Magnetothermal nanoparticle technology alleviates parkinsonian-like symptoms in mice methods
Aim. Evidence-backed execution summary for Magnetothermal nanoparticle technology alleviates parkinsonian-like symptoms in mice methods from Magnetothermal nanoparticle technology alleviates parkinsonian-like symptoms in mice.
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mouse
Subject model for the experiment.
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- confirm full cohort details in the source paper
Immunohistochemistry
reagent used in the protocol.
- Use
- For immunohistochemistry, sections were incubated overnight with polyclonal rabbit anti-c-Fos primary antibody (1:1000; K-25, Santa Cruz Biotechnology Inc, Santa Cruz, USA) or rabbit anti-tyrosine hydroxylase (TH, 1:500, H-196, Santa Cruz Biotechnology Inc, Santa Cruz, USA). C-Fos immunohistochemistry included bioti...
Introduction
Deep-brain stimulation (DBS) is an invasive treatment involving the implantation of electrodes, which deliver electrical impulses to specific parts of the brain. It has substantial therapeutic effects in a range of disorders, including Parkinson's disease (PD), essential tremor, and dystonia -. In parti...
- Use
- Deep-brain stimulation (DBS) is an invasive treatment involving the implantation of electrodes, which deliver electrical impulses to specific parts of the brain. It has substantial therapeutic effects in a range of disorders, including Parkinson's disease (PD), essential tremor, and dystonia -. In parti...
In vivo magnetothermal neuromodulation of mouse behavior
To further evaluate the biomedical potential of mDBS, we applied it to stimulate the STN due to its clinical relevance as a target for DBS treatment of movement disorders. It has been shown before that classical unilateral STN DBS can induce circling behavior in rodents and therefore we also aimed to assess rotation...
- Use
- To further evaluate the biomedical potential of mDBS, we applied it to stimulate the STN due to its clinical relevance as a target for DBS treatment of movement disorders. It has been shown before that classical unilateral STN DBS can induce circling behavior in rodents and therefore we also aimed to assess rotation...
Discussion
Our study relied on exogenously expressed TRPV1 to create uniform levels across mice, which might represent a major challenge for clinical applications. This channel, however, is also endogenously expressed in neurons and glia in certain regions of the mammalian central nervous system. Various studies have shown th...
- Use
- Our study relied on exogenously expressed TRPV1 to create uniform levels across mice, which might represent a major challenge for clinical applications. This channel, however, is also endogenously expressed in neurons and glia in certain regions of the mammalian central nervous system. Various studies have shown th...
Methods
Although TRPV1 is naturally expressed across the mammalian nervous system, we designed a transgene to establish sustained and uniform levels of TRPV1 expression for magnetothermal membrane depolarization. The preparation of pLenti-CaMKIIα-TRPV1-p2A-mCherry was performed by Boston Children's Hospital vir...
- Use
- Although TRPV1 is naturally expressed across the mammalian nervous system, we designed a transgene to establish sustained and uniform levels of TRPV1 expression for magnetothermal membrane depolarization. The preparation of pLenti-CaMKIIα-TRPV1-p2A-mCherry was performed by Boston Children's Hospital vir...
In vivo virus and nanoparticle injection
The stereotactic procedure is the same as mentioned above. This time, however, we injected Lenti-CaMKIIα::TRPV1-p2A-mCherry unilaterally into the left STN (1.5 µl per hemisphere). Seven weeks later, 1.5 µl of 80 mg/ml MNPs (SMG-25, Ocean Nanotech, San Diego, USA) were also injected i...
- Use
- The stereotactic procedure is the same as mentioned above. This time, however, we injected Lenti-CaMKIIα::TRPV1-p2A-mCherry unilaterally into the left STN (1.5 µl per hemisphere). Seven weeks later, 1.5 µl of 80 mg/ml MNPs (SMG-25, Ocean Nanotech, San Diego, USA) were also injected i...
Immunohistochemistry
For immunohistochemistry, sections were incubated overnight with polyclonal rabbit anti-c-Fos primary antibody (1:1000; K-25, Santa Cruz Biotechnology Inc, Santa Cruz, USA) or rabbit anti-tyrosine hydroxylase (TH, 1:500, H-196, Santa Cruz Biotechnology Inc, Santa Cruz, USA). C-Fos immunohistochemistry included bioti...
- Use
- For immunohistochemistry, sections were incubated overnight with polyclonal rabbit anti-c-Fos primary antibody (1:1000; K-25, Santa Cruz Biotechnology Inc, Santa Cruz, USA) or rabbit anti-tyrosine hydroxylase (TH, 1:500, H-196, Santa Cruz Biotechnology Inc, Santa Cruz, USA). C-Fos immunohistochemistry included bioti...
Introduction
Software used for acquisition, scoring, statistics, or reporting.
- Use
- Deep-brain stimulation (DBS) is an invasive treatment involving the implantation of electrodes, which deliver electrical impulses to specific parts of the brain. It has substantial therapeutic effects in a range of disorders, including Parkinson's disease (PD), essential tremor, and dystonia -. In parti...
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Introduction
Deep-brain stimulation (DBS) is an invasive treatment involving the implantation of electrodes, which deliver electrical impulses to specific parts of the brain. It has substantial therapeutic effects in a range of disorders, including Parkinson's disease (PD), essential tremor, and dystonia -. In particular, high-frequency DBS of the subthalamic nucleus (STN) is now considered a treatment of choice for patients suffering from severe forms of PD with motor fluctuations and major side effects of dopaminomimetic treatments. Nevertheless, the fundamentals of DBS hardware and software design have seen limited progress since its advent three decades ago and the main drawback of current DBS is that it requires a wired and chronically implanted system. As a result, many patients are reluctant to undergo DBS even when treatment is warranted. Recent advances in the field of neur...
In vivo magnetothermal neuromodulation of mouse behavior
To further evaluate the biomedical potential of mDBS, we applied it to stimulate the STN due to its clinical relevance as a target for DBS treatment of movement disorders. It has been shown before that classical unilateral STN DBS can induce circling behavior in rodents and therefore we also aimed to assess rotational behavior in response to unilateral STN mDBS. Rotational motor behavior offers an objective read-out and is also compatible with laboratory-scale magnetic field coils. We first heat-sensitized neurons in the STN through lentiviral delivery of TRPV1, which was followed by MNP injection ( n = 7) into the same region 6-8 weeks later (Fig. ). Control mice ( n = 10) were subjected to the same viral delivery and surgical procedures, but were injected with non-magnetic nanoparticles (non-MNPs) composed of wüstite (FeO). The mice were a...
Discussion
Here, we targeted the STN with mDBS and assessed its biomedical potential as wireless alternative to classical DBS. Although there is precedent for applying nanomaterials-mediated neuromodulation in clinically-inspired contexts, including photothermal approaches using gold nanoparticles,, there have been no prior reports of translational studies aiming to treat parkinsonian symptoms using mDBS.
In vivo virus and nanoparticle injection
Experiment 2: In the second experiment we used 40 male wild-type mice (C57BL/6, Jackson Laboratory), group-housed in standard ventilated cages (IVC) in a controlled environment (temperature 22 °C, humidity 59 (rH) using a 12/12-h reversed dark/light cycle (lights off 7 AM-7 PM). Food and water were given ad libitum. Animal procedures in experiment 2 were carried out under a protocol approved by the Institutional Animal Care Committee of Maastricht University in accordance to the Central Authority for Scientific Procedures on Animals (CCD; protocol # AVD1070020186046).
In vivo virus and nanoparticle injection
The stereotactic procedure is the same as mentioned above. This time, however, we injected Lenti-CaMKIIα::TRPV1-p2A-mCherry bilaterally into the STN (1.5 µl per hemisphere). Seven weeks later, 2 µl of 80 mg/ml MNPs (SMG-25, Ocean Nanotech, San Diego, USA) or non-MNPs were also injected bilaterally into the STN. Following a 1-week recovery period, MPTP (18 mg/kg in saline, intraperitoneally) or vehicle was administered two times in one day, each injection 2 h apart. We wanted to employ an acute MPTP treatment with 4 injections in one day, however, ~40% of the mice were found dead after the second injection and we decided to abort the following 2 injections. The final group numbers were naive control ( n = 8), naive mDBS ( n = 10), MPTP control ( n = 6), MPTP mDBS ( n = 7). Following 2 weeks...
In vivo virus and nanoparticle injection
Experiment 3: In the third experiment we used 22 male wild-type mice (C57BL/6, Jackson Laboratory), group-housed in standard ventilated cages (IVC) in a controlled environment (temperature 22 °C, humidity 59 (rH) using a 12/12-h reversed dark/light cycle (light off 7 AM-7 PM). Food and water were given ad libitum. Animal procedures in experiment 3 were carried out under a protocol approved by the Institutional Animal Care Committee of Maastricht University in accordance to the CCD (protocol # AVD1070020186046).
In vivo virus and nanoparticle injection
The stereotactic procedure is the same as mentioned above. This time, however, we injected Lenti-CaMKIIα::TRPV1-p2A-mCherry unilaterally into the left STN (1.5 µl per hemisphere). Seven weeks later, 1.5 µl of 80 mg/ml MNPs (SMG-25, Ocean Nanotech, San Diego, USA) were also injected into the STN. In the same surgery, 0.2 µl of 6-OHDA was also unilaterally injected into the ipsilateral median forebrain bundle (AP: -1.2 mm, ML: -1.1 mm, DV: -5 mm) at a rate of 0.1 µl/min (3 µg total). Two mice died during surgery and we experienced an additional 50% drop-out due to poor body condition of the 6-OHDA mice. We therefore switched to a within-subjects design and were able to test n = 10 mice before and after AMF stimulation in the rotarod, open field, and CatWalk gait analysis sy...
Measurement outputs
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Deep-brain stimulation (DBS) is an invasive treatment involving the implantation of electrodes, which deliver electrical impulses to specific parts of the brain. It has substant...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
To further evaluate the biomedical potential of mDBS, we applied it to stimulate the STN due to its clinical relevance as a target for DBS treatment of movement disorders. It ha...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
Although TRPV1 is naturally expressed across the mammalian nervous system, we designed a transgene to establish sustained and uniform levels of TRPV1 expression for magnetother...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
The stereotactic procedure is the same as mentioned above. This time, however, we injected Lenti-CaMKIIα::TRPV1-p2A-mCherry unilaterally into the left STN (1.5 µ...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
Analysis plan
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Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
inferred from protocolPreprocessing / cleaning
Deep-brain stimulation (DBS) is an invasive treatment involving the implantation of electrodes, which deliver electrical impulses to specific parts of the brain.
from paperScoring or quantification
Quantify the primary readouts for this experiment: Deep-brain stimulation (DBS) is an invasive treatment involving the implantation of electrodes, which deliver electrical impulses to specific parts of the brain. It has substant...; To further evaluate the biomedical potential of mDBS, we applied it to stimulate the STN due to its clinical relevance as a target for DBS treatment of movement disorders. It ha...; Although TRPV1 is naturally expressed across the mammalian nervous system, we designed a transgene to establish sustained and uniform levels of TRPV1 expression for magnetother...; The stereotactic procedure is the same as mentioned above. This time, however, we injected Lenti-CaMKIIα::TRPV1-p2A-mCherry unilaterally into the left STN (1.5 µ....
from paperStatistical comparison
Deep-brain stimulation (DBS) is an invasive treatment involving the implantation of electrodes, which deliver electrical impulses to specific parts of the brain. It has substant...; To further evaluate the biomedical potential of mDBS, we applied it to stimulate the STN due to its clinical relevance as a target for DBS treatment of movement disorders. It ha...
from paperReporting output
Report representative outputs alongside summary comparisons for Deep-brain stimulation (DBS) is an invasive treatment involving the implantation of electrodes, which deliver electrical impulses to specific parts of the brain. It has substant..., To further evaluate the biomedical potential of mDBS, we applied it to stimulate the STN due to its clinical relevance as a target for DBS treatment of movement disorders. It ha..., Although TRPV1 is naturally expressed across the mammalian nervous system, we designed a transgene to establish sustained and uniform levels of TRPV1 expression for magnetother..., The stereotactic procedure is the same as mentioned above. This time, however, we injected Lenti-CaMKIIα::TRPV1-p2A-mCherry unilaterally into the left STN (1.5 µ....
inferred from protocolStructured statistical methods
Deep-brain stimulation (DBS) is an invasive treatment involving the implantation of electrodes, which deliver electrical impulses to specific parts of the brain. It has substant...; To further evaluate the biomedical potential of mDBS, we applied it to stimulate the STN due to its clinical relevance as a target for DBS treatment of movement disorders. It ha...
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Evidence quotes (7)
Deep-brain stimulation (DBS) is an invasive treatment involving the implantation of electrodes, which deliver electrical impulses to specific parts of the brain. It has substantial therapeutic effects in a range of disorders, including Parkinson's disease (PD), essential tremor, and dystonia -. In particular, high-frequency DBS of the subthalamic nucleus (STN) is now considered a treatment of choice for patients suffering from severe forms of PD with motor fluctuations and major side effects of dopaminomimetic treatments. Nevertheless, the fundamentals of DBS hardware and software design have seen limited progress since its advent three decades ago and the main drawback of current DBS is that it requires a wired and chronically implanted system. As a result, many patients are reluctant to undergo DBS even when treatment is warranted. Recent advances in the field of neuromodulation stress the urgency of developing wireless deep-brain stimulation treatments. Existing non-invasive brain stimulation methods include repetitive transcranial magnetic stimulation (rTMS), transcranial direct current stimulation (tDCS), and focused ultrasound among others. In rTMS, high-in...
To further evaluate the biomedical potential of mDBS, we applied it to stimulate the STN due to its clinical relevance as a target for DBS treatment of movement disorders. It has been shown before that classical unilateral STN DBS can induce circling behavior in rodents and therefore we also aimed to assess rotational behavior in response to unilateral STN mDBS. Rotational motor behavior offers an objective read-out and is also compatible with laboratory-scale magnetic field coils. We first heat-sensitized neurons in the STN through lentiviral delivery of TRPV1, which was followed by MNP injection ( n = 7) into the same region 6-8 weeks later (Fig. ). Control mice ( n = 10) were subjected to the same viral delivery and surgical procedures, but were injected with non-magnetic nanoparticles (non-MNPs) composed of wüstite (FeO). The mice were allowed to recover for 1 week prior to behavioral testing. The circular arena was made out of plexiglass (Ø 9 mm, height 10 cm, fitting within the magnetic coil) and mice were stimulated continuously for 3 min at ƒ = 160 kHz, H 0 = 30 k...
Here, we targeted the STN with mDBS and assessed its biomedical potential as wireless alternative to classical DBS. Although there is precedent for applying nanomaterials-mediated neuromodulation in clinically-inspired contexts, including photothermal approaches using gold nanoparticles,, there have been no prior reports of translational studies aiming to treat parkinsonian symptoms using mDBS.
Experiment 2: In the second experiment we used 40 male wild-type mice (C57BL/6, Jackson Laboratory), group-housed in standard ventilated cages (IVC) in a controlled environment (temperature 22 °C, humidity 59 (rH) using a 12/12-h reversed dark/light cycle (lights off 7 AM-7 PM). Food and water were given ad libitum. Animal procedures in experiment 2 were carried out under a protocol approved by the Institutional Animal Care Committee of Maastricht University in accordance to the Central Authority for Scientific Procedures on Animals (CCD; protocol # AVD1070020186046).
The stereotactic procedure is the same as mentioned above. This time, however, we injected Lenti-CaMKIIα::TRPV1-p2A-mCherry bilaterally into the STN (1.5 µl per hemisphere). Seven weeks later, 2 µl of 80 mg/ml MNPs (SMG-25, Ocean Nanotech, San Diego, USA) or non-MNPs were also injected bilaterally into the STN. Following a 1-week recovery period, MPTP (18 mg/kg in saline, intraperitoneally) or vehicle was administered two times in one day, each injection 2 h apart. We wanted to employ an acute MPTP treatment with 4 injections in one day, however, ~40% of the mice were found dead after the second injection and we decided to abort the following 2 injections. The final group numbers were naive control ( n = 8), naive mDBS ( n = 10), MPTP control ( n = 6), MPTP mDBS ( n = 7). Following 2 weeks, mice were placed into the AMF coil and stimulated for 3 min. After 3 min, they were taken out of the coil and subsequently tested in the open field.
Experiment 3: In the third experiment we used 22 male wild-type mice (C57BL/6, Jackson Laboratory), group-housed in standard ventilated cages (IVC) in a controlled environment (temperature 22 °C, humidity 59 (rH) using a 12/12-h reversed dark/light cycle (light off 7 AM-7 PM). Food and water were given ad libitum. Animal procedures in experiment 3 were carried out under a protocol approved by the Institutional Animal Care Committee of Maastricht University in accordance to the CCD (protocol # AVD1070020186046).
The stereotactic procedure is the same as mentioned above. This time, however, we injected Lenti-CaMKIIα::TRPV1-p2A-mCherry unilaterally into the left STN (1.5 µl per hemisphere). Seven weeks later, 1.5 µl of 80 mg/ml MNPs (SMG-25, Ocean Nanotech, San Diego, USA) were also injected into the STN. In the same surgery, 0.2 µl of 6-OHDA was also unilaterally injected into the ipsilateral median forebrain bundle (AP: -1.2 mm, ML: -1.1 mm, DV: -5 mm) at a rate of 0.1 µl/min (3 µg total). Two mice died during surgery and we experienced an additional 50% drop-out due to poor body condition of the 6-OHDA mice. We therefore switched to a within-subjects design and were able to test n = 10 mice before and after AMF stimulation in the rotarod, open field, and CatWalk gait analysis system 2 weeks after surgery.
Machine-readable layer
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"text": "To further evaluate the biomedical potential of mDBS, we applied it to stimulate the STN due to its clinical relevance as a target for DBS treatment of movement disorders. It has been shown before that classical unilateral STN DBS can induce circling behavior in rodents and therefore we also aimed to assess rotational behavior in response to unilateral STN mDBS. Rotational motor behavior offers an objective read-out and is also compatible with laboratory-scale magnetic field coils. We first heat-sensitized neurons in the STN through lentiviral delivery of TRPV1, which was followed by MNP injection ( n = 7) into the same region 6-8 weeks later (Fig. ). Control mice ( n = 10) were subjected to the same viral delivery and surgical procedures, but were injected with non-magnetic nanoparticles (non-MNPs) composed of wüstite (FeO). The mice were a..."
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