Limits...
Brain microglia were activated in sporadic CJD but almost unchanged in fatal familial insomnia and G114V genetic CJD.

Shi Q, Xie WL, Zhang B, Chen LN, Xu Y, Wang K, Ren K, Zhang XM, Chen C, Zhang J, Dong XP - Virol. J. (2013)

Bottom Line: However, the situation of microglia in other genetic prion diseases such as fatal familial insomnia (FFI) and familial CJD remains less understood.The Iba1-levels maintained comparable in six different brain regions of FFI and G114V cases, including thalamus, cingulate gyrus, frontal cortex, parietal cortex, occipital cortex and temporal cortex.ELISA tests for inflammatory cytokines revealed significantly up-regulated IL-1β, IL-6 and TNF-α in the brain homogenates from sCJD, but not in those from FFI and G114V gCJD.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases (Hangzhou), National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Chang-Bai Rd 155, Beijing 102206, People's Republic of China.

ABSTRACT

Background: Microglial activations have been described in different subtypes of human prion diseases such as sporadic Creutzfeldt-Jakob disease (CJD), variant CJD, Kuru and Gerstmann-Sträussler-Scheinker disease (GSS). However, the situation of microglia in other genetic prion diseases such as fatal familial insomnia (FFI) and familial CJD remains less understood. The brain microglia was evaluated comparatively between the FFI, G114V and sCJD cases in the study.

Methods: Specific Western blots, immunohistochemical and immunofluorescent assays were used to detect the changes of microglia and ELISA tests were used for levels of inflammatory cytokines.

Results: Western blots, immunohistochemical and immunofluorescent assays illustrated almost unchanged microglia in the temporal lobes of FFI and G114V gCJD, but obviously increased in those of sCJD. The Iba1-levels maintained comparable in six different brain regions of FFI and G114V cases, including thalamus, cingulate gyrus, frontal cortex, parietal cortex, occipital cortex and temporal cortex. ELISA tests for inflammatory cytokines revealed significantly up-regulated IL-1β, IL-6 and TNF-α in the brain homogenates from sCJD, but not in those from FFI and G114V gCJD.

Conclusion: Data here demonstrates silent brain microglia in FFI and G114V gCJD but obviously increased in sCJD, which reflects various pathogenesis of different human prion diseases subtypes.

Show MeSH

Related in: MedlinePlus

Evaluation of the brain Iba1 levels of different human prion diseases by Western blots. A. Temporal lobes. Same amounts of 10% homogenates from three FFI, one sCJD (Case 1), one G114V gCJD and normal control were separated onto 15% SDS-PAGE and blotted with Iba1 or β-actin specific mAbs. The quantitative analyses of the relative gray values are showed on the right. The results are calculated from three blots and presented as mean ± SD. Statistical differences are illustrated as **P<0.01. B. Different brain regions. Same amounts of 10% homogenates of six regions, including thalamus, cingulate gyrus, frontal lobe, parietal lobe, occipital lobe and temporal lobe, from three FFI and the G114V gCJD cases were comparatively assayed. Same amounts of 10% homogenates of temporal lobe from one sCJD (Case 2) and normal one were loaded as control. Same amounts of 10% homogenates of frontal lobe, parietal lobe and occipital lobe from sCJD (Case 1) were also comparatively assayed with that of normal control (low panel). Molecular markers are indicated on the right.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3716817&req=5

Figure 2: Evaluation of the brain Iba1 levels of different human prion diseases by Western blots. A. Temporal lobes. Same amounts of 10% homogenates from three FFI, one sCJD (Case 1), one G114V gCJD and normal control were separated onto 15% SDS-PAGE and blotted with Iba1 or β-actin specific mAbs. The quantitative analyses of the relative gray values are showed on the right. The results are calculated from three blots and presented as mean ± SD. Statistical differences are illustrated as **P<0.01. B. Different brain regions. Same amounts of 10% homogenates of six regions, including thalamus, cingulate gyrus, frontal lobe, parietal lobe, occipital lobe and temporal lobe, from three FFI and the G114V gCJD cases were comparatively assayed. Same amounts of 10% homogenates of temporal lobe from one sCJD (Case 2) and normal one were loaded as control. Same amounts of 10% homogenates of frontal lobe, parietal lobe and occipital lobe from sCJD (Case 1) were also comparatively assayed with that of normal control (low panel). Molecular markers are indicated on the right.

Mentions: To see the potential change in microglia, the levels of Iba1 that was commonly used as the marker of total microglia including resting and activated state in temporal lobes were comparatively evaluated by Western blots. Compared with that of normal control, the Iba1 signal in sCJD (Case 1) was remarkably stronger (P<0.01), while that in the FFI and G114V gCJD maintained almost unchanged (Figure 2A). To test the Iba1 levels in other brain regions, six different regions including thalamus, callosal gyrus, frontal lobe, parietal lobe, occipital lobe and temporal lobe from three FFI cases and a G114V gCJD case were subjected into Iba1-specific Western blots, together with temporal lobe from sCJD Case 2 and normal control. It showed that the Iba1 signals in the tested six regions from FFI and G114V gCJD cases were fairly comparable, revealing similar signal intensities as that of normal control, whereas the intensities of Iba1 signals in sCJD Case 2 were significantly stronger (Figure 2B). Moreover, the Iba1 signals in three brain regions of sCJD (Case 1) were significantly stronger than that of normal control (Figure 2B). It indicates that the level of brain Iba1 increased in sCJD cases, but not in FFI and G114V gCJD cases.


Brain microglia were activated in sporadic CJD but almost unchanged in fatal familial insomnia and G114V genetic CJD.

Shi Q, Xie WL, Zhang B, Chen LN, Xu Y, Wang K, Ren K, Zhang XM, Chen C, Zhang J, Dong XP - Virol. J. (2013)

Evaluation of the brain Iba1 levels of different human prion diseases by Western blots. A. Temporal lobes. Same amounts of 10% homogenates from three FFI, one sCJD (Case 1), one G114V gCJD and normal control were separated onto 15% SDS-PAGE and blotted with Iba1 or β-actin specific mAbs. The quantitative analyses of the relative gray values are showed on the right. The results are calculated from three blots and presented as mean ± SD. Statistical differences are illustrated as **P<0.01. B. Different brain regions. Same amounts of 10% homogenates of six regions, including thalamus, cingulate gyrus, frontal lobe, parietal lobe, occipital lobe and temporal lobe, from three FFI and the G114V gCJD cases were comparatively assayed. Same amounts of 10% homogenates of temporal lobe from one sCJD (Case 2) and normal one were loaded as control. Same amounts of 10% homogenates of frontal lobe, parietal lobe and occipital lobe from sCJD (Case 1) were also comparatively assayed with that of normal control (low panel). Molecular markers are indicated on the right.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3716817&req=5

Figure 2: Evaluation of the brain Iba1 levels of different human prion diseases by Western blots. A. Temporal lobes. Same amounts of 10% homogenates from three FFI, one sCJD (Case 1), one G114V gCJD and normal control were separated onto 15% SDS-PAGE and blotted with Iba1 or β-actin specific mAbs. The quantitative analyses of the relative gray values are showed on the right. The results are calculated from three blots and presented as mean ± SD. Statistical differences are illustrated as **P<0.01. B. Different brain regions. Same amounts of 10% homogenates of six regions, including thalamus, cingulate gyrus, frontal lobe, parietal lobe, occipital lobe and temporal lobe, from three FFI and the G114V gCJD cases were comparatively assayed. Same amounts of 10% homogenates of temporal lobe from one sCJD (Case 2) and normal one were loaded as control. Same amounts of 10% homogenates of frontal lobe, parietal lobe and occipital lobe from sCJD (Case 1) were also comparatively assayed with that of normal control (low panel). Molecular markers are indicated on the right.
Mentions: To see the potential change in microglia, the levels of Iba1 that was commonly used as the marker of total microglia including resting and activated state in temporal lobes were comparatively evaluated by Western blots. Compared with that of normal control, the Iba1 signal in sCJD (Case 1) was remarkably stronger (P<0.01), while that in the FFI and G114V gCJD maintained almost unchanged (Figure 2A). To test the Iba1 levels in other brain regions, six different regions including thalamus, callosal gyrus, frontal lobe, parietal lobe, occipital lobe and temporal lobe from three FFI cases and a G114V gCJD case were subjected into Iba1-specific Western blots, together with temporal lobe from sCJD Case 2 and normal control. It showed that the Iba1 signals in the tested six regions from FFI and G114V gCJD cases were fairly comparable, revealing similar signal intensities as that of normal control, whereas the intensities of Iba1 signals in sCJD Case 2 were significantly stronger (Figure 2B). Moreover, the Iba1 signals in three brain regions of sCJD (Case 1) were significantly stronger than that of normal control (Figure 2B). It indicates that the level of brain Iba1 increased in sCJD cases, but not in FFI and G114V gCJD cases.

Bottom Line: However, the situation of microglia in other genetic prion diseases such as fatal familial insomnia (FFI) and familial CJD remains less understood.The Iba1-levels maintained comparable in six different brain regions of FFI and G114V cases, including thalamus, cingulate gyrus, frontal cortex, parietal cortex, occipital cortex and temporal cortex.ELISA tests for inflammatory cytokines revealed significantly up-regulated IL-1β, IL-6 and TNF-α in the brain homogenates from sCJD, but not in those from FFI and G114V gCJD.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases (Hangzhou), National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Chang-Bai Rd 155, Beijing 102206, People's Republic of China.

ABSTRACT

Background: Microglial activations have been described in different subtypes of human prion diseases such as sporadic Creutzfeldt-Jakob disease (CJD), variant CJD, Kuru and Gerstmann-Sträussler-Scheinker disease (GSS). However, the situation of microglia in other genetic prion diseases such as fatal familial insomnia (FFI) and familial CJD remains less understood. The brain microglia was evaluated comparatively between the FFI, G114V and sCJD cases in the study.

Methods: Specific Western blots, immunohistochemical and immunofluorescent assays were used to detect the changes of microglia and ELISA tests were used for levels of inflammatory cytokines.

Results: Western blots, immunohistochemical and immunofluorescent assays illustrated almost unchanged microglia in the temporal lobes of FFI and G114V gCJD, but obviously increased in those of sCJD. The Iba1-levels maintained comparable in six different brain regions of FFI and G114V cases, including thalamus, cingulate gyrus, frontal cortex, parietal cortex, occipital cortex and temporal cortex. ELISA tests for inflammatory cytokines revealed significantly up-regulated IL-1β, IL-6 and TNF-α in the brain homogenates from sCJD, but not in those from FFI and G114V gCJD.

Conclusion: Data here demonstrates silent brain microglia in FFI and G114V gCJD but obviously increased in sCJD, which reflects various pathogenesis of different human prion diseases subtypes.

Show MeSH
Related in: MedlinePlus