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. 2021 Mar 22;11(1):6551.
doi: 10.1038/s41598-021-85972-2.

Hericium erinaceus mycelium and its small bioactive compounds promote oligodendrocyte maturation with an increase in myelin basic protein

Hui-Ting Huang  1 , Chia-Hsin Ho  1 , Hsin-Yu Sung  1 , Li-Ya Lee  2 , Wan-Ping Chen  2 , Yu-Wen Chen  2 , Chin-Chu Chen  2 , Chung-Shi Yang  3 , Shun-Fen Tzeng  4
Affiliations

Hericium erinaceus mycelium and its small bioactive compounds promote oligodendrocyte maturation with an increase in myelin basic protein

Hui-Ting Huang et al. Sci Rep. .

Erratum in

Abstract

Oligodendrocytes (OLs), myelin-producing glia in the central nervous system (CNS), produce a myelin extension that enwraps axons to facilitate action potential propagation. An effective approach to induce oligodendrogenesis and myelination is important to foster CNS development and promote myelin repair in neurological diseases. Hericium (H.) erinaceus, an edible and culinary-medicinal mushroom, has been characterized as having neuroprotective activities. However, its effect on OL differentiation has not yet been uncovered. In this study using oligodendrocyte precursor cell (OPC) cultures and an ex vivo cerebellar slice system, we found that the extract from H. erinaceus mycelium (HEM) not only promoted the differentiation of OPCs to OLs in the differentiation medium, but also increased the level of myelin basic protein (MBP) on neuronal fibers. Moreover, daily oral administration of HEM into neonatal rat pups for 7 days enhanced MBP expression and OLs in the corpus callosum of the postnatal rat brain. The effect of HEM-derived bioactive compounds, the diterpenoid xylosides erinacine A (HeA) and HeC and a sesterterpene with 5 isoprene units called HeS, were further evaluated. The results showed that HeA and HeS more potently stimulated MBP expression in OLs and increased the number of OLs. Moreover, overlap between MBP immunoreactivity and neuronal fibers in cultured cerebellar tissue slices was significantly increased in the presence of HeA and HeS. In summary, our findings indicate that HEM extract and its ingredients HeA and HeS display promising functional effects and promote OL maturation, providing insights into their potential for myelination in neurodevelopmental disorders.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Schematic diagram of the experimental designs. (A) The structures of HeA, HeC, and HeS from the H. erinaceus mycelium (HEM) are depicted. (B) In an in vitro experiment, OPCs were seeded in GM for 2 days and then grown in DM for another 2 days. Then, crude HEM, HeA, HeC, or HeS at the indicated concentrations was added to the cultures maintained in DM for 3 consequential days. The cultures were subjected to immunofluorescence to detect GC and MBP, Western blot analysis to measure MBP and PLP, and qPCR to measure OL differentiation-associated genes. (C) The cell markers used to identify OL lineages are illustrated. In this study, immunostaining for NG2, APC, GC, and myelin proteins (MBP and PLP) was conducted to identify OPCs and OLs. Note that the detectable level of MBP and PLP immunoreactivity can be observed in immature OLs in culture.
Figure 2
Figure 2
Crude HEM enhanced OL maturation. Crude HEM at the indicated concentrations was added to the cultures for 3 days after OPCs had been maintained in DM for 2 days (Fig. 1B). (A) The cultures were subjected to GC and MBP staining (red), followed by DAPI nuclear counterstaining (blue). GC+- and MBP+-OLs in the cultures were quantified as described in the “Materials and methods” section. Each spot represents as the data quantified from one photo captured from the cultures. Arrows shown in the representative images indicate OLs with the shape of an extended membrane. (B) Total proteins were isolated from cultures treated with crude HEM for 3 days and subjected to Western blot analysis to examine MBP and PLP protein levels. The intensity of the immunoreactive bands shown in the immunoblots was quantified by ImageJ software version 1.52a (https://imagej.nih.gov/ij/) and normalized to the level of GAPDH, which was used as a loading control. The immunoblot images used for quantification are provided in Fig. S5. Data are presented as the means ± SEMs of the three independent experiments. The raw immunoblot images are shown in Fig. S5. *p < 0.05, **p < 0.01 compared with the control culture. Scale bar in (A) 50 μm.
Figure 3
Figure 3
Ex vivo and in vivo experiments reveal the effect of crude HEM in stimulating MBP production and OL maturation. (A) Cerebellar tissue slices taken from the vermis (as indicated by red lines) were prepared from P7 rats and then cultured for 3 days. Crude HEM at concentrations of 0.1 and 1 μg/ml was added to the cultures for 7 days. The cultures were subjected to immunofluorescence for NF200 and MBP staining. (B) Confocal images show overlap between the immunoreactive intensity of MBP with NF200+-fibers (arrows), which was quantified using ImageJ software. The data were obtained from four random fields per culture (right panel). The results are presented as the means ± SEMs of the four tissue slices for each treatment. Each dot denotes data obtained from one tissue slice. (C) For the in vivo study, crude HEM was administered to rat pups at the age of P3 orally daily for 7 consecutive days. After brains were removed and cryostat sectioned, followed by immunofluorescence for MBP or APC. MBP intensity and APC+-cells were quantified in the body portion of the corpus callosum using ImageJ software. (D). After immunofluorescence by anti-MBP (red) or anti-APC (red), the brain tissue sections were subjected to DAPI nuclear counterstaining (blue). The representative images in the body portion of the corpus callosum were captured by confocal microscopy, and the immunoreactive intensity of MBP and APC was then quantified as described in “Materials and methods” section. The results are presented as the means ± SEMs of 6 animals in each group. *p < 0.05, ***p < 0.001 versus the vehicle-treated control culture. Scale bar in (B) 20 μm; in (D) 50 μm.
Figure 4
Figure 4
Three HEM-derived compounds promote OL maturation. After OPCs were maintained in DM for 2 days, as described in Fig. 1B, three known HEM-derived compounds (HeA, HeC, HeS) were added to the cultures at the indicated concentrations and incubated for another 3 days. The cultures were processed for GC (A) and MBP (B) immunofluorescence (red) and DAPI nuclear counterstaining (blue). The OLs with a membranous extension are indicated by arrows in the representative images. GC+- and MBP+-OLs in the cultures were quantified as described in “Materials and methods” section. Each spot represents as the data quantified from one photo captured from the cultures. The experiments were repeated by the three independent cell preparations. *p < 0.05, **p < 0.01, ***p < 0.001 versus the vehicle-treated control culture. Scale bar, 50 μm.
Figure 5
Figure 5
HeA and HeS increased MBP protein production in OL cultures. OPCs were first maintained in DM for 2 days and then exposed to HeA, HeC, or HeS at the indicated concentrations for another 3 days (Fig. 1B). Total proteins were extracted from the cultures and subjected to Western blot analysis to examine MBP (A) and PLP (B) protein levels. The GAPDH level was used as a loading control. The intensity of the immunoreactive bands shown in the immunoblots was quantified by ImageJ software version 1.52a (https://imagej.nih.gov/ij/), and normalized to the level of GAPDH, which was used as a loading control. The results are presented as the means ± SEMs of three repeated experiments. The raw immunoblot images are shown in Figs. S6, S7, and S8. *p < 0.05, **p < 0.01 compared with the control culture.
Figure 6
Figure 6
Exposure to HeS increased the number of NG2+-OPCs in cerebellar tissue slices. (A) The schematic diagram depicts that HeA, HeC, and HeS at the indicated concentrations were added to cerebellar slice cultures and incubated for 7 days. (B) The cultures were subjected to NG2 immunofluorescence (red) and DAPI nuclear counterstaining (blue). The representative images captured from confocal microscopy (left panel) and NG2+-OPCs in 4 random fields per culture were quantified as described in “Materials and methods” section (right panel). The results are presented as the means ± SEMs of the total tissue slices (n = 4–5) for each treatment. Each dot denotes data obtained from one tissue slice. **p < 0.01 compared with the corresponding control culture. Scale bar, 50 μm.
Figure 7
Figure 7
HeA and HeS enhanced the ensheathment of neuronal fibers with MBP+-cell processes. As described in Fig. 6A, cerebellar tissue slices were treated with HeA, HeC, or HeS at the indicated concentrations for 7 days. NF200 (green) and MBP (red) double immunofluorescence was conducted. Representative images were captured by confocal microscopy, and the overlap between the MBP signal and NF200+-neuronal fibers in 4 random fields per culture was quantified as described in “Materials and methods” section (right panel). The results are presented as the means ± SEMs of three repeated experiments. *p < 0.05, **p < 0.01 compared with the corresponding control culture. Scale bar, 20 μm.
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References

    1. Bradl M, Lassmann H. Oligodendrocytes: biology and pathology. Acta Neuropathol. 2010;119:37–53. doi: 10.1007/s00401-009-0601-5. - DOI - PMC - PubMed
    1. Lee Y, et al. Oligodendroglia metabolically support axons and contribute to neurodegeneration. Nature. 2012;487:443–448. doi: 10.1038/nature11314. - DOI - PMC - PubMed
    1. Mayoral SR, Chan JR. The environment rules: spatiotemporal regulation of oligodendrocyte differentiation. Curr. Opin. Neurobiol. 2016;39:47–52. doi: 10.1016/j.conb.2016.04.002. - DOI - PMC - PubMed
    1. Zuchero JB, Barres BA. Intrinsic and extrinsic control of oligodendrocyte development. Curr. Opin. Neurobiol. 2013;23:914–920. doi: 10.1016/j.conb.2013.06.005. - DOI - PMC - PubMed
    1. van Tilborg E, et al. Origin and dynamics of oligodendrocytes in the developing brain: implications for perinatal white matter injury. Glia. 2018;66:221–238. doi: 10.1002/glia.23256. - DOI - PMC - PubMed

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