Dynamic changes in eosinophilic inclusion bodies in lactacystin-incubated PC12 cells
10.3760/cma.j.issn.1671-8925.2009.11.002
- VernacularTitle:Lactacystin诱导PC12细胞胞内嗜酸性包涵体动态变化的研究
- Author:
Hai-Na ZHANG
1
;
Xin-Yu HU
;
Guo-Hua HU
;
Qiu-Hui CHEN
;
Yan-Qiu HAN
Author Information
1. 吉林大学第二临床学院
- Keywords:
Proteasome;
Parkinson's disease;
PC12eell;
Inclusion;
α-synuclein
- From:
Chinese Journal of Neuromedicine
2009;8(11):1086-1089
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the development of eosinophilic inclusion bodies in PC12 cells incubated with proteasome inhibitor lactacystin. Methods PC12 cells were incubated for 24 h with 0, 5, 10 and 20 μmol/L lactacystin, and HE staining and α-synuclein immunohistochemistry was used to observe the changes of the inclusion bodies under light microscope. The changes in the inclusion bodies in the cytoplasm were also observed after treatment of PC12 cells with 10 μmol/l lactacystin for 24, 48, 72, 96 and 120 h. Results After incubation with lactacystin for 24 h, P12 cells showed increased number of eosinophilic inclusion bodies in the cytoplasm, and the increment appeared dose dependent. No inclusion bodies were observed in the cells without lactacystin treatment, and a few (3.33%±1.15%) occurred at lactacystin concentration of 5 μmol/L, numerous (71.33%±4.16%) at 10 μmol/l, and almost in every cell (90.33%±3.21%) at 20 μmol/L; some of the inclusion bodies were released from the cells. With the passage of time, inclusion bodies in the cells treated with 10 ?mol/l laetacystin began to dissociate with the cells, and the cytoplasm was reduced to contain only inclusion bodies and nucleus at 96 and 120 h. Immunohistochemistry showed that a -synuclein immunoreaetive granules concentrated and formed inclusion bodies close to the nucleus at 24 h. As the concentration increased and time prolonged, α-synuclein-immunoreactive inclusion bodies were released from the cells, leaving only inclusion bodies and cell nuclei presented at 96 and 120 h. Conclusion Incubation with laetacystin can induce cytoplasmic eosinophilic and α-synuclein-immunoreaetive inclusion bodies in PC12 cells. The development of inclusion bodies in this model is consistent with that in human Parkinson's disease and diffused Lewy body disease. This model is helpful for further study of the mechanism of neurodegenerative diseases.
