S100B is selectively expressed by gray matter protoplasmic astrocytes and myelinating oligodendrocytes in the developing CNS

Studies on the development of central nervous system (CNS) primarily rely on the use of specific molecular markers for different types of neural cells. S100B is widely being used as a specific marker for astrocytes in the CNS. However, the specificity of its expression in astrocyte lineage has not been systematically investigated and thus has remained a lingering issue. In this study, we provide several lines of molecular and genetic evidences that S100B is expressed in both protoplasmic astrocytes and myelinating oligodendrocytes. In the developing spinal cord, S100B is first expressed in the ventral neuroepithelial cells, and later in ALDH1L1+/GS+ astrocytes in the gray matter. Meanwhile, nearly all the S100B+ cells in the white matter are SOX10+/MYRF+ oligodendrocytes. Consistent with this observation, S100B expression is selectively lost in the white matter in Olig2-null mutants in which oligodendrocyte progenitor cells (OPCs) are not produced, and dramatically reduced in Myrf-conditional knockout mutants in which OPCs fail to differentiate. Similar expression patterns of S100B are observed in the developing forebrain. Based on these molecular and genetic studies, we conclude that S100B is not a specific marker for astrocyte lineage; instead, it marks protoplasmic astrocytes in the gray matter and differentiating oligodendrocytes.

Architecture of the Neuro-Glia-Vascular System

Astrocytes connect the vasculature to neurons and mediate the supply of nutrients and biochemicals. They also remove metabolites from the neurons and extracellular environment. They are involved in a growing number of physiological and pathophysiological processes. Understanding the biophysical, physiological, and molecular interactions in this neuro-glia-vascular ensemble (NGV) and how they support brain function is severely restricted by the lack of detailed cytoarchitecture. To address this problem, we used data from multiple sources to create a data-driven digital reconstruction of the NGV at micrometer anatomical resolution. We reconstructed 0.2 mm3 of rat somatosensory cortical tissue with approximately 16000 morphologically detailed neurons, its microvasculature, and approximately 2500 morphologically detailed protoplasmic astrocytes. The consistency of the reconstruction with a wide array of experimental measurements allows novel predictions of the numbers and locations of astrocytes and astrocytic processes that support different types of neurons. This allows anatomical reconstruction of the spatial microdomains of astrocytes and their overlapping regions. The number and locations of end-feet connecting each astrocyte to the vasculature can be determined as well as the extent to which they cover the microvasculature. The structural analysis of the NGV circuit showed that astrocytic shape and numbers are constrained by vasculature’s spatial occupancy and their functional role to form NGV connections. The digital reconstruction of the NGV is a resource that will enable a better understanding of the anatomical principles and geometric constraints which govern how astrocytes support brain function.Table of contentsMain pointsThe Blue Brain Project digitally reconstructs a part of neocortical Neuro-Glia-Vascular organizationInterdependencies and topological methods allow dense in silico reconstruction from sparse experimental dataThe polarized role of protoplasmic astrocytes constrains their shapes and numbersTable of contents image

Cell type-specific predictive models for prioritizing genes and gene sets associated with autism spectrum disorder

Background Autism spectrum disorder (ASD) is characterized by substantial phenotypic and genetic heterogeneity. Although bulk transcriptomic analyses revealed convergence of disease pathology on common pathways, the brain cell type-specific molecular pathology of ASD is still needed to study. Different gene functions may be dysregulated and causal genes may be distinct among different brain cells in ASD. Gene expression profiling-based machine learning studies can be conducted for the diagnosis of ASD, prioritizing high-confidence gene candidates and promoting the design of effective interventions.Methods To characterize the cell type heterogeneity of ASD and to take advantage of the potential of gene expression signature as diagnostic biomarkers for ASD, we construct multiple kinds of classification models for ASD based on the recently available human brain nucleus gene expression data of ASD and controls. Firstly, we construct cell type-specific predictive models based on individual genes to screen cell type-specific genes associated with ASD. Then from the view of gene set, we construct cell type-specific gene set-based predictive models to screen cell type-specific gene sets associated with ASD. These two kinds of predictive models can be applied to predict the diagnosis of a given nucleus with known cell type. Lastly, we further construct a multi-label predictive model for predicting the cell type and diagnosis of a given nucleus at the same time.Results It is found that the functions of genes with predictive power for ASD are not consistent and the top important genes are distinct among different cells, demonstrating the cell type heterogeneity of ASD. Our findings suggest that layer 2/3 and layer 4 excitatory neurons, layer 5/6 cortico-cortical projection neurons, parvalbumin interneurons, and protoplasmic astrocytes are preferentially affected in ASD. Gene BCYRN1 and CCK are prioritized in excitatory neurons, and HSPA1A is of note in protoplasmic astrocytes.Limitations Our study utilized methods of machine learning to identify biomarkers of ASD, while it is more convincing if subsequent experiments could be conducted to validate the results.Conclusions The results show that it may be feasible to use single cell/nucleus gene expression for ASD detection and the constructed predictive models can promote the diagnosis of ASD. Our analytical pipeline prioritizes ASD-associated cell type-specific genes and gene sets, which may be used as potential biomarkers of ASD.

Single-Cell Analysis for Glycogen Localization and Metabolism in Cultured Astrocytes

Cerebral glycogen is principally localized in astrocytes rather than in neurons. Glycogen metabolism has been implicated in higher brain functions, including learning and memory, yet the distribution patterns of glycogen in different types of astrocytes have not been fully described. Here, we applied a method based on the incorporation of 2-NBDG, a d-glucose fluorescent derivative that can trace glycogen, to investigate glycogen’s distribution in the brain. We identified two types of astrocytes, namely, 2-NBDGI (glycogen-deficient) and 2-NBDGII (glycogen-rich) cells. Whole-cell patch-clamp and fluorescence-activated cell sorting (FACS) were used to separate 2-NBDGII astrocytes from 2-NBDGI astrocytes. The expression levels of glycogen metabolic enzymes were analyzed in 2-NBDGI and 2-NBDGII astrocytes. We found unique glycogen metabolic patterns between 2-NBDGI and 2-NBDGII astrocytes. We also observed that 2-NBDGII astrocytes were mainly identified as fibrous astrocytes but not protoplasmic astrocytes. Our data reveal cell type-dependent glycogen distribution and metabolism patterns, suggesting diverse functions of these different astrocytes.

Ultrastructural Remodeling of the Neurovascular Unit in the Female Diabetic db/db Model—Part I: Astrocyte

Obesity, insulin resistance, and type 2 diabetes mellitus are associated with cognitive impairment, known as diabetic cognopathy. In this study, we tested the hypothesis that neurovascular unit(s) (NVU) within cerebral cortical gray matter regions display abnormal cellular remodeling. The monogenic (Leprdb) female diabetic db/db (BKS.CgDock7m +/+Leprdb/J; DBC) mouse model was utilized for this ultrastructural study. Upon sacrifice (at 20 weeks of age), left-brain hemispheres of the DBC and age-matched non-diabetic wild-type control C57BL/KsJ (CKC) mice were immediately immersion-fixed. We found attenuation/loss of endothelial blood–brain barrier tight/adherens junctions and pericytes, thickening of the basement membrane, aberrant mitochondria, and pathological remodeling of protoplasmic astrocytes. Additionally, there were adherent red blood cells and NVU microbleeds (cortical layer III) in DBC mice, which were not observed in CKC animals. While this study represents only a “snapshot in time”, it does allow for cellular remodeling comparisons between DBC and CKC. In this paper, the first of a three-part series, we report the observational ultrastructural remodeling changes of the NVU and its protoplasmic astrocytes in relation to the surrounding neuropil. Having identified multiple abnormal cellular remodeling changes in the DBC as compared to CKC models, we will design future experiments to evaluate various treatment modalities in DBC mice.

The Special Case of Human Astrocytes

In this first issue of Neuroglia, it is highly appropriate that Professor Jorge A. Colombo at the Unit of Applied Neurobiology (UNA, CEMIC-CONICET) in Buenos Aires, Argentina, writes a perspective of idiosyncrasies of astrocytes in the human brain. Much of his work has been focused on the special case of interlaminar astrocytes, so-named because of their long straight processes that traverse the layers of the human cerebral cortex. Notably, interlaminar astrocytes are primate-specific and their evolutionary development is directly related to that of the columnar organization of the cerebral cortex in higher primates. The human brain also contains varicose projection astrocytes or polarized astrocytes which are absent in lower animals. In addition, classical protoplasmic astrocytes dwelling in the brains of humans are ≈15-times larger and immensely more complex than their rodent counterparts. Human astrocytes retain their peculiar morphology even after grafting into rodent brains; that is, they replace the host astrocytes and confer certain cognitive advantages into so-called ‘humanised’ chimeric mice. Recently, a number of innovative studies have highlighted the major differences between human and rodent astrocytes. Nonetheless, these differences are not widely recognized, and we hope that Jorge Colombo’s Perspective and our associated Commentary will help stimulate appreciation of human astrocytes by neuroscientists and glial cell biologists alike.

Effects of FTY720 (Fingolimod) on Proliferation, Differentiation, and Migration of Brain-Derived Neural Stem Cells

Insufficient proliferation, differentiation, and migration are the main pitfalls of neural stem cells (NSCs) in reparative therapeutics for the central nervous system (CNS) diseases. The potent lipid mediator sphingosine-1-phosphate (S1P) regulates cells’ biological behavior broadly in the CNS. However, the effects of activating S1P on NSCs are not quite clear. In the current study, FTY720 (Fingolimod), an analog of S1P, was employed to induce the proliferation, differentiation, and migration of cultured brain-derived NSCs. The results indicated that proliferation and migration ability of NSCs were promoted by FTY720. Though we observed no obvious neuron prefers differentiation of NSCs, there were more protoplasmic astrocytes developed in the presence of certain concentration of FTY720. This work gives more comprehensive understanding of how FTY720 affects NSCs.