Planar cell polarity zebrafish models of congenital scoliosis reveal novel underlying defects in notochord morphogenesis.

Congenital scoliosis (CS) is a type of vertebral malformation whose etiology remains elusive. The notochord is pivotal for vertebrae development but its role in CS is still understudied. Zebrafish knockout of ptk7a, a planar cell polarity (PCP) gene that is essential for convergence and extension (C&E) of the notochord, developed congenital scoliosis-like vertebral malformations (CVM). Maternal zygotic ptk7a mutants displayed severe C&E defects of the notochord. Excessive apoptosis occurred in the malformed notochord, causing a significantly reduced number of vacuolated cells, and compromising the mechanical properties of the notochord. The latter manifested as a less stiff extracellular matrix along with a significant reduction in the number of the caveolae and severely loosened intercellular junctions in the vacuolated region. These defects led to focal kinks, abnormal mineralization, and CVM exclusively at the anterior spine. Loss of function of another PCP gene, vangl2, also revealed excessive apoptosis in the notochord associated with CVM. This study suggests a new model for CS pathogenesis that is associated with defects in notochord C&E and highlights an essential role of PCP signaling in vertebrae development.

The risk of contact between visitors and Borrelia burgdorferi-infected ticks is associated with fine-scale landscape features in a southeastern Canadian nature park.

BACKGROUND: Infectious diseases are emerging across temperate regions of the world, and, for some, links have been made between landscapes and emergence dynamics. For tick-borne diseases, public parks may be important exposure sites for people living in urbanized areas of North America and Europe. In most cases, we know more about the ecological processes that determine the hazard posed by ticks as disease vectors than we do about how human population exposure varies in urban natural parks. METHODS: In this study, infrared counters were used to monitor visitor use of a public natural park in southern Quebec, Canada. A risk index representing the probability of encounters between humans and infected vectors was constructed. This was done by combining the intensity of visitor trail use and the density of infected nymphs obtained from field surveillance. Patterns of risk were examined using spatial cluster analysis. Digital forest data and park infrastructure data were then integrated using spatially explicit models to test whether encounter risk levels and its components vary with forest fragmentation indicators and proximity to park infrastructure. RESULTS: Results suggest that, even at a very fine scales, certain landscape features and infrastructure can be predictors of risk levels. Both visitors and Borrelia burgdorferi-infected ticks concentrated in areas where forest cover was dominant, so there was a positive association between forest cover and the risk index. However, there were no associations between indicators of forest fragmentation and risk levels. Some high-risk clusters contributed disproportionately to the risk distribution in the park relative to their size. There were also two high-risk periods, one in early summer coinciding with peak nymphal activity, and one in early fall when park visitation was highest. CONCLUSIONS: Here, we demonstrate the importance of integrating indicators of human behaviour visitation with tick distribution data to characterize risk patterns for tick-borne diseases in public natural areas. Indeed, understanding the environmental determinants of human-tick interactions will allow organisations to deploy more effective risk reduction interventions targeted at key locations and times, and improve the management of public health risks associated with tick-borne diseases in public spaces.

Site occupancy by American martens and fishers in temperate deciduous forests of Québec.

Interspecific interactions can mediate site occupancy of sympatric species and can be a key factor in habitat use patterns. American martens (Martes americana) and Fishers (Pekania pennanti) are two sympatric mesocarnivores in eastern North American forests. Due to their larger size, fishers have a competitive advantage over martens. We investigated site occupancy of martens and fishers in temperate deciduous forests of Québec, an environment modified by forest management and climate change. We formulated hypotheses on the spatial distribution of the studied species based on the knowledge of local trappers and on the scientific literature regarding forest cover composition, habitat fragmentation, and competitive relationships. We used a network of 49 camera traps monitored over two fall seasons to document site occupancy by both species. We used two-species site occupancy models to assess habitat use and the influence of fishers on martens at spatial grains of different sizes. None of the habitat variables that we considered explained site occupancy by fishers. Availability of dense old coniferous stands explained the spatial distribution of martens both at the home range grain size and at the landscape grain size. We identified the characteristics of habitat hotspots based on the knowledge of trappers, which highlighted the importance of stand composition, height, age, and canopy closure. The characteristics of habitat hotspots for martens in temperate deciduous forests refine the habitat suitability model for American martens that was originally developed for boreal forests of Québec.

Les interactions interspécifiques peuvent affecter l'occupation de sites par des espèces sympatriques et jouer un rôle clé dans leur utilisation des habitats. La martre d'Amerique (Martes americana) et le pékan (Pekania pennanti) sont deux mésocarnivores sympatriques des forêts de l'est de l'Amérique du Nord. En raison de sa grande taille, le pékan est un compétiteur dominant de la martre. Nous avons étudié l'occupation des sites par la martre et le pékan dans la forêt tempérée feuillue du Québec, un environnement modifié par l'aménagement forestier et les changements climatiques. Nous avons formulé des hypothèses sur la répartition spatiale des espèces étudiées en nous basant sur les connaissances des trappeurs locaux et sur la littérature scientifique en ce qui a trait à la composition du couvert forestier, à la fragmentation de l'habitat, et aux relations de compétition. Nous avons utilisé un réseau de 49 appareils photo à déclenchement automatique pendant deux automnes pour documenter l'occupation des sites par les deux espèces. Nous avons utilisé des modèles d'occupation de sites à deux espèces afin d'évaluer l'effet de la présence du pékan sur l'utilisation de l'habitat par la martre à des échelles spatiales de résolutions variables. Aucune des variables d'habitat que nous avons prises en compte n'explique l'occupation des sites par les pékans. La disponibilité de vieux peuplements denses de conifères explique la répartition spatiale de la martre aux échelles spatiales du domaine vital et du paysage. Nous avons développé un indice d'habitat potentiel basé sur les connaissances des trappeurs, qui a mis en évidence l'importance de la composition, de la hauteur, de l'âge et de la densité des peuplements. Cet indice affine, pour les forêts tempérées feuillues du Québec, le modèle de qualité de l'habitat de la martre d'Amérique originellement élaboré pour la forêt boréale.

Lowering the rate of timber harvesting to mitigate impacts of climate change on boreal caribou habitat quality in eastern Canada.

Many boreal populations of woodland caribou (Rangifer tarandus caribou) have declined in Canada, a trend essentially driven by the increasing footprint of anthropogenic disturbances and the resulting habitat-mediated apparent competition that increases predation pressure. However, the influence of climate change on these ecological processes remains poorly understood. We evaluated how climate change will affect boreal caribou habitat over the 2030-2100 horizon and in a 9.94 Mha study area, using a climate-sensitive simulation ensemble that integrates climate-induced changes in stand dynamics, fire regime, and different levels of commercial timber harvesting. We assessed the relative importance of these three drivers under projections made using different radiative forcing scenarios (RCP 2.6, 4.5, 8.5). Habitat quality was estimated from resource selection functions built with telemetry data collected from 121 caribou between 2004 and 2011 in 7 local populations. At the beginning of our simulations, caribou habitat was already structured along a south-to-north increasing quality gradient. Simulations revealed changes in forest cover that are driven by climate-induced variations in fire regime and scenarios of harvesting levels, resulting in the loss of older coniferous forests and an increase in deciduous stands. These changes induced a generalized decrease in the average habitat quality and in the percentage of high-quality habitat for caribou, and in a northward recession of suitable habitat. Timber harvesting was the most important agent of change for the 2030-2050 horizon, although it was slowly replaced by changes in fire regime until 2100. Our results clearly showed that it is possible to maintain the current average habitat quality for caribou in future scenarios that consider a reduction in harvested volumes, the only lever under our control. This suggests that we still have the capacity to conciliate socioeconomic development and caribou conservation imperatives in the face of climate change, an important issue debated throughout the species distribution range.

Transmission patterns of tick-borne pathogens among birds and rodents in a forested park in southeastern Canada.

Ixodes scapularis ticks are expanding their range in parts of northeastern North America, bringing with them pathogens of public health concern. While rodents like the white-footed mouse, Peromyscus leucopus, are considered the primary reservoir of many emerging tick-borne pathogens, the contribution of birds, as alternative hosts and reservoirs, to local transmission cycles has not yet been firmly established. From 2016 to 2018, we collected host-seeking ticks and examined rodent and bird hosts for ticks at 48 sites in a park where blacklegged ticks are established in Quebec, Canada, in order to characterize the distribution of pathogens in ticks and mammalian and avian hosts. We found nearly one third of captured birds (n = 849) and 70% of small mammals (n = 694) were infested with I. scapularis. Five bird and three mammal species transmitted Borrelia burgdorferi to feeding larvae (n larvae tested = 2257) and we estimated that about one fifth of the B. burgdorferi-infected questing nymphs in the park acquired their infection from birds, the remaining being attributable to mice. Ground-foraging bird species were more parasitized than other birds, and species that inhabited open habitat were more frequently infested and were more likely to transmit B. burgdorferi to larval ticks feeding upon them. Female birds were more likely to transmit infection than males, without age differentiation, whereas in mice, adult males were more likely to transmit infection than juveniles and females. We also detected Borrelia miyamotoi in larvae collected from birds, and Anaplasma phagocytophilum from a larva collected from a white-footed mouse. This study highlights the importance of characterising the reservoir potential of alternative reservoir hosts and to quantify their contribution to transmission dynamics in different species assemblages. This information is key to identifying the most effective host-targeted risk mitigation actions.

Interpericyte tunnelling nanotubes regulate neurovascular coupling.

Signalling between cells of the neurovascular unit, or neurovascular coupling, is essential to match local blood flow with neuronal activity. Pericytes interact with endothelial cells and extend processes that wrap capillaries, covering up to 90% of their surface area1,2. Pericytes are candidates to regulate microcirculatory blood flow because they are strategically positioned along capillaries, contain contractile proteins and respond rapidly to neuronal stimulation3,4, but whether they synchronize microvascular dynamics and neurovascular coupling within a capillary network was unknown. Here we identify nanotube-like processes that connect two bona fide pericytes on separate capillary systems, forming a functional network in the mouse retina, which we named interpericyte tunnelling nanotubes (IP-TNTs). We provide evidence that these (i) have an open-ended proximal side and a closed-ended terminal (end-foot) that connects with distal pericyte processes via gap junctions, (ii) carry organelles including mitochondria, which can travel along these processes, and (iii) serve as a conduit for intercellular Ca2+ waves, thus mediating communication between pericytes. Using two-photon microscope live imaging, we demonstrate that retinal pericytes rely on IP-TNTs to control local neurovascular coupling and coordinate light-evoked responses between adjacent capillaries. IP-TNT damage following ablation or ischaemia disrupts intercellular Ca2+ waves, impairing blood flow regulation and neurovascular coupling. Notably, pharmacological blockade of Ca2+ influx preserves IP-TNTs, rescues light-evoked capillary responses and restores blood flow after reperfusion. Our study thus defines IP-TNTs and characterizes their critical role in regulating neurovascular coupling in the living retina under both physiological and pathological conditions.

The Novel Small Molecule TRVA242 Stabilizes Neuromuscular Junction Defects in Multiple Animal Models of Amyotrophic Lateral Sclerosis.

Amyotrophic lateral sclerosis (ALS) is a debilitating neurodegenerative disorder in which the neuromuscular junction progressively degenerates, leading to movement difficulties, paralysis, and eventually death. ALS is currently being treated by only two FDA-approved drugs with modest efficacy in slowing disease progression. Often, the translation of preclinical findings to bedside terminates prematurely as the evaluation of potential therapeutic compounds focuses on a single study or a single animal model. To circumscribe these issues, we screened 3,765 novel small molecule derivatives of pimozide, a recently identified repurposed neuroleptic for ALS, in Caenorhabditis elegans, confirmed the hits in zebrafish and validated the most active compounds in mouse genetic models. Out of the 27 small molecules identified from the high-throughput screen in worms, 4 were found to recover locomotor defects in C. elegans and genetic zebrafish models of ALS. TRVA242 was identified as the most potent compound as it significantly improved efficiency in rescuing locomotor, motorneuron, and neuromuscular junction synaptic deficits in a C. elegans TDP-43 model and in multiple zebrafish genetic (TDP-43, SOD1, and C9ORF72) models of ALS. The actions of TRVA242 were also conserved in a mammalian model as it also stabilized neuromuscular junction deficits in a mouse SOD1 model of ALS. Compounds such as TRVA242 therefore represent new potential therapeutics for the treatment of ALS.

Corrigendum: The Dietary Flavonoid, Luteolin, Negatively Affects Neuronal Differentiation.

[This corrects the article DOI: 10.3389/fnmol.2019.00041.].

secCl is a cys-loop ion channel necessary for the chloride conductance that mediates hormone-induced fluid secretion in Drosophila.

Organisms use circulating diuretic hormones to control water balance (osmolarity), thereby avoiding dehydration and managing excretion of waste products. The hormones act through G-protein-coupled receptors to activate second messenger systems that in turn control the permeability of secretory epithelia to ions like chloride. In insects, the chloride channel mediating the effects of diuretic hormones was unknown. Surprisingly, we find a pentameric, cys-loop chloride channel, a type of channel normally associated with neurotransmission, mediating hormone-induced transepithelial chloride conductance. This discovery is important because: 1) it describes an unexpected role for pentameric receptors in the membrane permeability of secretory epithelial cells, and 2) it suggests that neurotransmitter-gated ion channels may have evolved from channels involved in secretion.

Individual knock out of glycine receptor alpha subunits identifies a specific requirement of glra1 for motor function in zebrafish.

Glycine receptors (GlyRs) are ligand-gated chloride channels mediating inhibitory neurotransmission in the brain stem and spinal cord. They function as pentamers composed of alpha and beta subunits for which 5 genes have been identified in human (GLRA1, GLRA2, GLRA3, GLRA4, GLRB). Several in vitro studies showed that the pentameric subtype composition as well as its stoichiometry influence the distribution and the molecular function of the receptor. Moreover, mutations in some of these genes are involved in different human conditions ranging from tinnitus to epilepsy and hyperekplexia, suggesting distinct functions of the different subunits. Although the beta subunit is essential for synaptic clustering of the receptor, the specific role of each alpha subtype is still puzzling in vivo. The zebrafish genome encodes for five glycine receptor alpha subunits (glra1, glra2, glra3, glra4a, glra4b) thus offering a model of choice to investigate the respective role of each subtype on general motor behaviour. After establishing a phylogeny of GlyR subunit evolution between human and zebrafish, we checked the temporal expression pattern of these transcripts during embryo development. Interestingly, we found that glra1 is the only maternally transmitted alpha subunit. We also showed that the expression of the different GlyR subunits starts at different time points during development. Lastly, in order to decipher the role of each alpha subunit on the general motor behaviour of the fish, we knocked out individually each alpha subunit by CRISPR/Cas9-targeted mutagenesis. Surprisingly, we found that knocking out any of the alpha2, 3, a4a or a4b subunit did not lead to any obvious developmental or motor phenotype. However, glra1-/- (hitch) embryos depicted a strong motor dysfunction from 3 days, making them incapable to swim and thus leading to their premature death. Our results infer a strong functional redundancy between alpha subunits and confirm the central role played by glra1 for proper inhibitory neurotransmission controlling locomotion. The genetic tools we developed here will be of general interest for further studies aiming at dissecting the role of GlyRs in glycinergic transmission in vivo and the hitch mutant (hic) is of specific relevance as a new model of hyperekplexia.

Single and Synergistic Effects of Cannabidiol and Δ-9-Tetrahydrocannabinol on Zebrafish Models of Neuro-Hyperactivity.

In this study, we aimed to investigate the effect of the two main active cannabinoids extracted from cannabis: Δ-9-tetrahydrocannabinol (THC) and cannabidiol (CBD) on two distinct behavioral models of induced neuro-hyperactivity. We have taken advantage of two previously developed zebrafish models of neuro-hyperactivity: a chemically induced pentylenetetrazole model and a genetic model caused by loss-of-function mutations in the GABA receptor subunit alpha 1 (GABRA1-/-). Both CBD and THC have a significant effect on the behavioral changes induced by both models. Importantly, we have also shown that when applied together at different ratios of THC to CBD (1:1, 1:5, and 1:10), there was a synergistic effect at a ratio of 1:1. This was particularly important for the genetically induced neuro-hyperactivity as it brought the concentrations of THC and CBD required to oppose the induced behavioral changes to levels that had much less of an effect on baseline larval behavior. The results of this study help to validate the ability of THC and CBD to oppose neuro-hyperactivity linked to seizure modalities. Additionally, it appears that individually, each cannabinoid may be more effective against the chemically induced model than against the GABRA1-/- transgenic model. However, when applied together, the concentration of each compound required to oppose the GABRA1-/- light-induced activity was lowered. This is of particular interest since the use of cannabinoids as therapeutics can be dampened by their side-effect profile. Reducing the level of each cannabinoid required may help to prevent off target effects that lead to side effects. Additionally, this study provides a validation of the complimentary nature of the two zebrafish models and sets a platform for future work with cannabinoids, particularly in the context of neuro-hyperactivity disorders such as epilepsy.

Glycine Regulates Neural Stem Cell Proliferation During Development via Lnx1-Dependent Notch Signaling.

During development of the zebrafish embryo, glycine signaling promotes the differentiation of neural stem cells (NSCs). We found that glycine signaling suppresses the expression of Ligand of Numb X1 (lnx1, Ligand of numb protein-x1), a gene of unknown function during NSC differentiation that is selectively expressed in the embryonic central nervous system (CNS). As a consequence, Numb levels were stabilized and Notch activity (measured as her4.1 expression) was reduced, promoting NSC differentiation. These consequent actions were blocked by knockdown of lnx1. In contrast, lnx1 overexpression increased NSC proliferation and led to defects of neural tube closure at the early stages of development. Thus, our data provide evidence that glycine/lnx1 signaling modulates NSC proliferation by regulation of Notch signaling.

The Dietary Flavonoid, Luteolin, Negatively Affects Neuronal Differentiation.

Luteolin, a polyphenolic plant flavonoid, has been attributed with numerous beneficial properties like anti-cancer, antioxidant, and anti-inflammatory action. Luteolin has been reported earlier to be neuroprotective in models of spinal cord injury and traumatic brain injury and also induces neurite outgrowth in PC12 cells. However, the effect of luteolin on early differentiation, which might be important for its beneficial effects, is unknown. In this report, we show that luteolin negatively affects early differentiation of embryonic stem cells, hampering the formation of embryoid bodies. At later stages of differentiation, luteolin specifically inhibits neuronal differentiation, where the expression of early neuronal markers is suppressed, whereas luteolin treatment does not inhibit expression of meso- and endodermal markers. Further, in a developing zebrafish model, luteolin treatment leads to fewer numbers of mitotic cells in the brain. These specific effects of luteolin on neuronal differentiation could possibly be due to its ability to inhibit the lysine acetyltransferase, p300, since the structurally closely related p300 non-inhibitor flavonoid, apigenin, does not inhibit neuronal differentiation. These results show that luteolin perturbs neuronal differentiation of embryonic stem cells.

Neuromuscular junction abnormalities in a zebrafish loss-of-function model of TDP-43.

Almost 90% of amyotrophic lateral sclerosis (ALS) cases are characterized by the presence of aggregates of insoluble, misfolded cytoplasmic TAR DNA binding protein of 43 kDa (TDP-43). Distal axonopathy with impaired neuromuscular junctions (NMJs) before motor neuron degeneration or clinical onset of symptoms has been hypothesized as an early pathology in ALS. However, synaptic defects at the NMJ caused by TDP-43 mutations have not been characterized. In this study, we examined a previously reported zebrafish line expressing the tardbpY220X/Y220X variant, which results in an unstable and degraded protein. These tardbp-/- larvae, however, mature normally due to the upregulated expression of an alternative splice variant of the tardbp paralog tardbp-like, or tardbpl. We generated a mutant line with a CRISPR/Cas9-mediated 5-base pair deletion encompassing the ATG start codon of tardbpl and in-crossed these with tardbp-/- mutants to obtain tardbp-/- and tardbpl-/- double mutants, herein referred to as hom/hom. We subsequently characterized morphological, coiling, locomotor, synaptic, and NMJ structural abnormalities in the hom/hom mutants and in their genotypic controls. We observed that hom/hom mutants displayed gross morphological defects, early lethality, reduced locomotor function, aberrant quantal transmission, and perturbed synapse architecture at the NMJ. We further employed pharmacological manipulations in an effort to rescue phenotypic defects and observed that tardbp+/-; tardbpl-/- (herein referred to as het/hom) mutants, but not hom/hom mutants, were sensitive to chronic treatments of BAY K 8644, an L-type calcium channel agonist. This result highlights the importance of partial vs. complete loss of allelic functions of TDP-43. NEW & NOTEWORTHY This study highlights the importance of partial vs. complete loss of allelic functions of TDP-43 in a zebrafish loss of function model, thus making it an attractive tool for drug screening approaches.

Glycine decarboxylase deficiency-induced motor dysfunction in zebrafish is rescued by counterbalancing glycine synaptic level.

Glycine encephalopathy (GE), or nonketotic hyperglycinemia (NKH), is a rare recessive genetic disease caused by defective glycine cleavage and characterized by increased accumulation of glycine in all tissues. Here, based on new case reports of GLDC loss-of-function mutations in GE patients, we aimed to generate a zebrafish model of severe GE in order to unravel the molecular mechanism of the disease. Using CRISPR/Cas9, we knocked out the gldc gene and showed that gldc-/- fish recapitulate GE on a molecular level and present a motor phenotype reminiscent of severe GE symptoms. The molecular characterization of gldc-/- mutants showed a broad metabolic disturbance affecting amino acids and neurotransmitters other than glycine, with lactic acidosis at stages preceding death. Although a transient imbalance was found in cell proliferation in the brain of gldc-/- zebrafish, the main brain networks were not affected, thus suggesting that GE pathogenicity is mainly due to metabolic defects. We confirmed that the gldc-/- hypotonic phenotype is due to NMDA and glycine receptor overactivation, and demonstrated that gldc-/- larvae depict exacerbated hyperglycinemia at these synapses. Remarkably, we were able to rescue the motor dysfunction of gldc-/- larvae by counterbalancing pharmacologically or genetically the level of glycine at the synapse.

γ-Aminobutyric acid receptor alpha 1 subunit loss of function causes genetic generalized epilepsy by impairing inhibitory network neurodevelopment.

OBJECTIVE: In humans, mutations of the γ-aminobutyric acid receptor subunit 1 (GABRA1) cause either mild or severe generalized epilepsy. Although these epilepsy-causing mutations have been shown to disrupt the receptor activity in vitro, their in vivo consequences on brain development and activity are not known. Here, we aim at unraveling the epileptogenesis mechanisms of GABRA1 loss of function. METHODS: We generated a gabra1-/- zebrafish mutant line displaying highly penetrant epileptic seizures. We sought to identify the underlying molecular mechanisms through unbiased whole transcriptomic assay of gabra1-/- larval brains. RESULTS: Interestingly, mutant fish show fully penetrant seizures at juvenile stages that accurately mimic tonic-clonic generalized seizures observed in patients. Moreover, highly penetrant seizures can be induced by light stimulation, thus providing us with the first zebrafish model in which evident epileptic seizures can be induced by nonchemical agents. Our transcriptomic assay identified misregulated genes in several pathways essential for correct brain development. More specifically, we show that the early development of the brain inhibitory network is specifically affected. Although the number of GABAergic neurons is not altered, we observed a drastic reduction in the number of inhibitory synapses and a decreased complexity of the GABAergic network. This is consistent with the disruption in expression of many genes involved in axon guidance and synapse formation. SIGNIFICANCE: Together with the role of GABA in neurodevelopment, our data identify a novel aspect of epileptogenesis, suggesting that the substratum of GABRA1-deficiency epilepsy is a consequence of early brain neurodevelopmental defects, in particular at the level of inhibitory network wiring.

Glycine Promotes the Survival of a Subpopulation of Neural Stem Cells.

Glycine is mainly known as an inhibitory neurotransmitter in adult mature neurons, regulating neuronal network activity in the central nervous system. In contrast, during embryogenesis glycine can act as an excitatory neurotransmitter and generates the first electrical signal in immature neurons. The roles and functional significance of this excitatory glycinergic activity during neurodevelopment are still unclear. Using the zebrafish embryo as a model, we previously showed that glycine regulates proliferation and differentiation of neural stem cells (NSCs) to interneurons. Moreover, we identified that glycine signaling in NSCs is associated with several common developmental pathways and surprisingly also the p53-related apoptosis. Here we investigated how glycine signaling regulates NSC survival. First, we showed by two approaches, acridine orange staining and active caspase 3 immunostaining that defects in glycine signaling induce an early and transient cell death, which was suppressed by knockdown of p53. Then, we developed an NSC transplantation strategy to directly assess NSC-autonomous development upon perturbing glycine signaling. In vivo time-lapse imaging showed that disruption of glycine signaling disturbed the normal NSC interkinetic nuclear migration, leading to cell cycle arrest and apoptosis. Finally, we analyzed two main subpopulations of NSCs, expressing either nestin or GFAP, by in situ labeling and in transgenic lines expressing GFP in either population. We found that disruption of glycine signaling induced a drastic and selective loss of nestin-positive (nestin+) NSCs, which was only partially rescued upon p53 knockdown. Taken together, our findings support a role of glycine signaling in promoting survival of the nestin+ NSC subpopulation early during development.

Hindcasting the impacts of land-use changes on bird communities with species distribution models of Bird Atlas data.

Habitat loss and degradation induced by human development are among the major threats to biodiversity worldwide. In this study, we tested our ability to predict the response of bird communities (128 species) to land-use changes in southern Quebec (~483,100 km2 ) over the last 30 yr (between 1984-1989 and 2010-2014) by using species distribution models (299,302 occurrences in 30,408 locations) from a hindcasting perspective. Results were grouped by functional guilds to infer potential impacts on ecosystem services, and to relate model transferability (i.e., ability of our models to be generalized to other times and scales) to specific functional and life-history traits. Overall, our models were able to accurately predict, both in space and time, habitat suitability for 69% of species, especially for granivorous, nonmigrant, tree-nesting species, and species that are tied to agricultural areas under intensive use. These findings indicate that model transferability depends upon specific functional and life-history traits, providing further evidence that species' ecologies affect the ability of models to accurately predict bird distributions. Declining bird species were mostly short-distance migrants that were associated with open habitats (agricultural and nonproductive forest) with aerial insectivorous or granivorous diets, which may be related to agricultural intensification and land abandonment. Land-use changes were positive for some forest bird species that were mainly associated with mixed and deciduous forests, generalist diets and tree-nesting strategies. Yet cavity-nesting birds have suffered substantial reductions in their distributions, suggesting that cumulative effects of intensive logging and wildfires on mature forests pose a threat for forest-specialist species. Habitat suitability changes predicted by our coarse-scale species distribution models partially agreed with the long-term trends reported by the North American Breeding Bird Survey. Our findings confirm land-use change as a key driving force for shaping bird communities in southern Quebec, together with the need to explicitly incorporate it into global change scenarios that better inform decision-makers on conservation and management.

The positive carbon stocks-biodiversity relationship in forests: co-occurrence and drivers across five subclimates.

Carbon storage in forests and its ability to offset global greenhouse gas emissions, as well as biodiversity and its capacity to support ecosystem functions and services, are often considered separately in landscape planning. However, the potential synergies between them are currently poorly understood. Identifying the spatial patterns and factors driving their co-occurrence across different climatic zones is critical to more effectively conserve forest ecosystems at the regional level. Here, we integrated information of National Forest Inventories and Breeding Bird Atlases across Europe and North America (Spain and Quebec, respectively), covering five subclimates (steppe, dry Mediterranean, humid Mediterranean, boreal, and temperate). In particular, this study aimed to (1) determine the spatial patterns of both forest carbon stocks and biodiversity (bird richness, tree richness, and overall biodiversity) and the factors that influence them; (2) establish the relationships between forest carbon stocks and biodiversity; and (3) define and characterize the areas of high (hotspots) and low (coldspots) values of carbon and biodiversity, and ultimately quantify their spatial overlap. Our results show that the factors affecting carbon and biodiversity vary between regions and subclimates. The highest values of carbon and biodiversity were found in northern Spain (humid Mediterranean subclimate) and southern Quebec (temperate subclimate) where there was more carbon as climate conditions were less limiting. High density and structural diversity simultaneously favored carbon stocks, tree, and overall biodiversity, especially in isolated and mountainous areas, often associated with steeper slopes and low accessibility. In addition, the relationship between carbon stocks and biodiversity was positive in both regions and all subclimates, being stronger where climate is a limiting factor for forest growth. The spatial overlap between hotspots of carbon and biodiversity provides an excellent opportunity for landscape planning to maintain carbon stocks and conserve biodiversity. The variables positively affecting carbon and biodiversity were also driving the hotspots of both carbon and biodiversity, emphasizing the viability of "win-win" solutions. Our results highlight the need to jointly determine the spatial patterns of ecosystem services and biodiversity for an effective and sustainable planning of forest landscapes that simultaneously support conservation and mitigate climate change.