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25 protocols using xylocain

1

Neuropharmacological Intervention in Rats

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The surgery was conducted as earlier described,25 where two guides (stainless steel, length 10 mm, with an o.d./i.d. of 0.6/0.45 mm) targeting the NAc shell were implanted 2 days prior to the drug challenge. During surgery, the rats were kept on a heating pad to prevent hypothermia. The rats were anaesthetized with isoflurane (Isofluran Baxter, Kronans Apotek) and a local anaesthetic mixture [Xylocain (10 mg/ml) together with adrenalin (5 μg/ml), Astra Zeneca, Kronans Apotek] was applied on the skull surface. After incision, the skull bone was exposed, and three holes were drilled: two for the guides35(Table S2) and one for the anchoring screw. The tips of the guides were inserted 1 mm below the skull bone. The guides were anchored to the screw and the skull bone with dental cement (DENTALON®plus; Agntho's AB, Lidingö, Sweden). Carprofen (5 mg/kg, SC, Rimadyl®; Zoetis, Kronans Apotek) was used to relieve pain following surgery.
At the experimental day, a dummy cannula was carefully inserted and retracted into the guide to remove clotted blood and hamper spreading depression within NAc shell. One hour later, a cannula delivered the drug into the NAc shell (Table S2), the drug was delivered over 1 min, and after an additional minute, the cannula was retracted.
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2

Muscle Biopsy Sampling Protocol

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After 15 min of rest in the supine position, two 3-mm incisions were made over the lateral portion of the left thigh under local anesthesia (2 mL lidocaine without epinephrine, 20 mg•mL -1 Xylocain, AstraZeneca). Muscle biopsies of the vastus lateralis muscle were collected before, during, and immediately after exercise (Fig. 1) using a percutaneous Bergstrom needle with suction. Specifically, muscle samples were obtained at rest and immediately after the first sprint from the distal incision as well as 10 s before and immediately after the last sprint from the proximal incision. Muscle samples were snap-frozen in liquid nitrogen and stored at -80°C until further analysis.
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3

Muscle Biopsy Procedure for Protein Analysis

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Humans: After resting for 30 min, a muscle biopsy was obtained from m. vastus lateralis under local anaesthesia (Xylocain 20 mg/ml; AstraZeneca) using a modified Bergström needle with suction. The muscle tissue was rapidly frozen (<30 s) in liquid nitrogen and stored at −80°C until further analysis.
Rats: Only white vastus lateralis muscles (fast twitch fibres) were used for the present experiments, but it has previously been demonstrated that purinergic activation of the Na,K-ATPase exists in slow twitch muscles [4] (link).
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4

Vastus Lateralis Muscle Biopsy

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Subjects reported to the laboratory in the morning after an overnight fast. After 15 min of rest in the supine position, a 3 mm incision was made over the lateral portion of the thigh under local anesthesia (2 ml lidocaine without epinephrine, 20 mg/ml Xylocain; AstraZeneca Pharmaceuticals), and a biopsy was obtained from m. vastus lateralis using a percutaneous Bergstrom needle with suction.
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5

Neuronal Activity Modulation in Temporal Muscle

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When the ongoing (unstimulated) neuronal activity of a unit was visibly stable, it was recorded for a control period of 30 min without any treatment (baseline activity). Subsequently, saline (0.9 % NaCl, 50 μl) was injected with a U-100 insulin syringe (BD, Franklin Lakes, NJ, USA) deep into the temporal muscle close to the crista temporalis within one minute. After another 30 min of recording, 50 μl of 10 mM α,β-meATP (α,β-methylene adenosine 5′-triphosphate disodium salt hydrate, Sigma-Aldrich, Taufkirchen, Germany) was injected into the temporal muscle and the recording continued for 60 min. After another 10 min without further treatment (new baseline), in most of the experiments 60 μl of 2 % lidocaine (Xylocain, Astra Zeneca, Wedel, Germany) was injected into the ipsilateral medial (semispinalis and rectus capitis) neck muscles followed by a recording period of 20 min. Next the temporal muscle was injected with 60 μl lidocaine and the activity recorded for another 20 min. Finally, 60 μl of lidocaine was applied onto the exposed cranial dura mater followed by a final recording period of 20 min. In some experiments 60 μl of lidocaine was injected only into the ipsilateral temporal muscle immediately followed by 50 μl of α,β-meATP, then the recording was continued for 60 min.
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6

Viral Injections in Mouse Hippocampus

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Viral injections were performed under aseptic conditions in Prox1::cre or C57BL/6 mice up to 10 months old. The mice were anesthetized with a mixture of Fentanyl (Rotexmedica, Germany), Midazolam (Rotexmedica, Germany), and Domitor (Orion Pharma, Finland) via intraperitoneal injection ( 0.05/5.0/0.5  mg/kg ). Analgesia ( 0.05  mg/kg of buprenorphine; Buprenovet, Bayer, Germany) was administered intraperitoneally prior to the injection, and Xylocain (AstraZeneca, Germany) was used for local anesthesia. Stereotaxic injections were performed using an injection frame (WPI Benchmark/Kopf) and a microprocessor-controlled minipump (World Precision Instruments, Sarasota, Florida), 1000 nl of the viral solution was injected bilaterally into the hippocampus (rAAV-DIO-eGFP; rAAV-syn-GFP; coordinates: rostrocaudal: 2.1  mm from the Bregma; mediolateral: ±1.2  mm from the midline; dorsoventral: 2.1  mm ). After the injection, the scalp was sutured with PERMA-HAND Silk Suture (Ethicon), and an antibacterial ointment (Refobacin®, Almirall, Germany) was applied, followed by the intraperitoneal injection of a mixture of Naloxon (B. Braun, Germany), Flumazenil (B. Braun, Germany), and Antisedan (Orion Pharma, Finland) ( 1.2/0.5/2.5  mg/kg ). To prevent the wound pain, analgesia was administered on the 3 following days.
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7

Bovine Mammary Tissue Biopsy

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On the last day of experiment (d 28) and after milking, the left front quarter of the udder was shaved, and an incision site without visual blood vessels was localized and anaesthetized subcutaneously with 2 mL of Xylocain (20 mg/mL lidocaine, AstraZeneca, Albertslund, Denmark) and cleaned with Hibitane (70% alcohol solution 1:10, Sigma-Aldrich, St. Louis, MO). Biopsies were taken with an automatic biopsy instrument (Manan Pro-Mag 2.2, Manan Medical Products Inc., Wheeling, IL) loaded with a 14 gauge × 10 cm needle (Manan ACN II, Manan Medical Products Inc.). Approximately 20 mg of tissue was collected per sampling, and a maximum of 2 samples was taken per cow. Tissue was snap frozen in liquid nitrogen and stored in Cryotubes (Nunc GmbH & Co. KG, Langenselbold, Germany) at -80°C until further analysis as described by Sorensen et al. (2006) (link).
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8

Subcutaneous Fat Metabolism Assay

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Subcutaneous fat on fasting and after 3 h oral glucose tolerance test (OGTT) was obtained by needle aspiration after administration of local anesthetic, lidocaine (Xylocain; AstraZeneca, Sweden) from the lower part of the abdomen. Tissue was rinsed with a sterile saline solution and part of it was snap frozen within 2–3 minutes after biopsy and thereafter stored at −80°C for analysis of gene expression. The other part collected into a tube containing Hank’s medium (Invitrogen Corporation, Paisley, UK) comprising 5.6 mM glucose, 4% bovine serum albumin (BSA, Sigma, St Louis, Missouri, USA), 150 nM adenosine (Sigma) maintained at 37°C and used to perform metabolic assays. AT was first digested with collagenase (Roche, Indianapolis, USA) in Hank’s medium in a shaking water bath for 1 hour. Isolated adipocytes were then filtered through a 250 µm pore size nylon mesh and used to perform glucose uptake, lipolysis, and cell size measurement.
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9

Cranial Window Surgery in Mice

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7-8 week old mice of either sex were used for cranial window surgeries. Animals were anesthetized with a mixture of 40 µl fentanyl (1 mg/ml; Janssen), 160 µl midazolam (5 mg/ml; Hameln) and 60 µl medetomidin (1 mg/ml; Pfizer), dosed in 3.1 µl/g body weight and injected i.p.. After loss of pain reflexes, the fur over the scalp was removed with hair removal cream, eye ointment was applied (Bepanthen, Bayer) and 1% xylocain (AstraZeneca) was injected under the scalp as preincisional local anesthesia. The mouse was then placed in a stereotaxic apparatus (David Kopf Instruments, model 963) equipped with a heating pad (37°C) to preserve body temperature. The dorsal cranium was exposed by removing the scalp and periosteum with fine forceps and scissors to prepare the mouse for M1 ablation or cranial window implantation surgeries. For post surgical care, mice received pain relief (Metacam, Boehringer Ingelheim) and antibiotic (Baytril, Bayer) s.c. injections (0.1 and 0.5 mg/ml respectively, dosed 10μl/g body weight).
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10

Alzet Pump Implantation for PEDF Delivery

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We primed micro-osmotic Alzet pumps (model 1007D) under sterile conditions one day before implantation and left them in an incubator at 37.0°C overnight. We anesthetized the animals with a ketamine (Ketavet, Pfizer, Germany, 150 mg/kg) and xylazine (Rompun, Bayer, Germany, 15 mg/kg) mixture dissolved in 8.5 ml of sterile 0.9% saline. Head skin incision was performed exposing the skull.
We implanted the cannula 0.2 mm to the right from Bregma corresponding to the location of the right ventricle. A drop of cyanoacrylate clay (Weicon, Germany) was introduced between the cannula cap and the skull. We slowly moved the cannula down until the cap touched the skull. We left the animals for 5 min to ensure drying of the clay. Then, we carefully sutured the skin above the cannula with a 5–0 polypropilen thread (Ethicon, USA).
Animals received lidocaine gel (Xylocain, AstraZeneca, Germany) locally and 0.5 ml of ringer lactate solution (B Braun Melsungen, Germany) intraperitoneally to substitute the liquid loss. We placed the cage with operated animals onto the 37°C warm bed for 2 hours.
The animals received a total amount of 84±16.8 µl of PEDF (20 µg/ml in CSF) or CSF at a pumping rate of 0.5±0.1 µl/hour.
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