Survival assay of transiently transfected dopaminergic neurons

Journal of Neuroscience Methods
Li-Ying Yu, Urmas Arumae

Abstract

Death pathways in the apoptotic neurons are mostly studied by manipulating the levels of apoptosis-related proteins and counting the survival/death of affected neurons. Such assays are, however, technically complicated. We developed a transfection-survival assay for cultured embryonic dopaminergic (DA) neurons induced to die by deprivation of glial cell line-derived neurotrophic factor (GDNF). The calcium phosphate co-precipitation technique was used to transfect DA neurons. Microisland cultures and co-transfected enhanced green fluorescent protein allowed direct counting of transfected neurons from the same cultures at the beginning and the end of GDNF deprivation, whereas post hoc subtraction of tyrosine hydroxylase-negative neurons allowed exclusion of transfected non-DA neurons. Overexpression of dominant-negative mutant of caspase-6 significantly blocked the death of GDNF-deprived DA neurons. Thus, we have found a tool not only to transfect the neurons dissociated from midbrain, but also to analyze the apoptotic proteins particularly in DA neurons.

References

Jan 1, 1991·Annual Review of Neuroscience·R W Oppenheim
Jun 1, 1989·Neuron·Y A Barde
Jan 1, 1993·Annual Review of Neuroscience·E M Johnson, T L Deckwerth
Jul 1, 1996·Journal of Neuroscience Methods·T TakeshimaJ W Commissiong
Jul 25, 2000·The Journal of Comparative Neurology·V Jackson-LewisS Przedborski
May 17, 2001·Proceedings of the National Academy of Sciences of the United States of America·K SawamotoH Okano
Jun 23, 2001·BMJ : British Medical Journal·R M Gibson
Apr 12, 2002·Movement Disorders : Official Journal of the Movement Disorder Society·Hagai Bergman, Günther Deuschl
May 4, 2002·Nature Reviews. Neuroscience·Matti S Airaksinen, Mart Saarma
Jan 14, 2003·Annals of Medicine·Marja Jäättelä
May 3, 2003·Nature Reviews. Neuroscience·Miquel Vila, Serge Przedborski
Nov 26, 2003·Cellular Signalling·Andrew Thorburn
Dec 6, 2003·The Journal of Cell Biology·Li-Ying YuUrmas Arumae
Jan 24, 2004·Apoptosis : an International Journal on Programmed Cell Death·P Mehlen, D E Bredesen
Jan 28, 2004·Cell·Nika N Danial, Stanley J Korsmeyer
Aug 25, 2004·The International Journal of Biochemistry & Cell Biology·Richard A Lockshin, Zahra Zakeri
Jul 15, 2005·Cell Death and Differentiation·D E BredesenS Rabizadeh
Nov 30, 2005·Journal of Internal Medicine·B Fadeel, S Orrenius
May 24, 2006·Neurology·Peter Jenner, C Warren Olanow
Sep 8, 2006·Advances in Experimental Medicine and Biology·Kerstin Krieglstein
Oct 20, 2006·Nature·Dale E BredesenPatrick Mehlen
Jan 16, 2007·Trends in Pharmacological Sciences·Maxim M Bespalov, Mart Saarma
Mar 23, 2007·Nature Reviews. Molecular Cell Biology·Stefan J Riedl, Guy S Salvesen

❮ Previous
Next ❯

Citations

Mar 30, 2010·Journal of Neural Transmission·David B O'SullivanAideen M Sullivan
Jan 10, 2013·PloS One·Daniel Hernandez-BaltazarLouis-Eric Trudeau
Sep 15, 2020·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Prasannakumar DeshpandeEleanor T Coffey
Sep 21, 2018·Frontiers in Cell and Developmental Biology·Valentina BožokUrmas Arumäe
Oct 31, 2020·Molecular Pharmaceutics·Francisco E Lopez-SalasDaniel Martinez-Fong

❮ Previous
Next ❯

Related Concepts

Related Feeds

Apoptotic Caspases

Apoptotic caspases belong to the protease enzyme family and are known to play an essential role in inflammation and programmed cell death. Here is the latest research.

Astrocytes in Parkinson Disease

Parkinson's disease (PD) is a neurodegenerative disorder caused by the progressive loss of dopaminergic neurons. Some PD-genes may be associated with astrocyte dysfunction. Discover the latest research on astrocytes in Parkinson's disease here.

Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis

Amygdala and Midbrain Dopamine

The midbrain dopamine system is widely studied for its involvement in emotional and motivational behavior. Some of these neurons receive information from the amygdala and project throughout the cortex. When the circuit and transmission of dopamine is disrupted symptoms may present. Here is the latest research on the amygdala and midbrain dopamine.