The L5 vertebra of rats was laminectomized under adequate anesthesia with sevoflurane (2.5%–5.0% inhalation administration, in air), and a 7-cm soft tube (SILASTIC laboratory tubing, Dow Corning Corporation, Auburn, MI, USA; outer diameter, 0.64 mm) filled with saline was inserted into the subarachnoid space at a length of ∼0.5 cm. After the muscle incision was closed, the tube was laid under the skin and the cut end was ligated. Then, the incision was closed. Ten microliters of group III mGluR agonist L-(+)-2-amino-4-phosphonobutyric acid (L-AP4) (Tocris Bioscience) and group III mGluR antagonist (RS)-α-cyclopropyl-4-phosphonophenylglycine (CPPG) (Tocris Bioscience) were carefully administered, followed by 5 µl of saline. According to previous studies,19 (link),26 (link) the doses of L-AP4 and CPPG were 10 and 50 nmol/µl diluted in phosphate-buffered saline (PBS), respectively. One dose of each drug was given 5 min prior to formalin injection. Rats were anesthetized, and drugs were administered using a Hamilton syringe.
Protocol detail
Intrathecal Injection of mGluR Agonist and Antagonist in Rats
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SILASTIC laboratory tubing
L-(+)-2-amino-4-phosphonobutyric acid (L-AP4)
RS)-α-cyclopropyl-4-phosphonophenylglycine (CPPG)
2 amino 4 phosphonobutyric acid
Anesthesia
Cyclopropyl 4 phosphonophenylglycine
Drugs
Formalin
Inhalation administration
L ap4
Muscle incision
Phosphate
Rats
Saline
Sevoflurane
Silastic
Skin
Subarachnoid space
Syringe
Vertebra
Corresponding Organization : Hyogo Medical University
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Variable analysis
independent variables
- Dose of group III mGluR agonist L-(+)-2-amino-4-phosphonobutyric acid (L-AP4)
- Dose of group III mGluR antagonist (RS)-α-cyclopropyl-4-phosphonophenylglycine (CPPG)
- Not explicitly mentioned
- Anesthesia with sevoflurane (2.5%–5.0% inhalation administration, in air)
- Length of the SILASTIC laboratory tubing (7-cm soft tube, outer diameter 0.64 mm) inserted into the subarachnoid space (∼0.5 cm)
- Timing of drug administration (one dose of each drug given 5 min prior to formalin injection)
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