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Whole-mount Retinal Organoid Visualization with Cellular Resolution

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The Barcelona Institute of Science and Technology

Non-centrosymmetric polar compounds have important technological properties. Reported perovskite oxynitrides show centrosymmetric structures, and for some of them high permittivities have been observed and ascribed to local dipoles induced by partial order of nitride and oxide. Reported here is the first hexagonal perovskite oxynitride BaWON , which shows a polar 6H polytype. Synchrotron X-ray and neutron powder diffraction, and annular bright-field in scanning transmission electron microscopy indicate that it crystalizes in the non-centrosymmetric space group P6 mc, with a total order of nitride and oxide at two distinct coordination environments in cubic and hexagonal packed BaX layers. A synergetic second-order Jahn-Teller effect, supported by first principle calculations, anion order, and electrostatic repulsions between W cations, induce large distortions at two inequivalent face-sharing octahedra that lead to long-range ordered dipoles and spontaneous polarization along the c axis. The new oxynitride is a semiconductor with a band gap of 1.1 eV and a large permittivity.

Engineering Polar Oxynitrides: Hexagonal Perovskite BaWON.

 I-1582 is approved in Europe for the management of  on vegetable crops. However, little information about its modes of action and temperature requirements is available, despite the effect of these parameters in its efficacy. The cardinal temperatures for bacterial growth and biofilm formation were determined. The bacteria was transformed with GFP to study its effect on nematode eggs and root colonization of tomato () and cucumber () by laser-scanning confocal microscopy. Induction of plant resistance was determined in split-root experiments and the dynamic regulation of genes related to jasmonic acid (JA) and salicylic acid (SA) by RT-qPCR at three different times after nematode inoculation. The bacteria was able to grow and form biofilms between 15 and 45°C; it degraded egg-shells and colonized eggs; it colonized tomato roots more extensively than cucumber roots; it induced systemic resistance in tomato, but not in cucumber; SA and JA related genes were primed at different times after nematode inoculation in tomato, but only the SA-related gene was up-regulated at 7 days after nematode inoculation in cucumber. In conclusion,  I-1582 is active at a wide range of temperatures; its optimal growth temperature is 35°C; it is able to degrade  eggs, and to colonize plant roots, inducing systemic resistance in a plant dependent species manner.

 Strain I-1582, a Nematode Antagonist by Itself and Through the Plant.

Four rotation sequences consisting of ungrafted tomato cv. Durinta - melon cv. Paloma or tomato grafted onto the resistant rootstock 'Aligator' - melon grafted onto the resistant  accession BGV11135, and in reverse order, were conducted from 2015 to 2017 in a plastic greenhouse infested or not with  to determine the plant tolerance (), the minimum relative crop yield () and fruit quality. The relationship between  densities in soil at transplanting () of each crop and the crop yield was assessed and  and  were estimated by the Seinhorst's damage model. In addition, the volume and the number of nuclei of single giant cells and the number of giant cells, its volume and the number of nuclei per feeding site in susceptible tomato and melon were compared to those in the resistant tomato and  15 days after nematode inoculation in pot test. The relationship between the  and the relative crop yield fitted the Seinhorst's damage model in both ungrafted and grafted tomato and melon, but not for all years and cropping seasons. The estimated  for ungrafted and grafted tomato did not differ but  was lower in the former (34%) than the latter (67%). Sodium concentration in fruits from ungrafted but not from grafted tomato increased with nematode densities in spring 2015 and 2016. The estimated ungrafted melon  did not differ from the grafted melon cultivated in spring, but it did when it was cultivated in summer. The relative crop yield of ungrafted melon was lower (2%) than the grafted cultivated in spring (62%) and summer (20%). Sodium concentration in melon fruits from ungrafted plants increased with nematode densities. No variations in fruit quality from grafted melon cultivated in spring were found, although less dry matter and soluble solid content at highest nematode densities were registered when it was cultivated in summer. Lower number of giant cells per feeding site was observed in both susceptible tomato germplasms compared to the resistant ones but they were more voluminous and held higher number of nuclei per giant cell and per feeding site.

Tomato and Melon  Resistant Rootstocks Improve Crop Yield but Melon Fruit Quality Is Influenced by the Cropping Season.

A rotation sequence of ungrafted and grafted tomato-melon-pepper-watermelon on resistant rootstocks 'Brigeor', , 'Oscos' and , respectively, was carried out in a plastic greenhouse, ending with a susceptible or resistant tomato crop. The rotation was conducted in plots infested by an avirulent (Avi) or a partially virulent (Vi)  population to the  gene. At the beginning of the study, the reproduction index (RI, relative reproduction in the resistant respect susceptible tomato) of Avi and Vi populations was 1.3% and 21.6%, respectively. Soil nematode density at transplanting () and at the end () of each crop, disease severity and crop yield were determined. Moreover, the putative virulence selection and fitness cost were determined at the end of each crop in pot tests. In addition, a histopathological study was carried out 15 days after nematode inoculation in pot test. The volume and number of nuclei per giant cell (GC) and the number of GC, their volume and the number of nuclei per feeding site in susceptible watermelon and pepper were compared with  and resistant pepper. At the beginning of the study, the  of Avi and Vi plots did not differ between susceptible and resistant germplasm. At the end of the rotation, the  of Avi was 1.2 the  in susceptible and 0.06 in resistant, the cumulative yield of grafted crops was 1.82 times higher than that of the ungrafted susceptible ones, and the RI in resistant tomato less than 10% irrespective of the rotation sequence. Concerning the Vi,  was below the detection level at the end of the rotation in resistant and 3 times  in the susceptible. The cumulative yield of grafted crops was 2.83 times higher than that of the ungrafted and the RI in resistant tomato was 7.6%, losing the population's virulence. In the histopathological study, no differences in number of GC per feeding site were observed in watermelon compared to , but they were more voluminous and contained higher number of nuclei per GC and per feeding site. Regarding pepper, Avi population did not penetrate resistant rootstock.

Crop rotation with -resistant germplasm is useful to manage and revert the (a)virulent populations of  gene and reduce yield losses.

To interrogate neural circuits and crack their codes, in vivo brain activity imaging must be combined with spatiotemporally precise stimulation in three dimensions using genetic or pharmacological specificity. This challenge requires deep penetration and focusing as provided by infrared light and multiphoton excitation, and has promoted two-photon photopharmacology and optogenetics. However, three-photon brain stimulation in vivo remains to be demonstrated. We report the regulation of neuronal activity in zebrafish larvae by three-photon excitation of a photoswitchable muscarinic agonist at 50 pM, a billion-fold lower concentration than used for uncaging, and with mid-infrared light of 1560 nm, the longest reported photoswitch wavelength. Robust, physiologically relevant photoresponses allow modulating brain activity in wild-type animals with spatiotemporal and pharmacological precision. Computational calculations predict that azobenzene-based ligands have high three-photon absorption cross-section and can be used directly with pulsed infrared light. The expansion of three-photon pharmacology will deeply impact basic neurobiology and neuromodulation phototherapies.

Three-Photon Infrared Stimulation of Endogenous Neuroreceptors in Vivo.

Uncontrolled secretion of ECM proteins, such as collagen, can lead to excessive scarring and fibrosis and compromise tissue function. Despite the widespread occurrence of fibrotic diseases and scarring, effective therapies are lacking. A promising approach would be to limit the amount of collagen released from hyperactive fibroblasts. We have designed membrane permeant peptide inhibitors that specifically target the primary interface between TANGO1 and cTAGE5, an interaction that is required for collagen export from endoplasmic reticulum exit sites (ERES). Application of the peptide inhibitors leads to reduced TANGO1 and cTAGE5 protein levels and a corresponding inhibition in the secretion of several ECM components, including collagens. Peptide inhibitor treatment in zebrafish results in altered tissue architecture and reduced granulation tissue formation during cutaneous wound healing. The inhibitors reduce secretion of several ECM proteins, including collagens, fibrillin and fibronectin in human dermal fibroblasts and in cells obtained from patients with a generalized fibrotic disease (scleroderma). Taken together, targeted interference of the TANGO1-cTAGE5 binding interface could enable therapeutic modulation of ERES function in ECM hypersecretion, during wound healing and fibrotic processes.

TANGO1 inhibitors reduce collagen secretion and limit tissue scarring.

Neuroelectronic prostheses are being developed for restoring vision at the retinal level in patients who have lost their sight due to photoreceptor loss. The core component of these devices is the electrode array, which enables interfacing with retinal neurons. Generating the perception of meaningful images requires high-density microelectrode arrays (MEAs) capable of precisely activating targeted retinal neurons. Achieving this precision necessitates the downscaling of electrodes to micrometer dimensions. However, miniaturization increases electrode impedance, which poses challenges by limiting the amount of current that can be delivered, thereby impairing the electrode's capability for effective neural modulation. Additionally, it elevates noise levels, reducing the signal quality of the recorded neural activity. This report focuses on evaluating reduced graphene oxide (rGO) based devices for interfacing with the retina, showcasing their potential in vision restoration. Our findings reveal low impedance and high charge injection limit for microscale rGO electrodes, confirming their suitability for developing next-generation high-density retinal devices. We successfully demonstrated bidirectional interfacing with cell cultures and explanted retinal tissue, enabling the identification and modulation of multiple cells' activity. Additionally, calcium imaging allowed real-time monitoring of retinal cell dynamics, demonstrating a significant reduction in activated areas with small-sized electrodes. Overall, this study lays the groundwork for developing advanced rGO-based MEAs for high-acuity visual prostheses.

ArticleTotal : 3	Year
Publication title Cited by 1	2022
Publication title Cited by 2	2023
Publication title	2025

Article	Year
Engineering Polar Oxynitrides: Hexagonal Perovskite BaWON. Angewandte Chemie (International ed. in English)\| PubMed ID: 32649790	2020
Strain I-1582, a Nematode Antagonist by Itself and Through the Plant. Frontiers in plant science\| PubMed ID: 32765537	2020
Tomato and Melon Resistant Rootstocks Improve Crop Yield but Melon Fruit Quality Is Influenced by the Cropping Season. Frontiers in plant science\| PubMed ID: 33224158	2020
Crop rotation with -resistant germplasm is useful to manage and revert the (a)virulent populations of gene and reduce yield losses. Frontiers in plant science\| PubMed ID: 37008463	2023
Three-Photon Infrared Stimulation of Endogenous Neuroreceptors in Vivo. Angewandte Chemie (International ed. in English)\| PubMed ID: 37823736	2023
TANGO1 inhibitors reduce collagen secretion and limit tissue scarring. Nature communications\| PubMed ID: 38658535	2024
Graphene-based microelectrodes with bidirectional functionality for next-generation retinal electronic interfaces. Nanoscale horizons\| PubMed ID: 39229772	2024

Marina Cunquero

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