पार्किंसंस रोग के लिए murine मॉडल: 6-OH Dopamine घाव से व्यवहार टेस्ट
Summary
पार्किंसंस रोग डोपामिनर्जिक innervation striatum, जो प्रयोगात्मक 6-OH-डोपामाइन द्वारा प्रेरित किया जा सकता है नुकसान के कारण होता है. हम वर्णन कैसे एक stereotaxic घाव प्रदर्शन करने के लिए और apomorphine प्रेरित चूहों में घूर्णी व्यवहार की निगरानी. यह मॉडल पार्किंसंस रोग के लिए नए उपचारों के परीक्षण के लिए उपयोगी और विश्वसनीय है.
Abstract
Parkinson’s disease (PD) affects at least 6.5 million people worldwide, irrespective of gender, social, ethnic, economic, or geographic boundaries. Key symptoms, such as tremor, rigidity and bradikinesia, develop when about 3/4 of dopaminergic cells are lost in the substantia nigra, and fail to provide for the smooth, coordinated regulation of striatal motor circuits. Depression and hallucinations are common, and dementia eventually occurs in 20% of patients. At this time, there is no treatment to delay or stop the progression of PD. Rather, the medications currently available aim more towards the alleviation of these symptoms. New surgical strategies may reversibly switch on the functionally damaged circuits through the electrical stimulation of deep brain structures, but although deep brain stimulation is a major advance, it is not suitable for all patients. It remains therefore necessary to test new cell therapy approaches in preclinical models.
Selective neurotoxic disruption of dopaminergic pathways can be reproduced by injection of 6-hydroxydopamine (6-OHDA) or MPTP (1-methyl-4-phenyl-1,2,3,6-tertahydropyridine) whereas depleting drugs and oxidative-damaging chemicals may also reproduce specific features of PD in rodents. Unlike MPTP, 6-OHDA lesions cause massive irreversible neuronal loss, and can be uni- or bilateral. The 6-OHDA lesion model is reliable, leads to robust motor deficits, and is the most widely used after 40 years of research in rats1. As interactions between grafted cells and host can now be studied more thoroughly in mice rather than in rats, the model has been transposed to mice2,3, where it has been recently characterized4.
In this video, we demonstrate how to lesion the left nigro-striatal pathway of anesthetized mice by slowly delivering 2.0 μL of 6-OHDA through a stereotaxically inserted micro-syringe needle. The loss of dopaminergic input occurs within days, and the functional impairments can be monitored over post-operative weeks and months by rating animal rotations induced by dopaminergic agents5. Here, we show full-body contralateral rotations occurring 10 minutes after a single subcutaneous administration of apomorphine, measured one month after the lesion. Outcomes and drawbacks are discussed below.
Protocol
Discussion
Nigro-striatal मार्ग का व्यापक एकतरफा घाव चूहों में मज़बूती से कर सकते हैं striatum में एक 6-OHDA stereotaxic इंजेक्शन द्वारा हासिल की. घाव की हद तक immunohistochemistry द्वारा पोस्टमार्टम सत्यापित Tyrosine hydroxylase के लिए कर सकते हैं (जो डीए के संश्ले…
Disclosures
The authors have nothing to disclose.
Acknowledgements
Fundação डे Amparo द्वारा समर्थित Pesquisa राज्य रियो डी जनेरियो (FAPERJ) और Conselho Nacional de Desenvolvimento Científico ई Tecnologico (CNPq). हम अनाम रेफरी टिप्पणियों जिसका हमारी मदद पांडुलिपि और वीडियो दोनों में सुधार के लिए आभारी हैं.
Materials
| Material Name | Type | Company | Catalogue Number | Comment |
|---|---|---|---|---|
| Ketamine hydrochloride | König | 90-120 mg/Kg | ||
| Xylazine hydrochloride | König | 10 mg/Kg | ||
| Atropine Sulfate | Isofarma | Isofarma | ||
| 6 – Hydroxydopamine Hydrochloride | Sigma-Aldrich | 5 μg/μL | ||
| L – Ascorbic Acid | USB Corporation | 0.2% | ||
| Xylocaine (Lidocaine Chloridrate) |
ASTRA Chemistry | 5% 50 mg |
||
| Nebacetin (Neomycin Sulfate + Bacitracin) | Altana Pharma | 5 mg/g + 250 Ul/g | ||
| R-(-)-Apomorphine hydrochloride | Sigma-Aldrich | 0.5 mg/Kg | ||
| GenTeal lOphthalmic ointment Hypromellose |
NOVARTIS | 0.33% |
||
| Ibuprofen | Abbott | 4 mg/ 100 ml drinking water | ||
| Povidine | JohnsonDiversey | |||
| R-(-)-Apomorphine hydrochloride | Sigma-Aldrich | 0.5 mg/Kg |
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