An Automated Imaging-Based Screen for Genetic Modulators of ER Organisation in Cultured Human Cells

Hereditary spastic paraplegias (HSPs) are a heterogeneous group of mono-genetic inherited neurological disorders, whose primary manifestation is the disruption of the pyramidal system, observed as a progressive impaired gait and leg spasticity in patients. Despite the large list of genes linked to t...

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Bibliographic Details
Published in:Cells (Basel, Switzerland) Switzerland), 2024-04, Vol.13 (7), p.577
Main Authors: Garcia-Pardo, M Elena, Simpson, Jeremy C, O'Sullivan, Niamh C
Format: Article
Language:English
Subjects:
Analysis
automated image analysis
Automation
Care and treatment
Cell culture
Diagnosis
Endoplasmic reticulum
Genes
Genetic aspects
Health aspects
Hereditary spastic paraplegia
high-content imaging
Homeostasis
human cells
Invoices
Lipids
Morphogenesis
Morphology
Motor neurons
Mutation
Neurodegeneration
Neurological diseases
Neuromodulation
Paralysis, Spastic
Plasma
Proteins
siRNA
siRNA screen
Spasticity
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Summary:Hereditary spastic paraplegias (HSPs) are a heterogeneous group of mono-genetic inherited neurological disorders, whose primary manifestation is the disruption of the pyramidal system, observed as a progressive impaired gait and leg spasticity in patients. Despite the large list of genes linked to this group, which exceeds 80 loci, the number of cellular functions which the gene products engage is relatively limited, among which endoplasmic reticulum (ER) morphogenesis appears central. Mutations in genes encoding ER-shaping proteins are the most common cause of HSP, highlighting the importance of correct ER organisation for long motor neuron survival. However, a major bottleneck in the study of ER morphology is the current lack of quantitative methods, with most studies to date reporting, instead, on qualitative changes. Here, we describe and apply a quantitative image-based screen to identify genetic modifiers of ER organisation using a mammalian cell culture system. An analysis reveals significant quantitative changes in tubular ER and dense sheet ER organisation caused by the siRNA-mediated knockdown of HSP-causing genes and . This screen constitutes the first attempt to examine ER distribution in cells in an automated and high-content manner and to detect genes which impact ER organisation.
ISSN:2073-4409
2073-4409
DOI:10.3390/cells13070577