DOI: 10.5061/dryad.7pvmcvf59
title: Data from: Neurodegenerative and neurodevelopmental roles for bulk lipid transporters VPS13A and BLTP2

description: Background: Bridge‐like lipid transfer proteins (BLTPs) mediate bulk lipid
 transport at membrane contact sites. Mutations in BLTPs are linked to both
 early‐onset neurodevelopmental and later‐onset neurodegenerative diseases,
 including movement disorders. The tissue specificity and temporal
 requirements of BLTPs in disease pathogenesis remain poorly understood.
 Objective: The objective of this study was to determine tissue‐specific
 and aging‐dependent roles for VPS13A and BLTP2 using Drosophila models.
 Methods: We generated tissue‐specific knockdowns of the VPS13A ortholog
 (Vps13) and the BLTP2 ortholog (hobbit) in neurons and muscles of
 Drosophila. We analyzed age‐dependent locomotor behavior,
 neurodegeneration, and synapse development and function. Results:
 Neuron‐specific loss of the VPS13A ortholog caused neurodegeneration
 followed by aging‐dependent movement deficits and reduced lifespan,
 whereas muscle‐specific loss affected only lifespan. In contrast, neuronal
 loss of the BLTP2 ortholog resulted in severe early‐onset locomotor
 defects without neurodegeneration, whereas muscle loss impaired
 synaptogenesis and neurotransmission at the neuromuscular junction.
 Conclusions: VPS13A maintains neuronal survival, whereas BLTP2
 orchestrates synaptic development. The phenotypic specificity of BLTP
 function provides mechanistic insights into distinct disease trajectories
 for BLTP‐associated disorders. 
description: <strong>Adult climbing assays</strong>
 For adult climbing, groups of <em>n</em>~10 flies
 were collected at 1-2 days old and allowed to age for the appropriate
 time. Then, each group of flies of the appropriate age and genotype was
 placed into a 50 mL graduated cylinder and allowed to acclimate for 3 min.
 All flies were then tapped to the bottom of the cylinder, and the number
 of flies climbing 8 cm in 12 s was counted. This procedure was repeated
 three times for each group of flies, and the average of the three trials
 is reported on the graphs. Three independent biological replicates with
 approximately equal numbers of male and female flies were analyzed for
 each genotype. Note that the same <em>elav&gt;luc</em>,
 <em>elav&gt;Vps13(i)</em>,
 <em>24B&gt;luc</em>, and
 <em>24B&gt;Vps13(i)</em> flies were used for climbing
 assays during aging and for lifespan analysis. During these experiments,
 flies were transferred to fresh food several times weekly, and the number
 of living flies was recorded prior to obtaining climbing data
 weekly. <strong>Histology and neurodegeneration
 index (NI) quantification</strong> Fly heads were
 severed and fixed overnight at 4<sup>o</sup>C in a 6:3:1
 solution of ethanol:chloroform:glacial acetic acid, then washed and stored
 in 70% ethanol for histology. Heads were processed into paraffin using
 standard histological procedures; serial 5 μm sections spanning the whole
 fly head were obtained and stained with hematoxylin and eosin
 (H&E) (UW-Madison Translational Research Initiatives in Pathology
 lab (TRIP)). Images were obtained using a BioTek Lionheart FX Automated
 Microscope (10x objective) with Gen5 v.3.11 software, then rotated and
 cropped post-acquisition in Adobe Photoshop CS6. NI were quantified using
 the criteria described in Loewen & Ganetzky, 2018. The number and
 location of vacuoles in the brain determined the NI: 0 = no vacuoles; 1 =
 a few, small vacuoles mainly in optic lobes in only a few sections; 2 =
 many vacuoles in many sections, mainly in optic lobes, but possibly some
 in central brain; 3 = vacuoles become prominent in central brain and are
 abundant in optic lobes; 4 = many vacuoles in central brain, and some
 vacuoles in optic lobes and central brain are large. NI scoring was
 performed blind to age and genotype. Loewen CA,
 Ganetzky B. Mito-nuclear interactions affecting lifespan and
 neurodegeneration in a drosophila model of leigh syndrome. Genetics. 2018
 Apr 1;208(4):1535–52. <strong>Body size and
 lethal phase analysis</strong> Pupa imaging and
 body size and lethal phase analysis were performed as previously described
 (Neuman & Bashirullah, 2018). Neuman SD,
 Bashirullah A. Hobbit regulates intracellular trafficking to drive
 insulin-dependent growth during Drosophila development. Development.
 2018;145(11):dev161356.
description: # Data from: Neurodegenerative and neurodevelopmental roles for bulk lipid
 transporters VPS13A and BLTP2 Dataset DOI:
 [10.5061/dryad.7pvmcvf59](10.5061/dryad.7pvmcvf59) ## Description of the
 data and file structure This file contains the raw data used to generate
 the figures published by the Bashirullah Lab, published in: Neuman SD,
 Thakur RS, Gratz SJ, O'Connor-Giles KM, Bashirullah A.
 Neurodegenerative and Neurodevelopmental Roles for Bulk Lipid Transporters
 VPS13A and BLTP2. Mov Disord. 2025 Mar 28. doi: 10.1002/mds.30178. PMID:
 40152532. This manuscript used *Drosophila* as a model system to examine
 tissue-specific functions for the bridge-like lipid transfer proteins
 (BLTPs) *VPS13A* and *BLTP2* (*Vps13* and *hobbit* in *Drosophila*,
 respectively). This dataset includes lethal phase data, climbing data,
 lifespan data, neurodegeneration index data, and body size/pupa volume
 data. Each file name contains a reference to the figure where the data is
 published, as well as a description of the type of data included in the
 file. ### Files and variables #### Figure1BLethalPhaseData.csv  This file
 contains the lethal phase data for control and *Vps13-RNAi* (*Vps13(i)*)
 knockdown in specific tissues: ubiquitous (*act-GAL4*), neuron-specific
 (*elav-GAL4*), and muscle-specific (*24 B-GAL4*). Note that “&gt;” in
 genotypes is shorthand for *GAL4, UAS* (*e.g., act&gt;Luc* is short for
 *act-GAL4 UAS-Luc*). Number of variables: 5 Number of header rows: 1
 Number of rows: 6 Variable list: * Genotype (alphanumeric): the genotype
 analyzed - Number arrested as prepupa (PP) (numeric): animals that
 arrested development as prepupa * Number arrested as pupa/incomplete pupa
 (P/iP) (numeric): animals that arrested development as pupa/incomplete
 pupa - Number arrested as pharate adults (PA) (numeric): animals that
 arrested development as pharate adults * Numberenclosedd as adult (A)
 (numeric): animals that survived to adulthood Data type: alphanumeric,
 numeric #### **Figure1DClimbingData.csv** This file contains the climbing
 data for 3-4-da4-day-oldld control (*elav&gt;Luc*) and neuron-specific
 *Vps13* knockdown (*elav&gt;Vps13(i)*) flies. Data was initially collected
 for males and females separately; however, because no sex-specific
 differences were observed, the data reported in the associated published
 figure bins both sexes together. Note that “&gt;” in genotypes is
 shorthand for *GAL4, UAS* (*e.g., elav&gt;Luc* is short for *elav-GAL4
 UAS-Luc*). Number of variables: 5 Number of header rows: 1 Number of rows:
 12 Variable list: * Genotype and replicate number (alphanumeric): the sex,
 genotype, and replicate number analyzed - Total n (numeric): Total number
 of flies analyzed for the listed genotype * Trial 1 (total number
 climbing) (numeric): number of flies successfully climbing for the listed
 genotype - Trial 2 (total number climbing) (numeric): number of flies
 successfully climbing for the listed genotype * Trial 3 (total number
 climbing) (numeric): number of flies successfully climbing for the listed
 genotype Data type: alphanumeric, numeric #### Figure1EClimbingData.csv
 This file contains the climbing data for 3-4-day 4-day-old control
 (*24B&gt;Luc*) and muscle-specific *Vps13* knockdown (*24B&gt;Vps13(i)*)
 flies. Data was initially collected for males and females separately;
 however, because no sex-specific differences were observed, the data
 reported in the associated published figure bins both sexes together. Note
 that “&gt;” in genotypes is shorthand for *GAL4, UAS* (*e.g., 24B&gt;Luc*
 is short for 24 B-GAL4* UAS-Luc*). Number of variables: 5 Number of header
 rows: 1 Number of rows: 12 Variable list: * Genotype and replicate number
 (alphanumeric): the sex, genotype, and replicate number analyzed - Total n
 (numeric): Total number of flies analyzed for the listed genotype * Trial
 1 (total number climbing) (numeric): number of flies successfully climbing
 for the listed genotype - Trial 2 (total number climbing) (numeric):
 number of flies successfully climbing for the listed genotype * Trial 3
 (total number climbing) (numeric): number of flies successfully climbing
 for the listed genotype Data type: alphanumeric, numeric ####
 Figure1FClimbingData.csv This file contains the climbing data for control
 (*elav&gt;Luc*) and neuron-specific *Vps13* knockdown (*elav&gt;Vps13(i)*)
 in aging flies. Data was initially collected for males and females
 separately; however, because no sex-specific differences were observed,
 the data reported in the associated published figure bins both sexes
 together. Note that “&gt;” in genotypes is shorthand for *GAL4, UAS*
 (*e.g., elav&gt;Luc* is short for *elav-GAL4 UAS-Luc*). Number of
 variables: 6 Number of header rows: 1 Number of rows: 72 Variable list: *
 Genotype and replicate number (alphanumeric): the sex, genotype, and
 replicate number analyzed - Age (alphanumeric): the age of the flies
 analyzed * Total n (numeric): Total number of flies analyzed for the
 listed genotype - Trial 1 (total number climbing) (numeric): number of
 flies successfully climbing for the listed genotype * Trial 2 (total
 number climbing) (numeric): number of flies successfully climbing for the
 listed genotype - Trial 3 (total number climbing) (numeric): number of
 flies successfully climbing for the listed genotype Data type:
 alphanumeric, numeric #### Figure1GClimbingData.csv This file contains the
 climbing data for control (*24B&gt;Luc*) and muscle-specific *Vps13*
 knockdown (*24B&gt;Vps13(i)*) flies during aging. Data was initially
 collected for males and females separately; however, because no
 sex-specific differences were observed, the data reported in the
 associated published figure bins both sexes together. Note that “&gt;” in
 genotypes is shorthand for *GAL4, UAS* (*e.g., 24B&gt;Luc* is short for24
 B-GAL4*4 UAS-Luc*). Number of variables: 6 Number of header rows: 1 Number
 of rows: 84 Variable list: * Genotype and replicate number (alphanumeric):
 the sex, genotype, and replicate number analyzed - Age (alphanumeric): the
 age of the flies analyzed * Total n (numeric): Total number of flies
 analyzed for the listed genotype - Trial 1 (total number climbing)
 (numeric): number of flies successfully climbing for the listed genotype *
 Trial 2 (total number climbing) (numeric): number of flies successfully
 climbing for the listed genotype - Trial 3 (total number climbing)
 (numeric): number of flies successfully climbing for the listed genotype
 Data type: alphanumeric, numeric ####
 Figure2DNeurodegenerationIndexData.csv This file contains the raw data for
 neurodegeneration index scoring in control (*elav&gt;Luc* or *24B&gt;Luc*)
 and *Vps13-RNAi* (*elav&gt;Vps13(i)* or *24B&gt;Vps13(i)*) brains during
 aging. Neurodegeneration indices are scored numerically from 0 to 4,
 depending on the abundance and size of vacuoles present in brain sections.
 Note that “&gt;” in genotypes is shorthand for *GAL4, UAS* (*e.g.,
 elav&gt;Luc* is short for *elav-GAL4 UAS-Luc*). Number of variables: 4
 Number of header rows: 1 Number of rows: 12 Variable list: * Genotype
 (alphanumeric): the genotype being analyzed - Age (alphanumeric): the age
 of the flies analyzed * Total n brains (numeric): number of brains
 analyzed for a given genotype/age - Neurodegeneration Index Scores
 (alphanumeric): individual neurodegeneration index numerical scores for
 each brain analyzed; each brain separated by a comma Data type:
 alphanumeric, numeric #### Figure2ELifespanData.csv This file contains the
 lifespan data for control (*elav&gt;Luc*) and neuron-specific *Vps13*
 knockdown (*elav&gt;Vps13(i)*) in aging flies. Data was initially
 collected for males and females separately; however, because no
 sex-specific differences were observed, the data reported in the
 associated published figure bins both sexes together. Note that “&gt;” in
 genotypes is shorthand for *GAL4, UAS* (*e.g., elav&gt;Luc* is short for
 *elav-GAL4 UAS-Luc*). Number of variables: 4 Number of header rows: 1
 Number of rows: 84 Variable list: * Genotype and replicate number
 (alphanumeric): the sex, genotype, and replicate number analyzed - Age
 (alphanumeric): the age of the flies analyzed * n alive (numeric): number
 of flies alive - n dead (numeric): number of flies dead Data type:
 alphanumeric, numeric #### Figure2FLifespanData.csv This file contains the
 lifespan data for control (*24B&gt;Luc*) and muscle-specific *Vps13*
 knockdown (*24B&gt;Vps13(i)*) in aging flies. Data was initially collected
 for males and females separately; however, because no sex-specific
 differences were observed, the data reported in the associated published
 figure bins both sexes together. Note that “&gt;” in genotypes is
 shorthand for *GAL4, UAS* (*e.g., 24B&gt;Luc* is short for 24 B-GAL4*L4
 UAS-Luc*). Number of variables: 4 Number of header rows: 1 Number of rows:
 108 Variable list: * Genotype and replicate number (alphanumeric): the
 sex, genotype, and replicate number analyzed - Age (alphanumeric): the age
 of the flies analyzed * n alive (numeric): number of flies alive - n dead
 (numeric): number of flies dead Data type: alphanumeric, numeric ####
 Figure3ALethalPhaseData.csv This file contains the lethal phase data for
 control and *hob-RNAi *(*hob(i)*) knockdown in specific tissues:
 ubiquitous (*act-GAL4*), neuron-specific (*elav-GAL4*), and
 muscle-specific (*24B-GAL4*). Note that “&gt;” in genotypes is shorthand
 for *GAL4, UAS* (*e.g., act&gt;Luc* is short for *act-GAL4 UAS-Luc*).
 Number of variables: 5 Number of header rows: 1 Number of rows: 6 Variable
 list: * Genotype (alphanumeric): the genotype analyzed - Number arrested
 as prepupa (PP) (numeric): animals that arrested development as prepupa *
 Number arrested as pupa/incomplete pupa (P/iP) (numeric): animals that
 arrested development as pupa/incomplete pupa - Number arrested as pharate
 adults (PA) (numeric): animals that arrested development as pharate adults
 * Numbenclosedsed as adult (A) (numeric): animals that survived to
 adulthood Data type: alphanumeric, numeric #### Figure3BClimbingData.csv
 This file contains the climbing data for 3-4-day 4-day-old control
 (*elav&gt;Luc*) and neuron-specific *RNAi* knockdown (*elav&gt;hob(i)*)
 flies. Data was initially collected for males and females separately;
 however, because no sex-specific differences were observed, the data
 reported in the associated published figure bins both sexes together. Note
 that “&gt;” in genotypes is shorthand for *GAL4, UAS* (*e.g., elav&gt;Luc*
 is short for *elav-GAL4 UAS-Luc*). Number of variables: 5 Number of header
 rows: 1 Number of rows: 12 Variable list: * Genotype and replicate number
 (alphanumeric): the sex, genotype, and replicate number analyzed - Total n
 (numeric): Total number of flies analyzed for the listed genotype * Trial
 1 (total number climbing) (numeric): number of flies successfully climbing
 for the listed genotype - Trial 2 (total number climbing) (numeric):
 number of flies successfully climbing for the listed genotype * Trial 3
 (total number climbing) (numeric): number of flies successfully climbing
 for the listed genotype Data type: alphanumeric, numeric ####
 Figure3ENeurodegenerationIndexData.csv This file contains the raw data for
 neurodegeneration index scoring in control (*elav&gt;Luc*) and *hob-RNAi*
 (*elav&gt;hob(i)*) brains during aging. Neurodegeneration indices are
 scored numerically from 0 to 4, depending on the abundance and size of
 vacuoles present in brain sections. Note that “&gt;” in genotypes is
 shorthand for *GAL4, UAS* (*e.g., elav&gt;Luc* is short for *elav-GAL4
 UAS-Luc*). Number of variables: 4 Number of header rows: 1 Number of rows:
 6 Variable list: * Genotype (alphanumeric): the genotype being analyzed -
 Age (alphanumeric): the age of the flies analyzed * Total n brains
 (numeric): number of brains analyzed for a given genotype/age -
 Neurodegeneration Index Scores (alphanumeric): individual
 neurodegeneration index numerical scores for each brain analyzed; each
 brain separated by a comma Data type: alphanumeric, numeric ####
 FigureS1CBodySizeQuantification.csv This file contains the raw data used
 to calculate body size/pupa volume in control (*w1118*), *hob* mutant
 (*hob2/hob3*), and rescue (*hob2/hob3, end hob-GFP*) animals. The length
 and width of each pupa (in mm) were measured in Adobe Photoshop CS6, and
 pupa volume was calculated using a published formula. Number of variables:
 3 Number of header rows: 2 Number of rows: 50 Variable list: * Genotype
 (alphanumeric): the genotype being analyzed - pupa width (mm): the
 measured width of each pupa * pupa length (mm): the measured length of
 each pupa Data type: alphanumeric, numeric ####
 FigureS1DLethalPhaseData.csv This file contains the lethal phase data for
 control (*w1118*), *hob* mutant (*hob2/hob3*), and rescue (*hob2/hob3, end
 hob-GFP*) animals. Number of variables: 5 Number of header rows: 1 Number
 of rows: 3 Variable list: * Genotype (alphanumeric): the genotype analyzed
 - Number arrested as prepupa (PP) (numeric): animals that arrested
 development as prepupa * Number arrested as pupa/incomplete pupa (P/iP)
 (numeric): animals that arrested development as pupa/incomplete pupa -
 Number arrested as pharate adults (PA) (numeric): animals that arrested
 development as pharate adults * Numbeencloseded as adult (A) (numeric):
 animals that survived to adulthood Data type: alphanumeric, numeric ##
 Code/software All data is provided in a .csv file format, which can be
 opened in any free text editing software.