Novel Roles for mTORC1-dependent Translational Control during Synaptic Homeostasis.

The mechanistic target of rapamycin complex 1 (mTORC1), a kinase involved in regulating translation initiation, has recently emerged as a critical player responsible for orchestrating dynamic changes in local protein synthesis in response to altered synaptic activity. Here we identify a novel mode of synaptic regulation conferred by mTORC1-dependent signaling in dendrites, wherein mTORC1 activation gates a local retrograde signaling mechanism that drives changes in presynaptic function from apposed postsynaptic terminals. This unique role for dendritic mTORC1 signaling is critically dependent on BDNF release, which couples loss of excitatory synaptic drive with retrograde compensation of presynaptic function. Acute activation of mTORC1 signaling using the lipid second messenger phosphatidic acid (PA) or overexpression of the endogenous mTOR activator RhebGTPase exerts a powerful influence on network function, which is also dependent on dendritic synthesis of BDNF as a retrograde signal. We identify an additional feature of the putative postsynaptic homeostatic sensor mechanism, showing that phospholipase D (PLD)-mediated hydrolysis of the lipid second messenger phosphatidic acid (PA) is a crucial component of the signaling pathway which relays homeostatic signals to postsynaptic mTORC1 after loss of excitatory input. Lastly, we find that mTORC1-dependent retrograde signaling acts in coordination with dynamic relocalization of the ubiquitin proteasome system to and from axonal boutons to regulate presynaptic function in the expression of synaptic homeostasis. Increasing neuronal firing rates enriches proteasome accumulation at synaptic terminals, whereas inhibiting neuronal firing results in a dramatic redistribution away from synaptic terminals. This altered localization is due, at least in part, to an activity-dependent sequestration mechanism at presynaptic terminals. Moreover, activity-dependent phosphorylation of the Rpt6 subunit of the 19S proteasome is necessary and sufficient for axonal proteasome redistribution, and this altered localization plays a critical role in establishing mTORC1-dependent retrograde homeostatic changes in presynaptic function after loss of postsynaptic drive. Several monogenic neurodevelopmental conditions related to Autism Spectrum Disorder and Intellectual Disability share the common molecular phenotype of increased mTORC1 signaling. As such, a more thorough understanding of how mTORC1 regulates synaptic function may provide insights for targeting this signaling pathway as a therapeutic option for cognitive dysfunction.PhDNeuroscienceUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/107207/1/fehenry_1.pd

Attenuation of morphine antinociceptive tolerance by a CB1 receptor agonist and an NMDA receptor antagonist: Interactive effects

CB1 cannabinoid (CB1) receptor agonists and N-Methyl-d-Aspartate (NMDA) receptor antagonists attenuate the development of morphine antinociceptive tolerance. The present study used dose-addition analysis to evaluate CB1/NMDA receptor interactions on this endpoint. Chronic morphine administration (5 days, 100 mg/kg, twice daily) resulted in a 2.8-fold rightward shift in the morphine dose-effect curve. Co-administration of either the CB1 receptor agonist CP-55940 (5-(1,1-Dimethylheptyl)-2-[5-hydroxy-2-(3-hydroxypropyl)cyclohexyl]phenol; 0.32-1.0 mg/kg) or the NMDA receptor antagonist (−)-6-phosphonomethyl-deca-hydroisoquinoline-3-carboxylic acid (LY235959; 1.0-3.2 mg/kg) with morphine dose-dependently attenuated morphine tolerance. The relative potency of each drug alone was quantified using a defined level of effect (one-quarter log shift in the morphine dose-effect curve), resulting in equieffective doses of 0.42 mg/kg and 1.1 mg/kg for CP-55940 and LY235959, respectively. Subsequent experiments assessed CP-55940/LY235959 interactions using a fixed-proportion design. Co-administration of CP-55940/LY235959 mixtures (1:1, 1:3.2, or 1:10 CP-55940/LY235959) with morphine dose-dependently attenuated morphine tolerance. Isobolographic and dose-addition analysis were used to statistically compare the experimentally determined potency for each mixture (zmix) with predicted additive potency (zadd). Mixtures of 1:1 and 1:3.2 CP-55940/LY235959 produced additive effects (zadd = zmix), while the mixture of 1:10 CP-55940/LY235959 produced a supra-additive effect (zadd > zmix). These results suggest that CP-55940 and LY235959 produce additive or supra-additive attenuation of morphine antinociceptive tolerance after repeated morphine administration, depending on their relative concentrations

Mechanistic target of rapamycin is necessary for changes in dendritic spine morphology associated with long-term potentiation

Abstract Alterations in the strength of excitatory synapses in the hippocampus is believed to serve a vital function in the storage and recall of new information in the mammalian brain. These alterations involve the regulation of both functional and morphological features of dendritic spines, the principal sites of excitatory synaptic contact. New protein synthesis has been implicated extensively in the functional changes observed following long-term potentiation (LTP), and changes to spine morphology have similarly been documented extensively following synaptic potentiation. However, mechanistic links between de novo translation and the structural changes of potentiated spines are less clear. Here, we assess explicitly the potential contribution of new protein translation under control of the mechanistic target of rapamycin (mTOR) to LTP-associated changes in spine morphology. Utilizing genetic and pharmacological manipulations of mTORC1 function in combination with confocal microscopy in live dissociated hippocampal cultures, we demonstrate that chemically-induced LTP (cLTP) requires do novo protein synthesis and intact mTORC1 signaling. We observed a striking diversity in response properties across morphological classes, with mushroom spines displaying a particular sensitivity to altered mTORC1 signaling across varied levels of synaptic activity. Notably, while pharmacological inhibition of mTORC1 signaling significantly diminished glycine-induced changes in spine morphology, transient genetic upregulation of mTORC1 signaling was insufficient to produce spine enlargements on its own. In contrast, genetic upregulation of mTORC1 signaling promoted rapid expansion in spine head diameter when combined with otherwise sub-threshold synaptic stimulation. These results suggest that synaptic activity-derived signaling pathways act in combination with mTORC1-dependent translational control mechanisms to ultimately regulate changes in spine morphology. As several monogenic neurodevelopmental disorders with links to Autism and Intellectual Disability share a common feature of dysregulated mTORC1 signaling, further understanding of the role of this signaling pathway in regulating synapse function and morphology will be essential in the development of novel therapeutic interventions.https://deepblue.lib.umich.edu/bitstream/2027.42/139019/1/13041_2017_Article_330.pd

Metabotropic glutamate antagonists alone and in combination with morphine: comparison across two models of acute pain and a model of persistent, inflammatory pain

The present study examined the effects of the mGluR1 antagonist JNJ16259685 (JNJ) and the mGluR5 antagonist MPEP alone and in combination with morphine in two acute pain models (hotplate, warm water tail-withdrawal), and a persistent, inflammatory pain model (capsaicin). In the hotplate and warm water tail-withdrawal procedures, JNJ and MPEP were ineffective when administered alone. In both procedures, JNJ potentiated morphine antinociception. In the hot plate procedure, MPEP potentiated morphine antinociception at the highest dose examined, whereas in the warm water tail-withdrawal procedure MPEP attenuated morphine antinociception at a moderate dose and potentiated morphine antinociception at a high dose. For both JNJ and MPEP, the magnitude of this morphine potentiation was considerably greater in the hotplate procedure. In the capsaicin procedure, the highest dose of MPEP produced intermediate levels of antihyperalgesia and also attenuated the effects of a dose of morphine that produced intermediate levels of antihyperalgesia. In contrast, JNJ had no effect when administered alone in the capsaicin procedure and did not alter morphine-induced antihyperalgesia. The present findings suggest that the effects produced by mGluR1 and mGluR5 antagonists alone and in combination with morphine can be differentiated in models of both acute and persistent pain

Opioid antinociception, tolerance and dependence: interactions with the N-methyl-D-aspartate system in mice

This study explored the involvement of N-methyl-D-aspartate (NMDA) in the effects of mu opioid agonists. A hot plate procedure was used to assess antinociception and tolerance in mice in which the NR1 subunit of the NMDA receptor was reduced (knocked down, KD) to approximately 10% and in mice treated with the NMDA antagonist, LY235959 [(−)-6-phosphonomethyl-deca-hydroisoquinoline-3-carboylic acid]. The mu opioid agonists, morphine, l-methadone and fentanyl, were approximately 3-fold less potent in the NR1 KD mice than in wild type (WT) controls; however, the development of morphine tolerance and dependence did not differ markedly in the NR1 KD and WT mice. Acute administration of the NMDA antagonist, LY235959 produced dose-dependent, leftward shifts in the morphine dose-effect curve in WT mice, but not in the NR1 KD mice. Chronic administration of LY235959 during the morphine tolerance regimen did not attenuate the development of tolerance in the NR1 KD nor the WT mice. These results indicate that the NR1 subunit of the NMDA receptor does not play a prominent role in mu opioid tolerance

Precise Masses for Wolf 1062 AB from Hubble Space Telescope Interferometric Astrometry and McDonald Observatory Radial Velocities

We present an analysis of astrometric data from FGS 3, a white-light interferometer on {\it HST}, and of radial velocity data from two ground-based campaigns. We model the astrometric and radial velocity measurements simultaneously to obtain parallax, proper motion and component masses for Wolf 1062 = Gl 748 AB (M3.5V). To derive the mass fraction, we relate FGS 3 fringe scanning observations of the science target to a reference frame provided by fringe tracking observations of a surrounding star field. We obtain an absolute parallax $\pi_{abs} = 98.0 \pm 0.4$ milliseconds of arc, yielding {\cal M}_A = 0.379 \pm 0.005{\cal M}_{\sun} and {\cal M}_B= 0.192 \pm 0.003 {\cal M}_{\sun}, high quality component masses with errors of only 1.5%.Comment: 13 pages, 7 figures. To appear in AJ March 200

Immunochip analysis identifies multiple susceptibility loci for systemic sclerosis

In this study, 1,833 systemic sclerosis (SSc) cases and 3,466 controls were genotyped with the Immunochip array. Classical alleles, amino acid residues, and SNPs across the human leukocyte antigen (HLA) region were imputed and tested. These analyses resulted in a model composed of six polymorphic amino acid positions and seven SNPs that explained the observed significant associations in the region. In addition, a replication step comprising 4,017 SSc cases and 5,935 controls was carried out for several selected non-HLA variants, reaching a total of 5,850 cases and 9,401 controls of European ancestry. Following this strategy, we identified and validated three SSc risk loci, including DNASE1L3 at 3p14, the SCHIP1-IL12A locus at 3q25, and ATG5 at 6q21, as well as a suggested association of the TREH-DDX6 locus at 11q23. The associations of several previously reported SSc risk loci were validated and further refined, and the observed peak of association in PXK was related to DNASE1L3. Our study has increased the number of known genetic associations with SSc, provided further insight into the pleiotropic effects of shared autoimmune risk factors, and highlighted the power of dense mapping for detecting previously overlooked susceptibility loci

TOI-3984 A b and TOI-5293 A b: two temperate gas giants transiting mid-M dwarfs in wide binary systems

We confirm the planetary nature of two gas giants discovered by TESS to transit M dwarfs with stellar companions at wide separations. TOI-3984 A ($J=11.93$) is an M4 dwarf hosting a short-period ($4.353326 \pm 0.000005$ days) gas giant ($M_p=0.14\pm0.03~\mathrm{M_{J}}$ and $R_p=0.71\pm0.02~\mathrm{R_{J}}$) with a wide separation white dwarf companion. TOI-5293 A ($J=12.47$) is an M3 dwarf hosting a short-period ($2.930289 \pm 0.000004$ days) gas giant ($M_p=0.54\pm0.07~\mathrm{M_{J}}$ and $R_p=1.06\pm0.04~\mathrm{R_{J}}$) with a wide separation M dwarf companion. We characterize both systems using a combination of ground-based and space-based photometry, speckle imaging, and high-precision radial velocities from the Habitable-zone Planet Finder and NEID spectrographs. TOI-3984 A b ($T_{eq}=563\pm15$ K and $\mathrm{TSM}=138_{-27}^{+29}$) and TOI-5293 A b ($T_{eq}=675_{-30}^{+42}$ K and $\mathrm{TSM}=92\pm14$) are two of the coolest gas giants among the population of hot Jupiter-sized gas planets orbiting M dwarfs and are favorable targets for atmospheric characterization of temperate gas giants and three-dimensional obliquity measurements to probe system architecture and migration scenarios.Comment: Submitted to AJ, 42 pages, 14 figures. arXiv admin note: substantial text overlap with arXiv:2201.0996

Strong gravitational lensing probes of the particle nature of dark matter

There is a vast menagerie of plausible candidates for the constituents of dark matter, both within and beyond extensions of the Standard Model of particle physics. Each of these candidates may have scattering (and other) cross section properties that are consistent with the dark matter abundance, BBN, and the most scales in the matter power spectrum; but which may have vastly different behavior at sub-galactic "cutoff" scales, below which dark matter density fluctuations are smoothed out. The only way to quantitatively measure the power spectrum behavior at sub-galactic scales at distances beyond the local universe, and indeed over cosmic time, is through probes available in multiply imaged strong gravitational lenses. Gravitational potential perturbations by dark matter substructure encode information in the observed relative magnifications, positions, and time delays in a strong lens. Each of these is sensitive to a different moment of the substructure mass function and to different effective mass ranges of the substructure. The time delay perturbations, in particular, are proving to be largely immune to the degeneracies and systematic uncertainties that have impacted exploitation of strong lenses for such studies. There is great potential for a coordinated theoretical and observational effort to enable a sophisticated exploitation of strong gravitational lenses as direct probes of dark matter properties. This opportunity motivates this white paper, and drives the need for: a) strong support of the theoretical work necessary to understand all astrophysical consequences for different dark matter candidates; and b) tailored observational campaigns, and even a fully dedicated mission, to obtain the requisite data.Comment: Science white paper submitted to the Astro2010 Decadal Cosmology & Fundamental Physics Science Frontier Pane

Annual cycles are the most common reproductive strategy in African tropical tree communities

We present the first cross continental comparison of the flowering and fruiting phenology of tropical forests across Africa. Flowering events of 5,446 trees from 196 species across 12 sites, and fruiting events of 4,595 trees from 191 species, across 11 sites were monitored over periods of 6 to 29 years, and analysed to describe phenology at the continental level. To study phenology we used Fourier analysis to identify the dominant cycles of flowering and fruiting for each individual tree and we identified the time of year African trees bloom and bear fruit and their relationship to local seasonality. Reproductive strategies were diverse and no single regular cycle was found in >50% of individuals across all 12 sites. Additionally, we found annual flowering and fruiting cycles to be the most common. Sub-annual cycles were the next most common for flowering whereas supra-annual patterns were the next most common for fruiting. We also identify variation in different subsets of species, with species exhibiting mainly annual cycles most common in West and West-Central African tropical forests, while more species at sites in East-Central and Eastern African forests showed cycles ranging from sub-annual to supra-annual. Despite many trees showing strong seasonality, at most sites some flowering and fruiting occurred all year round. Environmental factors with annual cycles are likely to be important drivers of seasonal periodicity in trees across Africa, but proximate triggers are unlikely to be constant across the continen