Journal: bioRxiv

Article Title: Fan cells in layer 2 of lateral entorhinal cortex are critical for episodic-like memory

doi: 10.1101/543777

Figure Lengend Snippet: A) Quantification of cells positive for reelin and calbindin in LEC L2a (left) and L2b (right; n = 4 mice). Note that the majority of cells in LEC L2a and L2b are positive for reelin and calbindin, respectively, as reported by . Grey dots indicate percentage values calculated for each section of tissue. Error bars represnt SEM. B) Immunolabelling against reelin (green) and calbindin (red) in L2a and L2b of LEC. C) Quantification of cells back-labelled by the retrograde tracer which were positive for reelin and calbindin in LEC L2a (left) and L2b (right; n = 4 mice). Note that the majority of back-labelled cells are positive for reelin in both sub-layers. Grey dots indicate percentage values calculated for each section of tissue. Error bars represent SEM. D) Immunolabelling against reelin (green) and calbindin (red) overlaid with neurons that were back-labelled by the injection of the retrograde tracer FastBlue (FB) into the dentate gyrus (blue). White arrows indicate back-labelled neurons in LEC L2b which are positive for reelin. Scale bars represent 100 µm.

Article Snippet: Primary antibodies were mouse anti-reelin (MBL, 1:200, Catalogue #: D351-3) and rabbit anti-calbindin D-28K (SWANT, 1:2500, Catalog #: CB-38).

Techniques: Injection

Journal: bioRxiv

Article Title: Fan cells in layer 2 of lateral entorhinal cortex are critical for episodic-like memory

doi: 10.1101/543777

Figure Lengend Snippet: A) Schematic for targeting superficial LEC with AAV-FLEX-GFP and images of a horizontal brain section from a Sim1:Cre mouse showing GFP expression (green) counterstained with Neurotrace (violet). The dashed boxes in the upper left image indicate regions shown in the insets. Axonal labelling is found in the outer molecular layer of the dentate gyrus (upper right) and L2 of LEC (lower). Abbreviations: gl, granule cell layer; oml, outer molecular layer. The scale bar is 250 µm. B) Percentage of neurons that expressed the reporter gene that were in L2a (left, n = 4 mice, 87.4 ± 1.6%, 1282/1490 cells, 14 sections) and L2b (right, 12.6 ± 1.6%, 207/1490 cells) of the LEC. Grey dots indicate percentage values calculated for each section of tissue. Error bars represent SEM. C) Examples of AAV-FLEX-GFP labelled cells which are positive for reelin (red, L2a: 98.3 ± 0.4%, 1260/1282 cells; L2b: 52.0 ± 5.7%, 97/207 cells), but not calbindin (purple, L2a: 0.1 ± 0.1%, 1/1282 cells; L2b: 7.1 ± 2.8%, 17/207 cells). Scale bars represent 100 μm. D) Proportion of neurons expressing the reporter gene (GFP) that were labelled by staining with antibodies against reelin and calbindin. Grey dots indicate percentage values calculated for each section of tissue. Error bars represent SEM. E) Cells labelled by injection of retrograde tracer (blue) overlap with cells labelled by injections of AAV-FLEX-GFP. Scale bar represents 100 μm. Schematic shows the injection strategy used to target superficial LEC with AAV-FLEX-GFP (green) and the dorsal dentate gyrus with a retrograde tracer (Fastblue, blue). F) Proportion of neurons expressing the reporter gene (GFP) that were back-labelled by the injection of retrograde tracer into the dentate gyrus (74.3 ± 4.4%, 331/441 cells, 8 sections). Grey dots indicate percentage values calculated for each tissue section. Error bar represents SEM.

Article Snippet: Primary antibodies were mouse anti-reelin (MBL, 1:200, Catalogue #: D351-3) and rabbit anti-calbindin D-28K (SWANT, 1:2500, Catalog #: CB-38).

Techniques: Expressing, Staining, Injection

Journal: bioRxiv

Article Title: Fan cells in layer 2 of lateral entorhinal cortex are critical for episodic-like memory

doi: 10.1101/543777

Figure Lengend Snippet: Cells triple-labelled by reelin (red), calbindin (purple) and the reporter gene (GFP, green) are indicated by white arrows. A small population of cells was triple-labelled in L2a (top, 0.3 ± 0.2%, 4/1282 cells) and L2b (bottom, 4.6 ± 1.9%, 13/207 cells). Scale bars represent 100 µm.

Article Snippet: Primary antibodies were mouse anti-reelin (MBL, 1:200, Catalogue #: D351-3) and rabbit anti-calbindin D-28K (SWANT, 1:2500, Catalog #: CB-38).

Techniques: