Glucocorticoids rescue cell surface trafficking of R451C Neuroligin3 and enhance synapse formation

Neuroligins are synaptic cell adhesion proteins with a role in synaptic function, implicated in neurodevelopmental disorders. The autism spectrum disorder-associated substitution Arg451Cys (R451C) in NLGN3 promotes a partial misfolding of the extracellular domain of the protein leading to retention in the endoplasmic reticulum (ER) and the induction of the unfolded protein response (UPR). The reduced trafficking of R451C NLGN3 to the cell surface leads to altered synaptic function and social behavior. A screening in HEK-293 cells overexpressing NLGN3 of 2662 compounds (FDA-approved small molecule drug library), led to the identification of several glucocorticoids such as alclometasone dipropionate, desonide, prednisolone sodium phosphate, and dexamethasone (DEX), with the ability to favor the exit of full-length R451C NLGN3 from the ER. DEX improved the stability of R451C NLGN3 and trafficking to the cell surface, reduced the activation of the UPR, and increased the formation of artificial synapses between HEK-293 and hippocampal primary neurons. The effect of DEX was validated on a novel model system represented by neural stem progenitor cells and differentiated neurons derived from the R451C NLGN3 knock-in mouse, expressing the endogenous protein. This work shows a potential rescue strategy for an autism-linked mutation affecting cell surface trafficking of a synaptic protein

Biosynthesis of HLA-C heavy chains in melanoma cells with multiple defects in the expression of HLA-A, -B, -C molecules

Recent investigations have shown that malignant transformation may down-regulate the expression of class I HLA molecules, beta(2)-microglobulin (beta(2)m) and members of the antigen-processing machinery. In the present study, we HLA-genotyped and identified at a biochemical level the three (HLA-A25, -B8, -Cw7) class I alleles expressed by the previously described [D'Urso CM et al (1992) J Clin Invest 87: 284-292] beta(2)m-defective human melanoma FO-1 cell line and tested their ability to interact with calnexin, calreticulin and the TAP (transporter associated with antigen processing) complex. Ail these alleles were found to bind calnexin, but not calreticulin or the poorly expressed TAP complex, both in parental and beta(2)m-transfected FO-1 cells, demonstrating a complex defect of class I expression in FO-1 cells. In these conditions, Cw7 heavy chains interacted with calnexin more strongly than A25 and B8, and preferentially accumulated in the endoplasmic reticulum, in both a calnexin-associated and a calnexin-free form. In addition, they could be transported to the cell surface at low levels even in the absence of beta(2)m, without undergoing terminal glycosylation. These results establish a parallel between HLA-C and the murine D-b and L-d molecules which have been found to be surface expressed and functional in beta(2)m-defective cells. They also demonstrate distinctive features of HLA-C molecules. We propose that the accumulation of several assembly intermediates of HLA-C might favour the binding of peptide antigens not readily bound by HLA-A and -B molecules in neoplastic cells with suboptimal class I expression

HLA-C and HIV-1: friends or foes?

The major histocompatibility complex class I protein HLA-C plays a crucial role as a molecule capable of sending inhibitory signals to both natural killer (NK) cells and cytotoxic T lymphocytes (CTL) via binding to killer cell Ig-like receptors (KIR). Recently HLA-C has been recognized as a key molecule in the immune control of HIV-1. Expression of HLA-C is modulated by a microRNA binding site. HLA-C alleles that bear substitutions in the microRNA binding site are more expressed at the cell surface and associated with the control of HIV-1 viral load, suggesting a role of HLA-C in the presentation of antigenic peptides to CTLs. This review highlights the role of HLA-C in association with HIV-1 viral load, but also addresses the contradiction of the association between high cell surface expression of an inhibitory molecule and strong cell-mediated immunity. To explore additional mechanisms of control of HIV-1 replication by HLA-C, we address specific features of the molecule, like its tendency to be expressed as open conformer upon cell activation, which endows it with a unique capacity to associate with other cell surface molecules as well as with HIV-1 proteins

HIV-1 Env associates with HLA-C free-chains at the cell membrane modulating viral infectivity

HLA-C has been demonstrated to associate with HIV-1 envelope glycoprotein (Env). Virions lacking HLA-C have reduced infectivity and increased susceptibility to neutralizing antibodies. Like all others MHC-I molecules, HLA-C requires \u3b22-microglobulin (\u3b22m) for appropriate folding and expression on the cell membrane but this association is weaker, thus generating HLA-C free-chains on the cell surface. In this study, we deepen the understanding of HLA-C and Env association by showing that HIV-1 specifically increases the amount of HLA-C free chains, not bound to \u3b22m, on the membrane of infected cells. The association between Env and HLA-C takes place at the cell membrane requiring \u3b22m to occur. We report that the enhanced infectivity conferred to HIV-1 by HLA-C specifically involves HLA-C free chain molecules that have been correctly assembled with \u3b22m. HIV-1 Env-pseudotyped viruses produced in the absence of \u3b22m are less infectious than those produced in the presence of \u3b22m. We hypothesize that the conformation and surface expression of HLA-C molecules could be a discriminant for the association with Env. Binding stability to \u3b22m may confer to HLA-C the ability to preferentially act either as a conventional immune-competent molecule or as an accessory molecule involved in HIV-1 infectivity

First evidence of microplastics ingestion in benthic amphipods from Svalbard

The present paper provides the first record of ingestion of microplastics in natural context by Gammarus setosus from Svalbard Archipelago. The plastic particles were identified both by Nile Red staining and Micro FT-IR spectroscopy. The species studied ingests microplastic particles in natural conditions if present in its habitat, probably mistaking them as food. The microplastic particles ingested may be available for uptake to predators that consume this Arctic amphipod, producing consequences to the food web

Genotoxicity biomarkers in the amphipod Gammarus elvirae exposed in vivo to mercury and lead and basal levels of DNA damage in two cell types

The present work deals with the application of genotoxicity biomarkers by means of the Comet assay in haemocytes and spermatozoa of the crustacean Gammarus elvirae exposed in vivo to heavy metals. Furthermore, a basal levels (BLs) study of DNA damage in the two cell types considered for two different gammarids species, G. elvirae and Echinogammarus veneris, was carried out. It is important to identify factors that influence the outcome of the assay in order to obtain reliable and reproducible results usable for risk assessment purposes. Our results highlight that the Italian legal limits for Hg and Pb, respectively, 0.5 and 50 μg/L, are inadequate for establishing safety thresholds in the aquatic environment. Furthermore, the freshwater invertebrate G. elvirae, used for the first time to measure the effect of genotoxicants, is a good candidate for evaluating the genotoxicity damage induced by heavy metals. Our results concerning spermatozoa show excessively variable responses and high BLs

Preliminary observations on the effect of light and temperature on the hatching success and rate of Lepidurus arcticus eggs

Dormancy, which arrests development, is a well-known survival strategy among animals living in the Arctic to overcome harsh periods. It is not clear if the dormant state in notostracans is controlled endogenously (diapause) or exogenously (quiescence). For Lepidurus arcticus, it is unknown how it responds to the photoperiod entrainment, if it has a biological clock and if it has a rhythmic expression of the clock genes. We studied the hatching success of resting eggs at four constant temperatures (5, 10, 15 and 25 °C) and under different illumination regimes [continuous light (LL) and continuous dark (DD)]. It was assumed that light and temperature are both important triggers, with temperature having the most pronounced effect. In our experiment, hatching occurred only at 5 and 10 °C, while we did not observe hatching at 15 and 25 °C. The highest percentage of eggs hatched was at 10 °C in LL (60%); the lowest was at 5 °C in DD (18%). The percentages hatched at 5 °C in LL (24%) and at 10 °C in DD (26%) were similar. Our results indicate that both temperature and light had a significant and interacting effect on hatching in L. arcticus, with temperature being the dominant factor controlling the process. This suggests that changes in temperature affecting the Arctic may significantly impact phenology of this key species in the region. Given that no hatching was observed at 15 °C or above, the persistence of this species may be at risk in areas were arctic lakes are expected to warm to such levels during the summer months