Ultrastructural Characterization of the Lower Motor System in a Mouse Model of Krabbe Disease

Krabbe disease (KD) is a neurodegenerative disorder caused by the lack of β- galactosylceramidase enzymatic activity and by widespread accumulation of the cytotoxic galactosyl-sphingosine in neuronal, myelinating and endothelial cells. Despite the wide use of Twitcher mice as experimental model for KD, the ultrastructure of this model is partial and mainly addressing peripheral nerves. More details are requested to elucidate the basis of the motor defects, which are the first to appear during KD onset. Here we use transmission electron microscopy (TEM) to focus on the alterations produced by KD in the lower motor system at postnatal day 15 (P15), a nearly asymptomatic stage, and in the juvenile P30 mouse. We find mild effects on motorneuron soma, severe ones on sciatic nerves and very severe effects on nerve terminals and neuromuscular junctions at P30, with peripheral damage being already detectable at P15. Finally, we find that the gastrocnemius muscle undergoes atrophy and structural changes that are independent of denervation at P15. Our data further characterize the ultrastructural analysis of the KD mouse model, and support recent theories of a dying-back mechanism for neuronal degeneration, which is independent of demyelination

Size and specimen-dependent strategy for x-ray micro-ct and tem correlative analysis of nervous system samples

Correlative approaches are a powerful tool in the investigation of biological samples, but require specific preparation procedures to maintain the strength of the employed methods. Here we report the optimization of the embedding protocol of nervous system samples for a correlative synchrotron X-ray computed microtomography (micro-CT) and transmission electron microscopy (TEM) approach. We demonstrate that it is possible to locate, with the micrometric resolution of micro-CT, specific volumes of interest for a further ultrastructural characterization to be performed with TEM. This approach can be applied to samples of different size and morphology up to several cm. Our optimized method represents an invaluable tool for investigating those pathologies in which microscopic alterations are localized in few confined regions, rather than diffused in entire tissues, organs or systems. We present a proof of concept of our method in a mouse model of Globoid Cells Leukodistrophy

Aquatic Organic Matter in the Seine Basin: Sources, Spatio-Temporal Variability, Impact of Urban Discharges and Influence on Micro-pollutant Speciation

International audienc

GA-NIFS: co-evolution within a highly star-forming galaxy group at z=3.7 witnessed by JWST/NIRSpec IFS

We present NIRSpec IFS observations of a galaxy group around the massive GS_4891 galaxy at z=3.7 in GOODS-South that includes two other two systems, GS_4891_n to the north and GS_28356 to the east. These observations, obtained as part of the GTO GA-NIFS program, allow for the first time to study the spatially resolved properties of the interstellar medium (ISM) and ionized gas kinematics of a galaxy at this redshift. Leveraging the wide wavelength range spanned with the high-dispersion grating (with resolving power R=2700) observations, covering from [OII]$\lambda$$\lambda$3726,29 to [SII]$\lambda$$\lambda$6716,31, we explore the spatial distribution of star-formation rate, nebular attenuation and gas metallicity, together with the mechanisms responsible for the excitation of the ionized gas. GS_4891 presents a clear gradient of gas metallicity (as traced by 12 + log(O/H)) by more than 0.2dex from the south-east (where a star-forming clump is identified) to the north-west. The gas metallicity in the less-massive northern system, GS_4891_n, is also higher by 0.2 dex than at the center of GS_4891, suggesting that inflows of lower-metallicity gas might be favoured in higher-mass systems. The kinematic analysis shows that GS_4891 presents velocity gradients in the ionized gas consistent with rotation. The region between GS_4891 and GS_4891_n does not present high gas turbulence which, together with the difference in gas metallicities, suggests that these two systems might be in a pre-merger stage. Finally, GS_4891 hosts an ionized outflow that extends out to r_out=1.2 kpc from the nucleus and reaches maximum velocities v_out of approximately 400 km/s. Despite entraining an outflowing mass rate of M_out$\sim$2Msun/yr, the low associated mass-loading factor, $\eta$=0.05, implies that the outflow does not have a significant impact on the star-formation activity of the galaxy.Comment: Submitted to Astronomy & Astrophysics on September 25th, 202

Distinct mechanisms for aerenchyma formation in leaf sheaths of rice genotypes displaying a quiescence or escape strategy for flooding tolerance

Background and Aims Rice is one of the few crops able to withstand periods of partial or even complete submergence. One of the adaptive traits of rice is the constitutive presence and further development of aerenchyma which enables oxygen to be transported to submerged organs. The development of lysigenous aerenchyma is promoted by ethylene accumulating within the submerged plant tissues, although other signalling mechanisms may also co-exist. In this study, aerenchyma development was analysed in two rice (Oryza sativa) varieties, ‘FR13A’ and ‘Arborio Precoce’, which show opposite traits in flooding response in terms of internode elongation and survival. Methods The growth and survival of rice varieties under submergence was investigated in the leaf sheath of ‘FR13A’ and ‘Arborio Precoce’. The possible involvement of ethylene and reactive oxygen species (ROS) was evaluated in relation to aerenchyma formation. Cell viability and DNA fragmentation were determined by FDA/FM4-64 staining and TUNEL assay, respectively. Ethylene production was monitored by gas chromatography and by analysing ACO gene expression. ROS production was measured by using Amplex Red assay kit and the fluorescent dye DCFH2-DA. The expression of APX1 was also evaluated. AVG and DPI solutions were used to test the effect of inhibiting ethylene biosynthesis and ROS production, respectively. Key Results Both the varieties displayed constitutive lysigenous aerenchyma formation, which was further enhanced when submerged. ‘Arborio Precoce’, which is characterized by fast elongation when submerged, showed active ethylene biosynthetic machinery associated with increased aerenchymatous areas. ‘FR13A’, which harbours the Sub1A gene that limits growth during oxygen deprivation, did not show any increase in ethylene production after submersion but still displayed increased aerenchyma. Hydrogen peroxide levels increased in ‘FR13A’ but not in ‘Arborio Precoce’. Conclusions While ethylene controls aerenchyma formation in the fast-elongating ‘Arborio Precoce’ variety, in ‘FR13A’ ROS accumulation plays an important role

Removal of Uracil by Uracil DNA Glycosylase Limits Pemetrexed Cytotoxicity: Overriding the Limit with Methoxyamine to Inhibit Base Excision Repair

Uracil DNA glycosylase (UDG) specifically removes uracil bases from DNA, and its repair activity determines the sensitivity of the cell to anticancer agents that are capable of introducing uracil into DNA. In the present study, the participation of UDG in the response to pemetrexed-induced incorporation of uracil into DNA was studied using isogenic human tumor cell lines with or without UDG (UDG+/+/UDG−/−). UDG−/− cells were very sensitive to pemetrexed. Cell killing by pemetrexed was associated with genomic uracil accumulation, stalled DNA replication, and catastrophic DNA strand breaks. By contrast, UDG+/+ cells were \u3e10 times more resistant to pemetrexed due to the rapid removal of uracil from DNA by UDG and subsequent repair of the resultant AP sites (abasic sites) via the base excision repair (BER). The resistance to pemetrexed in UDG+/+ cells could be reversed by the addition of methoxyamine (MX), which binds to AP sites and interrupts BER pathway. Furthermore, MX-bound AP sites induced cell death was related to their cytotoxic effect of dual inactivation of UDG and topoisomerase IIα, two genes that are highly expressed in lung cancer cells in comparison with normal cells. Thus, targeting BER-based therapy exhibits more selective cytotoxicity on cancer cells through a synthetic lethal mechanism

JADES: the emergence and evolution of Lyα emission and constraints on the intergalactic medium neutral fraction

The rest-frame UV recombination emission line Lyα can be powered by ionising photons from young massive stars in star-forming galaxies, but the fact that it can be resonantly scattered by neutral gas complicates its interpretation. For reionisation-era galaxies, a neutral intergalactic medium will scatter Lyα from the line of sight, making Lyα a useful probe of the neutral fraction evolution. Here, we explore Lyα in JWST/NIRSpec spectra from the ongoing JADES programme, which targets hundreds of galaxies in the well-studied GOODS-S and GOODS-N fields. These sources are UV-faint (−20.4  5.6 (as derived with optical lines) with line and continuum models to search for significant line emission. Through exploration of the R100 data, we find evidence for Lyα in 17 sources. This sample allowed us to place observational constraints on the fraction of galaxies with Lyα emission in the redshift range 5.6

JADES: Probing interstellar medium conditions at z∼5.5−9.5z\sim5.5-9.5 with ultra-deep JWST/NIRSpec spectroscopy

We present emission line ratios from a sample of 26 Lyman break galaxies from $z\sim5.5-9.5$ with $-17.0<M_{1500}<-20.4$, measured from ultra-deep JWST/NIRSpec MSA spectroscopy from JADES. We use 28 hour deep PRISM/CLEAR and 7 hour deep G395M/F290LP observations to measure, or place strong constraints on, ratios of widely studied rest-frame optical emission lines including H$\alpha$, H$\beta$, [OII] $\lambda\lambda$3726,3729, [NeIII] $\lambda$3869, [OIII] $\lambda$4959, [OIII] $\lambda$5007, [OI] $\lambda$6300, [NII] $\lambda$6583, and [SII] $\lambda\lambda$6716,6731 in individual $z>5.5$ spectra. We find that the emission line ratios exhibited by these $z\sim5.5-9.5$ galaxies occupy clearly distinct regions of line-ratio space compared to typical z~0-3 galaxies, instead being more consistent with extreme populations of lower-redshift galaxies. This is best illustrated by the [OIII]/[OII] ratio, tracing interstellar medium (ISM) ionisation, in which we observe more than half of our sample to have [OIII]/[OII]>10. Our high signal-to-noise spectra reveal more than an order of magnitude of scatter in line ratios such as [OII]/H$\beta$ and [OIII]/[OII], indicating significant diversity in the ISM conditions within the sample. We find no convincing detections of [NII] in our sample, either in individual galaxies, or a stack of all G395M/F290LP spectra. The emission line ratios observed in our sample are generally consistent with galaxies with extremely high ionisation parameters (log $U\sim-1.5$), and a range of metallicities spanning from $\sim0.1\times Z_\odot$ to higher than $\sim0.3\times Z_\odot$, suggesting we are probing low-metallicity systems undergoing periods of rapid star-formation, driving strong radiation fields. These results highlight the value of deep observations in constraining the properties of individual galaxies, and hence probing diversity within galaxy population.Comment: 20 pages, 9 figures, submitted to Astronomy & Astrophysics, updated values in table

JADES NIRSpec Spectroscopy of GN-z11: Lyman-α\alpha emission and possible enhanced nitrogen abundance in a z=10.60z=10.60 luminous galaxy

We present JADES JWST/NIRSpec spectroscopy of GN-z11, the most luminous candidate $z>10$ Lyman break galaxy in the GOODS-North field with $M_{UV}=-21.5$. We derive a redshift of $z=10.603$ (lower than previous determinations) based on multiple emission lines in our low and medium resolution spectra over $0.8-5.3\,\mu$m. We significantly detect the continuum and measure a blue rest-UV spectral slope of $\beta=-2.4$. Remarkably, we see spatially-extended Lyman-$\alpha$ in emission (despite the highly-neutral IGM expected at this early epoch), offset 555 km/s redward of the systemic redshift. From our measurements of collisionally-excited lines of both low- and high-ionization (including [O II] $\lambda3727$, [Ne III] $\lambda 3869$ and C III] $\lambda1909$) we infer a high ionization parameter ($\log U\sim -2$). We detect the rarely-seen N IV] $\lambda1486$ and N III]$\lambda1748$ lines in both our low and medium resolution spectra, with other high ionization lines seen in low resolution spectrum such as He II (blended with O III]) and C IV (with a possible P-Cygni profile). Based on the observed rest-UV line ratios, we cannot conclusively rule out photoionization from AGN. The high C III]/He II ratios, however, suggest a likely star-formation explanation. If the observed emission lines are powered by star formation, then the strong N III] $\lambda1748$ observed may imply an unusually high $N/O$ abundance. Balmer emission lines (H$\gamma$, H$\delta$) are also detected, and if powered by star formation rather than an AGN we infer a star formation rate of $\sim 20-30 M_{\odot}\,\rm yr^{-1}$ (depending on the IMF) and low dust attenuation. Our NIRSpec spectroscopy confirms that GN-z11 is a remarkable galaxy with extreme properties seen 430 Myr after the Big Bang.Comment: Submitted to Astronomy & Astrophysics, 14 pages, 9 figure