6-OHDA assay was performed as previously described.54 (link) Metabolite- and EtAc-treated L4 worms (BY200) were washed with ddH2O three times, and treated with 30 mM 6-OHDA (Tocris Bioscience, Bristol, UK) containing 1 mM ascorbic acid, followed by gentle agitation for 1 h. Subsequently, the worms were again washed and put onto freshly made metabolite or EtAc plates until analysis. For rotenone assay, BY200 worms were exposed to the metabolite or EtAc until L4 and then transferred to plates containing metabolite, along with 5 μM of rotenone or DMSO (as a solvent control for rotenone).
6 ohda
Manufactured by Bio-Techne
Sourced in United Kingdom
6-OHDA is a chemical compound commonly used in research applications. It functions as a neurotoxin, specifically targeting and damaging dopaminergic neurons. This product is often utilized in studies related to Parkinson's disease and other neurological conditions.
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Lab products found in correlation
6ohda, by World Precision Instruments (1 mentions)
Nf33bl 2, by World Precision Instruments (1 mentions)
Stereotaxic frame, by Kopf Instruments (1 mentions)
D1507, by Merck Group (1 mentions)
B7283, by Merck Group (1 mentions)
6 ohda hydrobromide, by Bio-Techne (1 mentions)
6 ohda, by Merck Group (1 mentions)
Desferrioxamine, by Merck Group (1 mentions)
Erastin, by Bio-Techne (1 mentions)
Ml210, by Bio-Techne (1 mentions)
Reduced glutathione gsh, by Merck Group (1 mentions)
Rotenone, by Merck Group (1 mentions)
6 protocols using 6 ohda
1
6-OHDA and Rotenone Assays in C. elegans
2
Unilateral 6-OHDA Striatal Lesion Model
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Mice were anesthetized with 2% isoflurane and secured on a stereotactic frame. 6-Hydroxydopamine (6OHDA; 2547; Tocris) was dissolved in a 0.2% ascorbic acid saline solution. A 0.2% ascorbic acid saline solution was used as vehicle control. WT mice received a unilateral injection of 6OHDA or vehicle control in the striatum. The striatum coordinate used for stereotaxic injection was AP = 0.7 ML = − 2.0 DV = − 2.0 mm. A total of 5 μg of 6OHDA in 2 μL was injected in the striatum at a rate of 0.5 μL/min with a 33G nanofil blunt needle (NF33BL-2, World Precision Instruments) connected to FEP tubbing and a 25 μL Hamilton syringe. After 6OHDA infusion, the nanofil blunt needle was left in place for 5 min. Four weeks after injection, mice were PBS and 4% PFA perfused for immunohistochemistry analysis.
3
Unilateral 6-OHDA Lesioning and Neuronal Tracing
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Mice (3 to 4 mo old, 25 to 30 g) were anesthetized with sodium pentobarbital and stabilized in a stereotaxic frame (RWD Life Science). They were then subjected to unilateral 6-OHDA (0.3 µL, 15 µg/µL; Tocris) microinjection into the MFB, viral vector delivery (0.3 µL), optical fiber implantation, and transsynaptic neuronal tracing (WGA 0.1 µL, 1 µg/µL; Thermo Fisher Scientific). Details are provided in SI Appendix .
4
6-OHDA Lesion and L-Dopa Treatment Protocol
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Under isoflurane anesthesia, the mice were placed in a stereotaxic frame (David Kopf Instruments) and maintained throughout surgery using 1–2% isoflurane. Bilateral internal cannulas (Plastics One), for delivery of 6-OHDA to the median forebrain bundle, were positioned ±1.1 mm lateral and −5.0 mm ventral and were implanted 0.8 mm posterior to Bregma). 6-OHDA (Tocris 2547) was prepared at a concentration of 5 μg/μL in 0.9% NaCl containing 0.01% ascorbate (w/v) for bilateral depletions. Injections were performed using a 33-gauge cannula (Plastics One) attached to a 10 μL Hamilton syringe within a syringe pump running at 0.5 μL/min, to a total volume of 1 μL/side. The injection cannula was left in place for 5 min following the injection. Following recovery, beginning the day following surgery, mice were administered either L-Dopa (6.25 mg/kg, i.p, Sigma, D1507), a combination of L-Dopa (6.25 mg/kg, i.p.) and benserazide hydrochloride (10 mg/kg, i.p. Sigma, B7283), or saline alone once a day for 5 days. All compounds were dissolved in 0.9% saline.
5
6-OHDA Lesioning in C57BL/6J Mice
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In a separate group C57BL/6J (n = 11), and one Vgat-IRES-Cre mouse (Slc32a1 were injected with 1 μl of 6-OHDA into the medial forebrain bundle (−1.2 AP, 1.2 ML, −4.75 DV) using the procedure described in Lundblad et al. (2004) (link). Briefly, 0.02% ascorbic acid was added to a sterile saline solution (NaCl, 0.9% w/v) and 6-OHDA HCl powder (6-OHDA from Sigma or 6-OHDA hydrobromide from Tocris) was then dissolved to produce a final 6-OHDA concentration of 4.44 mg/ml. This method produced strong nigrostriatal lesioning as verified histologically through TH antibody staining (see immunohistochemistry below).
6
Compound Preparation for Cell Studies
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