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. 2016 Sep 28;6:34370. doi: 10.1038/srep34370

Figure 1. Depletion of primary cilia from mature dentate granule cells.

Figure 1

(a) Schematic diagram of the AAV injection in the dentate gyrus. (b) Representative confocal images of the hippocampus of IFT20 fl/fl mice transduced with AAV-CaMKII-eGFP or AAV-CaMKII-eGFP-Cre virus at 28 days post AAV labeling. Scale bar: 200 μm. (c) Representative confocal images of the granule cell layer of AAV-CaMKII-eGFP or AAV-CaMKII-eGFP-Cre 28 days post injection. Immunostaining with anti DCX (red) and ACIII (white) reveals that primary cilia were selectively ablated in mature DGCs by AAV-CaMKII-eGFP-Cre transduction, whereas no ciliary disruption was found by AAV-CaMKII-eGFP transduction. Scale bars: 30 μm (d) AAV-CaMKII-eGFP-Cre transduced cells exhibited a significantly depressed rate of ACIII expression versus AAV-CaMKII-eGFP controls. (Cre: 11.7 ± 2.1% ACIII+, eGFP: 82.6 ± 7.4% ACIII+; two-tailed un-paired t-test p = 8.5 × 10−7). (e) Cells in the granule cell layer that did not take up virus expressed primary cilia at comparable rates between eGFP control and Cre groups (CTRL: 82.6 ± 7.0%; Cre: 77.3 ± 7.2%; two-tailed unpaired t-test p = 0.634; n = 3,3). (f) Cre transduction resulted in significant gross depletion of primary cilia. (Averaged number of ACIII positive cells per field in CTRL and IFT20(−/−)mDGCs: 47.23 ± 2.78 for CTRL and 1.8 ± 0.28 for IFT20(−/−)mDGCs; p = 5.985 × 10−15; n = 4,4). (g) Expression of ACIII in DCX+ cells did not differ significantly between eGFP-Cre and eGFP controls (CTRL: 9.5 ± 3.6% co-localization; IFT20(−/−)mDGCs : 12.8 ± 2.9% co-localization; two-tailed unpaired t-test p = 0.50). Fields size: 320 μm × 320 μm × 36 μm. ***p < 0.001; ****p < 0.0001; n is the number of animals.