Journal: Frontiers in Neuroanatomy

Article Title: Multiple Origins of Secretagogin Expressing Cortical GABAergic Neuron Precursors in the Early Human Fetal Telencephalon

doi: 10.3389/fnana.2020.00061

Figure Lengend Snippet: Primary antibodies used in this study.

Article Snippet: Calretinin , Recombinant rat calretinin , Mouse Mncl , 1/500 , Merk Millipore , AB_94259.

Techniques: Recombinant, Purification

Journal: Frontiers in Neuroanatomy

Article Title: Multiple Origins of Secretagogin Expressing Cortical GABAergic Neuron Precursors in the Early Human Fetal Telencephalon

doi: 10.3389/fnana.2020.00061

Figure Lengend Snippet: Secretagogin (SCGN) expression 6.5–8 post-conceptional weeks (PCW). (A) Rostral coronal section of 6.5 PCW brain; intense SCGN expression in ganglionic eminences (GE) preoptic area (POA) basal telencephalon and hypothalamus (HTh) and (B) Caudal thalamus (Th) and several locations in the hypothalamus. (C) Horizontal section of 7.5 PCW brain; SCGN immunoreactivity in the medial ganglionic eminence (MGE), lateral ganglionic eminence (LGE), caudal ganglionic eminence (CGE), septum (SEP), and POA. (D) A high magnification view of the boxed area in (C) shows many SCGN+ cells that appeared to enter MGE from POA. (E) Double labeling for SCGN and specificity protein 8 (SP8) in LGE showing most SCGN+ cells were double-labeled with SP8, although there is a population of SP8+ only cells in the dorsal most LGE. (F) Double labeling for SCGN and calretinin (CalR) in LGE showing only a proportion of SCGN+ cells co-expressed CalR. (G–I) The rostral coronal section at 8 PCW showing intense SCGN expression in LGE, septum, and rostral migratory stream (RMS, G ). Few SCGN+ cells enter cortex from LGE (H) . Cells from LGE and septum appeared to be mainly migrating into RMS at this stage (I) . Scale bars: 1 mm in (C) (and for A , B ) and (G) , 50 μm in (D,F) (and for E ), and (I) (and for H ).

Article Snippet: Calretinin , Recombinant rat calretinin , Mouse Mncl , 1/500 , Merk Millipore , AB_94259.

Techniques: Expressing, Labeling

Journal: Cell reports

Article Title: Inappropriate Intrusion of an Axonal Mitochondrial Anchor into Dendrites Causes Neurodegeneration

doi: 10.1016/j.celrep.2019.09.012

Figure Lengend Snippet: (A–F) Immunohistochemistry technique for capturing SNPH intrusion in PC dendrites. Shown is SNPH (green), Syt2 (red), and Calbindin (blue) labeling in 3.5-month-old WT (A, C, and E) and Shi (B, D, and F) mice. Scale bar, 10 μm. (G and H) High magnification of the maximum intensity projection image from the z stack through dendritic regions of WT (G) and Shi (H). (I and J) Orthogonal (slice) view of SNPH punctum (indicated by arrows in panels G and H) in the dendritic region from WT (I) and Shi (J) in x-z and y-z orientations, respectively. (K) Quantification of percentage area occupied by SNPH within the dendritic volume from 3 mice of each group. Data are shown as mean ± SEM. *p < 0.05. (L–N) Capturing SNPH intrusion by pre-tagging dendritic mitochondria in vivo using viral transduction. (L) Technique to selectively transduce PCs with AAV-Mito-mCherry. (M) Demonstration of successful pre-tagging of mitochondria in dendrites of a single PC by Calbindin staining. Scale bar, 10 μm. (N) Demonstration of how pre-tagged dendritic mitochondria in PCs are used to capture SNPH intrusion by 3D rotation. (O and P) Single dendritic tree in WT (O) or Shi (P) pre-tagged with Mito-mCherry (red) and SNPH intrusion (green) captured by co-rotation with Mito-mCherry. Merged images show the fraction of dendritic mitochondria anchored by intruded SNPH (yellow). (Q and R) Percentage of PCs with SNPH intrusions (Q) and percentage of SNPH bound to mitochondria per dendritic tree (R) from WT (n = 73) and Shi (n = 106) PCs. Data are shown as mean ± SEM. ***p < 0.005.

Article Snippet: Sections were then incubated with primary antibodies against SNPH (1:250; Abcam), Synaptotagmin2 (1:250; Developmental Studies Hybridoma Bank) and Calbindin (1:250; BosterBio) overnight in PBS containing 0.3% Triton X-100 and 5% normal goat serum.

Techniques: Immunohistochemistry, Labeling, In Vivo, Transduction, Staining

Journal: Cell reports

Article Title: Inappropriate Intrusion of an Axonal Mitochondrial Anchor into Dendrites Causes Neurodegeneration

doi: 10.1016/j.celrep.2019.09.012

Figure Lengend Snippet: (A and B) Representative images of lentivirally transduced GFP-SNPH (1–469) (A) and GFP-SNPH (B) in PCs of SNPH-KO mice injected with saline (no harmaline, vehicle only). (C-H) Effect of harmaline on GFP-SNPH (1–469)-transduced (C) and GFP-SNPH-transduced (F) PC dendrites. Degenerating dendrites in GFP-SNPH-transduced PCs can be seen in (F). Also shown is Calbindin labeling of GFP SNPH (1–469) (D) and GFP-SNPH (G) from (C) and (F). Merged images of GFP SNPH (1–469) and GFP-SNPH with Calbindin are shown in (E) and (H), respectively. (I–K) Representative image of a harmaline-induced degenerating PC (white arrow in I) transduced with GFP-SNPH. Calbindin staining from the same section is shown in (J), whereas a merged image is shown in (K). Scale bars, 20 μm. (L) Quantification of dendritic shrinkage in GFP-SNPH (1–469)- and GFP-SNPH-transduced PCs in the absence (n = 3 mice, vehicle only) or presence of harmaline (n = 5 mice). Data are shown as mean ± SEM. ***p < 0.001.

Article Snippet: Sections were then incubated with primary antibodies against SNPH (1:250; Abcam), Synaptotagmin2 (1:250; Developmental Studies Hybridoma Bank) and Calbindin (1:250; BosterBio) overnight in PBS containing 0.3% Triton X-100 and 5% normal goat serum.

Techniques: Injection, Saline, Labeling, Transduction, Staining

Journal: Cell reports

Article Title: Inappropriate Intrusion of an Axonal Mitochondrial Anchor into Dendrites Causes Neurodegeneration

doi: 10.1016/j.celrep.2019.09.012

Figure Lengend Snippet:

Article Snippet: Sections were then incubated with primary antibodies against SNPH (1:250; Abcam), Synaptotagmin2 (1:250; Developmental Studies Hybridoma Bank) and Calbindin (1:250; BosterBio) overnight in PBS containing 0.3% Triton X-100 and 5% normal goat serum.

Techniques: Virus, Plasmid Preparation, Recombinant, Software, Imaging

Journal: bioRxiv

Article Title: YAP is involved in replenishment of granule cell progenitors following injury to the neonatal cerebellum

doi: 10.1101/558742

Figure Lengend Snippet: (A) Schematic showing experimental design. (B-E) Representative images from lobule 4/5 showing IF staining for TUNEL, SOX2, and DAPI on midsagittal sections from IR and Non-IR Nes-mYap cKOs and controls one day after IR at P1. Grey shadow highlights the PCL of the lobule. Arrows indicate TUNEL+ particles in the PCL. Scale bar, 100 µm. (F) Graph of the densities of TUNEL+ particles in the PCL of the midline CB from IR and Non-IR Nes-mYap cKOs (Non-IR, n = 3; IR, n = 3) and controls (Non-IR, n = 3; IR, n = 3) at P2. (G-J) Representative images from lobule 4/5 showing IF staining of GFP and DAPI on midsagittal CB sections from IR and Non-IR Nes-m controls and Nes-mYap cKOs at P8. Grey shadow highlights the EGL of the lobule. Scale bar, 100 µm. (K-L) Graphs of the numbers of GFP+ cells normalized to total area measured in the EGL (K) and the percentages of GFP+ cells in the EGL among the total number of GFP+ cells (L) from Nes-m controls (Non-IR, n = 3; IR, n = 3) and Nes-mYap cKOs (Non-IR, n = 4; IR, n = 3) at P8. Data are presented as mean ± S.D., and statistical analysis by two-way ANOVA. Each data point represents one animal.

Article Snippet: Cryosections were stained overnight at 4 °C with the following primary antibodies: mouse anti-YAP (Abcam, AB56701), rabbit anti-TAZ (Santa Cruz, sc-48805), rat anti-GFP (1:1,000; Nacalai Tesque; 0440484), mouse anti-NeuN (Millipore, MAB377), rabbit anti-Calbindin D-28K (Swant, CB38), rabbit anti-GFAP (Dako, Z0334), rabbit anti-S100β (Dako, Z0311), rabbit anti-PAX2 (Invitrogen, 71600), and goat anti-SOX2 (R&D System, AF2018).

Techniques: Staining, TUNEL Assay