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Characteristic response of striatal astrocytes to dopamine depletion.

Neural regeneration research (2019-10-23)
Yao-Feng Zhu, Wei-Ping Wang, Xue-Feng Zheng, Zhi Chen, Tao Chen, Zi-Yun Huang, Lin-Ju Jia, Wan-Long Lei
ABSTRACT

Astrocytes and astrocyte-related proteins play important roles in maintaining normal brain function, and also regulate pathological processes in brain diseases and injury. However, the role of astrocytes in the dopamine-depleted striatum remains unclear. A rat model of Parkinson's disease was therefore established by injecting 10 μL 6-hydroxydopamine (2.5 μg/μL) into the right medial forebrain bundle. Immunohistochemical staining was used to detect the immunoreactivity of glial fibrillary acidic protein (GFAP), calcium-binding protein B (S100B), and signal transducer and activator of transcription 3 (STAT3) in the striatum, and to investigate the co-expression of GFAP with S100B and STAT3. Western blot assay was used to measure the protein expression of GFAP, S100B, and STAT3 in the striatum. Results demonstrated that striatal GFAP-immunoreactive cells had an astrocytic appearance under normal conditions, but that dopamine depletion induced a reactive phenotype with obvious morphological changes. The normal striatum also contained S100B and STAT3 expression. S100B-immunoreactive cells were uniform in the striatum, with round bodies and sparse, thin processes. STAT3-immunoreactive cells presented round cell bodies with sparse processes, or were darkly stained with a large cell body. Dopamine deprivation induced by 6-hydroxydopamine significantly enhanced the immunohistochemical positive reaction of S100B and STAT3. Normal striatal astrocytes expressed both S100B and STAT3. Striatal dopamine deprivation increased the number of GFAP/S100B and GFAP/STAT3 double-labeled cells, and increased the protein levels of GFAP, S100B, and STAT3. The present results suggest that morphological changes in astrocytes and changes in expression levels of astrocyte-related proteins are involved in the pathological process of striatal dopamine depletion. The study was approved by Animal Care and Use Committee of Sun Yat-sen University, China (Zhongshan Medical Ethics 2014 No. 23) on September 22, 2014.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Mouse IgG (whole molecule) F(ab′)2 fragment–Cy3 antibody produced in sheep, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
6-Hydroxydopamine hydrochloride, ≥97% (titration), powder
Sigma-Aldrich
3,3′-Diaminobenzidine tetrahydrochloride hydrate, ≥96%
Sigma-Aldrich
Anti-Rabbit IgG (H+L), F(ab′)2 fragment, CF640R antibody produced in goat, ~2 mg/mL, affinity isolated antibody, buffered aqueous solution