Type

Journal Article

Authors

Jochen H M Prehn
Niamh M C Connolly
Heiko Düssmann
Heinrich J Huber
Tytus Bernas
Jasmin Schmid
Petronela Weisova
Ujval Anilkumar

Subjects

Biochemistry

Topics
apoptosis culture media conditioned drug effects animals calcium mice neurons cell survival cells cultured algorithms metabolism necrosis cerebellum cytology

Defining external factors that determine neuronal survival, apoptosis and necrosis during excitotoxic injury using a high content screening imaging platform. (2017)

Abstract Cell death induced by excessive glutamate receptor overactivation, excitotoxicity, has been implicated in several acute and chronic neurological disorders. While numerous studies have demonstrated the contribution of biochemically and genetically activated cell death pathways in excitotoxic injury, the factors mediating passive, excitotoxic necrosis are less thoroughly investigated. To address this question, we developed a high content screening (HCS) based assay to collect high volumes of quantitative cellular imaging data and elucidated the effects of intrinsic and external factors on excitotoxic necrosis and apoptosis. The analysis workflow consisted of robust nuclei segmentation, tracking and a classification algorithm, which enabled automated analysis of large amounts of data to identify and quantify viable, apoptotic and necrotic neuronal populations. We show that mouse cerebellar granule neurons plated at low or high density underwent significantly increased necrosis compared to neurons seeded at medium density. Increased extracellular Ca2+ sensitized neurons to glutamate-induced excitotoxicity, but surprisingly potentiated cell death mainly through apoptosis. We also demonstrate that inhibition of various cell death signaling pathways (including inhibition of calpain, PARP and AMPK activation) primarily reduced excitotoxic apoptosis. Excitotoxic necrosis instead increased with low extracellular glucose availability. Our study is the first of its kind to establish and implement a HCS based assay to investigate the contribution of external and intrinsic factors to excitotoxic apoptosis and necrosis.
Collections Ireland -> Royal College of Surgeons in Ireland -> PubMed

Full list of authors on original publication

Jochen H M Prehn, Niamh M C Connolly, Heiko Düssmann, Heinrich J Huber, Tytus Bernas, Jasmin Schmid, Petronela Weisova, Ujval Anilkumar

Experts in our system

1
Jochen H M Prehn
Royal College of Surgeons in Ireland
Total Publications: 206
 
2
Niamh M C Connolly
Royal College of Surgeons in Ireland
Total Publications: 14
 
3
Heiko Düssmann
Royal College of Surgeons in Ireland
Total Publications: 45
 
4
Heinrich J Huber
Royal College of Surgeons in Ireland
Total Publications: 39
 
5
Tytus Bernas
Royal College of Surgeons in Ireland
Total Publications: 8
 
6
Jasmin Schmid
Royal College of Surgeons in Ireland
Total Publications: 13
 
7
Petronela Weisova
Royal College of Surgeons in Ireland
Total Publications: 10
 
8
Ujval Anilkumar
Royal College of Surgeons in Ireland
Total Publications: 14