Identification of novel cellular clusters define a specialized area in the cerebellar periventricular zone

Abstract

The periventricular zone of cerebellum is a germinative niche during the embryonic development, nevertheless its structural organization and functional implications in adult have not been widely studied. Here we disclose the presence of two novel clusters of cells in that area. The first one was named the subventricular cellular cluster (SVCC) and is composed of cells that express glial and neuronal markers. The second was named the ventromedial cord (VMC) and appears as a streak of biciliated cells with microvillosities facing the ventricle, that includes GFAP+ and nestin+ cells organized along the periventricular vasculature. The dorsal limit of the SVCC is associated with myelinated axons of neurons of unknown origin. This paper describes the characteristics and organization of these groups of cells. They can be observed from late embryonic development in the transgenic mouse line GFAP-GFP. The SVCC and VMC expand during early postnatal development but are restricted to the central area of the ventricle in adulthood. We did not find evidence of cell proliferation, cell migration or the presence of fenestrated blood vessels. These findings provide new insights into the knowledge of the cellular composition and structural organization of the periventricular zone of cerebellum.

Human brain transcriptome analysis finds region- and subject-specific expression signatures of GABAAR subunits

“…Noise-level expression of transcripts for ρ1, ρ2, and ρ3 across the brain in microarray datasets is consistent with studies reporting a limited and cell-specific expression for these genes in animal models 25,69–72 . Transcripts for π were at noise levels in microarray datasets and extremely low, if any, in RNA-Seq of the hippocampus and the temporal lobe.…”

Morphological and Calcium Signaling Alterations of Neuroglial Cells in Cerebellar Cortical Dysplasia Induced by Carmustine

“…This procedure confirmed the prevalence of cortical dysplasia at the roof of the fourth ventricle and revealed an increase in the size of the soma and an atrophy in the complexity of BG cells and astrocyte processes (Figure 3). The morphology of BG and astrocytes showed changes in their soma and processes, which were retracted and diffuse (Figure 3C,D), similar to that reported previously at postnatal age P5 [20], therefore, we measured different characteristics of these cells: BG showed significant differences for soma diameter (Ctrl 10.20 ± 0.68 µm, Carmustine 13.55 ± 0.72 µm) **, area (Ctrl 63.99 ± 7.95 µm, Carmustine 109.60 ± 9.33 µm) * and perimeter of the soma (Ctrl 26.14 ± 1.78 µm, Carmustine 39.20 ± 1.33 µm) **, as well as for process length (Ctrl 58.11 ± 2.56 µm, Carmustine 48.92 ± 3.09 µm) * and process protrusion length (Ctrl 3.88 ± 0.30 µm, Carmustine 2.24 ± 0.26 µm) **; while no significant difference was found in the number of processes (Ctrl 4.36 ± 0.43, Carmustine 3.45 ± 0.36) (1 to 3 BG cells were analyzed from each experiment, n = 6 for control mice and n = 6 for carmustine-treated mice). In astrocytes, the analysis showed that there is a significant difference for the soma diameter (Ctrl 16.42 ± 0.69 µm, Carmustine 23.48 ± 0.55 µm) **, area (Ctrl 142.65 ± 7.61 µm, Carmustine 253.79 ± 8.37 µm) ** and perimeter (Ctrl 45.76 ± 1.58 µm, Carmustine 58.95 ± 1.17 µm) **, as well as for the number (Ctrl 7.68 ± 0.26 µm, Carmustine 5.31 ± 0.14 µm) ** and length of the processes (Ctrl 61.04 ± 1.30 µm, Carmustine 33.29 ± 0.74 µm) ** (six to eight astrocyte cells were analyzed from each experiment, n = 6 for control mice and n = 6 for carmustine-treated mice).…”

“…The hydrogel solution was kept on ice and prepared by dissolving 40 mL of acrylamide (40%, Bio-Rad Laboratories, Hercules, CA, USA), 10 mL of bis-acrylamide (2%, Bio-Rad Laboratories), 1 g of initiator VA-044 (0.25%, Alpha Laboratories, Toronto, ON, Canada), 40 mL of 10X PBS, 100 mL of 16% PFA in 1 L of dH 2 O. After perfusion, brains were removed and kept in hydrogel solution for 3 days at 4 °C [ 20 ]. The tissue was placed in a 15 mL Falcon tube that was completely filled with fresh hydrogel solution at 37 °C in a water bath for 3 h to induce the polymerization of hydrogel.…”

“…The clarifying solution was prepared by dissolving 12.36 g of boric acid (Sigma Aldrich, Saint-Louis, MO, USA), 40 g of sodium dodecyl sulfate (Sigma Aldrich, Saint-Louis, MO, USA) and made up to 1 L with dH 2 O, pH 7.5. Then, the tissue was placed on a shaker at 100 RPM at 37 °C for passive clarification [ 20 ]. Once clarified, the sample was washed four times with PBST for 30 min at 37 °C on a shaker at 80 RPM.…”

Hydrogel-Tissue Chemistry: Principles and Applications

“…Passive CLARITY on sections of medial prefrontal cortex (mPFC) established the presence of nonoverlapping corticotropin-releasing factor and corticotropin-releasing factor receptor-1 circuits relevant to acute stress (138) and was used to map brain-wide viral expression in mice inoculated with western equine encephalitis virus in the foot pad (101). The distribution of microglia within the subventricular zone (a neurogenic region of the adult central nervous system) was mapped using passive CLARITY (38), and in the periventricular zone of the cerebellum, passive CLARITY was employed to analyze the organization of astrocytes during development (43). Passive CLARITY was used to show increased dendritic complexity in hippocampal pyramidal neurons of transgenic mice that exhibit enhanced learning (114) and to observe the localization of cells expressing neuromedin B, a bombesin-like neuropeptide that influences sighing behavior, around the facial nucleus, including the retrotrapezoid nucleus (a control center for breathing) (76).…”