DOI: 10.5061/dryad.76hdr7sxx
title: Pathogenic LRRK2 control of primary cilia and Hedgehog signaling in neurons and astrocytes of mouse brain

description: 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.
description: <b>Figure 1 zip
 file:</b> <b>Figure
 1AB</b> - 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<b> Figure 1 supplement 1C</b>. 
 <b>Figure 1CD</b> - 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.   
 <strong>Figure 1 Supplement 1 zip
 file:</strong> 
 <b>Figure 1 Supplement 1AB </b>- Images from 13 WT and G2019S
 KI animals. G2019S KI Brains: B1-B4 WT Brains: B5-B8.
 <strong>Figure 2 zip
 file:</strong> 
 <b>Figure 2AB</b> - Images captured from 13-month WT and
 G2019S KI animals. G2019S KI Brains: #1-4: WT Brains: 5-8. 
 <b>Figure 2CD</b> - 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.   
 <strong>Figure 3 zip
 file:</strong> 
 <b>Figure 3ACD</b> - 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. <b>Figure 3B</b> - Images for
 ChAT+ and ChaT- striatal neurons. Brains 1, 2, 3, 4 are WT. Brains 5, 6, 7
 are mutant. <b>Figure 3EFG</b> - 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. 
 <strong>Figure 4 zip
 file:</strong> 
 <b>Figure 4</b> - 700 and 800 channel blots for 4A, 4B, and
 4C. <b>Figure 5 zip
 file:</b> <b>Figure
 5AB</b> - ChAT+ neuron images from WT and 2.5 month PPM1H homozygous
 KO animals, WT and 5.5 PPM1H heterozygous KO animals.  
 <b>Figure 5CD</b> - Astrocyte images from 2.5 month
 old WT, PPM1H, PPM1H KO animals. <b>Figure 5
 Supplement 1</b> - 700 and 800 channel blots. 
 <strong>Figure 6 zip
 file:</strong> 
 <b>Figure 6A</b> - Control Images used in Figure
 6A.  <b>Figure 6B</b> - 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. <strong>Figure 7
 zip file:</strong> 
 <b>Figure 7A - GFAP+ Astrocyte Images used for 7A, 7C, and 7E.
 </b>WT brains are 7, 12, 22, and 34. RC brains are 9, 23, and
 38. <b>Figure 7B - S100B Astrocyte Images used
 for 7B and 7D. </b>WT brains are 7, 12, 22, and 34. RC brains are 9,
 23, and 38. <strong>Figure
 8 zip file:</strong> 
 <b>Figure 8ABCD</b> - ChAT+ neuron images from the dorsal
 striatum of 5.5 month old WT and G2019S KI animals. 4 WT and 4 G2019S
 brains. <strong>Figure 9
 zip file:</strong> 
 <b>Figure 9</b> - Images of immune-panned untreated and MLi-2
 treated G2019S LRRK2 rat astrocytes. 
 <strong>Raw Figure TIF Files
 zip:</strong> - Raw Image Files used to generate
 Figures.
 <strong>Data_Fig5suppl.xlsx</strong>: - Tables and data used to generate Figure 5 supplement 1 graph. <strong>Data_Graphs_Fig4.xlsx:</strong> - Tables and data used to generate Figure 4B graphs. <strong>Tables_and_Graphs_for_Figures.pzfx: </strong>- Tabular data and graphs used to generate graphs shown in all figures except Fig4. Fig4 data is uploaded separately as an excel file.