Upregulation of <i>App</i>, <i>Adam10</i> and <i>Adam17</i> mRNAs in the dentate gyrus at 7 days after entorhinal denervation.

<p>Expression levels of <i>App, Aplp1</i> and <i>Aplp2</i> mRNAs were determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) in the denervated outer molecular layer (A) and in the granule cell layer (B). Only <i>App</i> mRNA was found to be upregulated in the granule cell layer. Similarly, mRNA expression levels of <i>Adam10</i>, <i>Adam17</i> and <i>Bace1</i> were determined by RT-qPCR in the denervated outer molecular layer (C) and in the granule cell layer (D). Significant upregulation of <i>Adam10</i> and <i>Adam17</i> mRNAs were seen in the denervated outer molecular layer (C). In addition, <i>Adam10</i> mRNA was found to be increased in the granule cell layer (D). (n = 4–6 animals each; t-test (two-tailed), * = p≤0.05).</p

TaqMan assays used in the present study.

<p>TaqMan assays used in the present study.</p

Entorhino-hippocampal denervation model.

<p>(A) Schematic of a horizontal brain section illustrating the entorhino-hippocampal denervation model. The perforant path (green) originates from stellate neurons in the entorhinal cortex (EC) and terminates on distal dendritic segments of granule cells (gray) in the outer molecular layer (oml) of the dentate gyrus (DG). The transection site of the perforant path is indicated by a dashed line. (B, C) Layer-specific denervation of the dentate oml following unilateral transection of the perforant path. Fluoro-Jade C staining of the hippocampus contralateral (B) and ipsilateral (C) to the lesion side reveals degenerating axons in denervated areas of the dentate molecular layer at 7 days post lesion. Arrowheads point to the border between the denervated oml and non-denervated inner molecular layer (iml). Nuclei were counterstained with Hoechst 33342. gcl: granule cell layer; h: hilar region. Scale bar: 200 µm.</p

Reactive astrocytes upregulate ADAM10 in the dentate gyrus 7 days after entorhinal denervation.

<p>ADAM10 immunofluorescence of the hippocampus contralateral (A, C) and ipsilateral (B, D) to the lesion side revealed an upregulation of ADAM10 in the denervated outer molecular layer (oml) of the dentate gyrus. Arrowheads (in B) point to the border between the denervated oml and the non-denervated inner molecular layer (iml). Confocal analysis of sections double-stained for ADAM10 (E, F; red), the astrocytic marker GFAP (E; in yellow/green) and the microglia marker IBA1 (F; green) revealed that increased ADAM10 immunofluorescence (D–F, arrows) is associated with reactive astrocytes (E, arrows), but not with activated microglia (F, stars) following entorhinal denervation. Nuclei were counterstained with DRAQ5. h: hilar region. Scale bars: (A) 200 µm; (C) 25 µm.</p

Microarray analysis of candidate genes in the dentate outer molecular layer and granule cell layer after entorhinal denervation.

<p>Microarray analysis of candidate genes in the dentate outer molecular layer and granule cell layer after entorhinal denervation.</p

Additional file 1: of A general homeostatic principle following lesion induced dendritic remodeling

Supplementary Material. (PDF 580 kb

Additional file 1: of Rewiring neuronal microcircuits of the brain via spine head protrusions-a role for synaptopodin and intracellular calcium stores

SI Materials and Methods. (DOC 71 kb

Electrically evoked asynchronous EPSCs confirmed layer-specific increase in excitatory synaptic strength.

<p>(<b>A</b>) Asynchronous EPSCs (aEPSC) were recorded from identified dentate granule cells in control and denervated cultures upon local electrical stimulation of the OML and IML (recordings performed in Sr²⁺-containing bath solution; red, pipette containing the stimulating electrode and Alexa568; green, patch-clamp electrode containing Alexa488). Scale bar: 40 µm. (<b>B</b>) An increase in mean aEPSC amplitude was observed in response to electrical stimulations of the OML but not in response to electrical stimulations of the IML of denervated dentate granule cells (n = 4 cultures each; 1–3 neurons per culture).</p

Additional file 1: of Rewiring neuronal microcircuits of the brain via spine head protrusions-a role for synaptopodin and intracellular calcium stores

SI Materials and Methods. (DOC 71 kb

Additional file 2: of Rewiring neuronal microcircuits of the brain via spine head protrusions-a role for synaptopodin and intracellular calcium stores

SI Figure Legend. Figure S1 CA1 neurons in slices cultures from synaptopodin-knockout (SP-KO) mice have comparable morphological and functional properties to wild type (WT) slices. a, Examples of CA1 dendrites rendered in 3D from WT and SP-KO slices. Scale bar, 2 μm. b, Quantification of spine densities, lengths and volumes from WT and SP-KO slices, expressed as percent of WT values. For spine densities, WT, n = 23 branches, 599 μm of dendrite from 22 slices; SP-KO, n = 23 branches, 645 μm of dendrite from 18 slices were analyzed. For spine lengths and volumes, WT, n = 1,021 spines from 22 slices; SP-KO, n = 1,121 spines from 18 slices were studied. c, Cumulative probability distributions of spine lengths (left) and spine volumes (right) in WT and SP-KO slice cultures. d, Example traces of mEPSCs recorded from CA1 pyramidal cells in WT (left) and SP-KO (right) slice cultures. e, Quantification of mean mEPSC amplitude (left; WT, 13.9 ± 0.38 pA and SP-KO, 13.6 ± 0.67 pA) and mean inter-mEPSC interval (right; WT, 228.37 ± 25.97 ms; SP-KO, 328.06 ± 49.14 ms). WT, n = 17 cells from 10 cultures; SP-KO, n = 15 cells from 8 cultures. f, Cumulative probability distributions of mEPSC amplitudes (left) and inter-mEPSC intervals (right) from WT and SP-KO CA1 neurons. (TIF 360 kb