Retinal aging in 3× Tg-AD mice model of Alzheimer's disease

The retina, as part of the central nervous system (CNS), can be the perfect target for in vivo, in situ, and noninvasive neuropathology diagnosis and assessment of therapeutic efficacy. It has long been established that several age-related brain changes are more pronounced in Alzheimer’s disease (AD). Nevertheless, in the retina such link is still under-explored. This study investigates the differences in the aging of the CNS through the retina of 3×Tg-AD and wild-type mice. A dedicated optical coherence tomograph imaged mice’s retinas for 16 months. Two neural networks were developed to model independently each group’s ages and were then applied to an independent set containing images fromboth groups. Our analysis shows amean absolute error of 0.875±1.1×10−2 and 1.112 ± 1.4 × 10−2 months, depending on training group. Our deep learning approach appears to be a reliable retinal OCT aging marker. We show that retina aging is distinct in the two classes: the presence of the three mutated human genes in the mouse genome has an impact on the aging of the retina. For mice over 4 months-old, transgenic mice consistently present a negative retina age-gap when compared to wildtype mice, regardless of training set. This appears to contradict AD observations in the brain. However, the ‘black-box” nature of deep-learning implies that one cannot infer reasoning. We can only speculate that some healthy age-dependent neural adaptations may be altered in transgenic animals.This study was supported by The Portuguese Foundation for Science and Technology (FCT) through PTDC/EMD-EMD/28039/2017, UIDB/04950/2020, PestUID/NEU/04539/2019, and by FEDER-COMPETE through POCI-01-0145-FEDER-028039.info:eu-repo/semantics/publishedVersio

Retinal imaging in animal models: searching for biomarkers of neurodegeneration

There is a pressing need for novel diagnostic and progression biomarkers of neurodegeneration. However, the inability to determine disease duration and stage in patients with Alzheimer’s disease (AD) hinders their discovery. Because animal models of disease allow us to circumvent some of these limitations, they have proven to be of paramount importance in clinical research. Due to the clear optics of the eye, the retina combined with optical coherence tomography (OCT) offers the perfect opportunity to image neurodegeneration in the retina in vivo, non-invasively, directly, quickly, and inexpensively. Based on these premises, our group has worked towards uncovering neurodegeneration-associated changes in the retina of the triple-transgenic mouse model of familial AD (3×Tg-AD). In this work, we present an overview of our work on this topic. We report on thickness variations of the retina and retinal layers/layer aggregates caused by healthy aging and AD-like conditions and discuss the implications of focusing research efforts solely on retinal thickness. We explore what other information is embedded in the OCT data, extracted based on texture analysis and deep-learning approaches, to further identify biomarkers that could be used for early detection and diagnosis. We were able to detect changes in the retina of the animal model of AD as early as 1 month of age. We also discuss our work to develop an optical coherence elastography system to measure retinal elasticity, which can be used in conjunction with conventional OCT. Finally, we discuss the potential application of these technologies in human patients and the steps needed to make OCT a helpful screening tool for the detection of neurodegeneration.This study was supported by The Portuguese Foundation for Science and Technology (FCT) through PTDC/EMD-EMD/28039/2017, UIDB/04950/2020, Pest-UID/NEU/04539/2019, PTDC/EMD-EMD/32162/2017, and by FEDER-COMPETE through POCI-01-0145-FEDER-028039.info:eu-repo/semantics/publishedVersio

Normative mice retinal thickness: 16-month longitudinal characterization of wild-type mice and changes in a model of Alzheimer's disease

Animal models of disease are paramount to understand retinal development, the pathophysiology of eye diseases, and to study neurodegeneration using optical coherence tomography (OCT) data. In this study, we present a comprehensive normative database of retinal thickness in C57BL6/129S mice using spectral-domain OCT data. The database covers a longitudinal period of 16 months, from 1 to 16 months of age, and provides valuable insights into retinal development and changes over time. Our findings reveal that total retinal thickness decreases with age, while the thickness of individual retinal layers and layer aggregates changes in different ways. For example, the outer plexiform layer (OPL), photoreceptor inner segments (ILS), and retinal pigment epithelium (RPE) thickened over time, whereas other retinal layers and layer aggregates became thinner. Additionally, we compare the retinal thickness of wild-type (WT) mice with an animal model of Alzheimer's disease (3×Tg-AD) and show that the transgenic mice exhibit a decrease in total retinal thickness compared to age-matched WT mice, with statistically significant differences observed at all evaluated ages. This normative database of retinal thickness in mice will serve as a reference for future studies on retinal changes in neurodegenerative and eye diseases and will further our understanding of the pathophysiology of these conditions.The authors would like to acknowledge the support by the Portuguese Foundation for Science and Technology (FCT) through PTDC/EMD-EMD/28039/2017, FCT/UIDB/4950/Base/2020 and FCT/UIDP/4950/Programatico/2020, Pest-UID/NEU/04539/2019, and by FEDER-COMPETE through POCI-01-0145-FEDER-028039.info:eu-repo/semantics/publishedVersio

Stage-independent biomarkers for Alzheimer’s disease from the living retina: an animal study

The early diagnosis of neurodegenerative disorders is still an open issue despite the many efforts to address this problem. In particular, Alzheimer’s disease (AD) remains undiagnosed for over a decade before the first symptoms. Optical coherence tomography (OCT) is now common and widely available and has been used to image the retina of AD patients and healthy controls to search for biomarkers of neurodegeneration. However, early diagnosis tools would need to rely on images of patients in early AD stages, which are not available due to late diagnosis. To shed light on how to overcome this obstacle, we resort to 57 wild-type mice and 57 triple-transgenic mouse model of AD to train a network with mice aged 3, 4, and 8 months and classify mice at the ages of 1, 2, and 12 months. To this end, we computed fundus images from OCT data and trained a convolution neural network (CNN) to classify those into the wild-type or transgenic group. CNN performance accuracy ranged from 80 to 88% for mice out of the training group’s age, raising the possibility of diagnosing AD before the first symptoms through the non-invasive imaging of the retina.Tis study was supported by Te Portuguese Foundation for Science and Technology (FCT) through PTDC/EMD-EMD/28039/2017, UIDB/04950/2020, UIDB/04539/2020, Pest-UID/NEU/04539/2019, and by FEDERCOMPETE through POCI-01-0145-FEDER-028039.info:eu-repo/semantics/publishedVersio

Longitudinal normative OCT retinal thickness data for wild-type mice, and characterization of changes in the 3×Tg-AD mice model of Alzheimer's disease

Mice are widely used as models for many diseases, including eye and neurodegenerative diseases. However, there is a lack of normative data for retinal thickness over time, especially at young ages. In this work, we present a normative thickness database from one to four-months-old, for nine layers/layer-aggregates, including the total retinal thickness, obtained from the segmentation of spectral-domain optical coherence tomography (SD-OCT) data from the C57BL6/129S mouse strain. Based on fifty-seven mice, this normative database provides an opportunity to study the ageing of control mice and characterize disease models' ageing, such as the triple transgenic mouse model of Alzheimer's disease (3×Tg-AD) used in this work. We report thickness measurements, the differences in thickness per layer, demonstrate a nasal-temporal asymmetry, and the variation of thickness as a function to the distance to the optic disc center. Significant differences were found between the transgenic group's thickness and the normative database for the entire period covered in this study. Even though it is well accepted that retinal nerve fiber layer (RNFL) thinning is a hallmark of neurodegeneration, our results show a thicker RNFL-GCL (RNFL-Ganglion cell layer) aggregate for the 3×Tg-AD mice until four-months-old.This study was supported by The Portuguese Foundation for Science and Technology (FCT) through PTDC/ EMD-EMD/28039/2017, UIDB/04950/2020, Pest-UID/ NEU/04539/2019, and by FEDER-COMPETE through POCI-01-0145-FEDER-028039.info:eu-repo/semantics/publishedVersio

Retinal imaging in animal models: Searching for biomarkers of neurodegeneration

There is a pressing need for novel diagnostic and progression biomarkers of neurodegeneration. However, the inability to determine disease duration and stage in patients with Alzheimer’s disease (AD) hinders their discovery. Because animal models of disease allow us to circumvent some of these limitations, they have proven to be of paramount importance in clinical research. Due to the clear optics of the eye, the retina combined with optical coherence tomography (OCT) offers the perfect opportunity to image neurodegeneration in the retina in vivo, non-invasively, directly, quickly, and inexpensively. Based on these premises, our group has worked towards uncovering neurodegeneration-associated changes in the retina of the triple-transgenic mouse model of familial AD (3×Tg-AD). In this work, we present an overview of our work on this topic. We report on thickness variations of the retina and retinal layers/layer aggregates caused by healthy aging and AD-like conditions and discuss the implications of focusing research efforts solely on retinal thickness. We explore what other information is embedded in the OCT data, extracted based on texture analysis and deep-learning approaches, to further identify biomarkers that could be used for early detection and diagnosis. We were able to detect changes in the retina of the animal model of AD as early as 1 month of age. We also discuss our work to develop an optical coherence elastography system to measure retinal elasticity, which can be used in conjunction with conventional OCT. Finally, we discuss the potential application of these technologies in human patients and the steps needed to make OCT a helpful screening tool for the detection of neurodegeneration

Normative mice retinal thickness: 16-month longitudinal characterization of wild-type mice and changes in a model of Alzheimer's disease

Animal models of disease are paramount to understand retinal development, the pathophysiology of eye diseases, and to study neurodegeneration using optical coherence tomography (OCT) data. In this study, we present a comprehensive normative database of retinal thickness in C57BL6/129S mice using spectral-domain OCT data. The database covers a longitudinal period of 16 months, from 1 to 16 months of age, and provides valuable insights into retinal development and changes over time. Our findings reveal that total retinal thickness decreases with age, while the thickness of individual retinal layers and layer aggregates changes in different ways. For example, the outer plexiform layer (OPL), photoreceptor inner segments (ILS), and retinal pigment epithelium (RPE) thickened over time, whereas other retinal layers and layer aggregates became thinner. Additionally, we compare the retinal thickness of wild-type (WT) mice with an animal model of Alzheimer's disease (3 × Tg-AD) and show that the transgenic mice exhibit a decrease in total retinal thickness compared to age-matched WT mice, with statistically significant differences observed at all evaluated ages. This normative database of retinal thickness in mice will serve as a reference for future studies on retinal changes in neurodegenerative and eye diseases and will further our understanding of the pathophysiology of these conditions

Metamitron and shade effects on leaf physiology and thinning efficacy of malus × domestica borkh

Funding Information: Funding: This study was supported by ADAMA-Israel, as well as by Fundação para a Ciência e a Tecnologia (FCT) through the research units UID/AGR/04129/2020 (LEAF). Publisher Copyright: © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).Thinning strategies, namely shade or photosynthetic inhibitors, rely on the reduction of carbon supply to the fruit below the demand, causing fruit abscission. In order to clarify the subject, seven field trials were carried out in Lleida, Girona, and Sint-Truiden (2017 + 2018), using orchards of ‘Golden’ and ‘Gala’ apple trees. At the stage of 9–14-mm fruit diameter, four treatments were implemented: (A) CTR-control, trees under natural environmental conditions; (B) SN-shaded trees, trees above which shading nets reducing 50% of irradiance were installed 24 h after metamitron application date—without application of metamitron—and removed after five days; (C) MET-trees sprayed with 247.5 ppm of metamitron; (D) MET + SN-trees submitted to the combined exposure to metamitron application and shading nets. Low radiation significantly increased metamitron absorption (36–53% in the three locations in 2018) and reduced its degradation. Net photosynthesis and stomatal conductance were strongly reduced in all treatments, with minimum values 2 days after spraying (DAS) and incomplete recovery 10 DAS in MET + SN. All treatments resulted in leaf sucrose and sorbitol decreases, leading to a negative carbon balance. SN and MET + SN promoted the highest thinning efficacy, increasing fruit weight and size, with MET + SN causing over-thinning in some trials. Leaf antioxidant enzymes showed moderate changes in activity increases under MET or MET + SN, accompanied by a rise of glutathione content and a reduction in ascorbate, however without lipid peroxidation. This work shows that environmental conditions, such as cloudy days, must be carefully considered upon metamitron application, since the low irradiance enhances metamitron efficacy and may cause over-thinning.publishersversionpublishe

Metamitron and Shade Effects on Leaf Physiology and Thinning Efficacy of Malus × domestica Borkh

Thinning strategies, namely shade or photosynthetic inhibitors, rely on the reduction of carbon supply to the fruit below the demand, causing fruit abscission. In order to clarify the subject, seven field trials were carried out in Lleida, Girona, and Sint-Truiden (2017 + 2018), using orchards of ‘Golden’ and ‘Gala’ apple trees. At the stage of 9–14-mm fruit diameter, four treatments were implemented: (A) CTR-control, trees under natural environmental conditions; (B) SN-shaded trees, trees above which shading nets reducing 50% of irradiance were installed 24 h after metamitron application date—without application of metamitron—and removed after five days; (C) MET-trees sprayed with 247.5 ppm of metamitron; (D) MET + SN-trees submitted to the combined exposure to metamitron application and shading nets. Low radiation significantly increased metamitron absorption (36–53% in the three locations in 2018) and reduced its degradation. Net photosynthesis and stomatal conductance were strongly reduced in all treatments, with minimum values 2 days after spraying (DAS) and incomplete recovery 10 DAS in MET + SN. All treatments resulted in leaf sucrose and sorbitol decreases, leading to a negative carbon balance. SN and MET + SN promoted the highest thinning efficacy, increasing fruit weight and size, with MET + SN causing over-thinning in some trials. Leaf antioxidant enzymes showed moderate changes in activity increases under MET or MET + SN, accompanied by a rise of glutathione content and a reduction in ascorbate, however without lipid peroxidation. This work shows that environmental conditions, such as cloudy days, must be carefully considered upon metamitron application, since the low irradiance enhances metamitron efficacy and may cause over-thinninginfo:eu-repo/semantics/publishedVersio