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Pathogenic LRRK2 control of primary cilia and Hedgehog signaling in neurons and astrocytes of mouse brain

Citation

Khan, Shahzad et al. (2021), Pathogenic LRRK2 control of primary cilia and Hedgehog signaling in neurons and astrocytes of mouse brain, Dryad, Dataset, https://doi.org/10.5061/dryad.76hdr7sxx

Abstract

Previously, we showed that cholinergic interneurons of the dorsal striatum lose cilia in mice harboring the Parkinson’s disease associated, kinase activating, R1441C LRRK2 mutation (Dhekne et al., 2018). Here we show that this phenotype is also seen in two mouse strains carrying the most common human G2019S LRRK2 mutation. Heterozygous loss of the PPM1H phosphatase that is specific for LRRK2-phosphorylated Rab GTPases (Berndsen et al., 2019) yields the same cilia loss phenotype, strongly supporting a connection between Rab GTPase phosphorylation and cilia loss. In addition, astrocytes throughout the striatum show a ciliation defect in LRRK2 and PPM1H-/+ mutant models. Hedgehog signaling requires cilia, and loss of cilia correlates here with a loss in induction of Hedgehog signaling as monitored by in situ hybridization of Gli1 transcripts. These data support a model in which LRRK2 and PPM1H mutant mice struggle to receive and respond to critical Hedgehog signals in the nigral-striatal pathway.

Usage Notes

Figure 1 zip file:

  • Figure 1AB - Images from 13-month WT and G2019S KI animals. G2019S KI Brains: #1-4: WT Brains: #5-8. Imaging sets including NeuN were used for graph generated in Figure 1 supplement 1C
  • Figure 1CD - Image from 10-month WT and G2019S BAC Tg animals. G2019S BAC Tg: 164, 186, 195, 161, 165. WT Brains: 173, 187, 198, 200, 185.  

Figure 1 Supplement 1 zip file:

  • Figure 1 Supplement 1AB - Images from 13 WT and G2019S KI animals. G2019S KI Brains: B1-B4 WT Brains: B5-B8.

Figure 2 zip file:

  • Figure 2AB - Images captured from 13-month WT and G2019S KI animals. G2019S KI Brains: #1-4: WT Brains: 5-8. 
  • Figure 2CD - Images captured from 10-month WT and G2019S BAC Tg animals. G2019S BAC Tg: 164, 186, 195, 161, 165. WT Brains: 173, 187, 198, 200, 185.  

Figure 3 zip file:

  • Figure 3ACD - Images from ChAT+ neurons in the dorsal striatum of MLi-2 Treated 10-week old R1441C mice. Brains 3, 5, 6, 11, and 13 are no drug controls. Brains 4, 7, 8, and 12 are MLi-2 treated.
  • Figure 3B - Images for ChAT+ and ChaT- striatal neurons. Brains 1, 2, 3, 4 are WT. Brains 5, 6, 7 are mutant.
  • Figure 3EFG - Images from Astrocytes in the dorsal striatum of MLi-2 Treated 10-week old R1441C mice. Brains 3, 5, 6, 11, and 13 are no drug controls. Brains 4, 7, 8, and 12 are MLi-2 treated.

Figure 4 zip file:

  • Figure 4 - 700 and 800 channel blots for 4A, 4B, and 4C.

Figure 5 zip file:

  • Figure 5AB - ChAT+ neuron images from WT and 2.5 month PPM1H homozygous KO animals, WT and 5.5 PPM1H heterozygous KO animals.  
  • Figure 5CD - Astrocyte images from 2.5 month old WT, PPM1H, PPM1H KO animals.
  • Figure 5 Supplement 1 - 700 and 800 channel blots.

Figure 6 zip file:

  • Figure 6A - Control Images used in Figure 6A. 
  • Figure 6B - Images from 10 month old WT and R1441C LRRK2 mice. WT brains are 7, 12, 22, and 34. RC brains are 9, 23, and 38. Images were also used for data generated in Figure 8E.

Figure 7 zip file:

  • Figure 7A - GFAP+ Astrocyte Images used for 7A, 7C, and 7E. WT brains are 7, 12, 22, and 34. RC brains are 9, 23, and 38.
  • Figure 7B - S100B Astrocyte Images used for 7B and 7D. WT brains are 7, 12, 22, and 34. RC brains are 9, 23, and 38.

Figure 8 zip file:

  • Figure 8ABCD - ChAT+ neuron images from the dorsal striatum of 5.5 month old WT and G2019S KI animals. 4 WT and 4 G2019S brains.

Figure 9 zip file:

  • Figure 9 - Images of immune-panned untreated and MLi-2 treated G2019S LRRK2 rat astrocytes.

Raw Figure TIF Files zip: - Raw Image Files used to generate Figures.

Data_Fig5suppl.xlsx: - Tables and data used to generate Figure 5 supplement 1 graph.

Data_Graphs_Fig4.xlsx: - Tables and data used to generate Figure 4B graphs.

Tables_and_Graphs_for_Figures.pzfx: - Tabular data and graphs used to generate graphs shown in all figures except Fig4. Fig4 data is uploaded separately as an excel file.

Funding

Aligning Science Across Parkinson’s, Award: ASAP-000463