TY - JOUR
T1 - Drosophila melanogaster
T2 - A new model to study cisplatin-induced neurotoxicity
AU - Podratz, Jewel L.
AU - Staff, Nathan P.
AU - Froemel, Dara
AU - Wallner, Anna
AU - Wabnig, Florian
AU - Bieber, Allan J.
AU - Tang, Amy
AU - Windebank, Anthony J.
N1 - Funding Information:
This work is supported by NIH-NS40471 . We would like to thank Jane Meyer for her administrative role in manuscript preparation as well as Maria Caruso and Scott Gamb for their excellent technical assistance with transmission electron microscopy of the Drosophila brain.
PY - 2011/8
Y1 - 2011/8
N2 - Platinum-based compounds are widely used and effective chemotherapeutic agents; however, sensory peripheral neuropathy is a dose-limiting and long term side effect for 20-30% of patients. A critical question is whether the mechanisms of cell death underlying clinical efficacy can be separated from the effects on neurons in order to develop strategies that prevent platinum-induced neuropathy. In rodent dorsal root ganglion neurons (DRG), cisplatin has been shown to bind and damage neuronal DNA, inducing apoptosis; however genetic manipulation in order to study mechanisms of this phenomenon in the rodent model system is costly and time-consuming. Drosophila melanogaster are commonly used to study neurological disorders, have DNA damage-apoptosis mechanisms homologous to mammalian systems, and have readily-available, inexpensive tools for rapid genetic manipulation. We therefore sought to develop adult Drosophila as a new model to study cisplatin-induced neurotoxicity. Adult Drosophila were exposed to 10, 25, 50, 100, 200 and 400μg/ml cisplatin for 3. days and observed for fly survival and geotactic climbing behavior, cisplatin-DNA binding and cellular apoptosis. On day 3, 50μg/ml cisplatin reduced the number of flies able to climb above 2. cm to 43% while fly survival was maintained at 92%. 100% lethality was observed at 400μg/ml cisplatin. Whole fly platinum-genomic DNA adducts were measured and found to be comparable to adduct levels previously measured in rat DRG neurons. Brain, ovaries, kidney and heart harvested from cisplatin treated flies were stained for active caspase 3. Apoptosis was found in ovaries and brain but not in heart and kidney. Brain apoptosis was confirmed by transmission electron microscopy. Expression of the anti-apoptotic baculoviral protein, p35, in neurons using the GAL4-UAS system prevented cisplatin-induced apoptosis in the brain and restored climbing behavior. In conclusion, cisplatin-induced behavioral and apoptotic changes in Drosophila resemble those seen in mammals. Furthermore, the use of lethality and climbing assays combined with powerful gene manipulation, make Drosophila a suitable model to study mechanisms of cisplatin neurotoxicity.
AB - Platinum-based compounds are widely used and effective chemotherapeutic agents; however, sensory peripheral neuropathy is a dose-limiting and long term side effect for 20-30% of patients. A critical question is whether the mechanisms of cell death underlying clinical efficacy can be separated from the effects on neurons in order to develop strategies that prevent platinum-induced neuropathy. In rodent dorsal root ganglion neurons (DRG), cisplatin has been shown to bind and damage neuronal DNA, inducing apoptosis; however genetic manipulation in order to study mechanisms of this phenomenon in the rodent model system is costly and time-consuming. Drosophila melanogaster are commonly used to study neurological disorders, have DNA damage-apoptosis mechanisms homologous to mammalian systems, and have readily-available, inexpensive tools for rapid genetic manipulation. We therefore sought to develop adult Drosophila as a new model to study cisplatin-induced neurotoxicity. Adult Drosophila were exposed to 10, 25, 50, 100, 200 and 400μg/ml cisplatin for 3. days and observed for fly survival and geotactic climbing behavior, cisplatin-DNA binding and cellular apoptosis. On day 3, 50μg/ml cisplatin reduced the number of flies able to climb above 2. cm to 43% while fly survival was maintained at 92%. 100% lethality was observed at 400μg/ml cisplatin. Whole fly platinum-genomic DNA adducts were measured and found to be comparable to adduct levels previously measured in rat DRG neurons. Brain, ovaries, kidney and heart harvested from cisplatin treated flies were stained for active caspase 3. Apoptosis was found in ovaries and brain but not in heart and kidney. Brain apoptosis was confirmed by transmission electron microscopy. Expression of the anti-apoptotic baculoviral protein, p35, in neurons using the GAL4-UAS system prevented cisplatin-induced apoptosis in the brain and restored climbing behavior. In conclusion, cisplatin-induced behavioral and apoptotic changes in Drosophila resemble those seen in mammals. Furthermore, the use of lethality and climbing assays combined with powerful gene manipulation, make Drosophila a suitable model to study mechanisms of cisplatin neurotoxicity.
KW - Apoptosis
KW - Behavior
KW - Chemotherapy
KW - Cisplatin
KW - Drosophila
KW - Genetics
KW - Model
KW - Neuropathy
KW - Neuroprotection
KW - Neurotoxicity
UR - http://www.scopus.com/inward/record.url?scp=79958222116&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79958222116&partnerID=8YFLogxK
U2 - 10.1016/j.nbd.2011.03.022
DO - 10.1016/j.nbd.2011.03.022
M3 - Article
C2 - 21514385
AN - SCOPUS:79958222116
SN - 0969-9961
VL - 43
SP - 330
EP - 337
JO - Neurobiology of Disease
JF - Neurobiology of Disease
IS - 2
ER -