millimeter wavelengths. To maximize the optical efficiency (and, thus, sensitivity) of such a sensor at a specific wavelength, resonant optical structures are created by placing the backshorts at a quarter wavelength behind the bolometer plane. The bolometer and backshort arrays are fabricated separately, then integrated to form a single unit denoted a backshort-under-grid (BUG) bolometer array. In a subsequent fabrication step, the BUG bolometer array is connected, by use of single-sided indium bump bonding, to a readout device that comprises mostly a superconducting quantum interference device (SQUID) multiplexer circuit. The resulting sensor unit comprising the BUG bolometer array and the readout device is operated at a temperature below 1 K. The concept of increasing optical efficiency by use of backshorts at a quarter wavelength behind the bolometers is not new. Instead, the novelty of the present development lies mainly in several features of the design of the BUG bolometer array and the fabrication sequence used to implement the design. Prior to joining with the backshort array, the bolometer array comprises, more specifically, a square grid of free-standing molybdenum/gold superconducting-transition-edge bolometer elements on a 1.4- m-thick top layer of silicon that is part of a silicon support frame made from a silicon-on-insulator wafer. The backshort array is fabricated separately as a frame structure that includes support beams and contains a correspond - ing grid of optically reflective patches on a single-crystal silicon substrate. The process used to fabricate the bolometer array includes standard patterning and etching steps that result in the formation of deep notches in the silicon support frame. These notches are designed to interlock with the support beams on the backshort-array structure to provide structural support and precise relative positioning. The backshort-array structure is inserted in the silicon support frame behind the bolometer array, and the notches in the frame serve to receive the support beams of the backshort-array structure and thus determine the distance between the backshort and bolometer planes. The depth of the notches and, thus, the distance between the backshort and bolometer planes, can be tailored to a value between 25 to 300 m adjusting only a few process steps. The backshort array is designed so as not to interfere with the placement of indium bumps for subsequent indium bump-bonding to the multiplexing readout circuitry

Miller, Timothy M.↗
NASA NTRS2014

Deployable Fresnel Rings

Deployable Fresnel rings (DFRs) significantly enhance the realizable gain of an antenna. This innovation is intended to be used in combination with another antenna element, as the DFR itself acts as a focusing or microwave lens element for a primary antenna. This method is completely passive, and is also completely wireless in that it requires neither a cable, nor a connector from the antenna port of the primary antenna to the DFR. The technology improves upon the previous NASA technology called a Tri-Sector Deployable Array Antenna in at least three critical aspects. In contrast to the previous technology, this innovation requires no connector, cable, or other physical interface to the primary communication radio or sensor device. The achievable improvement in terms of antenna gain is significantly higher than has been achieved with the previous technology. Also, where previous embodiments of the Tri-Sector antenna have been constructed with combinations of conventional (e.g., printed circuit board) and conductive fabric materials, this innovation is realized using only conductive and non-conductive fabric (i.e., "e-textile") materials, with the possible exception of a spring-like deployment ring. Conceptually, a DFR operates by canceling the out-of-phase radiation at a plane by insertion of a conducting ring or rings of a specific size and distance from the source antenna, defined by Fresnel zones. Design of DFRs follow similar procedures to those outlined for conventional Fresnel zone rings. Gain enhancement using a single ring is verified experimentally and through computational simulation. The experimental test setup involves a microstrip patch antenna that is directly behind a single-ring DFR and is radiating towards a second microstrip patch antenna. The first patch antenna and DFR are shown. At 2.42 GHz, the DFR improves the transmit antenna gain by 8.6 dB, as shown in Figure 2, relative to the wireless link without the DFR. A figure illustrates the relative strength of power coupling between the first and second microstrip antennas with and without the DFR. Typically, a DFR is designed for use at a particular frequency; however, testing of a DFR indicated a relatively wide operational bandwidth of approximately 8.2%. Wider bandwidth operation and multi-band operation are anticipated by extending the known art of conventional Fresnel rings to the DFRs. Increasing the number of rings used to construct a DFR antenna increases the gain, with the upper bound limited often by the largest practical dimensions that can be tolerated for a given application. The maximum theoretical improvement in gain for a single ring is 9.5 dB. Experimental results are within 0.9 dB of this theoretical value. Adding rings increases gain, and theoretically, improvements of 10 to 13 dB above that of the primary antenna gain can be achieved with two- and three-ring versions.

Kennedy, Timothy F.↗
NASA NTRS1999

The MSP 2001 Mars Environmental Compatibility Assessment (MECA)

The Mars Environmental Compatibility Assessment (MECA) will evaluate the Martian environment for soil and dust-related hazards to human exploration as part of the Mars Surveyor Program 2001 Lander. Sponsored by the Human Exploration and Development of Space (HEDS) enterprise, MECA's goal is to evaluate potential geochemical and environmental hazards that may confront future Martian explorers, and to guide HEDS scientists in the development of high fidelity Mars soil simulants. The integrated MECA payload contains a wet-chemistry laboratory, a microscopy station, an electrometer to characterize the electrostatics of the soil and its environment, and arrays of material patches to study the abrasive and adhesive properties of soil grains. The instrument will acquire soil samples with a robotic arm equipped with a camera. MECA will examine surface and subsurface soil and dust in order to characterize particle size, shape, hardness, and also physical characteristics that may provide clues to mineralogy. MECA will characterize soil/water mixtures with respect to pH, redox potential, total dissolved ions, and trace toxins. MECA will determine the nature of electrostatic charging associated with excavation of soil, and the influence of ionizing radiation on material properties. It will also observe natural dust accumulation on engineering materials. To accomplish these objectives, MECA is allocated a mass of 10 kg within an enclosure of 35 x 25 x 15 cm. The Wet Chemistry Laboratory (WCL) consists of four identical cells that will accept samples from surface and subsurface regions accessible to the Lander's robotic arm, mix them with water, and perform extensive analysis of the solution. Ion-selective electrodes and related sensors will evaluate total dissolved solids, redox potential, pH, and the concentration of many soluble ions and gases in wet Martian soil. These electrodes can detect potentially dangerous heavy-metal ions, emitted pathogenic gases, and the soil's corrosive potential. Experiments will include cyclic voltammetry and anodic stripping voltammetry. Complementary to the Viking experiments, the chemical laboratory will characterize the water-soil solution rather than emitted gases. Nonetheless, through analysis of dissolved gases it will be able to replicate many of the Viking observations related to oxidants. MECA's microscopy station combines optical and atomic-force microscopy (AFM) in an actively focused, controlled illumination environment to image particles from millimeters to nanometers in size. Careful selection of substrates allows controlled experiments in adhesion, abrasion, hardness, aggregation, magnetic and other properties. Special tools allow primitive manipulation (brushing and scraping) of samples. Soil particle properties including size, shape, color, hardness, adhesive potential (electrostatic and magnetic), will be determined using an array of sample receptacles and collection substrates. The simple, rugged atomic-force microscope will image in the submicron size range and has the capability of performing a particle-by-particle analysis of the dust and soil. On Earth, the earliest forms of life are preserved as microfossils. The atomic-force microscope will have the required resolution to image down to the scale of terrestrial microfossils and beyond. Mounted on the end of the robot arm, MECA's electrometer actually consists of four types of sensors: an electric field meter, several triboelectricity monitors, an ion gauge, and a thermometer. Tempered only by ultraviolet-light-induced ions and a low-voltage breakdown threshold, the dry, cold, dusty martian environment presents an imposing electrostatic hazard to both robots and humans. The field meter will measure the ambient field on nearby objects while the triboelectric sensors, using identical circuitry, will measure the charge accumulated on test substances as they are dragged through the soil by the arm. The ion chamber, open to the environment, will sense both charged dust and free ions in the air. Over and above the potential threat to electronics, the electrostatic environment holds one of the keys to transport of dust and, consequently, Martian meteorology. Viewed with the robot arm camera, the abrasion and adhesion plates are strategically placed to allow direct observation of the interaction between materials and soils on a macroscopic scale. Materials of graded hardness are placed directly under the robot arm scoop to sense wear and soil hardness. A second array, placed on the lander deck, is deployed after the dust plume of landing has settled. It can be manipulated in a primitive fashion by the arm, first having dirt deposited on it from the scoop and subsequently shaken clean. A third array will passively collect dust from the atmosphere. In addition to objectives related to human exploration, the MECA data set will be rich in information relevant to basic geology, paleoclimate, and exobiology issues. To understand both contemporaneous and ancient processes on Mars, the mineralogy, petrology, and reactivity of Martian surface materials should be constrained. The MECA experiment will shed light on these quantities through its combination of chemistry and microscopy. MECA will be capable of measuring the composition of ancient surface water environments, observing microscopic evidence of geological (and biological?) processes, inferring soil and dust transport, comminution and weathering mechanisms, and characterizing soil horizons that might be encountered during excavation.

Hecht, M. H.↗
NASA NTRS2018

CubeX: A Compact X-Ray Telescope Enables Both X-Ray Fluorescence Imaging Spectroscopy and Pulsar Timing Based Navigation

This paper describes the benefits of a miniaturized X-ray telescope payload in the context of a lunar mission. The first part describes the payload in detail, the second part summarizes a small satellite mission concept that utilizes its compact form factor and performance. The CubeX instrument can be used for both X-ray fluorescence (XRF) imaging spectroscopy and X-ray pulsar timing based navigation (XNAV). Using our recent technological advances in X-ray optics and sensors, CubeX combines high angular resolution (<1 arcminutes) Miniature Wolter-I X-ray optics (MiXO) with a common focal plane consisting of high spectral resolution (<150 eV at 1 keV) CMOS X-ray sensors and a high timing resolution (< 1 usec) SDD X-ray sensor. This novel combination of the instruments enables both XRF measurements and XNAV operations without moving parts. The high angular resolution of the MiXO opens a wide range of orbital configurations for observation. Given that performance, the instrument has unprecedented small volume (~1×1×6U), mass (<6 kg), and power (<9W) requirements and opens a wide range of applications for a variety of targets and missions including NEOs and Martian moons. In this paper we illustrate one potential application for a lunar mission concept: The elemental composition of the Moon holds keys to understanding the origin and evolution of both the Moon and the Earth. X-ray fluorescence (XRF), induced either by solar X-ray flux or energetic ions, carries decisive signatures of surface elemental composition. X-ray observations, therefore, give a unique, powerful diagnostic tool for remotely determining elemental abundances including major rock forming elements such as Mg, Al, Na, Si, Fe, and Ca. Through high-resolution XRF imaging spectroscopy, CubeX searches for small patches of elusive lower crust and mantle material excavated within and around impact craters. CubeX identifies regional compositional variations and allows straightforward comparison of elemental distributions with the surface topography from LRO and the gravity data from GRAIL. The elemental compositions of the lower crust and the mantle are sensitive to the conditions of the giant impact which led to the Moon's formation and the subsequent lunar magma ocean (LMO), and thus they are key missing pieces in understanding the formation and early evolution of the Moon. In between XRF observations, CubeX also leverages the technology of high resolution X-ray imaging and time series measurements to conduct XNAV operations and evaluate their performance. Deep space navigation is a critical issue for small planetary missions. XNAV can enable low-cost autonomous deep-space navigation, and has the potential to greatly assist, or even outperform, NASA's Deep Space Network (DSN) or ESA's European Space Tracking (ESTRACK). CubeX is designed to perform sequential observations of 3-4 millisecond pulsars (MSPs) to solve the spacecraft trajectory for absolute navigation, and explore the remote sensing capability of XNAV. In the presented mission concept, the Moon's relative proximity enables a straightforward evaluation of the XNAV performance through DSN.

x-ray spectroscopy↗
DOE OSTI2021

Generative Patch Priors for Practical Compressive Image Recovery

In this work, we propose the generative patch prior (GPP) that defines a generative prior for compressive image recovery, based on patch-manifold models. Unlike learned, image-level priors that are restricted to the range space of a pre-trained generator, GPP can recover a wide variety of natural images using a pre-trained patch generator. Additionally, GPP retains the benefits of generative priors like high reconstruction quality at extremely low sensing rates, while also being much more generally applicable. We show that GPP outperforms several unsupervised and supervised techniques on three different sensing models – linear compressive sensing with known, and unknown calibration settings, and the non-linear phase retrieval problem. Finally, we propose an alternating optimization strategy using GPP for joint calibration-and-reconstruction which performs favorably against several baselines on a real world, uncalibrated compressive sensing dataset. The code and models for GPP are available on github.

compressed sensing↗
NASA NTRS2004

An Overview of My 2004 Summer Internship

This summer I have been working with the Non-destructive evaluation (NDE) group and NASA Glenn Research Center. As this is my second summer with the group, I was able to begin working as soon as I arrived. My first task was to develop a system to acquire an impedance analyzer. The basic setup of the system is as follows: a piezo- electric patch is attached to a sample, and a lead is attached to that patch. Another lead is attached directly to the sample, and the leads are connected to the impedance analyzer. The system then puts a voltage through the material over a range of frequencies, and the corresponding impedances are measured for each frequency. After data is collected, it can be compared to another data set, and through a series of calculations a damage parameter is produced. For the time being, we are using a correlation calculation to find the damage parameter. The hope for this project is that a baseline measurement can be taken, and then sometime later another measurement could be taken, and the damage parameter would determine how much damage had been done to the sample. To test this hypothesis, we took baseline data from a sample, and then sent it out to have a notch cut into it. When it was returned, we again took measurements on the sample, and the damage parameter was significantly lower. Another project that I have been working on pertains to the group's newly acquired acoustography system. This system creates a full field ultrasonic signal on one side of a sample, and an acousto-optic sensor is placed on the other side of the sample.

Hertert, Lucien↗
DOE OSTI2026

Ecological Insights from Transferable Plant Biomass Mapping across the Arctic using High-resolution Structure-from-Motion and LiDAR Data

Warmer temperatures, permafrost thaw, and increased wildfire activity are driving rapid ecological change across the Arctic, significantly altering plant productivity and aboveground biomass (AGB). These rapid changes highlight the urgent need to improve monitoring of vegetation dynamics in the Earth’s northern ecosystems, where high spatiotemporal heterogeneity occurs at scales finer than those captured by traditional satellite observations. The growing use of Unoccupied Aerial Systems (UASs) presents an opportunity to overcome this limitation. Yet, the diversity of UAS platforms, sensors, and data collection and processing workflows presents challenges for developing standardized, generalizable approaches. To address this challenge, we compiled 672 AGB plots co-located with 183 UAS-based Structure-from-Motion (SfM) or Light Detection and Ranging (LiDAR) surveys collected across the Arctic. Here, we: (1) evaluated the generalizability of UAS-derived canopy structure derived from high-resolution SfM and LiDAR for estimating AGB, (2) assessed scaling errors and their sources in two recent satellite-based AGB products derived from Landsat and MODIS, and (3) demonstrated the use of high-resolution AGB maps to quantify biomass variation across tundra plant functional types (PFTs) and to monitor post-fire recovery. Our results show that both SfM and LiDAR accurately captured AGB and its variability across tundra PFTs using a Random Forest (RF) model (overall RMSE: 0.336 kg/m2), with mapping performance varying slightly by region and data source. Using UAS-derived AGB maps as a benchmark, we identified systematic biases in satellite-derived AGB products, largely attributable to the magnitude of AGB and structural heterogeneity within coarse-resolution pixels. Applying our model to repeat UAS surveys following a tundra fire on Seward Peninsula, we observed rapid AGB recovery in non-shrub patches, with biomass recovering to pre-fire levels within 2 years. In contrast, shrub patches recovered more slowly, with AGB gains continuing over 2–4 years through both in-patch growth and lateral expansion (via dispersal) into remaining burned areas. Overall, these findings demonstrate the generalizability of UAS-based SfM and LiDAR data for estimating tundra AGB and highlight the potential of our approach to be broadly applied to generate high-quality AGB data for ecological monitoring and model benchmarking across the Arctic.

Yang, Daryl [ORNL] (ORCID:0000000317057823)↗
NASA NTRS1999

Global Distribution of Aerosols Over the Open Ocean as Derived from the Coastal Zone Color Scanner

Climatological maps of monthly mean aerosol radiance levels derived from the coastal zone color scanner (CZCS) were constructed for the world's ocean basins. This is the first study to use the 7.5.-year CZCS data set to examine the distribution and seasonality of aerosols over the open ocean on a global scale. Examination of our satellite images found the most prominent large-scale patch of elevated aerosol radiances in each month off the coast of northwest Africa. The well-known, large-scale plumes of elevated aerosol levels in the Arabian Sea, the northwest Pacific, and off the east coast of North America were also successfully captured. Radiance data were extracted from 13 major open-ocean zones, ranging from the subpolar to equatorial regions. Results from these extractions revealed the aerosol load in both subpolar and subtropical zones to be higher in the Northern Hemisphere than in the Southern Hemisphere. Aerosol radiances in the subtropics of both hemispheres were about 2 times higher in summer than in winter. In subpolar regions, aerosol radiances in late spring/early summer were almost 3 times that observed in winter. In general, the aerosol signal was higher during the warmer months and lower during the cooler months, irrespective of location. A comparison between our mean monthly aerosol radiance maps with mean monthly chlorophyll maps (also from CZCS) showed similar seasonality between aerosol and chlorophyll levels in the subpolar zones of both hemispheres, i.e., high levels in summer, low levels in winter. In the subtropics of both hemispheres, however, chlorophyll levels were higher in winter months which coincided with a depressed aerosol signal. Our results indicate that the near-IR channel on ocean color sensors can be used to successfully capture well-known, large-scale aerosol plumes on a global scale and that future ocean color sensors may provide a platform for long-term synoptic studies of combined aerosol-phytoplankton productivity interactions.

Stegmann, P. M.↗
NASA NTRS2014

Features of Point Clouds Synthesized from Multi-View ALOS/PRISM Data and Comparisons with LiDAR Data in Forested Areas

LiDAR waveform data from airborne LiDAR scanners (ALS) e.g. the Land Vegetation and Ice Sensor (LVIS) havebeen successfully used for estimation of forest height and biomass at local scales and have become the preferredremote sensing dataset. However, regional and global applications are limited by the cost of the airborne LiDARdata acquisition and there are no available spaceborne LiDAR systems. Some researchers have demonstrated thepotential for mapping forest height using aerial or spaceborne stereo imagery with very high spatial resolutions.For stereo imageswith global coverage but coarse resolution newanalysis methods need to be used. Unlike mostresearch based on digital surface models, this study concentrated on analyzing the features of point cloud datagenerated from stereo imagery. The synthesizing of point cloud data from multi-view stereo imagery increasedthe point density of the data. The point cloud data over forested areas were analyzed and compared to small footprintLiDAR data and large-footprint LiDAR waveform data. The results showed that the synthesized point clouddata from ALOSPRISM triplets produce vertical distributions similar to LiDAR data and detected the verticalstructure of sparse and non-closed forests at 30mresolution. For dense forest canopies, the canopy could be capturedbut the ground surface could not be seen, so surface elevations from other sourceswould be needed to calculatethe height of the canopy. A canopy height map with 30 m pixels was produced by subtracting nationalelevation dataset (NED) fromthe averaged elevation of synthesized point clouds,which exhibited spatial featuresof roads, forest edges and patches. The linear regression showed that the canopy height map had a good correlationwith RH50 of LVIS data with a slope of 1.04 and R2 of 0.74 indicating that the canopy height derived fromPRISM triplets can be used to estimate forest biomass at 30 m resolution.

LiDARD↗
DOE OSTI2020

Coupling of Ca 2+ and voltage activation in BK channels through the αB helix/voltage sensor interface

Large-conductance Ca 2+ and voltage-activated K + (BK) channels control membrane excitability in many cell types. BK channels are tetrameric. Each subunit is composed of a voltage sensor domain (VSD), a central pore-gate domain, and a large cytoplasmic domain (CTD) that contains the Ca 2+ sensors. While it is known that BK channels are activated by voltage and Ca 2+ , and that voltage and Ca 2+ activations interact, less is known about the mechanisms involved. In this work we explore these mechanisms by examining the gating contribution of an interface formed between the VSDs and the αB helices located at the top of the CTDs. Proline mutations in the αB helix greatly decreased voltage activation while having negligible effects on gating currents. Analysis with the Horrigan, Cui, and Aldrich model indicated a decreased coupling between voltage sensors and pore gate. Proline mutations decreased Ca 2+ activation for both Ca 2+ bowl and RCK1 Ca 2+ sites, suggesting that both high-affinity Ca 2+ sites transduce their effect, at least in part, through the αB helix. Mg 2+ activation also decreased. The crystal structure of the CTD with proline mutation L390P showed a flattening of the first helical turn in the αB helix compared to wild type, without other notable differences in the CTD, indicating that structural changes from the mutation were confined to the αB helix. These findings indicate that an intact αB helix/VSD interface is required for effective coupling of Ca 2+ binding and voltage depolarization to pore opening and that shared Ca 2+ and voltage transduction pathways involving the αB helix may be involved.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS2000

First-Generation Jet Propulsion Laboratory "Hockey-Puck" Free-Flying Magnetometers for Distributed In-Situ Multiprobe Measurement of Current Density Filamentation in the Northern Auroral Zone: Enstrophy Mission

The sub-orbital rocket mission was a collaborative project between the University of New Hampshire, Cornell University, and the Jet Propulsion Laboratory (JPL) to study filamentation phenomena in the northern Auroral zone. The Enstrophy mission test flies the JPL Free-Flying Magnetometer (FFM) concept. The FFM technology development task has been funded by NASA develop miniaturized, low-power, integrated "sensorcrafts". JPL's role was to design, integrate, test, and deliver four FFMs for deployment from the sounding rocket, allowing a unique determination of curl-B. This provides a direct measurement of magnetic-field-aligned current density along the rocket trajectory. A miniaturized three-axis fluxgate magnetometer was integrated with a 4-channel 22-bit sigma-delta Analog to Digital Converter (ADC), four temperature sensors, digital control electronics, seven (Li-SOCl2) batteries, two (4 deg x 170 deg field of view) sun-sensors, a fan-shaped-beam laser diode beacon, a (16 MHz) stable Temperature Compensated Crystal Oscillator (TCXO) clock, Radio Frequency (RF) communication subsystem, and an antenna for approximately 15 minutes of operation where data was collected continuously and transmitted in three (3) bursts (approximately 26 seconds each) to ground station antennas at Poker Flat, Alaska. FFMs were stowed within two trays onboard the rocket during the rocket launch and were released simultaneously using the spinning action of the rocket at approximately 300 km altitude (approximately 100 sec. into the flight). FFMs were deployed with spin rate of approximately 17 Hz and approximately 3 m/sec linear velocity with respect to the rocket. For testing purposes while the rocket was in the launch pad and during flight prior to release of FFMs from the rocket, commands (such as "power on", "test", "flight", "power off', and clock "Reset" signal) were transmitted via a infrared Light Emitting Diode to an infrared detector in the FFM. Special attention was paid to low magnetic signature electronic design and choice of materials in packaging. The miniaturized fluxgate magnetometers had a range of 1-60000 nT with 0.1% full-scale linearity. The frequency range of interest for magnetic measurement was 10 mHz - 50 Hz. Digital data from the magnetometer's three axes were placed in a 4MB Static Random Access Memory (SRAM) in data packages (frames) formatted together with time tags and frame ID. After a specified time was elapsed, the data were Viterbi encoded and transmitted at a rate of 100 kbps (BPSK). Each of the four FFMs transmitted at different frequency. These carrier frequencies were in the range of 2200-2300 MHz. The antenna was a single patch on a high dielectric constant substrate covering one end-plate of the hockey-puck-sized unit. The local clocks aboard the FFMs were reset at the start of the mission and stayed synchronized within 3 msec during the mission. Position of each FFM with respect to the rocket is calculated by the knowledge of its release velocity (measured at exit point of the FFM launcher tract) providing an accuracy of 1 m over the maximum range of 3 km. Spatial and temporal nature of observants can be separated to within 3 m in space or 3 msec time interval.

Javadi, H.↗
NASA NTRS2003

Physiological Maturation of Regenerating Hair Cells

The bullfrog saccule, a sensor of gravity and substrate-borne vibration, is a model system for hair cell transduction. Saccular hair cells also increase in number throughout adult life and rapidly recover after hair cell damage, making this organ an ideal system for studying hair cell development, repair, and regeneration. We have used of hair cell and supporting cell immunocytochemical markers to identify damaged hair cells and hair cell precursors in organotypic cultures of the bullfrog saccule. We then used an innovative combination of confocal, electron, and time-lapse microscopy to study the fate of damaged hair cells and the origin of new hair cells after gentamicin ototoxicity in normal and mitotically blocked saccular cultures. These studies have shown that gentamicin ototoxicity produces both lethal and sublethal hair cell damage. They have also shown that hair cell recovery in this organ takes place by both the repair of sublethally damaged hair cells and by the replacement of lost hair cells by mitotic regeneration. In parallel studies, we have used biophysical and molecular biological techniques to study the differentiation and innervation of developing, repairing, and regenerating hair cells. More specifically, we have used RT-PCR to obtain the bullfrog homologues of L-type voltage- gated calcium (L-VGCC) and large-conductance Ca(2+)-activated potassium (BK) channel genes. We have then obtained probes for these genes and, using in situ hybridization, begun to examine their expression in the bullfrog saccule and amphibian papilla. We have also used fluorescent-labeled channel toxins and channel toxin derivatives to determine the time of appearance of L-type voltage-gated calcium (L-VGCC) and Ca(2+)-activated potassium (BK) channels and to study dynamic changes in the number, distribution, and co-localization of these proteins in developing, repairing, and regenerating hair cells. Using time-lapse microscopy, we are also studying the dynamic relationship between ion channel clustering and synaptic formation in hair cells and afferent neurons. In future studies, we will determine when hair cell precursors acquire electrical tuning, and, using whole-cell patch-clamp techniques, identify and characterize their L-VGCC and BK currents. We will also use biophysical techniques to determine the number of L-VGCC and BK channels and the size and gating kinetics of their underlying L-VGCC and BK conductances, correlating these variables with the amplitude and frequency of membrane oscillations produced by intracellular current steps. We expect these studies to determine how hair cells regulate ion channel expression to achieve specific physiological responses.

Baird, Richard A.↗