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Smaller Absolute Quantities but Greater Relative Densities of Microvessels Are Associated with Cerebellar Degeneration in Lurcher Mice.

Kolinko Y, Cendelin J, Kralickova M, Tonar Z - Front Neuroanat (2016)

Bottom Line: The greatest number and length of vessels were found in the granular layer; the number of vessels was lower in the molecular layer, and the lowest number of vessels was observed in the cerebellar nuclei corresponding with their low volume.Nevertheless, the nuclei had the greatest density of blood vessels.The complete primary morphometric data, in the form of continuous variables, is included as a supplement.

View Article: PubMed Central - PubMed

Affiliation: Biomedical Center, Faculty of Medicine in Pilsen, Charles University in Prague Pilsen, Czech Republic.

ABSTRACT
Degenerative affections of nerve tissues are often accompanied by changes of vascularization. In this regard, not much is known about hereditary cerebellar degeneration. In this study, we compared the vascularity of the individual cerebellar components and the mesencephalon of 3-month-old wild type mice (n = 5) and Lurcher mutant mice, which represent a model of hereditary olivocerebellar degeneration (n = 5). Paraformaldehyde-fixed brains were processed into 18-μm thick serial sections with random orientation. Microvessels were visualized using polyclonal rabbit anti-laminin antibodies. Then, the stacks comprised of three 5-μm thick optical sections were recorded using systematic uniform random sampling. Stereological assessment was conducted based on photo-documentation. We found that each of the cerebellar components has its own features of vascularity. The greatest number and length of vessels were found in the granular layer; the number of vessels was lower in the molecular layer, and the lowest number of vessels was observed in the cerebellar nuclei corresponding with their low volume. Nevertheless, the nuclei had the greatest density of blood vessels. The reduction of cerebellum volume in the Lurcher mice was accompanied by a reduction in vascularization in the individual cerebellar components, mainly in the cortex. Moreover, despite the lower density of microvessels in the Lurcher mice compared with the wild type mice, the relative density of microvessels in the cerebellar cortex and nuclei was greater in Lurcher mice. The complete primary morphometric data, in the form of continuous variables, is included as a supplement. Mapping of the cerebellar and midbrain microvessels has explanatory potential for studies using mouse models of neurodegeneration.

No MeSH data available.


Related in: MedlinePlus

Histological comparison of microcirculation in the separated cerebellar layers and the midbrain in the wild type (A–C) and Lurcher (D–F) mice. (A,D)—The complete absence of Purkinje cells (red arrow) with intensive degeneration of the molecular (ML) and granular (GL) layers in Lurcher mice; (B,E)—Morphological features of vascularity in the white matter (WM) and nuclei (N). The boundary between the molecular and granular layers marked by the purple dotted lines; between the granular layer and white matter by the green and between the white matter and the nuclei by the blue dotted lines. (C,D)—No significant difference in the midbrain vascularity. Immunohistochemical detection of the microvessel laminin outlines, visualization with horseradish peroxidase/diaminobenzidine (brown), counterstaining with haematoxylin. Scale bar: 50 μm, uniform magnification in (A–F).
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Figure 4: Histological comparison of microcirculation in the separated cerebellar layers and the midbrain in the wild type (A–C) and Lurcher (D–F) mice. (A,D)—The complete absence of Purkinje cells (red arrow) with intensive degeneration of the molecular (ML) and granular (GL) layers in Lurcher mice; (B,E)—Morphological features of vascularity in the white matter (WM) and nuclei (N). The boundary between the molecular and granular layers marked by the purple dotted lines; between the granular layer and white matter by the green and between the white matter and the nuclei by the blue dotted lines. (C,D)—No significant difference in the midbrain vascularity. Immunohistochemical detection of the microvessel laminin outlines, visualization with horseradish peroxidase/diaminobenzidine (brown), counterstaining with haematoxylin. Scale bar: 50 μm, uniform magnification in (A–F).

Mentions: The total volume of the Lc cerebellum was reduced more than three times compared with WT mice. The most apparent reductions in volume were observed in the cortex and were uniform throughout the separated layers of the cortex. The volume of white matter was also reduced, but not as drastically as the cortex layers. On the other hand, the cerebellar nuclei were the only cerebellar structures that were not significantly reduced in Lc (Figures 2A, 4). The Lc nuclei also showed significantly lower mean vessel length than WT, while this parameter remained unchanged in the rest of the cerebellum compared with WT (Figure 3B). The CE of the total volume of the cerebellum was 0.041 ± 0.002%.


Smaller Absolute Quantities but Greater Relative Densities of Microvessels Are Associated with Cerebellar Degeneration in Lurcher Mice.

Kolinko Y, Cendelin J, Kralickova M, Tonar Z - Front Neuroanat (2016)

Histological comparison of microcirculation in the separated cerebellar layers and the midbrain in the wild type (A–C) and Lurcher (D–F) mice. (A,D)—The complete absence of Purkinje cells (red arrow) with intensive degeneration of the molecular (ML) and granular (GL) layers in Lurcher mice; (B,E)—Morphological features of vascularity in the white matter (WM) and nuclei (N). The boundary between the molecular and granular layers marked by the purple dotted lines; between the granular layer and white matter by the green and between the white matter and the nuclei by the blue dotted lines. (C,D)—No significant difference in the midbrain vascularity. Immunohistochemical detection of the microvessel laminin outlines, visualization with horseradish peroxidase/diaminobenzidine (brown), counterstaining with haematoxylin. Scale bar: 50 μm, uniform magnification in (A–F).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4835681&req=5

Figure 4: Histological comparison of microcirculation in the separated cerebellar layers and the midbrain in the wild type (A–C) and Lurcher (D–F) mice. (A,D)—The complete absence of Purkinje cells (red arrow) with intensive degeneration of the molecular (ML) and granular (GL) layers in Lurcher mice; (B,E)—Morphological features of vascularity in the white matter (WM) and nuclei (N). The boundary between the molecular and granular layers marked by the purple dotted lines; between the granular layer and white matter by the green and between the white matter and the nuclei by the blue dotted lines. (C,D)—No significant difference in the midbrain vascularity. Immunohistochemical detection of the microvessel laminin outlines, visualization with horseradish peroxidase/diaminobenzidine (brown), counterstaining with haematoxylin. Scale bar: 50 μm, uniform magnification in (A–F).
Mentions: The total volume of the Lc cerebellum was reduced more than three times compared with WT mice. The most apparent reductions in volume were observed in the cortex and were uniform throughout the separated layers of the cortex. The volume of white matter was also reduced, but not as drastically as the cortex layers. On the other hand, the cerebellar nuclei were the only cerebellar structures that were not significantly reduced in Lc (Figures 2A, 4). The Lc nuclei also showed significantly lower mean vessel length than WT, while this parameter remained unchanged in the rest of the cerebellum compared with WT (Figure 3B). The CE of the total volume of the cerebellum was 0.041 ± 0.002%.

Bottom Line: The greatest number and length of vessels were found in the granular layer; the number of vessels was lower in the molecular layer, and the lowest number of vessels was observed in the cerebellar nuclei corresponding with their low volume.Nevertheless, the nuclei had the greatest density of blood vessels.The complete primary morphometric data, in the form of continuous variables, is included as a supplement.

View Article: PubMed Central - PubMed

Affiliation: Biomedical Center, Faculty of Medicine in Pilsen, Charles University in Prague Pilsen, Czech Republic.

ABSTRACT
Degenerative affections of nerve tissues are often accompanied by changes of vascularization. In this regard, not much is known about hereditary cerebellar degeneration. In this study, we compared the vascularity of the individual cerebellar components and the mesencephalon of 3-month-old wild type mice (n = 5) and Lurcher mutant mice, which represent a model of hereditary olivocerebellar degeneration (n = 5). Paraformaldehyde-fixed brains were processed into 18-μm thick serial sections with random orientation. Microvessels were visualized using polyclonal rabbit anti-laminin antibodies. Then, the stacks comprised of three 5-μm thick optical sections were recorded using systematic uniform random sampling. Stereological assessment was conducted based on photo-documentation. We found that each of the cerebellar components has its own features of vascularity. The greatest number and length of vessels were found in the granular layer; the number of vessels was lower in the molecular layer, and the lowest number of vessels was observed in the cerebellar nuclei corresponding with their low volume. Nevertheless, the nuclei had the greatest density of blood vessels. The reduction of cerebellum volume in the Lurcher mice was accompanied by a reduction in vascularization in the individual cerebellar components, mainly in the cortex. Moreover, despite the lower density of microvessels in the Lurcher mice compared with the wild type mice, the relative density of microvessels in the cerebellar cortex and nuclei was greater in Lurcher mice. The complete primary morphometric data, in the form of continuous variables, is included as a supplement. Mapping of the cerebellar and midbrain microvessels has explanatory potential for studies using mouse models of neurodegeneration.

No MeSH data available.


Related in: MedlinePlus