Journal: EBioMedicine

Article Title: Potent and Selective BACE-1 Peptide Inhibitors Lower Brain Aβ Levels Mediated by Brain Shuttle Transport

doi: 10.1016/j.ebiom.2017.09.004

Figure Lengend Snippet: Aβ inhibition by BACE1 peptide inhibitors and small molecule β- and γ-secretase inhibitors in vitro . (a) Human stem-cell derived neurons were cultured for six weeks and neuronal identity verified by staining for markers like Map2 (green) and HuC/D (white). The level of astrocytes in fully differentiated cells was verified with anti-GFAP (red) labeling. As reference, cell nuclei are stained with DAPI (blue). (b) Aβ production inhibition in human stem cell-derived neurons. Fully differentiated cells were incubated with the different compounds for 48 h at doses ranging from 10 to 0.001 μM, decreasing in half-logarithmic steps. Experimental setup included cells from three independent differentiations and two technical replicates each dose (0.128–10,000 nM; solid colors), and cells from one differentiation and two technical replicates each dose (0.001024–0.0256 nM; shaded colors). Both small molecule and BACE1 peptide inhibitors blocked Aβ production to a comparable extend, SM GSI (black bars), SM BACE1 (red bars), Pep#16 BACE1 (blue bars), Pep#15 BACE1 palm (green bars) and Pep#14 BACE1 chol (orange bars). All % values represent Aβ40 level changes in treated cells relative to respective vehicle-treated cells (means ± SEM). n.d. signal below detection limit.

Article Snippet: The neuronal identity of the culture was verified by immunostaining for GFAP (Dako, Z033401), HuC/D (Invitrogen, A21271), and anti-Map2 (Neuromics, CH22103).

Techniques: Inhibition, In Vitro, Derivative Assay, Cell Culture, Staining, Labeling, Incubation

Journal: Frontiers in Cellular Neuroscience

Article Title: Optogenetic Control of Neural Circuits in the Mongolian Gerbil

doi: 10.3389/fncel.2018.00111

Figure Lengend Snippet: AAV8(Y733F) drives neuron-specific EYFP expression in auditory nuclei. (A 1 –C 1 ) Low-magnification images of EYFP fluorescence (green) showing correct targeting of the control vector AAV8(Y733F).hSyn.EYFP to the IC (A 1 ) , DNLL (B 1 ) , and MNTB (C 1 ) . The pictures show scans of fixed coronal brain sections taken with the virtualSlide epi-fluorescence microscope. The white frames mark the transduction sites shown enlarged in (A 2 –C 2 ) , respectively. (A 2 –C 2 ) Maximum projection of a confocal z-stack taken at 20× optical magnification from the IC (A 2 ) , DNLL (B 2 ) , and MNTB (C 2 ) . The white frames mark the areas shown at higher magnification in (A 3 –C 3 ) , respectively. (A 3 –C 3 ) Further magnification (63× objective) reveals neuron-specific expression in IC (A 3 ) , DNLL (B 3 ) , and MNTB (C 3 ) . All EYFP-expressing cells are co-labeled with neuronal markers (Map2, red or NeuN, cyan) as seen in the overlay (top left) and the indicated single channels. Note that not all neurons in the field of view are transduced. m, medial; v, ventral.

Article Snippet: Standard immunohistochemistry procedures were used to stain free-floating slices with primary antibodies for NeuN (Millipore, MAB377), S100B (SWANT, 37A), and MAP2 (Neuromics, CH22103).

Techniques: Expressing, Fluorescence, Control, Plasmid Preparation, Microscopy, Transduction, Labeling