The Role of Extracellular Vesicles in the Progression of ALS and Their Potential as Biomarkers and Therapeutic Agents with Which to Combat the Disease

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that impairs motor neuron function, leading to severe muscular atrophy. The non-cell autonomous and heterogeneous nature of the disease has so far hindered attempts to define ALS etiology, leaving the disease incurable and without effective treatments. Recent studies have focused on the pathologic role of intercellular communication between nerve cells to further our understanding of ALS pathophysiology. In this chapter, we summarize recent works investigating the role of extracellular vesicles (EVs) as a means of cellular crosstalk for ALS disease propagation, diagnosis, and treatment. There is growing evidence that EVs secreted by the majority of mammalian cells serve as effective biomolecule carriers to modulate recipient cell behavior. This underscores the need to understand the EV-mediated interplay that occurs within irreversibly degenerating nervous tissue in ALS patients. Additionally, we highlight current gaps in EV-ALS research, especially in terms of the pathologic role and responsibilities of specific EV cargos in diseased cells, specificity issues associated with the use of EVs in ALS diagnosis, and the efficacy of EV-mediated treatments for the restoration of diseased neuromuscular tissue. Finally, we provide suggestions for future EV-ALS research to better understand, diagnose, and cure this inveterate disease

Effects of K+ on the catalytic and regulatory properties of homoserine dehydrogenase of Pseudomonas fluorescens

Partially purified homoserine dehydrogenase (-homoserine; NADP+ oxidoreductase, EC 1.1.1.3) isolated from Pseudomonas fluorescens requires K+ for its stability. At 1 mM K+, the apparent first-order rate constant for enzyme inactivation at 25[deg] was 25-fold higher than that observed at 20 mM K+. A dissociation constant, KD, of 3 mM of the ion-enzyme complex was calculated.Following inactivation of the enzyme by depletion of K+, the activity could be regenerated to a large extent by incubation with KCl and NADP+; no other cation was nearly as effective as K+.Binding of K+ on the enzyme molecule also plays an important part in the regulation of enzyme activity by the allosteric modifier -threonine. At 1 mM K+, the catalytic activity was strongly inhibited by 10 mM -threonine, whereas, at 10 mM K+ concentration in the assay mixture, -threonine inhibition was almost completely abolished; threonine and K+ were kinetically competitive. It is proposed that K+ can induce specific conformational changes in the protein molecule that are either sensitive or insensitive to feedback inhibition control; in addition, the cation is necessary for the maintenance of the protein structure to ensure catalytic activity.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/33671/1/0000181.pd

Phenylbutyric Acid Rescues Endoplasmic Reticulum Stress-Induced Suppression of APP Proteolysis and Prevents Apoptosis in Neuronal Cells

BACKGROUND: The familial and sporadic forms of Alzheimer's disease (AD) have an identical pathology with a severe disparity in the time of onset [1]. The pathological similarity suggests that epigenetic processes may phenocopy the Familial Alzheimer's disease (FAD) mutations within sporadic AD. Numerous groups have demonstrated that FAD mutations in presenilin result in 'loss of function' of gamma-secretase mediated APP cleavage [2], [3], [4], [5]. Accordingly, ER stress is prominent within the pathologically impacted brain regions in AD patients [6] and is reported to inhibit APP trafficking through the secretory pathway [7], [8]. As the maturation of APP and the cleaving secretases requires trafficking through the secretory pathway [9], [10], [11], we hypothesized that ER stress may block trafficking requisite for normal levels of APP cleavage and that the small molecular chaperone 4-phenylbutyrate (PBA) may rescue the proteolytic deficit. METHODOLOGY/PRINCIPAL FINDINGS: The APP-Gal4VP16/Gal4-reporter screen was stably incorporated into neuroblastoma cells in order to assay gamma-secretase mediated APP proteolysis under normal and pharmacologically induced ER stress conditions. Three unrelated pharmacological agents (tunicamycin, thapsigargin and brefeldin A) all repressed APP proteolysis in parallel with activation of unfolded protein response (UPR) signaling-a biochemical marker of ER stress. Co-treatment of the gamma-secretase reporter cells with PBA blocked the repressive effects of tunicamycin and thapsigargin upon APP proteolysis, UPR activation, and apoptosis. In unstressed cells, PBA stimulated gamma-secretase mediated cleavage of APP by 8-10 fold, in the absence of any significant effects upon amyloid production, by promoting APP trafficking through the secretory pathway and the stimulation of the non-pathogenic alpha/gamma-cleavage. CONCLUSIONS/SIGNIFICANCE: ER stress represses gamma-secretase mediated APP proteolysis, which replicates some of the proteolytic deficits associated with the FAD mutations. The small molecular chaperone PBA can reverse ER stress induced effects upon APP proteolysis, trafficking and cellular viability. Pharmaceutical agents, such as PBA, that stimulate alpha/gamma-cleavage of APP by modifying intracellular trafficking should be explored as AD therapeutics

Activation of the p75 neurotrophin receptor through conformational rearrangement of disulphide-linked receptor dimers

Ligand-mediated dimerization has emerged as a universal mechanism of growth factor receptor activation. Neurotrophins interact with dimers of the p75 neurotrophin receptor (p75(NTR)), but the mechanism of receptor activation has remained elusive. Here, we show that p75(NTR) forms disulphide-linked dimers independently of neurotrophin binding through the highly conserved Cys(257) in its transmembrane domain. Mutation of Cys(257) abolished neurotrophin-dependent receptor activity but did not affect downstream signaling by the p75(NTR)/NgR/Lingo-1 complex in response to MAG, indicating the existence of distinct, ligand-specific activation mechanisms for p75(NTR). FRET experiments revealed a close association of p75(NTR) intracellular domains that was transiently disrupted by conformational changes induced upon NGF binding. Although mutation of Cys(257) did not alter the oligomeric state of p75(NTR), the mutant receptor was no longer able to propagate conformational changes to the cytoplasmic domain upon ligand binding. We propose that neurotrophins activate p75(NTR) by a mechanism involving rearrangement of disulphide-linked receptor subunits

Survival of Star-Forming Giant Clumps in High-Redshift Galaxies

We investigate the effects of radiation pressure from stars on the survival of the star-forming giant clumps in high-redshift massive disc galaxies, during the most active phase of galaxy formation. The clumps, typically of mass ~10^8-10^9 Msun and radius ~0.5-1, are formed in the turbulent gas-rich discs by violent gravitational instability and then migrate into a central bulge in ~10 dynamical times. We show that the survival or disruption of these clumps under the influence of stellar feedback depends critically on the rate at which they form stars. If they convert a few percent of their gas mass to stars per free-fall time, as observed for all local star-forming systems and implied by the Kennicutt-Schmidt law, they cannot be disrupted. Only if clumps convert most of their mass to stars in a few free-fall times can feedback produce significant gas expulsion. We consider whether such rapid star formation is likely in high-redshift giant clumps.Comment: 11 pages, 1 figure, accepted to MNRA

COLD GASS, an IRAM legacy survey of molecular gas in massive galaxies: I. Relations between H2, HI, stellar content and structural properties

We are conducting COLD GASS, a legacy survey for molecular gas in nearby galaxies. Using the IRAM 30m telescope, we measure the CO(1-0) line in a sample of ~350 nearby (D=100-200 Mpc), massive galaxies (log(M*/Msun)>10.0). The sample is selected purely according to stellar mass, and therefore provides an unbiased view of molecular gas in these systems. By combining the IRAM data with SDSS photometry and spectroscopy, GALEX imaging and high-quality Arecibo HI data, we investigate the partition of condensed baryons between stars, atomic gas and molecular gas in 0.1-10L* galaxies. In this paper, we present CO luminosities and molecular hydrogen masses for the first 222 galaxies. The overall CO detection rate is 54%, but our survey also uncovers the existence of sharp thresholds in galaxy structural parameters such as stellar mass surface density and concentration index, below which all galaxies have a measurable cold gas component but above which the detection rate of the CO line drops suddenly. The mean molecular gas fraction MH2/M* of the CO detections is 0.066+/-0.039, and this fraction does not depend on stellar mass, but is a strong function of NUV-r colour. Through stacking, we set a firm upper limit of MH2/M*=0.0016+/-0.0005 for red galaxies with NUV-r>5.0. The average molecular-to-atomic hydrogen ratio in present-day galaxies is 0.3, with significant scatter from one galaxy to the next. The existence of strong detection thresholds in both the HI and CO lines suggests that "quenching" processes have occurred in these systems. Intriguingly, atomic gas strongly dominates in the minority of galaxies with significant cold gas that lie above these thresholds. This suggests that some re-accretion of gas may still be possible following the quenching event.Comment: Accepted for publications in MNRAS. 32 pages, 25 figure

COLD GASS, an IRAM Legacy Survey of Molecular Gas in Massive Galaxies: II. The non-universality of the Molecular Gas Depletion Timescale

We study the relation between molecular gas and star formation in a volume-limited sample of 222 galaxies from the COLD GASS survey, with measurements of the CO(1-0) line from the IRAM 30m telescope. The galaxies are at redshifts 0.025<z<0.05 and have stellar masses in the range 10.0<log(M*/Msun)<11.5. The IRAM measurements are complemented by deep Arecibo HI observations and homogeneous SDSS and GALEX photometry. A reference sample that includes both UV and far-IR data is used to calibrate our estimates of star formation rates from the seven optical/UV bands. The mean molecular gas depletion timescale, tdep(H2), for all the galaxies in our sample is 1 Gyr, however tdep(H2) increases by a factor of 6 from a value of ~0.5 Gyr for galaxies with stellar masses of 10^10 Msun to ~3 Gyr for galaxies with masses of a few times 10^11 Msun. In contrast, the atomic gas depletion timescale remains contant at a value of around 3 Gyr. This implies that in high mass galaxies, molecular and atomic gas depletion timescales are comparable, but in low mass galaxies, molecular gas is being consumed much more quickly than atomic gas. The strongest dependences of tdep(H2) are on the stellar mass of the galaxy (parameterized as log tdep(H2)= (0.36+/-0.07)(log M* - 10.70)+(9.03+/-0.99)), and on the specific star formation rate. A single tdep(H2) versus sSFR relation is able to fit both "normal" star-forming galaxies in our COLD GASS sample, as well as more extreme starburst galaxies (LIRGs and ULIRGs), which have tdep(H2) < 10^8 yr. Normal galaxies at z=1-2 are displaced with respect to the local galaxy population in the tdep(H2) versus sSFR plane and have molecular gas depletion times that are a factor of 3-5 times longer at a given value of sSFR due to their significantly larger gas fractions.Comment: Accepted for publication in MNRAS. 19 pages, 11 figure

The physical scale of the far-infrared emission in the most luminous submillimetre galaxies II: evidence for merger-driven star formation

We present high-resolution 345 GHz interferometric observations of two extreme luminous (L_{IR}>10^{13} L_sun), submillimetre-selected galaxies (SMGs) in the COSMOS field with the Submillimeter Array (SMA). Both targets were previously detected as unresolved point-sources by the SMA in its compact configuration, also at 345 GHz. These new data, which provide a factor of ~3 improvement in resolution, allow us to measure the physical scale of the far-infrared in the submillimetre directly. The visibility functions of both targets show significant evidence for structure on 0.5-1 arcsec scales, which at z=1.5 translates into a physical scale of 5-8 kpc. Our results are consistent with the angular and physical scales of two comparably luminous objects with high-resolution SMA followup, as well as radio continuum and CO sizes. These relatively compact sizes (<5-10 kpc) argue strongly for merger-driven starbursts, rather than extended gas-rich disks, as the preferred channel for forming SMGs. For the most luminous objects, the derived sizes may also have important physical consequences; under a series of simplifying assumptions, we find that these two objects in particular are forming stars close to or at the Eddington limit for a starburst.Comment: 9 pages, 3 Figures, submitted to MNRA

Constraints on the Redshift Evolution of the L_X-SFR Relation from the Cosmic X-Ray Backgrounds

Observations of local star forming galaxies have revealed a correlation between the rate at which galaxies form stars and their X-Ray luminosity. We combine this correlation with the most recent observational constraints on the integrated star formation rate density, and find that star forming galaxies account for 5-20% of the total soft and hard X-ray backgrounds, where the precise number depends on the energy band and the assumed average X-ray spectral energy distribution of the galaxies below ~20 keV. If we combine the L_X-SFR relation with recently derived star formation rate function, then we find that star forming galaxies whose X-ray flux falls well (more than a factor of 10) below the detection thresholds of the Chandra Deep Fields, can fully account for the unresolved soft X-ray background, which corresponds to ~6% of its total. Motivated by this result, we put limits on the allowed redshift evolution of the parameter c_X \equiv L_X/SFR, and/or its evolution towards lower and higher star formation rates. If we parametrize the redshift evolution of c_X ~ (1+z)^b, then we find that b \leq 1.3 (95% CL). On the other hand, the observed X-ray luminosity functions (XLFs) of star forming galaxies indicate that c_X may be increasing towards higher redshifts and/or higher star formation rates at levels that are consistent with the X-ray background, but possibly at odds with the locally observed L_X-SFR relation.Comment: Accepted to MNRAS after minor revisions. 12 pages, 8 figure