Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2016 Mar 8:10:86.
doi: 10.3389/fnins.2016.00086. eCollection 2016.

Corpora Amylacea of Brain Tissue from Neurodegenerative Diseases Are Stained with Specific Antifungal Antibodies

Diana Pisa  1 , Ruth Alonso  1 , Alberto Rábano  2 , Luis Carrasco  1
Affiliations

Corpora Amylacea of Brain Tissue from Neurodegenerative Diseases Are Stained with Specific Antifungal Antibodies

Diana Pisa et al. Front Neurosci. .

Abstract

The origin and potential function of corpora amylacea (CA) remains largely unknown. Low numbers of CA are detected in the aging brain of normal individuals but they are abundant in the central nervous system of patients with neurodegenerative diseases. In the present study, we show that CA from patients diagnosed with Alzheimer's disease (AD) contain fungal proteins as detected by immunohistochemistry analyses. Accordingly, CA were labeled with different anti-fungal antibodies at the external surface, whereas the central portion composed of calcium salts contain less proteins. Detection of fungal proteins was achieved using a number of antibodies raised against different fungal species, which indicated cross-reactivity between the fungal proteins present in CA and the antibodies employed. Importantly, these antibodies do not immunoreact with cellular proteins. Additionally, CNS samples from patients diagnosed with amyotrophic lateral sclerosis (ALS) and Parkinson's disease (PD) also contained CA that were immunoreactive with a range of antifungal antibodies. However, CA were less abundant in ALS or PD patients as compared to CNS samples from AD. By contrast, CA from brain tissue of control subjects were almost devoid of fungal immunoreactivity. These observations are consistent with the concept that CA associate with fungal infections and may contribute to the elucidation of the origin of CA.

Keywords: Alzheimer's disease; amyotrophic lateral sclerosis; corpora amylacea; fungal infection; neurodegenerative disease.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Corpora amylacea are detected in different regions of the CNS. Tissue sections (5 μm) from different CNS regions of patient AD1 were tested as follows: lateral frontal cortex (LFC), cerebellar cortex (CEC) and entorhinal cortex/hippocampus (ERH). Immunohistochemistry analysis was carried out by double immunofluorescence staining employing a rabbit polyclonal anti-C. glabrata antibody (green) and a mouse monoclonal anti-tau antibody (red). Sections were mounted and observed by confocal microscopy after incubation with the corresponding secondary antibodies, as described in Materials and Methods. Overlapping red and green pixels appear as orange/yellow. DAPI appears in blue. Scale bar: 10 μm.
Figure 2
Figure 2
Immunoreactivity of corpora amylacea against different antifungal antibodies. Tissue sections analyzed from patient AD1 are indicated in the figure. Immunohistochemistry analysis was carried out using a monoclonal antibody to human neurofilaments (red) and rabbit polyclonal antibodies raised against the following fungi: C. famata, C. albicans. S. racemosum, and P. betae (green). Sections were mounted and examined by confocal microscopy after incubation with the corresponding secondary antibodies. Orange/yellow corresponds to red and green pixels. DAPI (blue) and scale bar: 10 μm.
Figure 3
Figure 3
Detection of fungal proteins in corpora amylacea from different AD patients. ERH sections from 10 AD patients (AD2-AD11) were incubated with rabbit polyclonal antibodies against C. albicans and P. betae (green) and a monoclonal antibody against α-tubulin (red). Sections were mounted and examined by confocal microscopy after incubation with the corresponding secondary antibodies. DAPI and scale bar as in Figure 1.
Figure 4
Figure 4
Fungal proteins in corpora amylacea from ALS patients. Tissue sections of patient ALS1 were obtained from the following regions: primary motor cortex (MC), medulla (MD) and different levels of the spinal cord (SC1, SC2, and SC3) (upper panels). Different regions of the CNS from five additional ALS patients (ALS2–ALS6) were also analyzed (lower panels). Sections were incubated with rabbit polyclonal antibodies against C. albicans and P. betae (green) and a monoclonal antibody against α-tubulin (red). Sections were mounted and examined by confocal microscopy after incubation with the corresponding secondary antibodies. DAPI and scale bar as in Figure 1.
Figure 5
Figure 5
Detection of fungal proteins in corpora amylacea from PD patients. Tissue sections of patient PD1 were obtained from the following regions: pons (PN), mesencephalon (MSP), hypothalamus (HT), callosal body (CB), and caudate and lenticular nuclei (CLN) (upper panels). Different regions of the CNS from five additional PD patients (PD2-PD6) were also analyzed (lower panels). Sections were incubated with rabbit polyclonal antibodies against C. albicans and P. betae (green) and a monoclonal antibody against α-tubulin (red). Sections were mounted and examined by confocal microscopy after incubation with the corresponding secondary antibodies. Overlapping red and green pixels appear as orange/yellow. DAPI and scale bar as in Figure 1.
Figure 6
Figure 6
Analysis of corpora amylacea present in the CNS of control individuals. Tissue sections from the ERH of five control individuals (C1–C5) were analyzed by immunohistochemistry. Sections were incubated with rabbit polyclonal antibodies against C. albicans and P. betae (green) and a monoclonal antibody against α-tubulin (red). Sections were mounted and examined by confocal microscopy after incubation with the corresponding secondary antibodies. Overlapping red and green pixels appear as orange/yellow. Note the near absence of activity against the fungal antibodies. DAPI and scale bar as in Figure 1.
Figure 7
Figure 7
Quantitation of the number and positiveness of CA from different patients and control subjects. Brain sections from the patients indicated in the Figure were examined by confocal microscopy and the total number of CA in three different fields was estimated, as well as their staining with anti-P. betae antibodies. Wide fields were analyzed using 40x magnification. The sections examined were ERC from AD patients and control subjects, MD from ALS patients, PD3 and PD5 and MC from PD4. Red bar: total number of CA in the three fields; blue bar: positive CA with antifungal antibody.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Alonso R., Pisa D., Marina A. I., Morato E., Rabano A., Carrasco L. (2014a). Fungal infection in patients with Alzheimer's disease. J. Alzheimers. Dis. 41, 301–311. 10.3233/JAD-132681 - DOI - PubMed
    1. Alonso R., Pisa D., Marina A. I., Morato E., Rabano A., Rodal I., et al. . (2015). Evidence for fungal infection in cerebrospinal fluid and brain tissue from patients with amyotrophic lateral sclerosis. Int. J. Biol. Sci. 11, 546–558. 10.7150/ijbs.11084 - DOI - PMC - PubMed
    1. Alonso R., Pisa D., Rabano A., Carrasco L. (2014b). Alzheimer's disease and disseminated mycoses. Eur. J. Clin. Microbiol. Infect. Dis. 33, 1125–1132. 10.1007/s10096-013-2045-z - DOI - PubMed
    1. Badea P., Petrescu A., Moldovan L., Zarnescu O. (2015). Structural heterogenity of intraluminal content of the prostate: a histochemical and ultrastructural study. Microsc. Microanal. 21, 368–376. 10.1017/S1431927615000197 - DOI - PubMed
    1. Botez G., Rami A. (2001). Immunoreactivity for Bcl-2 and C-Jun/AP1 in hippocampal corpora amylacea after ischaemia in humans. Neuropathol. Appl. Neurobiol. 27, 474–480. 10.1046/j.1365-2990.2001.00362.x - DOI - PubMed