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At least 739 records · Page 41

Observations of Ocean Primary Productivity Using MODIS

Measuring the magnitude and variability of oceanic net primary productivity (NPP) represents a key advancement toward our understanding of the dynamics of marine ecosystems and the role of the ocean in the global carbon cycle. MODIS observations make two new contributions in addition to continuing the bio-optical time series begun with Orbview-2's SeaWiFS sensor. First, MODIS provides weekly estimates of global ocean net primary productivity on weekly and annual time periods, and annual empirical estimates of carbon export production. Second, MODIS provides additional insight into the spatial and temporal variations in photosynthetic efficiency through the direct measurements of solar-stimulated chlorophyll fluorescence. The two different weekly productivity indexes (first developed by Behrenfeld & Falkowski and by Yoder, Ryan and Howard) are used to derive daily productivity as a function of chlorophyll biomass, incident daily surface irradiance, temperature, euphotic depth, and mixed layer depth. Comparisons between these two estimates using both SeaWiFS and MODIS data show significant model differences in spatial distribution after allowance for the different integration depths. Both estimates are strongly dependence on the accuracy of the chlorophyll determination. In addition, an empirical approach is taken on annual scales to estimate global NPP and export production. Estimates of solar stimulated fluorescence efficiency from chlorophyll have been shown to be inversely related to photosynthetic efficiency by Abbott and co-workers. MODIS provides the first global estimates of oceanic chlorophyll fluorescence, providing an important proof of concept. MODIS observations are revealing spatial patterns of fluorescence efficiency which show expected variations with phytoplankton photo-physiological parameters as measured during in-situ surveys. This has opened the way for research into utilizing this information to improve our understanding of oceanic NPP variability. Deriving the ocean bio-optical properties places severe demands on instrument performance (especially band to band precision) and atmospheric correction. Improvements in MODIS instrument characterization and calibration over the first 16 mission months have greatly improved the accuracy of the chlorophyll input fields and FLH, and therefore the estimates of NPP and fluorescence efficiency. Annual estimates now show the oceanic NPP accounts for 40-50% of the global total NPP, with significant interannual variations related to large scale ocean processes. Spatial variations in ocean NPP, and exported production, have significant effects on exchange of CO2 between the ocean and atmosphere. Further work is underway to improve both the primary productivity model functions, and to refine our understanding of the relationships between fluorescence efficiency and NPP estimates. We expect that the MODIS instruments will prove extremely useful in assessing the time dependencies of oceanic carbon uptake and effects of iron enrichment, within the global carbon cycle.

Esaias, Wayne E.↗

Role of Growth Hormone, Exercise and Serum Phosphorus in Unloaded Bone of Young Rats

Growth hormone, known to be stimulated by exercise, is suppressed in rats after space flight and in a ground-based model in which the hind-limbs are unloaded (S). To determine the role of GH in the osteopenia of unloaded bones of S rats, young males were treated with GH combined with insulin-like growth factor-1 (IGF-1), a peptide that mediates the local actions of the hormone. 200 g rats, hypophysectomized (hypox) 17 d earlier, were treated with 1 mg/kg/d GH/IGF-1 (H) or saline (C) in 3 divided daily doses x10 d. Hind-limb bones were unloaded (S), ambulated (A) or exercised (X) by climbing a ladder while carrying a weight. Growth was monitored daily. Tibial growth plate (Tepi) was measured with a micrometer, and femoral (F) area, length, and mineral content (BMC) by DEXA. Parameters of calcium metabolism were measured by autoanalyzer and calciotropic hormones by radioimmunoassay. F bone density, g/square cm, (BMD) or BW were not affected by S in Hypox. However, FBMD was lower in S+H than A+H (p is less than 0.002) and H stimulated whole body growth in S (5.2 g/d) and SX (5.6 g/d) to a lesser extent than in A (6.6 g/d) (p is less than 0.05). Adjusted for BW, Tepi showed the greatest increase in S+H+X (64%), the next highest increase in S+H (50%) and no change in S+X. F area, length and BMC/100 g BW were lower in all H groups than respective C's. By multiple regression analysis, serum phosphorus (Pi) which correlated with Tepi (r = 0.88, p is less than 0.001) and was inversely related to FBMC (r = -0.68, p is less than 0.001) proved to be the most significant determinant of BMC. This illustrates the dependence of osteopenia in S on GH, the maximizing effect of X for epiphyseal growth and the major role of Pi metabolism on BMC in weight bearing bone during growth.

Arnnaud, Sara B.↗

Statistical Study of Auroral Kilometric Radiation Fine Structure Striations Observed by Polar

We have conducted a statistical survey of a semirandom sample of the auroral kilometric radiation (AKR) data observed by the plasma wave instrument wideband receiver on board the Polar spacecraft. We have determined that AKR fine structure patterns with very narrowband, negative drifting striations occur in approximately 6% of the high-resolution wideband spectrograms when AKR is present. Positive sloping striations are also observed, but at a much lower rate. More than 8200 AKR stripes have been scaled. The stripes are predominantly found in the 40 to 215-kHz frequency range and have a frequency extent of about 4 kHz and a duration of usually less than 2 s. The majority of the stripes have drift rates between -8 and -2 kHz/s, with a peak in the distribution between -6 and -4 kHz/s. There is also a much smaller group of striations with positive drift rates of up to about 5 or 6 kHz/s. We have further investigated the change of drift rate with frequency. Almost all striations are observed in the lowest two frequency bands of the wideband receiver (f < 215 kHz). There is an increase in the statistical drift rate with increasing frequency. The statistical slope of the striations increases with frequency from about -4.4 kHz/s at 75 kHz to about -5.7 kHz/s at 170 kHz. This frequency dependence of the drift rate is consistent, under certain conditions, with a production mechanism stimulated by an upward propagating electromagnetic ion cyclotron wave, as had been suggested earlier. However, such a changing drift rate is also compatible with a stimulated source region that propagates upward along the magnetic field line at the velocity of an ion beam accelerated by a local, upward directed electric field, as is typically observed in the auroral region. An explanation for this association is not apparent at this time.

Menietti, J. D.↗

The Semicircular Canal Microphonic

Present experiments were designed to quantify the alternating current (AC) component of the semicircular canal microphonic for angular motion stimulation as a function of stimulus frequency and amplitude. The oyster toadfish, Opsanus tau, was used as the experimental model. Calibrated mechanical indentation of the horizontal canal duct was used as a stimulus to generate hair-cell and afferent responses reproducing those present during head rotation. Sensitivity to polarization of the endolymph DC voltage re: perilymph was also investigated. Modulation of endolymph voltage was recorded using conventional glass electrodes and lock-in amplification over the frequency range 0.2-80 Hz. Access to the endolymph for inserting voltage recording and current passing electrodes was obtained by sectioning the anterior canal at its apex and isolating the cut ends in air. For sinusoidal stimulation below approx.10 Hz, the horizontal semicircular canal AC microphonic was nearly independent of stimulus frequency and equal to approximately 4 microV per micron indent (equivalent to approx. 1 microV per deg/s). A saturating nonlinearity decreasing the microphonic gain was present for stimuli exceeding approx.3 micron indent (approx. 12 deg/s angular velocity). The phase was not sensitive to the saturating nonlinearity. The microphonic exhibited a resonance near 30Hz consistent with basolateral current hair cell resonance observed previously in voltage-clamp records from semicircular canal hair cells. The magnitude and phase of the microphonic exhibited sensitivity to endolymphatic polarization consistent with electro-chemical reversal of hair cell transduction currents.

Rabbitt, R. D.↗

Commercial Research and Development: Power to Explore, Opportunities from Discovery

The technical and economic goals of commercial use of space are laudable, and are addressed as a high priority by almost every national space program and most major aerospace companies the world over. Yet, the focus of most organizational agendas and discussions tends to focus on one or two very narrow enabling aspects of this potentially large technological and economic opportunity. While government sponsored commercial launch activities and private space platforms are an integral part of efforts to leverage the commercial use of space, these activities are possibly one of the smallest parts of creating, a viable and sustainable market for the commercial use of space. Most of the current programs usually do not appropriately address some of the critical issues of the current, already interested, potential space user communities. Current programs place the focus of the majority of the user requirements on the vehicle payload weight and mass performance considerations as the primary payload economical factor in providing a commercial market with a stimulating price for gaining access to the space environment. The larger user challenges of transformation from Earth-based research and development approaches to space environment approaches are not addressed early enough in programs to impact the new business considerations of potential users. Currently, space-based research and development user activities require a large user investment in time, in development of new areas of support expertise, in development of new systems, in risk of schedule to completion, and in long term capital positioning. The larger opportunities for stimulating a strong market driven interest in commercial use of space that could result from the development of vehicle payload "leap ahead technologies" for users are being missed, and there is a real risk of limiting the potentially broader market base to support a more technologically advanced and economically lucrative outcome. A major driving force for strengthening the commercial space activities is not only the technological advances in launch vehicle, or newer satellites, but the myriad of enabling payloads technologies that could, as a goal, result in an almost transparent facilitation to regular CD a, -n access to space and microgravity environments by the future users from the existing Earth-based research and development organizations market segments. Rather than focusing only on developing high lift performance launch vehicles and then developing payloads to fit them, the real focus from a business model perspective should to be on the customer payloads requirements, and on designing launch vehicles and platforms systems for a space transportation and facility infrastructure to support all aspects of the business model for the user market. To harness the full potential of space commercialization, new efforts need to be made to comprehensively examine all the critical business model areas for commercial research, development, and manufacturing in space so as to identify specific products and efforts; to determine how such operations must be both similar to and different from current Earth-based activities; to evaluate the enabling technological devices, processes and efforts so that like efforts can be addressed in a synergistic fashion for maximum user cost effectiveness; to delineate the services that are both needed and can be provided by such activities; and to use this information to drive design and development of space commercialization efforts and policy.

Casas, Joseph C.↗

Relaxation System

Environ Corporation's relaxation system is built around a body lounge, a kind of super easy chair that incorporates sensory devices. Computer controlled enclosure provides filtered ionized air to create a feeling of invigoration, enhanced by mood changing aromas. Occupant is also surrounded by multidimensional audio and the lighting is programmed to change colors, patterns, and intensity periodically. These and other sensory stimulators are designed to provide an environment in which the learning process is stimulated, because research has proven that while an individual is in a deep state of relaxation, the mind is more receptive to new information.

Source record↗

The Use of NASA Light-Emitting Diode Near-Infrared Technology for Biostimulation

Studies on cells exposed to microgravity and hypergravity indicate that human cells need gravity to stimulate growth. As the gravitational force increases or decreases, the cell function responds in a linear fashion. This poses significant health risks for astronauts in long-term spaceflight. The application of light therapy with the use of NASA LEDs will significantly improve the medical care that is available to astronauts on long-term space missions. NASA LEDs stimulate the basic energy processes in the mitochondria (energy compartments) of each cell, particularly when near-infrared light is used to activate the color sensitive chemicals (chromophores, cytochrome systems) inside. Optimal LED wavelengths include 680, 730 and 880 nm and our laboratory has improved the healing of wounds in laboratory animals by using both NASA LED light and hyperbaric oxygen. Furthermore, DNA synthesis in fibroblasts and muscle cells has been quintupled using NASA LED light alone, in a single application combining 680, 730 and 880 nm each at 4 Joules per centimeter squared. Muscle and bone atrophy are well documented in astronauts, and various minor injuries occurring in space have been reported not to heal until landing on Earth. An LED blanket device may be used for the prevention of bone and muscle atrophy in astronauts. The depth of near-infrared light penetration into human tissue has been measured spectroscopically.

Whelan, Harry T.↗

Space Derived Health Aids (PRN, Neuropacemaker)

St. Jude Medical's (formerly known as Pacesetter Systems, Inc.) Programmable Rechargeable Neuropacemaker (PRN) is an implantable human tissue stimulator designed to provide relief to patients with disorders treatable by electrical stimulation. Examples are back, leg, and arm pain, cancer pain and multiple sclerosis.

Source record↗

Visual and Vestibular Determinants of Perceived Eye-Level

Both gravitational and optical sources of stimulation combine to determine the perceived elevations of visual targets. The ways in which these sources of stimulation combine with one another in operational aeronautical environments are critical for pilots to make accurate judgments of the relative altitudes of other aircraft and of their own altitude relative to the terrain. In a recent study, my colleagues and I required eighteen observers to set visual targets at their apparent horizon while they experienced various levels of G(sub z) in the human centrifuge at NASA-Ames Research Center. The targets were viewed in darkness and also against specific background optical arrays that were oriented at various angles with respect to the vertical; target settings were lowered as Gz was increased; this effect was reduced when the background optical array was visible. Also, target settings were displaced in the direction that the background optical array was pitched. Our results were attributed to the combined influences of otolith-oculomotor mechanisms that underlie the elevator illusion and visual-oculomotor mechanisms (optostatic responses) that underlie the perceptual effects of viewing pitched optical arrays that comprise the background. In this paper, I present a mathematical model that describes the independent and combined effects of G(sub z) intensity and the orientation and structure of background optical arrays; the model predicts quantitative deviations from normal accurate perceptions of target localization under a variety of conditions. Our earlier experimental results and the mathematical model are described in some detail, and the effects of viewing specific optical arrays under various gravitational-inertial conditions encountered in aeronautical environments are discussed.

Cohen, Malcolm Martin↗

Neural Control of the Cardiovascular System in Space

During the acute transition from lying supine to standing upright, a large volume of blood suddenly moves from the chest into the legs. To prevent fainting, the blood pressure control system senses this change immediately, and rapidly adjusts flow (by increasing heart rate) and resistance to flow (by constricting the blood vessels) to restore blood pressure and maintain brain blood flow. If this system is inadequate, the brain has a backup plan. Blood vessels in the brain can adjust their diameter to keep blood flow constant. If blood pressure drops, the brain blood vessels dilate; if blood pressure increases, the brain blood vessels constrict. This process, which is called autoregulation, allows the brain to maintain a steady stream of oxygen, even when blood pressure changes. We examined what changes in the blood pressure control system or cerebral autoregulation contribute to the blood pressure control problems seen after spaceflight. We asked: (1) does the adaptation to spaceflight cause an adaptation in the blood pressure control system that impairs the ability of the system to constrict blood vessels on return to Earth?; (2) if such a defect exists, could we pinpoint the neural pathways involved?; and (3) does cerebral autoregulation become abnormal during spaceflight, impairing the body s ability to maintain constant brain blood flow when standing upright on Earth? We stressed the blood pressure control system using lower body negative pressure, upright tilt, handgrip exercise, and cold stimulation of the hand. Standard cardiovascular parameters were measured along with sympathetic nerve activity (the nerve activity causing blood vessels to constrict) and brain blood flow. We confirmed that the primary cardiovascular effect of spaceflight was a postflight reduction in upright stroke volume (the amount of blood the heart pumps per beat). Heart rate increased appropriately for the reduction in stroke volume, thereby showing that changes in heart rate regulation alone cannot be responsible for orthostatic hypotension after spaceflight. All of the astronauts in our study had an increase in sympathetic nerve activity during upright tilting on Earth postflight. This increase was well calibrated for the reduction in stroke volume induced by the upright posture. The results obtained from stimulating the sympathetic nervous system using handgrip exercise or cold stress were also entirely normal during and after spaceflight. No astronaut had reduced cerebral blood flow during upright tilt, and cerebral autoregulation was normal or even enhanced inflight. These experiments show that the cardiovascular adaptation to spaceflight does not lead to a defect in the regulation of blood vessel constriction via sympathetic nerve activity. In addition, cerebral autoregulation is well-maintained. It is possible that despite the increased sympathetic nerve activity, blood vessels did not respond with a greater degree of constriction than occurred preflight, possibly uncovering a limit of vasoconstrictor reserve.

Levine, Benjamin D.↗

Geologic Mapping Applications Using THEMIS Data for the Medusae Fossae Formation, Mars

The Medusae Fossae Formation (MFF) is a regionally extensive deposit located along the equator of Mars between roughly 130 and 240 E longitude, the origin of which has stimulated a host of published hypotheses. A volcanic or aeolian origin appear most consistent with Viking and MGS data, but other hypotheses remain viable and new data, as from the Mars Odyssey spacecraft, is likely to stimulate additional hypotheses of origin. NASA is supporting geologic mapping of portions of the MFF deposits, but it is now quite clear that this on-going mapping will need considerable revision as data from the Thermal Emission Imaging System (THEMIS) on Mars Odyssey become available. The daytime IR THEMIS images hold particularly strong potential for providing a new base on which geologic mapping can be carried out, as illustrated by the examples discussed.

Zimbelman, J. R.↗

Cellular Mechanisms Underlying Bone-Forming Cell Proliferative Response to Hypergravity

Life on Earth has evolved under the continuous influence of gravity (1-g). As humans explore and develop space, however, we must learn to adapt to an environment with little or no gravity. Studies indicate that lack of weightbearing for vertebrates occurring with immobilization, paralysis, or in a microgravity environment may cause muscle and bone atrophy through cellular and subcellular level mechanisms. We hypothesize that gravity is needed for the efficient transduction of cell growth and survival signals from the extra-cellular matrix (ECM) (consisting of molecules such as collagen, fibronectin, and laminin) in mechanosensitive tissues. We test for the presence of gravity-sensitive pathways in bone-forming cells (osteoblasts) using hypergravity applied by a cell culture centrifuge. Stimulation of 50 times gravity (50-g) increased proliferation in primary rat osteoblasts for cells grown on collagen Type I and fibronectin, but not on laminin or uncoated surfaces. Survival was also enhanced during hypergravity stimulation by the presence of ECM. Bromodeoxyuridine incorporation in proliferating cells showed an increase in the number of actively dividing cells from about 60% at 1-g to over 90% at 25-g. Reverse transcription-polymerase chain reaction was used to test for all possible integrins. Our combined results indicate that beta1 and/or beta3 integrin subunits may be involved. These data indicate that gravity mechanostimulation of osteoblast proliferation involves specific matrix-integrin signalling pathways which are sensitive to g-level. Further research to define the mechanisms involved will provide direction so that we may better adapt and counteract bone atrophy caused by the lack of weightbearing.

Vercoutere, W.↗

Is there a common mechanism underlying genomic instability, bystander effects and other nontargeted effects of exposure to ionizing radiation?

A number of nontargeted and delayed effects associated with radiation exposure have now been described. These include radiation-induced genomic instability, death-inducing and bystander effects, clastogenic factors and transgenerational effects. It is unlikely that these nontargeted effects are directly induced by cellular irradiation. Instead, it is proposed that some as yet to be identified secreted factor can be produced by irradiated cells that can stimulate effects in nonirradiated cells (death-inducing and bystander effects, clastogenic factors) and perpetuate genomic instability in the clonally expanded progeny of an irradiated cell. The proposed factor must be soluble and capable of being transported between cells by cell-to-cell gap junction communication channels. Furthermore, it must have the potential to stimulate cellular cytokines and/or reactive oxygen species. While it is difficult to imagine a role for such a secreted factor in contributing to transgenerational effects, the other nontargeted effects of radiation may all share a common mechanism.

Non-NASA Center↗

Sensorimotor aspects of high-speed artificial gravity: III. Sensorimotor adaptation

As a countermeasure to the debilitating physiological effects of weightlessness, astronauts could live continuously in an artificial gravity environment created by slow rotation of an entire spacecraft or be exposed to brief daily "doses" in a short radius centrifuge housed within a non-rotating spacecraft. A potential drawback to both approaches is that head movements made during rotation may be disorienting and nauseogenic. These side effects are more severe at higher rotation rates, especially upon first exposure. Head movements during rotation generate aberrant vestibular stimulation and Coriolis force perturbations of the head-neck motor system. This article reviews our progress toward distinguishing vestibular and motor factors in side effects of rotation, and presents new data concerning the rates of rotation up to which adaptation is possible. We have studied subjects pointing to targets during constant velocity rotation, because these movements generate Coriolis motor perturbations of the arm but do not involve unusual vestibular stimulation. Initially, reaching paths and endpoints are deviated in the direction of the transient lateral Coriolis forces generated. With practice, subjects soon move in straighter paths and land on target once more. If sight of the arm is permitted, adaptation is more rapid than in darkness. Initial arm movement trajectory and endpoint deviations are proportional to Coriolis force magnitude over a range of rotation speeds from 5 to 20 rpm, and there is rapid, complete motor adaptation at all speeds. These new results indicate that motor adaptation to high rotation rates is possible. Coriolis force perturbations of head movements also occur in a rotating environment but adaptation gradually develops over the course of many head movements.

Non-NASA Center↗

Host resistance of CD18 knockout mice against systemic infection with Listeria monocytogenes

Mice with targeted mutations of CD18, the common beta2 subunit of CD11/CD18 integrins, have leukocytosis, impaired transendothelial neutrophil emigration, and reduced host defense to Streptococcus pneumoniae, a gram-positive extracellular bacterium. Previous studies using blocking monoclonal antibodies suggested roles for CD18 and CD11b in hepatic neutrophil recruitment and host innate response to Listeria monocytogenes, a gram-positive intracellular bacterium. We induced systemic listeriosis in CD18 knockout (CD18-ko) and wild-type (WT) mice by tail vein injection with Listeria. By 14 days postinjection (dpi), 8 of 10 WT mice died, compared with 2 of 10 CD18-ko mice (P < 0.01). Quantitative organ culture showed that numbers of Listeria organisms in livers and spleens were similar in both groups at 20 min postinfection. By 3, 5, and 7 dpi, however, numbers of Listeria organisms were significantly lower in livers and spleens of CD18-ko mice than in WT mice. Histopathology showed that following Listeria infection, CD18-ko mice had milder inflammatory and necrotizing lesions in both spleens and livers than did WT mice. Cytokine assays indicated that baseline interleukin-1beta and granulocyte colony-stimulating factor (G-CSF) levels were higher in CD18-ko mice than in WT mice and that CD18-ko splenocytes produced higher levels of interleukin-1beta and G-CSF than WT splenocytes under the same amount of Listeria stimulation. These findings show that CD18 is not an absolute requirement for antilisterial innate immunity or hepatic neutrophil recruitment. We propose that the absence of CD18 in the mice results in the priming of innate immunity, as evidenced by elevated cytokine expression, and neutrophilic leukocytosis, which augments antilisterial defense.

Non-NASA Center↗

The human respiratory gate

Respiratory activity phasically alters membrane potentials of preganglionic vagal and sympathetic motoneurones and continuously modulates their responsiveness to stimulatory inputs. The most obvious manifestation of this 'respiratory gating' is respiratory sinus arrhythmia, the rhythmic fluctuations of electrocardiographic R-R intervals observed in healthy resting humans. Phasic autonomic motoneurone firing, reflecting the throughput of the system, depends importantly on the intensity of stimulatory inputs, such that when levels of stimulation are low (as with high arterial pressure and sympathetic activity, or low arterial pressure and vagal activity), respiratory fluctuations of sympathetic or vagal firing are also low. The respiratory gate has a finite capacity, and high levels of stimulation override the ability of respiration to gate autonomic responsiveness. Autonomic throughput also depends importantly on other factors, including especially, the frequency of breathing, the rate at which the gate opens and closes. Respiratory sinus arrhythmia is small at rapid, and large at slow breathing rates. The strong correlation between systolic pressure and R-R intervals at respiratory frequencies reflects the influence of respiration on these two measures, rather than arterial baroreflex physiology. A wide range of evidence suggests that respiratory activity gates the timing of autonomic motoneurone firing, but does not influence its tonic level. I propose that the most enduring significance of respiratory gating is its use as a precisely controlled experimental tool to tease out and better understand otherwise inaccessible human autonomic neurophysiological mechanisms.

Review↗

4,4,4-trifluoro-3-(indole-3-)butyric acid promotes root elongation in Lactuca sativa independent of ethylene synthesis and pH

We studied the mode of action of 4,4,4-trifluoro-3- (indole-3-) butyric acid (TFIBA), a recently described root growth stimulator, on primary root growth of Lactuca sativa L. seedlings. TFIBA (100 micromoles) promoted elongation of primary roots by 40% in 72 h but inhibited hypocotyl growth by 35%. TFIBA induced root growth was independent of pH. TFIBA did not affect ethylene production, but reduced the inhibitory effect of ethylene on root elongation. TFIBA promoted root growth even in the presence of the ethylene biosynthesis inhibitor L-alpha-(2-aminoethoxyvinyl)glycine. TFIBA and the ethylene-binding inhibitor silver thiosulphate (STS) had a similar effect on root elongation. The results indicate that TFIBA-stimulated root elongation was neither pH-dependent nor related to inhibition of ethylene synthesis, but was possibly related to ethylene action.

Non-NASA Center↗

Up-regulation of phosphoinositide metabolism in tobacco cells constitutively expressing the human type I inositol polyphosphate 5-phosphatase

To evaluate the impact of suppressing inositol 1,4,5-trisphosphate (InsP(3)) in plants, tobacco (Nicotiana tabacum) cells were transformed with the human type I inositol polyphosphate 5-phosphatase (InsP 5-ptase), an enzyme which specifically hydrolyzes InsP(3). The transgenic cell lines showed a 12- to 25-fold increase in InsP 5-ptase activity in vitro and a 60% to 80% reduction in basal InsP(3) compared with wild-type cells. Stimulation with Mas-7, a synthetic analog of the wasp venom peptide mastoparan, resulted in an approximately 2-fold increase in InsP(3) in both wild-type and transgenic cells. However, even with stimulation, InsP(3) levels in the transgenic cells did not reach wild-type basal values, suggesting that InsP(3) signaling is compromised. Analysis of whole-cell lipids indicated that phosphatidylinositol 4,5-bisphosphate (PtdInsP(2)), the lipid precursor of InsP(3), was greatly reduced in the transgenic cells. In vitro assays of enzymes involved in PtdInsP(2) metabolism showed that the activity of the PtdInsP(2)-hydrolyzing enzyme phospholipase C was not significantly altered in the transgenic cells. In contrast, the activity of the plasma membrane PtdInsP 5 kinase was increased by approximately 3-fold in the transgenic cells. In vivo labeling studies revealed a greater incorporation of (32)P into PtdInsP(2) in the transgenic cells compared with the wild type, indicating that the rate of PtdInsP(2) synthesis was increased. These studies show that the constitutive expression of the human type I InsP 5-ptase in tobacco cells leads to an up-regulation of the phosphoinositide pathway and highlight the importance of PtdInsP(2) synthesis as a regulatory step in this system.

NASA Discipline Cell Biology↗