Postnatal Development of NPY and Somatostatin-28 Peptidergic Populations in the Human Angular Bundle

The angular bundle is a white matter fiber fascicle, which runs longitudinally along the parahippocampal gyrus. It is best known for carrying fibers from the entorhinal cortex (EC) to the hippocampus through the perforant and alvear pathways, as well as for carrying hippocampal output to the neocortex, and distributing fibers to polysensory cortex. The angular bundle is already present prenatally at the beginning of the fetal period. Connections between the EC and the hippocampus are established by the 20th gestational week (gw). In the postnatal period, it shows increasing myelination. The angular bundle, as well as other white matter portions of gyral surfaces in the brain, presents interstitial neurons, a remnant of subplate neurons. Those interstitial neurons show neurochemical phenotypes both prenatally and postnatally, among which, neuropeptide Y (NPY) and Somatostatin-28 (SOM-28) peptidergic populations are noticeable, and accompany the fiber connections in the maturation of the hippocampal formation. We sought to investigate the topography of the postnatal distribution and relative density of neurons immunoreactive for NPY or SOM in the angular bundle along the rostrocaudal axis of the hippocampus. The study was carried out in 15 cases, ranging from 35 gws, up to 14 year old. All cases showed positive neurons showing a polygonal or spindle shaped morphology for both peptides, scattered throughout the angular bundle. The highest number of positive neurons appeared around birth and the ensuing weeks. Up to one and a half years, the density of both peptidergic populations decreased slightly. However, cases older than 2 years of age showed a substantial decrease in density of immunolabeled neurons, density that did not showed a minor decrease in density of positive neurons in cases older than 2 years. In addition, a topography from caudal to rostral levels of the angular bundle was detected at all ages. The functional significance of interstitial cells is unknown, but the existence of SOM and NPY peptidergic neurons, presumably inhibitory, in the white matter of the angular bundle, could contribute to the basic wiring of the hippocampal formation, through which autobiographical and spatial memories can begin to be stored in the infant brain

Distribución de los Neuropéptidos Somatostatina-28, Somatostatina-28 (1-12) y Neuropéptido y en la formación del hipocampo humano durante el desarrollo postnatal

La formación del hipocampo (FH), junto a otras áreas corticales del lóbulo temporal medial, tiene un papel fundamental en el procesamiento y consolidación de la memoria declarativa. Desde el punto de vista anatómico, la FH engloba al giro dentado (GD), los campos amónicos CA3, CA2, CA1, el subículo (S), el presubículo (PrS), el parasubículo (PaS) y la corteza entorrinal (CE). Estas estructuras se agrupan principalmente por su estrecha relación hodológica, presentando un conjunto de conexiones que en gran medida forman un circuito cerrado y unidireccional. El presente trabajo se ha planteado para contribuir al conocimiento del desarrollo postnatal de la FH humano desde un enfoque morfológico y neuroquímico ya que son muy escasos los trabajos que abordan la maduración neuroquímica de las poblaciones neuronales de la FH humano. Para tal fin se ha analizado la distribución de los neuropéptidos somatostatina-28 (Som28), somatostatina-28 (1-12) (Som12) y neuropéptido Y (NPY), considerados marcadores de interneuronas, en esta región del cerebro humano a lo largo del desarrollo postnatal por medio de técnicas inmunohistoquímicas. Los resultados de este trabajo muestran que en todos los campos de la FH humano analizados (GD, CA3, CA2, CA1, S, PrS y PaS) se detectan células y prolongaciones inmunorreactivas frente a Som28, Som12 y NPY desde las 35 semanas de gestación, la menor edad analizada, en los tres niveles rostrocaudales analizados. La distribución topográfica y laminar de los cuerpos neuronales y prolongaciones inmunorreactivas frente a estos péptidos en estos campos de la FH no varía durante el desarrollo postnatal. En todos los campos los cuerpos neuronales inmunorreactivos se localizan principalmente en las capas piramidales y las capas que contienen células polimórficas. Las prolongaciones inmunorreactivas están presentes en todas las capas de todos los campos, aunque las fibras inmunorreactivas frente a Som28 y NPY son más abundantes en el S en su conjunto y en la capa polimórfica del GD. Sin embargo, en los casos de edades superiores a los 5 meses no se aprecian claras diferencias entre el S y el PrS y el PaS. El neuropéptido Som28 es el que presenta, de manera general, mayor marcaje celular y de puntos aislados en todas las edades analizadas. Este estudio contribuye al conocimiento del sustrato morfológico que subyace a la maduración postnatal de las diferentes poblaciones neuronales de la FH humano, conocimiento necesario para la completa comprensión de la maduración estructural y funcional de los distintos circuitos de la FH y por ende del desarrollo de la memoria declarativa

Nursing Student Satisfaction with the Teaching Methodology Followed during the COVID-19 Pandemic

Background: Halfway through the 2019–2020 academic year, the entire university system was affected by an exceptional situation caused by the COVID-19 pandemic. Online learning was globally implemented for all degrees to finish the course and to meet academic objectives. This unforeseen change in teaching and subsequent evaluations meant teachers and students had to invest significant effort. Student satisfaction is used to measure the evaluation of teaching/learning processes in higher education. Our objective was to know and compare the satisfaction of nursing students taught at a Spanish public university after making changes to the teaching methodology. Methods: A descriptive observational study that measures student satisfaction. Study population: 240 students registered in academic years 2019–2020 and 2020–2021 answered the survey. The survey contained 30 items answered on a Likert-type scale. The main variables: the learning methodology (online or blended) was the independent variable; student satisfaction was the dependent variable. Descriptive and bivariate analyses were performed. Results: A response rate between 37.4% and 41.2%. Overall satisfaction was 2.75 points (SD 0.56) and 2.94 points (SD 0.49) with online learning and bimodal learning, respectively (maximum score 4 points) (p < 0.004). Conclusions: Student satisfaction was moderate–high for both learning methodologies. Students found that the b-learning methodology was the most valued

Neuropeptides in the developing human hippocampus under hypoxic–ischemic conditions

The perinatal period, sensitive for newborn survival, is also one of the most critical moments in human brain development. Perinatal hypoxia due to reduced blood supply to the brain (ischemia) is one of the main causes of neonatal mortality. Brain damage caused by perinatal hypoxia–ischemia (HI) can lead to neuro- and psychological disorders. However, its impact seems to be region-dependent, with the hippocampus being one of the most affected areas. Among the neuronal populations of the hippocampus, some interneuron groups – such as somatostatin- or neuropeptide Y-expressing neurons – seem to be particularly vulnerable. The limited information available about the effects of HI in the hippocampus comes mainly from animal models and adult human studies. This article presents an immunohistochemical analysis of somatostatin (SOM) and neuropeptide Y (NPY) expression in the developing human hippocampus after perinatal HI. Two rostrocaudal sections of the body of the hippocampus were analysed, and the number of immunostained cells in the polymorphic layer of the dentate gyrus (DG) and the pyramidal cell layer and stratum oriens of the CA3, CA2 and CA1 fields of the hippocampus proper were quantified. The results showed a lower density of both neuropeptides in hypoxic compared to control cases. In the HI group, the number of SOM-immunoreactive cell bodies was statistically significantly lower in the pyramidal cell layer and stratum oriens of CA1, while the number of NPY-expressing neurons was statistically lower in the pyramidal cell layer of CA2. Besides, the number of SOM-expressing neurons was significantly higher in the stratum oriens of CA1 compared to that in CA2. In sum, we observed a different vulnerability of SOM- and NPY-containing neurons in the developing human hippocampus following perinatal HI damage. Our results could contribute to a better understanding of the behaviour of these neuronal populations under stressful conditions during the perinatal period

Ex vivo, in situ perfusion protocol for human brain fixation compatible with microscopy, MRI techniques, and anatomical studies

We present a method for human brain fixation based on simultaneous perfusion of 4% paraformaldehyde through carotids after a flush with saline. The left carotid cannula is used to perfuse the body with 10% formalin, to allow further use of the body for anatomical research or teaching. The aim of our method is to develop a vascular fixation protocol for the human brain, by adapting protocols that are commonly used in experimental animal studies. We show that a variety of histological procedures can be carried out (cyto- and myeloarchitectonics, histochemistry, immunohistochemistry, intracellular cell injection, and electron microscopy). In addition, ex vivo, ex situ high-resolution MRI (9.4T) can be obtained in the same specimens. This procedure resulted in similar morphological features to those obtained by intravascular perfusion in experimental animals, provided that the postmortem interval was under 10 h for several of the techniques used and under 4 h in the case of intracellular injections and electron microscopy. The use of intravascular fixation of the brain inside the skull provides a fixed whole human brain, perfectly fitted to the skull, with negligible deformation compared to conventional techniques. Given this characteristic of ex vivo, in situ fixation, this procedure can probably be considered the most suitable one available for ex vivo MRI scans of the brain. We describe the compatibility of the method proposed for intravascular fixation of the human brain and fixation of the donor’s body for anatomical purposes. Thus, body donor programs can provide human brain tissue, while the remainder of the body can also be fixed for anatomical studies. Therefore, this method of human brain fixation through the carotid system optimizes the procurement of human brain tissue, allowing a greater understanding of human neurological diseases, while benefiting anatomy departments by making the remainder of the body available for teaching purposes.This work was supported in part by the National Institute of Health (Grant R01 AG056014-R01), a UCLM travel grant (to RI), a UCLM research grant to Human Neuroanatomy Laboratory (2020-GRIN-28837), and project EQC2019-006341-P (AEI/FEDER UE)

Integrating Color Deconvolution Thresholding and Weakly Supervised Learning for Automated Segmentation of Neurofibrillary Tangle and Neuropil Threads

Abnormally phosphorylated tau proteins are known to be a major indicator of Alzheimer's Disease (AD) with strong association with memory loss and cognitive decline. Automated generation of pixel-wise accurate neurofibrillary tangles (NFTs) and neuropil threads (NTs) segmentation is a challenging task, due to lack of ground truth segmentation data of these abnormal tau pathology. This problem is most prominent in the case of segmenting NTs, where the small threadlike morphology makes pixel-wise labeling a laborious task and unrealistic for large-scale studies. Lack of ground truth data poses a significant limitation for many learning-based methods to generate accurate segmentations of NFTs and NTs. This work presents an automated pipeline for pixel level segmentation of NFTs and NTs that does not rely on ground truth segmentation data. The pipeline is composed of four main steps: (1) color deconvolution is used to separate histopathology images into staining channels (DAB, Hematoxylin, and Eosin), (2) Otsu's thresholding is used on the DAB stain channel to generate pixel level segmentation of abnormal tau proteins staining, (3) a weakly-supervised learning paradigm (WildCat), using only global descriptors of images, is used to generate density maps of potential regions of NFTs and NTs, and (4) density maps and segmentations are then integrated using connected component analysis to localize NFTs and NTs in the detected tau segmentations. Our results show high global classification accuracy for NFTs (Acc:0.96) and NTs (Acc:0.91), and statistically significant distinctions when evaluating the percent area occupied of the detected NTs relative to expert ratings of NTs severity. Qualitative assessment of the NFTs and NTs results showed accurate pixel-level segmentations of the NFTs, while modest performance for NTs

Unfolding the Medial Temporal Lobe Cortex to Characterize Neurodegeneration Due to Alzheimer’s Disease Pathology Using Ex vivo Imaging

Neurofibrillary tangle (NFT) pathology in the medial temporal lobe (MTL) is closely linked to neurodegeneration, and is the early pathological change associated with Alzheimer’s Disease (AD). In this work, we investigate the relationship between MTL morphometry features derived from high-resolution ex vivo imaging and histology-based measures of NFT pathology using a topological unfolding framework applied to a dataset of 18 human postmortem MTL specimens. The MTL has a complex 3D topography and exhibits a high degree of inter-subject variability in cortical folding patterns which poses a significant challenge for volumetric registration methods typically used during MRI template construction. By unfolding the MTL cortex, the proposed framework explicitly accounts for the sheet-like geometry of the MTL cortex and provides a two-dimensional reference coordinate space which can be used to implicitly register cortical folding patterns across specimens based on distance along the cortex despite large anatomical variability. Leveraging this framework in a subset of 15 specimens, we characterize the associations between NFTs and morphological features such as cortical thickness and surface curvature and identify regions in the MTL where patterns of atrophy are strongly correlated with NFT pathology

Ex vivo MRI atlas of the human medial temporal lobe : characterizing neurodegeneration due to tau pathology

Tau neurofibrillary tangle (NFT) pathology in the medial temporal lobe (MTL) is closely linked to neurodegeneration, and is the early pathological change associated with Alzheimer’s disease (AD). To elucidate patterns of structural change in the MTL specifically associated with tau pathology, we compared high-resolution ex vivo MRI scans of human postmortem MTL specimens with histology-based pathological assessments of the MTL. MTL specimens were obtained from twenty-nine brain donors, including patients with AD, other dementias, and individuals with no known history of neurological disease. Ex vivo MRI scans were combined using a customized groupwise diffeomorphic registration approach to construct a 3D probabilistic atlas that captures the anatomical variability of the MTL. Using serial histology imaging in eleven specimens, we labelled the MTL subregions in the atlas based on cytoarchitecture. Leveraging the atlas and neuropathological ratings of tau and TAR DNA-binding protein 43 (TDP-43) pathology severity, morphometric analysis was performed to correlate regional MTL thickness with the severity of tau pathology, after correcting for age and TDP-43 pathology. We found significant correlations between tau pathology and thickness in the entorhinal cortex (ERC) and stratum radiatum lacunosum moleculare (SRLM). When focusing on cases with low levels of TDP-43 pathology, we found strong associations between tau pathology and thickness in the ERC, SRLM and the subiculum/cornu ammonis 1 (CA1) subfields of the hippocampus, consistent with early Braak stages