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Amphotericin B induces glial cell line-derived neurotrophic factor in the rat brain.

Motoyoshi-Yamashiro A, Takano K, Kawabe K, Izawa T, Nakajima H, Moriyama M, Nakamura Y - J. Vet. Med. Sci. (2014)

Bottom Line: These results suggested that neurotrophic factors derived from glial cells might be involved in the therapeutic mechanism of AmB.In the present study, we examined immunohistochemically the effects of AmB on the expression of neurotrophic factors in the rat brain.We found that direct injection of AmB into the striatum significantly enhanced the expression of glial cell line-derived neurotrophic factor protein.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Integrative Physiology in Veterinary Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Osaka 598-8531, Japan.

ABSTRACT
Amphotericin B (AmB) is a polyene antifungal drug and is reported to be one of a few reagents having therapeutic effects on prion diseases, that is, a delay in the appearance of clinical signs and prolongation of the survival time in an animal model. In prion diseases, glial cells have been suggested to play important roles; however, the therapeutic mechanism of AmB on prion diseases remains elusive. We have previously reported that AmB changed the expression of neurotrophic factors in microglia and astrocytes (Motoyoshi et al., 2008, Neurochem. Int. 52, 1290-1296; Motoyoshi-Yamashiro et al., 2013, ibid. 63, 93-100). These results suggested that neurotrophic factors derived from glial cells might be involved in the therapeutic mechanism of AmB. In the present study, we examined immunohistochemically the effects of AmB on the expression of neurotrophic factors in the rat brain. We found that direct injection of AmB into the striatum significantly enhanced the expression of glial cell line-derived neurotrophic factor protein. Amphotericin B also increased the expressions of CD11b and glial fibrillary acidic protein, markers of microglia and astrocytes, respectively. Moreover, expressions of the two neurotrophic factors by AmB were co-localized with the expression of CD11b or glial fibrillary acidic protein. These results suggest that AmB in vivo might also activate glial cells and induce the production of neurotrophic factors protecting neurons in prion diseases.

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The picture shows the administration site, and the photo shows a representative brainsection stained with anti-GDNF antibody in the vehicle group. The circles with dashedline show the peripheral region of the right striatum around the injection site andthe contralateral region used in the densitometric analysis. Scale bar=1 mm.
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fig_001: The picture shows the administration site, and the photo shows a representative brainsection stained with anti-GDNF antibody in the vehicle group. The circles with dashedline show the peripheral region of the right striatum around the injection site andthe contralateral region used in the densitometric analysis. Scale bar=1 mm.

Mentions: For the determination of protein expression level, sections were rinsed with PBS for 5 minthree times and placed in PBS containing 0.3% hydrogen peroxide and 0.065% sodium azide for30 min to eliminate the activity of endogenous peroxidase. After being washed with PBS,sections were reacted with Simple Stain Max PO (G) or Simple Stain Max PO (Multi) as asecondary antibody at room temperature for 30 min. The sections were washed again with PBSand then incubated in freshly prepared DAB solution for the appropriate period of time.After being washed in distilled water, sections were photographed, and the expression ofeach protein immunoreactively positive against antibodies was assessed by densitometricanalysis using quantification software, Image J (NIH). The measurements were carried out inthe peripheral region of the striatum around the site of injection but avoiding the trace ofinjection. Similarly, the contralateral regions were also measured, and the ratios of thedensities were quantified (Fig. 1Fig. 1.


Amphotericin B induces glial cell line-derived neurotrophic factor in the rat brain.

Motoyoshi-Yamashiro A, Takano K, Kawabe K, Izawa T, Nakajima H, Moriyama M, Nakamura Y - J. Vet. Med. Sci. (2014)

The picture shows the administration site, and the photo shows a representative brainsection stained with anti-GDNF antibody in the vehicle group. The circles with dashedline show the peripheral region of the right striatum around the injection site andthe contralateral region used in the densitometric analysis. Scale bar=1 mm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4221168&req=5

fig_001: The picture shows the administration site, and the photo shows a representative brainsection stained with anti-GDNF antibody in the vehicle group. The circles with dashedline show the peripheral region of the right striatum around the injection site andthe contralateral region used in the densitometric analysis. Scale bar=1 mm.
Mentions: For the determination of protein expression level, sections were rinsed with PBS for 5 minthree times and placed in PBS containing 0.3% hydrogen peroxide and 0.065% sodium azide for30 min to eliminate the activity of endogenous peroxidase. After being washed with PBS,sections were reacted with Simple Stain Max PO (G) or Simple Stain Max PO (Multi) as asecondary antibody at room temperature for 30 min. The sections were washed again with PBSand then incubated in freshly prepared DAB solution for the appropriate period of time.After being washed in distilled water, sections were photographed, and the expression ofeach protein immunoreactively positive against antibodies was assessed by densitometricanalysis using quantification software, Image J (NIH). The measurements were carried out inthe peripheral region of the striatum around the site of injection but avoiding the trace ofinjection. Similarly, the contralateral regions were also measured, and the ratios of thedensities were quantified (Fig. 1Fig. 1.

Bottom Line: These results suggested that neurotrophic factors derived from glial cells might be involved in the therapeutic mechanism of AmB.In the present study, we examined immunohistochemically the effects of AmB on the expression of neurotrophic factors in the rat brain.We found that direct injection of AmB into the striatum significantly enhanced the expression of glial cell line-derived neurotrophic factor protein.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Integrative Physiology in Veterinary Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Osaka 598-8531, Japan.

ABSTRACT
Amphotericin B (AmB) is a polyene antifungal drug and is reported to be one of a few reagents having therapeutic effects on prion diseases, that is, a delay in the appearance of clinical signs and prolongation of the survival time in an animal model. In prion diseases, glial cells have been suggested to play important roles; however, the therapeutic mechanism of AmB on prion diseases remains elusive. We have previously reported that AmB changed the expression of neurotrophic factors in microglia and astrocytes (Motoyoshi et al., 2008, Neurochem. Int. 52, 1290-1296; Motoyoshi-Yamashiro et al., 2013, ibid. 63, 93-100). These results suggested that neurotrophic factors derived from glial cells might be involved in the therapeutic mechanism of AmB. In the present study, we examined immunohistochemically the effects of AmB on the expression of neurotrophic factors in the rat brain. We found that direct injection of AmB into the striatum significantly enhanced the expression of glial cell line-derived neurotrophic factor protein. Amphotericin B also increased the expressions of CD11b and glial fibrillary acidic protein, markers of microglia and astrocytes, respectively. Moreover, expressions of the two neurotrophic factors by AmB were co-localized with the expression of CD11b or glial fibrillary acidic protein. These results suggest that AmB in vivo might also activate glial cells and induce the production of neurotrophic factors protecting neurons in prion diseases.

Show MeSH
Related in: MedlinePlus