Identification of oligodendrocytes in experimental disease models

Abstract

The ability to identify oligodendrocytes in culture, in fixed tissue, and in vivo using unique markers is a requisite step to understanding their responses in any damage, recovery, or developmental process. Their nuclei are readily seen in histological preparations of healthy white and gray matter, and their cell bodies can be reliably identified with a variety of immunocytochemical markers. However, there is little consensus regarding optimal methods to assess oligodendrocyte survival or morphology under experimental injury conditions. We review common approaches for histological and immunocytochemical identification of these cells. Transgenic and viral methods for cell type-selective transfer of genes encoding fluorescent proteins offer promising new approaches for manipulating and visualizing oligodendrocytes in models of health and disease.

An Oligovascular Niche: Cerebral Endothelial Cells Promote the Survival and Proliferation of Oligodendrocyte Precursor Cells

“…OPCs were prepared as previously described with some modifications (Ness et al, 2005;van Leyen et al, 2008). Briefly, cerebral cortices from 1-to 2-d-old Sprague Dawley rats were dissected, minced, and digested.…”

“…After cells reached 70 -80% confluence, they were washed with ice-cold PBS, pH 7.4, followed by 4% paraformaldehyde for 30 min. After being further washed three times in PBS containing 0.1% Triton X-100, they were incubated with 1% bovine serum albumin in PBS for 1 h. Then cells were incubated with primary antibodies against the OPC markers (Ness et al, 2005) A2B5 (1:50), O4 (1:50), NG2 (1:50), or the astrocytic marker glial fibrillary acidic protein (GFAP; 1:400) at 4°C overnight. After washing with PBS, they were incubated with secondary antibodies conjugated with fluorescein isothiocyanate for 1 h at room temperature.…”

NMDA receptors expressed in oligodendrocytes

“…As with neurons, oligodendrocytes are highly sensitive to injury mediated by trophic factor deprivation, oxidative stress, excitatory amino acids and launching of apoptotic pathways. Hypoxic-ischemic damage to oligodendrocytes is a frequent feature in global ischemia (cardiac arrest), focal ischemia (stroke), cyanide intoxication and vascular dementia, (4) and oligodendrocytes are also injured in brain and spinal cord trauma, multiple sclerosis and even Alzheimer's disease. During the perinatal period, damage to oligodendrocyte progenitor cells results in periventricular leukomalacia and long-term demyelination, a key etiology of cerebral palsy.…”

“…In spinal cord or brain injury, damage to these cells is important in a growing list of acute and chronic conditions. (4) Much of our current knowledge about the oligodendrocyte concern their role in myelinated axons in the CNS. As with neurons, oligodendrocytes are highly sensitive to injury mediated by trophic factor deprivation, oxidative stress, excitatory amino acids and launching of apoptotic pathways.…”

Deleterious Role of IFNγ in a Toxic Model of Central Nervous System Demyelination

“…Paraffin-embedded sections were stained for the pi isoform of glutathione S-transferase (GST-pi, a marker of mature oligodendrocytes), 14,15 Ricinus communis agglutinin 1 (RCA-1, a marker for macrophages and microglia), 16,17 and glial fibrillary acidic protein (GFAP, a marker for astrocytes). Briefly, for GST-pi and GFAP staining, antigen retrieval was performed by boiling sections in citrate buffer, pH 6.0, for 30 minutes.…”