The glial cell-specific proteome
The function of the nervous system is to receive, process and execute the coordinated higher functions of perception, motion and cognition that signify human life. The nervous system is divided into a central nervous system (CNS), including the brain and the spinal cord, and a peripheral nervous system (PNS), including nerves branching out from the spinal cord to all parts of the body. Retina is an extension of the CNS responsible specifically for vision. The cellular components of this highly complex signal network include neurons and supportive glial cells.
Transcriptome analysis shows that 77% (n=15561) of all human proteins (n=20162) are detected in glial cells and 3758 of these genes show an elevated expression in any glial cells compared to other cell type groups. In-depth analysis of the elevated genes in glial cells using scRNA-seq and antibody-based protein profiling allowed us to visualize the expression patterns of these proteins in the following types of glial cells: astrocytes, oligodendrocyte precursor cells, oligodendrocytes, microglial cells, Muller glia cells and Schwann cells.
The glial cell transcriptome
The scRNA-seq-based glial cell transcriptome can be analyzed with regard to specificity, illustrating the number of genes with elevated expression in each specific glial cell type compared to other cell types (Table 1). Genes with an elevated expression are divided into three subcategories:
As shown in Table 1, 1251 genes are elevated in astrocytes compared to other cell types. Astrocytes are glial cells in the brain and spinal cord that fulfill many support functions like providing nutrients to the nerve cells and regulation of cerebral blood flow. One astrocyte enriched gene is the glial fibrillary acidic protein (GFAP) which is an intermediate filament protein specific for astrocytes. Another gene strongly enriched in astrocytes is glypican 5 (GPC5), a cell surface proteoglycan that may play a role in cell division and growth regulation.
Oligodendrocyte precursor cells
As shown in Table 1, 1622 genes are elevated in oligodendrocyte precursor cells compared to other cell types. PTPRZ1 negatively regulates oligodendrocyte precursor proliferation in the embryonic spinal cord and is required for normal differentiation of the precursor cells into mature, fully myelinating oligodendrocytes.
As shown in Table 1, 1630 genes are elevated in oligodendrocytes compared to other cell types. Oligodendrocytes are involved in myelination, such as the compact myelin proteins myelin basic protein (MBP), which contributes to stabilization and formation of the myelin throughout the CNS and the peripheral nervous system (PNS).
As shown in Table 1, 806 genes are elevated in microglial cells compared to other cell types. Microglial cells are a specialized type of macrophage only found in the central nervous system. Genes enriched in microglial cells are for example purinergic receptor P2RY12 required for platelet aggregation and blood coagulation. Another example of a protein expressed in microglia is the integrin subunit alpha M (ITGAM), which can be found in the immune system.
Muller glia cells
As shown in Table 1, 449 genes are elevated in Muller glia cells compared to other cell types. Muller glia cells are a type of glial cell found only in the retina that buffers potassium and neurotransmitters essential for the normal function of different types of neuronal cells of the retina, as well as maintains the structural integrity of the retina. An example of a protein with elevated expression in Muller glia cells is retinaldehyde binding protein 1 (RLBP1). It carries 11-cis-retinaldehyde, or 11-cis-retinal, molecules that are essential for the conversion of light into neuronal signals in the photoreceptor cells. The function of RLBP1 in Muller glia cells is yet to be characterized.
As shown in Table 1, 362 genes are elevated in Schwann cells compared to other cell types. Schwann cells are a type of glial cell found in the PNS surrounding and supporting the neurons throughout the body, involved in the production of the myelin sheaths that sometimes cover and insulate the nerves. An example of a protein with elevated expression in Schwann cells is myelin protein zero (MPZ). It is a large transmembrane protein that is necessary for the formation of normal myelination of nerves in the PNS. A second example of a protein with elevated expression in Schwann cells is S100 calcium-binding protein B (S100B), involved in the proliferation and differentiation of glial cells.
Glial cell function
Glial cells maintain the microenvironment essential for neuronal activity. An ion and water flow homeostasis is essential for the generation of the action potential by the neuronal cells. In the CNS this homeostasis is mainly managed by astrocytes and oligodendrocytes that form an intricate network, called panglial syncytium, while in the retina, Muller glial cells buffer potassium ions. The action potential is propagated along neuronal axons and to increase the speed of transmission, axons are insulated by myelin sheaths, which are produced by oligodendrocytes in the CNS, and by Schwann cells in the PNS.
Neuronal cells release neurotransmitters, e.g. glutamate, at the synapses and the neurotransmitters are recycled by glial cells (Mueller glia in retina and astrocytes in other parts of CNS), that maintain contact with the synapses. Neurotransmitters are captured by glial cells, transformed into inactive forms, and shuttled back to the synapses where they can be re-used by neuronal cells. This process requires a great amount of energy (ATP) and since the glial cells shoulder this task, the energy expenditure of neuronal cells is decreased. Certain molecules, e.g. glucose required for energy, pass through the blood-brain barrier, while harmful substances are prevented from entering the brain. Endothelial cells, pericytes, and astrocyte end-feet together comprise the blood-brain barrier. The end-feet ensheath the capillary and regulate the passage of molecules by affecting e.g. tight-junction formation and expression of different transporters. Astrocytes associated with capillaries also capture molecules, e.g. glucose, and process them into metabolites usable by neuronal cells. Pathogens that do pass the blood-brain barrier, as well as damaged neurons and harmful aggregations of proteins (plaques) are removed by microglia, a type of glial cell that is related to macrophages outside the brain.
The histology of organs that contain glial cells, including interactive images, is described in the Protein Atlas Histology Dictionary.
Here, the protein-coding genes expressed in glial cells are described and characterized, together with examples of immunohistochemically stained tissue sections that visualize corresponding protein expression patterns of genes with elevated expression in different glial cell types.
The transcript profiling was based on publicly available genome-wide expression data from scRNA-seq experiments covering 29 tissues and peripheral blood mononuclear cells (PBMCs). All datasets (unfiltered read counts of cells) were clustered separately using louvain clustering, resulting in a total of 557 different cell type clusters. The clusters were then manually annotated based on a survey of known tissue and cell type-specific markers. The scRNA-seq data from each cluster of cells was aggregated to mean normalized protein-coding transcripts per million (nTPM) and the normalized expression value (nTPM) across all protein-coding genes. A specificity and distribution classification was performed to determine the number of genes elevated in these single cell types, and the number of genes detected in one, several or all cell types, respectively.
It should be noted that since the analysis was limited to datasets from 29 tissues and PBMC only, not all human cell types are represented. Furthermore, some cell types are present only in low amounts, or identified only in mixed cell clusters, which may affect the results and bias the cell type specificity.
Relevant links and publications
Uhlén M et al., Tissue-based map of the human proteome. Science (2015)