Quantitative assessment of glial cells in the human and guinea pig enteric nervous system with an anti‐Sox8/9/10 antibody

Abstract

Quantitative changes of enteric glia (EGC) have been implicated in gastrointestinal disorders. To facilitate future studies of EGC in human pathology, we aimed to characterize thoroughly glial markers in the human enteric nervous system (ENS) and to compare EGC in man and guinea pig. Whole-mount preparations of the enteric nerve plexuses from human and guinea pig ileum and colon were labeled with antibodies against S100b, glial fibrillary acidic protein (GFAP), and p75NGFR and the transcription factors Sox8/9/10 and neuronally counterstained. Abundant immunoreactivity (IR) for S100b, GFAP, p75NGFR, and Sox8/9/10 was detected in EGC of all studied regions. Although the cytoplasmatic staining pattern of most markers did not permit glial quantification, the nuclear localization of Sox8/9/10-IR allowed to identify and count all EGC individually. In both man and guinea pig, myenteric ganglia were larger and contained more EGC and neurons than submucous ganglia. Furthermore, there were more EGC in the human than in the guinea pig myenteric plexus (MP), glial density was consistently higher in the human ENS, and the glia index (glia:neuron ratio) ranged from 1.3 to 1.9 and from 5.9 to 7.0 in the human submucous plexus (SMP) and MP, respectively, whereas, in guinea pig, the glia index was 0.8-1.0 in the SMP and 1.7 in the MP. The glia index was the most robust quantitative descriptor within one species. This is a comprehensive set of quantitative EGC measures in man and guinea pig that provides a basis for pathological assessment of glial proliferation and/or degeneration in the diseased gut.

Glial cells in the mouse enteric nervous system can undergo neurogenesis in response to injury

“…From that point onward, our Sox10-driven reporter system exclusively labels glial cells, as identified by the expression of enteric glia markers, such as GFAP and S100β. The total number of Sox10-expressing cells within the gut does not decrease over time, but the gradual reduction of neurogenesis is coupled to the increasing representation of gliogenic precursors and glial cells, which also express Sox10 (19,39). The neurogenic to gliogenic switch of Sox10 + progenitors could result from changes in the gut microenvironment, or the gradual accumulation of neuron-derived signals that reach levels sufficient to induce gliogenic fates.…”

Enteric glia and neuroprotection: basic and clinical aspects

“…A subtype of EGCs surrounds nerve processes in the mucosa, thereby establishing close contacts with the epithelial cells, while another population wraps around neuronal cell bodies in the ganglionated plexuses (20,32,37). The EGCs outnumber neurons with a ratio ranging from 1.3 to 1.9 and from 5.9 to 7.0 in the human submucosal and myenteric plexuses, respectively (23), and surround the neuronal cell bodies tightly (i.e., the remaining extracellular space consists of narrow 20-nm gaps) (20). As in the blood-brain barrier, ECGs possess several flat endfeet (or cytoplasmic expansions) that form a blood-enteric barrier protecting the enteric neurons from potentially harmful extraganglionic substances (41).…”

“…Available immunohistochemical markers for EGC labeling in the adult gut include GFAP, S100␤, and Sox8/9/10, the first two being the most frequently used (23,37) (Fig. 1).…”

“…Sox8/9 expression has been demonstrated in neural crest cells and in immature glial cells, whereas Sox10 appears to be restricted to EGCs in the mature ENS (51). Thus, by using antibodies targeting Sox8/9/10, it is possible to label EGC nuclei and thereby perform precise and quantitative analysis of EGC density (21,23).…”

Estudo do plexo mioentérico do cólon descendente de cães acometidos pela distrofia muscular (GRMD)

“…foram identificados e fotografados 10 gânglios, sob a objetiva de 10x, de cada animal para cada uma das 3 marcações (HOFF et al, 2008;SILVEIRA, 2013 A imunomarcação com HuC/D demonstrou muitos neurônios marcados dentro do gânglio. Observou-se muitos neurônios por gânglio e bem distribuídos dentro deste, (Figura 1).…”