Joana Ferreira
@cnc.uc.pt
Center for Neuroscience and Cell Biology - CNC / Institute of Interdisciplinary Research - IIIUC
University of Coimbra
Joana Ferreira, graduated in Biology in 2006 (University of Coimbra) and completed her PhD in Biosciences, specialty Neurosciences (University of Coimbra) in 2012, under the co-supervision of Ana Luísa Carvalho at the Center for Neuroscience and Cell Biology (University of Coimbra, Portugal) and Ann Marie Craig at the Center for Brain Health (University of British Columbia, Vancouver, Canada). After completing her PhD, she stayed at the Center for Neuroscience and Cell Biology (CNC) as a postdoctoral fellow to complete her studies on the molecular mechanisms of traffic of glutamate receptors. In 2016 she joined the group of Laurent Groc, at the Interdisciplinary Institute for Neuroscience (CNRS, University of Bordeaux, France), to explore the regulation of the nanoscale distribution of NMDA receptors (NMDARs). In 2021 she has joined the CNC-UC as an Assistant Investigator to start her independent line of research on the nanoscale architecture of the synapse and transsynaptic signaling.
EDUCATION
2012 PhD in Biosciences, specialization in Neurosciences, University of Coimbra (Portugal)
2006 Bachelor's in Biology, University of Coimbra (Portugal)
RESEARCH INTERESTS
Neuroscience, Glutamate Receptors, Super-resolution Microscopy, Transsynaptic signalling, Molecular Neuroscience
FUTURE PROJECTS
The study of the novel NMDA receptor-Neurexin-2 interaction and potential role in schizophrenia
Applications Invited
PhD students
Scopus Publications
Scholar Citations
Scholar h-index
Scholar i10-index
Scopus Publications
Joana Ferreira and Laurent Groc
Springer US
Inmaculada M. González-González, John A. Gray, Joana Ferreira, María Jose Conde-Dusman, Delphine Bouchet, Isabel Perez-Otaño, and Laurent Groc
Elsevier BV
Chiara Paviolo, Joana S. Ferreira, Antony Lee, Daniel Hunter, Ivo Calaresu, Somen Nandi, Laurent Groc, and Laurent Cognet
American Chemical Society (ACS)
We provide evidence of a local synaptic nano-environment in the brain extracellular space (ECS) lying within 500 nm of postsynaptic densities. To reveal this brain compartment, we developed a correlative imaging approach dedicated to thick brain tissue based on single-particle tracking of individual fluorescent single wall carbon nanotubes (SWCNTs) in living samples and on speckle-based HiLo microscopy of synaptic labels. We show that the extracellular space around synapses bears specific properties in terms of morphology at the nanoscale and inner diffusivity. We finally show that the ECS juxta-synaptic region changes its diffusion parameters in response to neuronal activity, indicating that this nano-environment might play a role in the regulation of brain activity.
Joana S. Ferreira, Blanka Kellermayer, Ana Luísa Carvalho, and Laurent Groc
Wiley
Proteasome activity at the excitatory synapse plays an important role in neuronal communication. The proteasome translocation to synapses is mediated by neuronal activity, in particular the activation of N‐methyl‐d‐aspartate receptors (NMDARs). These receptors are composed of different subunits with distinct trafficking properties that provide various signalling and plasticity features to the synapse. Yet whether the interplay between the proteasome and NMDAR relies on specific subunit properties remain unclear. Using a combination of single molecule and immunocytochemistry imaging approaches in rat hippocampal neurons, we unveil a specific interplay between GluN2B‐containing NMDARs (GluN2B‐NMDARs) and the synaptic proteasome. Sustained proteasome activation specifically increases GluN2B‐NMDAR (not GluN2A‐NMDAR) lateral diffusion. In addition, when GluN2B‐NMDAR expression is downregulated, the proteasome localization decreases at glutamatergic synapses. Collectively, our data fuel a model in which the cellular dynamics and location of GluN2B‐NMDARs and proteasome are intermingled, shedding new lights on the NMDAR‐dependent regulation of synaptic adaptation.
Morgane Rosendale, Jessica Flores, Chiara Paviolo, Paolo Pagano, Jonathan Daniel, Joana Ferreira, Jean‐Baptiste Verlhac, Laurent Groc, Laurent Cognet, and Mireille Blanchard‐Desce
Wiley
Fluorescent nanoparticles dedicated to bioimaging applications should possess specific properties that have to be maintained in the aqueous, reactive, and crowded biological environment. These include chemical and photostability, small size (on the scale of subcellular structures), biocompatibility, high brightness, and good solubility. The latter is a major challenge for inorganic nanoparticles, which require surface coating to be made water soluble. Molecular‐based fluorescent organic nanoparticles (FONs) may prove a promising, spontaneously water‐soluble alternative, whose bottom‐up design allows for the fine‐tuning of individual properties. Here, the critical challenge of controlling the interaction of nanoparticles with cellular membranes is addressed. This is a report on bright, size‐tunable, red‐emitting, naturally stealthy FONs that do not require the use of antifouling agents to impede interactions with cellular membranes. As a proof of concept, single FONs diffusing up to 150 µm deep in brain tissue are imaged and tracked.
Tomohisa Hosokawa, Pin-Wu Liu, Qixu Cai, Joana S. Ferreira, Florian Levet, Corey Butler, Jean-Baptiste Sibarita, Daniel Choquet, Laurent Groc, Eric Hosy,et al.
Springer Science and Business Media LLC
Amit Kumar Mandal, Xiaojian Wu, Joana S. Ferreira, Mijin Kim, Lyndsey R. Powell, Hyejin Kwon, Laurent Groc, YuHuang Wang, and Laurent Cognet
Springer Science and Business Media LLC
AbstractCellular and tissue imaging in the second near-infrared window (NIR-II, ~1000–1350 nm) is advantageous for in vivo studies because of low light extinction by biological constituents at these wavelengths. However, deep tissue imaging at the single molecule sensitivity has not been achieved in the NIR-II window due to lack of suitable bio-probes. Single-walled carbon nanotubes have emerged as promising near-infrared luminescent molecular bio-probes; yet, their inefficient photoluminescence (quantum yield ~1%) drives requirements for sizeable excitation doses (~1–10 kW/cm2) that are significantly blue-shifted from the NIR-II region (<850 nm) and may thus ultimately compromise live tissue. Here, we show that single nanotube imaging can be achieved in live brain tissue using ultralow excitation doses (~0.1 kW/cm2), an order of magnitude lower than those currently used. To accomplish this, we synthesized fluorescent sp3-defect tailored (6,5) carbon nanotubes which, when excited at their first order excitonic transition (~985 nm) fluoresce brightly at ~1160 nm. The biocompatibility of these functionalized nanotubes, which are wrapped by encapsulation agent (phospholipid-polyethylene glycol), is demonstrated using standard cytotoxicity assays. Single molecule photophysical studies of these biocompatible nanotubes allowed us to identify the optimal luminescence properties in the context of biological imaging.
Joana S. Ferreira, Julien P. Dupuis, Blanka Kellermayer, Nathan Bénac, Constance Manso, Delphine Bouchet, Florian Levet, Corey Butler, Jean-Baptiste Sibarita, and Laurent Groc
Proceedings of the National Academy of Sciences
Significance Glutamatergic synapses onto pyramidal neurons are diverse in shape, size, and NMDAR-dependent long-term plasticity according to their dendritic location, i.e., proximal or distal to the soma. Deciphering the fine organization of NMDAR subtypes along the dendritic arbor of hippocampal neurons is key to understand the functional diversity of these various synapses. We unveil that the nanoscale organization of NMDAR changes as a function of distance from the soma in a subunit-specific and CaMKII interaction-dependent manner at proximal dendritic segments.
Chiara Paviolo, Federico N. Soria, Joana S. Ferreira, Antony Lee, Laurent Groc, Erwan Bezard, and Laurent Cognet
Elsevier BV
Chiara Paviolo, Joana S. Ferreira, Antony Lee, Laurent Groc, and Laurent Cognet
SPIE
The brain extracellular space (ECS) is a complex network that constitutes a key microenvironment for cellular communication, homeostasis, and clearance of toxic metabolites1. Signaling molecules, neuromodulators, and nutrients transit via the ECS, therefore mediating the communication between cells. Despite the relevance of this important part of the brain, its dynamics and structural organization at the nanoscale is still mostly unknown2. We have recently demonstrated that single-walled carbon nanotubes (SWCNTs) can be used to image and probe live brain tissue, providing super-resolved maps of the brain ECS and quantitative information on the local diffusion environment3,4. Here, we propose an important refinement of this approach by implementing a structured illumination technique (named HiLo microscopy5) to image fluorescently labelled neuronal structures in parallel to SWCNT NIR imaging. This technique is based on speckle illumination and relies on the acquisition of one structured and one uniform illumination image to obtain images deep into tissues with good optical sectioning. Having access to spatially resolved SWCNT diffusivity around specific neuronal structures will provide more precise insights about the heterogeneity of the brain environment.
Jeanne Linarès-Loyez, Joana S. Ferreira, Olivier Rossier, Brahim Lounis, Gregory Giannone, Laurent Groc, Laurent Cognet, and Pierre Bon
Frontiers Media SA
Through the formation of fluorescent self-interference (SELFI), quantitative intensity and phase imaging enables the 3D localization of single fluorescent molecules inside a fixed tissue with an accuracy well beyond the diffraction limit.. Here we demonstrate that this concept can be extended to 3D super-resolution microscopy and 3D single particle tracking in various living samples ranging from adherent cells to organotypic brain slices, using diverse fluorescent emitters (fluorescent proteins, organic dyes or quantum dots). This basically covers the most popular single molecule imaging techniques used for live sample studies. We also show that SELFI can be used in combination with different illumination schemes including highly inclined illumination and total internal reflection.
Blanka Kellermayer, Joana S. Ferreira, Julien Dupuis, Florian Levet, Dolors Grillo-Bosch, Lucie Bard, Jeanne Linarès-Loyez, Delphine Bouchet, Daniel Choquet, Dmitri A. Rusakov,et al.
Elsevier BV
Joana S Ferreira, Thomas Papouin, Laurent Ladépêche, Andrea Yao, Valentin C Langlais, Delphine Bouchet, Jérôme Dulong, Jean-Pierre Mothet, Silvia Sacchi, Loredano Pollegioni,et al.
eLife Sciences Publications, Ltd
The subunit composition of synaptic NMDA receptors (NMDAR), such as the relative content of GluN2A- and GluN2B-containing receptors, greatly influences the glutamate synaptic transmission. Receptor co-agonists, glycine and D-serine, have intriguingly emerged as potential regulators of the receptor trafficking in addition to their requirement for its activation. Using a combination of single-molecule imaging, biochemistry and electrophysiology, we show that glycine and D-serine relative availability at rat hippocampal glutamatergic synapses regulate the trafficking and synaptic content of NMDAR subtypes. Acute manipulations of co-agonist levels, both ex vivo and in vitro, unveil that D-serine alter the membrane dynamics and content of GluN2B-NMDAR, but not GluN2A-NMDAR, at synapses through a process requiring PDZ binding scaffold partners. In addition, using FRET-based FLIM approach, we demonstrate that D-serine rapidly induces a conformational change of the GluN1 subunit intracellular C-terminus domain. Together our data fuels the view that the extracellular microenvironment regulates synaptic NMDAR signaling.
Marta M. Vieira, Jeannette Schmidt, Joana S. Ferreira, Kevin She, Shinichiro Oku, Miranda Mele, Armanda E. Santos, Carlos B. Duarte, Ann Marie Craig, and Ana Luísa Carvalho
Elsevier BV
Joana S. Ferreira, Jeannette Schmidt, Pedro Rio, Rodolfo Águas, Amanda Rooyakkers, Ka Wan Li, August B. Smit, Ann Marie Craig, and Ana Luisa Carvalho
Society for Neuroscience
NMDA receptors play a central role in shaping the strength of synaptic connections throughout development and in mediating synaptic plasticity mechanisms that underlie some forms of learning and memory formation in the CNS. In the hippocampus and the neocortex, GluN1 is combined primarily with GluN2A and GluN2B, which are differentially expressed during development and confer distinct molecular and physiological properties to NMDA receptors. The contribution of each subunit to the synaptic traffic of NMDA receptors and therefore to their role during development and in synaptic plasticity is still controversial. We report a critical role for the GluN2B subunit in regulating NMDA receptor synaptic targeting. In the absence of GluN2B, the synaptic levels of AMPA receptors are increased and accompanied by decreased constitutive endocytosis of GluA1-AMPA receptor. We used quantitative proteomic analysis to identify changes in the composition of postsynaptic densities from GluN2B−/− mouse primary neuronal cultures and found altered levels of several ubiquitin proteasome system components, in particular decreased levels of proteasome subunits. Enhancing the proteasome activity with a novel proteasome activator restored the synaptic levels of AMPA receptors in GluN2B−/− neurons and their endocytosis, revealing that GluN2B-mediated anchoring of the synaptic proteasome is responsible for fine tuning AMPA receptor synaptic levels under basal conditions.
Kevin She, Joana S. Ferreira, Ana Luisa Carvalho, and Ann Marie Craig
Elsevier BV
Background: Ligand binding is essential for surface delivery of non-NMDA-type glutamate receptors. Results: GluN2B ligand binding site mutants showed reduced surface and synaptic expression correlating with glutamate efficacy. Conclusion: Glutamate binding controls surface delivery of NMDA receptors. Significance: This work extends the receptor classes subject to glutamate regulation of surface delivery and reports parameters controlling synaptic delivery of NMDA receptors critical for neuron function. Trafficking of NMDA receptors to the surface of neurons and to synapses is critical for proper brain function and activity-dependent plasticity. Recent evidence suggests that surface trafficking of other ionotropic glutamate receptors requires ligand binding for exit from the endoplasmic reticulum. Here, we show that glutamate binding to GluN2 is required for trafficking of NMDA receptors to the cell surface. We expressed a panel of GluN2B ligand binding mutants in heterologous cells with GluN1 or in rat cultured neurons and found that surface expression correlates with glutamate efficacy. Such a correlation was found even in the presence of dominant negative dynamin to inhibit endocytosis and surface expression correlated with Golgi localization, indicating differences in forward trafficking. Co-expression of wild type GluN2B did not enhance surface expression of the mutants, suggesting that glutamate must bind to both GluN2 subunits in a tetramer and that surface expression is limited by the least avid of the two glutamate binding sites. Surface trafficking of a constitutively closed cleft GluN2B was indistinguishable from that of wild type, suggesting that glutamate concentrations are typically not limiting for forward trafficking. YFP-GluN2B expressed in hippocampal neurons from GluN2B−/− mice rescued synaptic accumulation at similar levels to wild type. Under these conditions, surface synaptic accumulation of YFP-GluN2B mutants also correlated with apparent glutamate affinity. Altogether, these results indicate that glutamate controls forward trafficking of NMDA receptors to the cell surface and to synapses and raise the intriguing idea that NMDA receptors may be functional at intracellular sites.
Sandra D. Santos, Olga Iuliano, Luís Ribeiro, Julien Veran, Joana S. Ferreira, Pedro Rio, Christophe Mulle, Carlos B. Duarte, and Ana Luísa Carvalho
Elsevier BV
Background: The strength of excitatory synapses is determined by the synaptic content of AMPA receptors. Results: We found that Contactin associated protein 1 (Caspr1) binds to AMPA receptors and regulates their neuronal cell surface and synaptic expression. Conclusion: Caspr1 is a binding partner for AMPA receptors, which regulates their traffic and synaptic targeting. Significance: Caspr1 is a new player in plasticity mechanisms in excitatory synapses. Glutamate receptors of the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) type mediate fast excitatory synaptic transmission in the CNS. Synaptic strength is modulated by AMPA receptor binding partners, which regulate receptor synaptic targeting and functional properties. We identify Contactin-associated protein 1 (Caspr1) as an AMPA receptor interactor. Caspr1 is present in synapses and interacts with AMPA receptors in brain synaptic fractions. Coexpression of Caspr1 with GluA1 increases the amplitude of glutamate-evoked currents. Caspr1 overexpression in hippocampal neurons increases the number and size of synaptic GluA1 clusters, whereas knockdown of Caspr1 decreases the intensity of synaptic GluA1 clusters. Hence, Caspr1 is a regulator of the trafficking of AMPA receptors to synapses.
Joana S. Ferreira, Amanda Rooyakkers, Kevin She, Luis Ribeiro, Ana Luísa Carvalho, and Ann Marie Craig
Elsevier BV
NMDA receptors are calcium-permeable ionotropic receptors that detect coincident glutamate binding and membrane depolarization and are essential for many forms of synaptic plasticity in the mammalian brain. The obligatory GluN1 subunit of NMDA receptors is alternatively spliced at multiple sites, generating forms that vary in N-terminal N1 and C-terminal C1, C2, and C2′ cassettes. Based on expression of GluN1 constructs in heterologous cells and in wild type neurons, the prevalent view is that the C-terminal cassettes regulate synaptic accumulation and its modulation by homeostatic activity blockade and by protein kinase C (PKC). Here, we tested the role of GluN1 splicing in regulated synaptic accumulation of NMDA receptors by lentiviral expression of individual GluN1 splice variants in hippocampal neurons cultured from GluN1 (−/−) mice. High efficiency transduction of GluN1 at levels similar to endogenous was achieved. Under control conditions, the C2′ cassette mediated enhanced synaptic accumulation relative to the alternate C2 cassette, whereas the presence or absence of N1 or C1 had no effect. Surprisingly all GluN1 splice variants showed >2-fold increased synaptic accumulation with chronic blockade of NMDA receptor activity. Furthermore, in this neuronal rescue system, all GluN1 splice variants were equally rapidly dispersed upon activation of PKC. These results indicate that the major mechanisms mediating homeostatic synaptic accumulation and PKC dispersal of NMDA receptors occur independently of GluN1 splice isoform.
André R. Gomes, Joana S. Ferreira, Ana V. Paternain, Juan Lerma, Carlos B. Duarte, and Ana Luísa Carvalho
Elsevier BV