Research Article: The origin of Rosenthal fibers and their contributions to astrocyte pathology in Alexander disease

Date Published: March 31, 2017

Publisher: BioMed Central

Author(s): Alexander A. Sosunov, Guy M. McKhann, James E. Goldman.

http://doi.org/10.1186/s40478-017-0425-9

Abstract

Rosenthal fibers (RFs) are cytoplasmic, proteinaceous aggregates. They are the pathognomonic feature of the astrocyte pathology in Alexander Disease (AxD), a neurodegenerative disorder caused by heterozygous mutations in the GFAP gene, encoding glial fibrillary acidic protein (GFAP). Although RFs have been known for many years their origin and significance remain elusive issues. We have used mouse models of AxD based on the overexpression of human GFAP (transgenic, TG) and a point mutation in mouse GFAP (knock-in, KI) to examine the formation of RFs and to find astrocyte changes that correlate with the appearance of RFs. We found RFs of various sizes and shapes. The smallest ones appear as granular depositions on intermediate filaments. These contain GFAP and the small heat shock protein, alphaB-crystallin. Their aggregation appears to give rise to large RFs. The appearance of new RFs and the growth of previously formed RFs occur over time. We determined that DAPI is a reliable marker of RFs and in parallel with Fluoro-Jade B (FJB) staining defined a high variability in the appearance of RFs, even in neighboring astrocytes. Although many astrocytes in AxD with increased levels of GFAP and with or without RFs change their phenotype, only some cells with large numbers of RFs show a profound reconstruction of cellular processes, with a loss of fine distal processes and the appearance of large, lobulated nuclei, likely due to arrested mitosis. We conclude that 1) RFs appear to originate as small, osmiophilic masses containing both GFAP and alphaB-crystallin deposited on bundles of intermediate filaments. 2) RFs continue to form within AxD astrocytes over time. 3) DAPI is a reliable marker for RFs and can be used with immunolabeling. 4) RFs appear to interfere with the successful completion of astrocyte mitosis and cell division.

Partial Text

Rosenthal fibers (RFs) are distinctly characteristic features of the astrocyte pathology in Alexander Disease (AxD), a neurodegenerative disorder caused by heterozygous mutations in the gene encoding glial fibrillary acidic protein (GFAP), the major intermediate filament in astrocytes [15, 21]. RFs are protein aggregates in the cytoplasm of astrocytes. They were originally identified by inspection with the light microscope as bright, eosinophilic profiles inside astrocytes. Ultrastructurally, RFs appear as amorphous, electron dense material, surrounded by skeins of intermediate filaments, and are not membrane bound [1, 10, 19, 23]. RFs are composed mainly of intermediate filaments, GFAP [36], vimentin, and synemin [20] in addition to other proteins including the small heat shock proteins alphaB-crystallin [36] and Hsp 27 [5], the filament binding protein plectin [35], and Cyclin D2 [9].

In this study we examined four different strains of AxD mutant mice at three different ages to observe stages of RF formation. At all times and in all lines of the mice we found a high degree of variability in the sizes and shapes of RFs. The appearance of RFs ranged from large, dense structures to small, less dense structures that appeared to be deposited on intermediate filaments. We also found that DAPI was an excellent marker for RFs, allowing us to view the distributions of RFs.

RFs appear to originate as small, osmiophilic masses containing both GFAP and alphaB-crystallin deposited on bundles of intermediate filaments.RFs continue to form within AxD astrocytes over time.DAPI is a reliable marker for RFs and can be used with immunolabeling.RFs and bundles of filaments appear to interfere with the successful completion of astrocyte mitosis.

 

Source:

http://doi.org/10.1186/s40478-017-0425-9