Source Paper
Blocking Soluble Tumor Necrosis Factor Signaling with Dominant-Negative Tumor Necrosis Factor Inhibitor Attenuates Loss of Dopaminergic Neurons in Models of Parkinson's Disease
Melissa K. McCoy, Terina N. Martinez, Kelly A. Ruhn, et al.
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Protocol Steps
Start here. The step list is optimized for running the experiment, with direct vendor links available inline when you need to source a cited item.
Striatal 6-OHDA injection
Inject 6-hydroxydopamine into the striatum to induce retrograde nigral degeneration
Note: This is the primary neurotoxic insult to model Parkinson's disease
View evidence from paper
“retrograde nigral degeneration induced by striatal injection of the oxidative neurotoxin 6-hydroxydopamine (6-OHDA)”
Nigral XENP345 infusion
Infuse XENP345 into the substantia nigra to neutralize soluble TNF
Note: XENP345 was neuroprotective only when infused into the nigra, not the striatum
View evidence from paper
“XENP345 was neuroprotective only when infused into the nigra, not the striatum”
Amphetamine-induced rotational behavior assessment
Administer amphetamine and measure rotational behavior to assess preservation of striatal dopamine levels
Note: Attenuated rotational behavior indicates preservation of dopamine
View evidence from paper
“XENP345/6-OHDA rats displayed attenuated amphetamine-induced rotational behavior, indicating preservation of striatal dopamine levels”
Chronic nigral coinfusion of XENP345 and LPS
Chronically coinfuse XENP345 with bacterial lipopolysaccharide into the substantia nigra
Note: Used to confirm role of solTNF-dependent neuroinflammation in nigral degeneration
View evidence from paper
“Similar protective effects were observed with chronic in vivo coinfusion of XENP345 with bacterial lipopolysaccharide (LPS) into the substantia nigra”
In vitro embryonic midbrain culture preparation
Prepare embryonic rat midbrain neuron/glia cell cultures for in vitro studies
Note: Cultures exposed to LPS to model neuroinflammation
View evidence from paper
“In embryonic rat midbrain neuron/glia cell cultures exposed to LPS, even delayed administration of XENP345 prevented selective degeneration of DA neurons”
Delayed XENP345 administration in vitro
Administer XENP345 with delayed timing in LPS-exposed midbrain cultures
Note: Demonstrates neuroprotection even with delayed treatment despite sustained microglia activation
View evidence from paper
“even delayed administration of XENP345 prevented selective degeneration of DA neurons despite sustained microglia activation and secretion of solTNF”
In vitro 6-OHDA toxicity assessment
Expose midbrain cultures to 6-OHDA and assess XENP345 neuroprotective effects
Note: Evaluates TNF role in 6-OHDA-induced dopaminergic neuron toxicity
View evidence from paper
“XENP345 also attenuated 6-OHDA-induced DA neuron toxicity in vitro”