Alternative Splicing in CaV2.2 Regulates Neuronal Trafficking via Adaptor Protein Complex-1 Adaptor Protein Motifs

UNLABELLED: N-type voltage-gated calcium (CaV2.2) channels are expressed in neurons and targeted to the plasma membrane of presynaptic terminals, facilitating neurotransmitter release. Here, we find that the adaptor protein complex-1 (AP-1) mediates trafficking of CaV2.2 from the trans-Golgi network to the cell surface. Examination of splice variants of CaV2.2, containing either exon 37a (selectively expressed in nociceptors) or 37b in the proximal C terminus, reveal that canonical AP-1 binding motifs, YxxΦ and [DE]xxxL[LI], present only in exon 37a, enhance intracellular trafficking of exon 37a-containing CaV2.2 to the axons and plasma membrane of rat DRG neurons. Finally, we identify differential effects of dopamine-2 receptor (D2R) and its agonist-induced activation on trafficking of CaV2.2 isoforms. D2R slowed the endocytosis of CaV2.2 containing exon 37b, but not exon 37a, and activation by the agonist quinpirole reversed the effect of the D2R. Our work thus reveals key mechanisms involved in the trafficking of N-type calcium channels. SIGNIFICANCE STATEMENT: CaV2.2 channels are important for neurotransmitter release, but how they are trafficked is still poorly understood. Here, we describe a novel mechanism for trafficking of CaV2.2 from the trans-Golgi network to the cell surface which is mediated by the adaptor protein AP-1. Alternative splicing of exon 37 produces CaV2.2-exon 37a, selectively expressed in nociceptors, or CaV2.2-exon 37b, which is the major splice isoform. Our study reveals that canonical AP-1 binding motifs (YxxΦ and [DE]xxxL[LI]), present in exon 37a, but not 37b, enhance intracellular trafficking of exon 37a-containing CaV2.2 to axons and plasma membrane of DRG neurons. Interaction of APs with CaV2.2 channels may also be key underlying mechanisms for differential effects of the dopamine D2 receptor on trafficking of CaV2.2 splice variants

Sediment dynamics of a nearshore sandbank: Results from TELEMAC-2D, TOMAWAC and SISYPHE modelling

Water Qualit

Tidal dwarfs in the M81 group: the second generation?

We derive quantitative star formation histories of the four suspected tidal dwarf galaxies in the M81 group, HolmbergIX, BK3N, Arp-loop (A0952+69), and Garland, using Hubble Space Telescope/Wide Field Planetary Camera2 images in F606W and F814W obtained as part of a Snapshot survey of dwarf galaxies in the Local Universe. We consider the spatial distribution and ages of resolved stellar populations in these dwarf irregular galaxies. We use synthetic color-magnitude diagrams to derive the ages of the major star formation episodes, star formation rates, and approximate metallicity ranges. All the galaxies show evidence of continuous star formation between about 20 and 200 Myr ago with star formation rates in the range 7.5*10^(-3)- 7.6*10^(-4) M(sun)/yr. The metallicity of the detected stars spans a wide range, and have lower than solar abundance. A possible scenario is that all four dwarf galaxies were formed from material in the metal-poor outer part of the giant spiral galaxy M81 after the tidal interaction between M81, M82, and NGC3077 about 200 Myr ago. While we do not directly detect pronounced old stellar populations, the photometric limits of our data are such that the presence of such a population is not entirely ruled out

The Hubble flow around the CenA / M83 galaxy complex

We present HST/ACS images and color-magnitude diagrams for 24 nearby galaxies in and near the constellation of Centaurus with radial velocities V_LG < 550 km/s. Distances are determined based on the luminosities of stars at the tip of the red giant branch that range from 3.0 Mpc to 6.5 Mpc. The galaxies are concentrated in two spatially separated groups around Cen A (NGC 5128) and M 83 (NGC 5236). The Cen A group itself has a mean distance of 3.76 +/-0.05 Mpc, a velocity dispersion of 136 km/s, a mean harmonic radius of 192 kpc, and an estimated orbital/virial mass of (6.4 - 8.1) x 10^12 M_sun. This elliptical dominated group is found to have a relatively high mass-to-light ratio: M/L_B = 125 M_sun/L_sun. For the M 83 group we derived a mean distance of 4.79 +/-0.10 Mpc, a velocity dispersion of 61 km/s, a mean harmonic radius of 89 kpc, and estimated orbital/virial mass of (0.8 - 0.9) x 10^12 M_sun. This spiral dominated group is found to have a relatively low M/L_B = 34 M_sun/L_sun. The radius of the zero-velocity surface around Cen A lies at R_0 = 1.40 +/-0.11 Mpc, implying a total mass within R_0 of M_T = (6.0 +/-1.4) x 10^12 M_sun. This value is in good agreement with the Cen A virial/orbital mass estimates and provides confirmation of the relatively high M/L_B of this elliptical-dominated group. The centroids of both the groups, as well as surrounding field galaxies, have very small peculiar velocities, < 25 km/s, with respect to the local Hubble flow with H_0 = 68 km/s/Mpc.Comment: 31 pages including 9 figures and 3 tables. Accepted for publication in Astronomical Journal, 133, N0. 2 (February), 200

Determinants of the voltage dependence of G protein modulation within calcium channel β subunits

CaVβ subunits of voltage-gated calcium channels contain two conserved domains, a src-homology-3 (SH3) domain and a guanylate kinase-like (GK) domain with an intervening HOOK domain. We have shown in a previous study that, although Gβγ-mediated inhibitory modulation of CaV2.2 channels did not require the interaction of a CaVβ subunit with the CaVα1 subunit, when such interaction was prevented by a mutation in the α1 subunit, G protein modulation could not be removed by a large depolarization and showed voltage-independent properties (Leroy et al., J Neurosci 25:6984–6996, 2005). In this study, we have investigated the ability of mutant and truncated CaVβ subunits to support voltage-dependent G protein modulation in order to determine the minimal domain of the CaVβ subunit that is required for this process. We have coexpressed the CaVβ subunit constructs with CaV2.2 and α2δ-2, studied modulation by the activation of the dopamine D2 receptor, and also examined basal tonic modulation. Our main finding is that the CaVβ subunit GK domains, from either β1b or β2, are sufficient to restore voltage dependence to G protein modulation. We also found that the removal of the variable HOOK region from β2a promotes tonic voltage-dependent G protein modulation. We propose that the absence of the HOOK region enhances Gβγ binding affinity, leading to greater tonic modulation by basal levels of Gβγ. This tonic modulation requires the presence of an SH3 domain, as tonic modulation is not supported by any of the CaVβ subunit GK domains alone

A Hubble View of Star Forming Regions in the Magellanic Clouds

The Magellanic Clouds (MCs) offer an outstanding variety of young stellar associations, in which large samples of low-mass stars (with masses less than 1 solar mass) currently in the act of formation can be resolved and explored sufficiently with the Hubble Space Telescope. These pre-main sequence (PMS) stars provide a unique snapshot of the star formation process, as it is being recorded for the last 20 Myr, and they give important information on the low-mass Initial Mass Function (IMF) of their host environments. We present the latest results from observations with the Advanced Camera for Surveys (ACS) of such star-forming regions in the MCs, and discuss the importance of Hubble}for a comprehensive collection of substantial information on the most recent low-mass star formation and the low-mass IMF in the MCs.Comment: To appear in the proceedings of the 41st ESLAB Symposium: The Impact of HST on European Astronomy, 4 pages, LaTeX ESA Publications style, 5 Figure

The nature of V39: an LBV candidate or LBV impostor in the very low metallicity galaxy IC 1613?

[abridged] Context: Very few examples of luminous blue variable (LBV) stars or LBV candidates (LBVc) are known, particularly at metallicities below the SMC. The LBV phase is crucial for the evolution of massive stars, and its behavior with metallicity is poorly known. V39 in IC 1613 is a well-known photometric variable, with B-band changes larger than 1mag. over its period. The star, previously proposed to be a projection of a Galactic W Virginis and an IC 1613 red supergiant, shows features that render it a possible LBVc. Method: We investigate mid-resolution blue and red VLT-VIMOS spectra of V39, covering a time span of 40 days, and perform a quantitative analysis of the combined spectrum using the model atmosphere code CMFGEN. Results: We identify strong Balmer and FeII P-Cygni profiles, and a hybrid spectrum resembling a B-A supergiant in the blue and a G-star in the red. No significant Vrad variations are detected, and the spectral changes are small over the photometric period. Our analysis places V39 in the low-luminosity part of the LBV and LBVc region, but it is also consistent with a sgB[e] star. Conclusions: The radial velocity indicates that V39 belongs to IC 1613. The lack of Vrad changes and spectroscopic variations excludes binary scenarios. The features observed are not consistent with a W Virginis star, and this possibility is also discarded. We propose that the star is a B-A LBVc or sgB[e] star surrounded by a thick disk precessing around it. If confirmed, V39 would be the lowest metallicity resolved LBV candidate known to date. Alternatively, it could represent a new transient phase of massive star evolution, an LBV impostor.Comment: In press at A&A. 10 pages, 11 figure

An HST/ACS View of the Inhomogeneous Outer Halo of M31

We present a high precision photometric view of the stellar populations in the outer halo of M31, using data taken with the Hubble Space Telescope Advanced Camera for Surveys (HST/ACS). We analyse the field populations adjacent to 11 luminous globular clusters which sample the galactocentric radial range 18 < R < 100 kpc and reach a photometric depth of ~2.5 magnitudes below the horizontal branch (m_F814W ~27 mag). The colour-magnitude diagrams (CMDs) are well populated out to ~60 kpc and exhibit relatively metal-rich red giant branches, with the densest fields also showing evidence for prominent red clumps. We use the Dartmouth isochrones to construct metallicity distribution functions (MDFs) which confirm the presence of dominant populations with = -0.6 to -1.0 dex and considerable metallicity dispersions of 0.2 to 0.3 dex (assuming a 10 Gyr population and scaled-Solar abundances). The average metallicity over the range 30 - 60 kpc is [Fe/H] = -0.8 +/- 0.14 dex, with no evidence for a significant radial gradient. Metal-poor stars ([Fe/H] <= -1.3) typically account for < 10-20 % of the population in each field, irrespective of radius. Assuming our fields are unbiased probes of the dominant stellar populations in these parts, we find that the M31 outer halo remains considerably more metal-rich than that of the Milky Way out to at least 60 kpc.Comment: Accepted for publication in MNRAS. 10 pages, 6 figure

ACS imaging of 25 galaxies in nearby groups and in the field

We present HST/ACS images and color-magnitude diagrams for 25 nearby galaxies with radial velocities V_LG < 500 km/s. Distances are determined based on the luminosities of stars at the tip of the red giant branch that range from 2 Mpc to 12 Mpc. Two of the galaxies, NGC 4163 and IC 4662, are found to be the nearest known representatives of blue compact dwarf (BCD) objects. Using high-quality data on distances and radial velocities of 110 nearby field galaxies, we derive their mean Hubble ratio to be 68 km/(s Mpc) with standard deviation of 15 km/(s Mpc). Peculiar velocities of most of the galaxies, V_pec = V_LG - 68 D, follow a Gaussian distribution with sigma_v = 63 km/s, but with a tail towards high negative values. Our data displays the known correlation between peculiar velocity and galaxy elevation above the Local Supercluster plane. The small observed fraction of galaxies with high peculiar velocities, V_pec < -500 km/s, may be understood as objects associated with nearby groups (Coma I, Eridanus) outside the Local volume.Comment: 17 pages, 8 figures, accepted to A