Biology subjects

Rosa, F.

Publications and source records attributed to Rosa, F..

7 recordsLinked to original sources

bioRxiv2024

Non-apoptotic caspase events and Atf3 expression underlie direct neuronal differentiation of adult neural stem cells

Neural stem cells (NSCs) are key physiological components of adult vertebrate brains, generating neurons over a lifetime. In the adult zebrafish pallium, NSCs persist at long term through balanced fate decisions that include direct neuronal conversions, i.e., delamination and neurogenesis without a division. The characteristics and mechanisms of these events remain unknown. Here we reanalyze intravital imaging data of adult pallial NSCs and observe shared delamination dynamics between NSCs and committed neuronal progenitors. In a candidate approach for mechanisms predicting NSC decisions, we build an NSC-specific genetic tracer of Caspase3/7 activation (Cas3*/Cas7*) in vivo and show that non-apoptotic Cas3*/7* events occur in adult NSCs and are biased towards neuronal conversion under physiological conditions. We further identify the transcription factor Atf3 as necessary to express this fate. Finally, we show that the Cas3*/7*/Atf3 pathways are part of the processes engaged when NSCs are recruited for neuronal regeneration. These results provide evidence for the non-apoptotic caspase events occurring in vertebrate adult NSCs and link these events with the NSC fate decision of direct conversion, important for long-term NSC population homeostasis.

developmental biology↗

bioRxiv2024

Electrophysiological signatures of a developmental delay in a stem cell model of KCNQ2 developmental and epileptic encephalopathy

BackgroundKCNQ2, encoding KV7.2 ion channels, has emerged as one of the prominent genes causing early onset seizures with developmental delay (KCNQ2 developmental and epileptic encephalopathy; KCNQ2-DEE). KCNQ2 de novo loss-of-function (LOF) and associated neuronal hyperexcitability have been accepted as mechanisms contributing to seizures. To investigate the developmental impact of KCNQ2 LOF, we generated patient iPSC-derived models for two previously reported de novo variants, p.(Arg325Gly) and p.(Gly315Arg), linked to severe congenital DEE. MethodsFunctional investigation of the two variants was initially performed in Xenopus laevis oocyte system. Patient-derived iPSC lines were differentiated using NGN2- and embryoid body-based protocols yielding neurons roughly corresponding to mid- and mid-late gestational stages, respectively. KV7- mediated M-current, passive neuronal properties, action potential generation and spontaneous oscillatory network activities were analysed with whole-cell patch clamping. FindingsStudied KCNQ2 variants showed LOF with a dominant-negative effect in the heterologous system. Reduced M-currents in patient iPSC-derived neurons corroborated a LOF as the main pathomechanism. Interestingly, this led to the reduced neuronal firing of the early neurons and to a disruption of complex oscillatory activity, with significantly reduced duration and amplitude of these events in patient iPSC-derived neurons. InterpretationWe provide experimental evidence for changing roles of the M-current throughout development and place disease variant-mediated M-current reduction in the context of the neuronal maturation in the prenatal brain. Based on the reduced neuronal firing and disrupted oscillatory activity seen in patient iPSC-derived neurons, we propose that a delayed/impaired maturation of neuronal and network properties underlies KCNQ-DEE caused by LOF variants.

neuroscience↗

bioRxiv2024

Land-use impacts on plant functional diversity throughout Europe

AimGlobal biodiversity loss resulting from anthropogenic land-use activities is a pressing concern, requiring precise assessments of impacts at large spatial extents. Existing models mainly focus on species richness and abundance, lacking insights into ecological mechanisms and species roles in ecosystem functioning. To bridge this gap, we conducted an extensive analysis of the impact of human land use on vascular plant functional diversity, across diverse land-use classes and bioregions in Europe, comparing it to traditional metrics. Location: Europe Time period: 1992-2019 Major taxa studied: Vascular plants MethodsIntegrating extensive databases of vegetation plots with spatial data on land use and land cover, we paired plots from areas actively used and modified by humans with plots from natural habitats under similar environmental conditions. Using species occurrences and traits, in each plot we computed three complementary functional diversity metrics (functional richness, evenness, and divergence), species richness and abundance. We assessed the impact of land use by comparing the metrics in the paired plots. ResultsOur findings revealed that, compared to natural habitats, anthropogenic land use exhibits lower functional richness and divergence but higher functional evenness across most land-use classes and bioregions. The response of functional richness was more marked than the other two metrics and especially pronounced in croplands and urban areas and in northern bioregions. Functional richness exhibited a pattern that did not fully overlap with the trend in species richness, providing useful complementary information. Main conclusionsWe provide a large-scale precise assessment of anthropogenic land-use impacts on functional diversity across Europe. Our findings indicate that: (i) human disturbance significantly alters plant functional diversity compared to natural habitats; (ii) this alteration goes in the direction of functional homogenization; (i) functional diversity metrics complement traditional metrics by offering deeper insights into the ecological mechanisms in response to anthropogenic land use.

ecology↗

bioRxiv2020

Using big data to estimate whale watching effort

The growing concerns about the negative effects caused by whale watching on wild The growing concerns about the negative effects caused by whale watching on wild cetacean populations are evincing the need to measure whale watching effort more precisely. The current alternatives do not provide sufficient information or imply time-consuming and staff intensive tasks that limit their effectiveness to establish the maximum carrying capacity for this touristic activity. A methodology based on big data analysis, using Automatic Information System (AIS) messages, can provide valuable vessel activity information, which is necessary to estimate whale watching effort on cetacean populations. We used AIS data to automatically detect whale watching operations and quantify whale watching effort with high spatial and temporal resolution in the Canary Islands. The results obtained in this study are very encouraging, proving that the methodology is able to estimate seasonal and annual trends in the whale watching effort. The methodology has also proved to be effective in providing detailed spatial information about the whale watching effort, which makes an interesting tool to manage spatial regulations and enforce exclusion zones. The widespread use of AIS devices in maritime navigation provides an enormous potential to easily extend this methodology to other regions worldwide. Any public strategy aimed at the sustainable use of the marine resources should enhance the use of this kind of information technologies, collecting and archiving detailed information on the activity of all the vessels, especially in marine protected areas.

ecology↗

bioRxiv2019

Manganese-enhanced MRI (MEMRI) in breast and prostate cancers: exploring the potential role of calcium receptors.

ProceduresTo assess the relationship between Manganese Enhanced Magnetic Resonance Imaging (MEMRI) and the expression of calcium receptors in human prostate and breast cancer animal models.\n\nMethodsNOD/SCID mice were inoculated with MDA-MB-231 breast cancer cells and prostate PC3 cancer cells to develop orthotopic or pseudometastatic cancer animal models. Mice were studied on a clinical 3T scanner by using a prototype birdcage coil before and after intravenous injection of MnCl2. Assessment of receptors status was carried out after the MR images acquisition by immunohistochemistry on excised tumours.\n\nResultsManganese contrast enhancement in breast or prostate cancer animal models well correlated with CaSR expression (p<0.01), whereas TRPV6 expression levels appeared not relevant to the Mn uptake.\n\nConclusionMEMRI appears an efficient tool to characterize human breast and prostate cancer animal models in the presence of different expression level of CaSR.

cancer biology↗

bioRxiv2019

Interplay between the human gut microbiome and host metabolism

The human gut is inhabited by a complex and metabolically active microbial ecosystem regulating host health. While many studies have focused on the effect of individual microbial taxa, the metabolic potential of the entire gut microbial ecosystem has been largely under-explored. We characterised the gut microbiome of 1,004 twins via whole shotgun metagenomic sequencing (average 39M reads per sample). We observed greater similarity, across unrelated individuals, for functional metabolic pathways (82%) than for taxonomic composition (43%). We conducted a microbiota-wide association study linking both taxonomic information and microbial metabolic pathways with 673 blood and 713 faecal metabolites (Metabolon, Inc.). Metabolic pathways associated with 34% of blood and 95% of faecal metabolites, with over 18,000 significant associations, while species-level results identified less than 3,000 associations, suggesting that coordinated action of multiple taxa is required to affect the metabolome. Finally, we estimated that the microbiome mediated a crosstalk between 71% of faecal and 15% of blood metabolites, highlighting six key species (unclassified Subdoligranulum spp., Faecalibacterium prausnitzii, Roseburia inulinivorans, Methanobrevibacter smithii, Eubacterium rectale, and Akkermansia muciniphila). Because of the large inter-person variability in microbiome composition, our results underline the importance of studying gut microbial metabolic pathways rather than focusing purely on taxonomy to find therapeutic and diagnostic targets.

microbiology↗

bioRxiv2018

Mycobacterium tuberculosis-specific CD4+ and CD8+ T cells differ in their capacity to recognize infected macrophages

Containment of Mycobacterium tuberculosis (Mtb) infection requires T cell recognition of infected macrophages. Mtb has evolved to tolerate, evade, and subvert host immunity. Despite a vigorous and sustained CD8+ T cell response during Mtb infection, CD8+ T cells make limited contribution to protection. Here, we ask whether the ability of Mtb-specific T cells to restrict Mtb growth is related to their capacity to recognize Mtb-infected macrophages.\n\nWe derived CD8+ T cell lines that recognized the Mtb immunodominant epitope TB10.44-11 and compared them to CD4+ T cell lines that recognized Ag85b240-254 or ESAT63-17. While the CD4+ T cells recognized Mtb-infected macrophages and inhibited Mtb growth in vitro, the TB10.4-specific CD8+ T cells neither recognized Mtb-infected macrophages nor restricted Mtb growth. TB10.4-specific CD8+ T cells recognized macrophages infected with Listeria monocytogenes expressing TB10.4. However, over-expression of TB10.4 in Mtb did not confer recognition by TB10.4-specific CD8+ T cells. Importantly, CD8+ T cells recognized macrophages pulsed with irradiated Mtb, indicating that macrophages can efficiently cross-present the TB10.4 protein and raising the possibility that viable bacilli might suppress cross-presentation. Importantly, polyclonal CD8+ T cells specific for Mtb antigens other than TB10.4 recognized Mtb-infected macrophages in a MHC-restricted manner.\n\nAs TB10.4 elicits a dominant CD8+ T cell response that poorly recognizes Mtb-infected macrophages, we propose that TB10.4 acts as a decoy antigen. Moreover, it appears that this response overshadows subdominant CD8+ T cell response that can recognize Mtb-infected macrophages. The ability of Mtb to subvert the CD8+ T cell response may explain why CD8+ T cells make a disproportionately small contribution to host defense compared to CD4+ T cells. The selection of Mtb antigens for vaccines has focused on antigens that generate immunodominant responses. We propose that establishing whether vaccine-elicited, Mtb-specific T cells recognize Mtb-infected macrophages could be a useful criterion for preclinical vaccine development.

immunology↗