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, David E. Szymkowski, Christine G. Smith et al.
Journal of Neuroscience • 2006
Amphetamine-Induced Rotational Behavior
Objective: Assessment of striatal dopamine levels and motor asymmetry through measurement of rotational behavior induced by amphetamine in rats with unilateral dopaminergic lesions
This is a Amphetamine-Induced Rotational Behavior protocol using rat as the model organism. The procedure involves 6 procedural steps, 1 equipment items, 4 materials. Extracted from a 2006 paper published in Journal of Neuroscience.
Model and subjects
rat • embryonic rat midbrain used in culture studies; specific strain not stated for in vivo studies • unknown • Not specified • Not specified
Study window
Estimated timing pending
Core workflow
Striatal injection of 6-OHDA • XENP345 infusion into substantia nigra • Amphetamine administration and rotational behavior assessment
Primary readouts
- Amphetamine-induced rotational behavior (number of rotations)
- Striatal dopamine levels (inferred from rotational behavior attenuation)
- Nigral dopaminergic neuron survival/degeneration
- Microglia activation status
Key equipment and reagents
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Protocol Steps
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Striatal injection of 6-OHDA
Rats received a striatal injection of the oxidative neurotoxin 6-OHDA to induce retrograde nigral degeneration
Note: This creates a unilateral dopaminergic lesion model
View evidence from paper
“retrograde nigral degeneration induced by a striatal injection of the oxidative neurotoxin 6-hydroxydopamine (6-OHDA)”
XENP345 infusion into substantia nigra
XENP345 was infused into the substantia nigra to neutralize soluble TNF and provide neuroprotection
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 administration and rotational behavior assessment
Rats received amphetamine and rotational behavior was measured to assess preservation of striatal dopamine levels
Note: Attenuated rotational behavior indicates preservation of striatal dopamine
View evidence from paper
“XENP345/6-OHDA rats displayed attenuated amphetamine-induced rotational behavior, indicating preservation of striatal dopamine levels”
Chronic coinfusion of XENP345 and LPS
Chronic in vivo coinfusion of XENP345 with bacterial lipopolysaccharide into the substantia nigra to model neuroinflammatory conditions
Note: This confirmed a role for solTNF-dependent neuroinflammation in nigral degeneration
View evidence from paper
“chronic in vivo coinfusion of XENP345 with bacterial lipopolysaccharide (LPS) into the substantia nigra”
Embryonic rat midbrain neuron/glia cell culture exposure to LPS
Embryonic rat midbrain neuron/glia cell cultures were exposed to LPS with delayed XENP345 administration
Note: Even delayed XENP345 administration prevented selective degeneration of dopaminergic neurons despite sustained microglia activation
View evidence from paper
“embryonic rat midbrain neuron/glia cell cultures exposed to LPS, even delayed administration of XENP345 prevented selective degeneration of DA neurons”
In vitro 6-OHDA toxicity assessment
XENP345 was tested for its ability to attenuate 6-OHDA-induced dopaminergic neuron toxicity in cell culture
Note: XENP345 demonstrated neuroprotective effects in vitro
View evidence from paper
“XENP345 also attenuated 6-OHDA-induced DA neuron toxicity in vitro”