J Korean Med Sci.  2014 Nov;29(11):1555-1561. 10.3346/jkms.2014.29.11.1555.

A Rat Model of Striatonigral Degeneration Generated by Simultaneous Injection of 6-Hydroxydopamine into the Medial Forebrain Bundle and Quinolinic Acid into the Striatum

• Yoon HH
• Kim YH
• Shin ES
• Jeon SR

• KMID: 2069939
• DOI: http://doi.org/10.3346/jkms.2014.29.11.1555

Abstract

A double toxin-double lesion strategy is well-known to generate a rat model of striatonigral degeneration (SND) such as multiple system atrophy-parkinsonian type. However, with this model it is difficult to distinguish SND from Parkinson's disease (PD). In this study, we propose a new rat model of SND, which is generated by simultaneous injection of 6-hydroxydopamine into the medial forebrain bundle and quinolinic acid into the striatum. Stepping tests performed 30 min after intraperitoneal L-dopa administration at 6 weeks post-surgery revealed an L-dopa response in the PD group but not the SND group. Apomorphine-induced rotation tests revealed no rotational bias in the SND group, which persisted for 2 months, but contralateral rotations in the PD group. MicroPET scans revealed glucose hypometabolism and dopamine transporter impairment on the lesioned striatum in the SND group. Tyrosine hydroxylase immunostaining in the SND group revealed that 74.7% of nigral cells on the lesioned side were lost after lesion surgery. These results suggest that the proposed simultaneous double toxin-double lesion method successfully created a rat model of SND that had behavioral outcomes, multitracer microPET evaluation, and histological aspects consistent with SND pathology. This model will be useful for future study of SND.

Keyword

FP-CIT; L-dopa Response; Multiple System Atrophy; Positron Emission Tomography; Simultaneous Injection; Striatonigral Degeneration

MeSH Terms

Animals
Apomorphine/pharmacology
Behavior, Animal/drug effects
Corpus Striatum/drug effects/pathology
Disease Models, Animal
Dopamine Plasma Membrane Transport Proteins/metabolism
Glucose/metabolism
Injections, Intraperitoneal
Levodopa/pharmacology
Male
Medial Forebrain Bundle/drug effects/pathology
Oxidopamine/*toxicity
Parkinson Disease/metabolism/pathology
Positron-Emission Tomography
Quinolinic Acid/*toxicity
Rats
Rats, Wistar
Striatonigral Degeneration/*chemically induced/metabolism/pathology
Touch/drug effects
Apomorphine
Dopamine Plasma Membrane Transport Proteins
Glucose
Levodopa
Oxidopamine
Quinolinic Acid

Figure

• 

  Fig. 1 Stepping tests. To evaluate a change in motor deficit and L-dopa response, these tests were performed during the intact state and at 2, 4, 6, 8, and 9 weeks post-surgery. Data are shown as contralateral paw touches rate (%) of the ipsilateral paw touches. The stepping test performed at 6 weeks post-surgery was preceded by intraperitoneal administration of L-dopa. Significant motor deficit in contralateral forelimb was examined after lesion surgery in both SND and PD group during the experiments. After L-dopa administration, the rats of SND group showed no response, while PD rats were improved that was equal to intact state, which showed a significant difference between SND and PD group (*P = 0.003).

• 

  Fig. 2 Apomorphine-induced rotation test. Net (contralateral-ipsilateral) rotations per min in the apomorphine-induced rotation test performed at 4 and 8 weeks post-surgery. There was a significant difference between groups at week 4 (*P = 0.006), which persisted at week 8 (*P < 0.001). PD group increased in contralateral rotations at week 8 compared to week 4 (†P = 0.015).

• 

  Fig. 3 [18F]-FDG uptake using microPET scanning. Ipsilateral [18F]-FDG uptake expressed relative to contralateral [18F]-FDG uptake in the volume of interest (VOI) at 5 and 10 weeks post-surgery. Arrows indicate lesion sites. The significant difference between groups persisted during 10 weeks after lesion surgery (At week 5; 77.1±7.0% vs. 94.8±6.3%;*P = 0.003, at week 10; 64.9±11.4% vs. 95.9±3.3%;*P = 0.003).

• 

  Fig. 4 [18F]-FP-CIT uptake using microPET scanning. Ipsilateral [18F]-FP-CIT uptake expressed relative to contralateral [18F]-FP-CIT uptake in the volume of interest (VOI) at 5 and 10 weeks post-surgery. There was no significant difference between groups at week 5 (54.9±16.4% vs. 45.6±6.6%; P = 0.246), which persisted at week 10 (49.6±6.2% vs. 46.8±4.2%; P = 0.441). Arrows indicate lesion sites.

• 

  Fig. 5 (A) Tyrosine hydroxylase (TH) staining in the SND group, showing decreased staining of dopaminergic axons and terminals in the lesioned striatum, and atrophy (a). Dopaminergic cell count in the SN revealed 74.7±19.4% losses of TH-positive cells in lesioned SN against contralateral SN (b, rostral; c, medial; d, caudal sections) (VTA, ventral tegmental area) (a, ×10; b, c, d, ×40 magnification). (B) TH staining in the PD group, showing decreased staining of dopaminergic axons and terminals in the lesioned striatum (a). Dopaminergic cell count in the SN revealed 92.8 ± 2.7% losses of TH-positive cells in lesioned SN against contralateral SN (b, rostral; c, medial; d, caudal sections) (VTA, ventral tegmental area) (a, ×10; b, c, d, ×40 magnification).

• 

  Fig. 6 Nissl staining of striatal tissue. Cresyl violet staining revealed enlarged ventricle and striatal atrophy on the lesioned side in the SND group (A) compared to the PD group (B) (A, B ×10 magnification).

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