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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

Quantification of cerebellar and midbrain microvessels. (A) —The dorsal view of the wild type (WT) and Lurcher (Lc) fixed brain contained the following parts: cerebral cortex (1), midbrain (2), cerebellum (3), pons and medulla (4); (B)—The preparations were processed into serial of equidistant randomly orientated sections (a1, a2, …ai …an) with constant step k; (C)—Sagittal section through the midbrain and cerebellum: nuclei (N), white matter (WM), granular (GL), and molecular layers (ML) of the cerebellum. The microscopic regions of interest (ROI) in the x–y plane were selected at a constant interval. This was performed for the cerebellum and midbrain separately. Only fields marked with the green dots were taken into account; (D–F)—One of the microscopic fields is shown as a stack with three 5 μm-thick optical sections in the z-axis that illustrates a disector volume probe. Vessel profiles (marked by dots) and valence of nodes (n) are marked with respective dots. Scale bars: (C) 500 μm; (D–F) 10 μm.
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Figure 1: Quantification of cerebellar and midbrain microvessels. (A) —The dorsal view of the wild type (WT) and Lurcher (Lc) fixed brain contained the following parts: cerebral cortex (1), midbrain (2), cerebellum (3), pons and medulla (4); (B)—The preparations were processed into serial of equidistant randomly orientated sections (a1, a2, …ai …an) with constant step k; (C)—Sagittal section through the midbrain and cerebellum: nuclei (N), white matter (WM), granular (GL), and molecular layers (ML) of the cerebellum. The microscopic regions of interest (ROI) in the x–y plane were selected at a constant interval. This was performed for the cerebellum and midbrain separately. Only fields marked with the green dots were taken into account; (D–F)—One of the microscopic fields is shown as a stack with three 5 μm-thick optical sections in the z-axis that illustrates a disector volume probe. Vessel profiles (marked by dots) and valence of nodes (n) are marked with respective dots. Scale bars: (C) 500 μm; (D–F) 10 μm.

Mentions: For imaging and further analysis, one brain section from each prepared glass (16 ± 2 sections per unit) was used. It was photo-documented in accordance with the systematic sampling rules (West et al., 1991; Burke et al., 2009; Boyce et al., 2010). In the first phase, a full section was imaged using a Plan N 2 × microscope objective with a numerical aperture of 0.06 to quantify the volume of the specified cerebellum layers and the midbrain (Figures 1B–C).


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)

Quantification of cerebellar and midbrain microvessels. (A) —The dorsal view of the wild type (WT) and Lurcher (Lc) fixed brain contained the following parts: cerebral cortex (1), midbrain (2), cerebellum (3), pons and medulla (4); (B)—The preparations were processed into serial of equidistant randomly orientated sections (a1, a2, …ai …an) with constant step k; (C)—Sagittal section through the midbrain and cerebellum: nuclei (N), white matter (WM), granular (GL), and molecular layers (ML) of the cerebellum. The microscopic regions of interest (ROI) in the x–y plane were selected at a constant interval. This was performed for the cerebellum and midbrain separately. Only fields marked with the green dots were taken into account; (D–F)—One of the microscopic fields is shown as a stack with three 5 μm-thick optical sections in the z-axis that illustrates a disector volume probe. Vessel profiles (marked by dots) and valence of nodes (n) are marked with respective dots. Scale bars: (C) 500 μm; (D–F) 10 μm.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 1: Quantification of cerebellar and midbrain microvessels. (A) —The dorsal view of the wild type (WT) and Lurcher (Lc) fixed brain contained the following parts: cerebral cortex (1), midbrain (2), cerebellum (3), pons and medulla (4); (B)—The preparations were processed into serial of equidistant randomly orientated sections (a1, a2, …ai …an) with constant step k; (C)—Sagittal section through the midbrain and cerebellum: nuclei (N), white matter (WM), granular (GL), and molecular layers (ML) of the cerebellum. The microscopic regions of interest (ROI) in the x–y plane were selected at a constant interval. This was performed for the cerebellum and midbrain separately. Only fields marked with the green dots were taken into account; (D–F)—One of the microscopic fields is shown as a stack with three 5 μm-thick optical sections in the z-axis that illustrates a disector volume probe. Vessel profiles (marked by dots) and valence of nodes (n) are marked with respective dots. Scale bars: (C) 500 μm; (D–F) 10 μm.
Mentions: For imaging and further analysis, one brain section from each prepared glass (16 ± 2 sections per unit) was used. It was photo-documented in accordance with the systematic sampling rules (West et al., 1991; Burke et al., 2009; Boyce et al., 2010). In the first phase, a full section was imaged using a Plan N 2 × microscope objective with a numerical aperture of 0.06 to quantify the volume of the specified cerebellum layers and the midbrain (Figures 1B–C).

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