Two kinds of ACSF were used in in vitro experiments: mACSF in the dissection dish before recording and nACSF in the recording chamber. mACSF was composed of (in mM) 118 NaCl, 24 NaHCO3, 1.5 CaCl2, 3 KCl, 5 MgCl2, 1.4 NaH2PO4, 1.3 MgSO4, 25 d -glucose, and 1 kynurenic acid. nACSF was composed of (in mM) 122 NaCl, 25 NaHCO3, 2.5 CaCl2, 3 KCl, 1 MgCl2, 0.5 NaH2PO4, and 12 d -glucose. Both types of ACSF were saturated with carbogen (95% O2-5% CO2) and maintained at pH 7.4. The drugs added to the nACSF were 5-HTP, clorgyline, pargyline, tryptamine, L-tryptophan, tyramine, L-tyrosine, 2-phenylethylamine, L-phenyalanine, NSD1015 (Sigma-Aldrich), α-methyl-5-HT, octopamine, SB206553, RX821002, GR127935, dobutamine, vasopressin (Tocris) and zolmitriptan (AstraZenica). All drugs were first dissolved as a 10 – 50 mM stock solution in water before final dilution in ACSF for in vitro ventral root reflexes recording and DIC microscopy or in saline for in vivo oxygen measurements and two photon microscopy, with the exception of zolmitriptan, which was dissolved in minimal amounts of DMSO (DMSO final concentration in nACSF was 0.04%). DMSO alone had no effect on in vitro LLR in vehicle controls, as compare to nACSF control state.
Gr127935
Manufactured by Bio-Techne
Sourced in Macao, United States
GR127935 is a laboratory instrument designed for general experimental and analytical purposes. It is a multi-functional device capable of performing various tasks in a research setting. The core function of this product is to provide a platform for conducting scientific experiments and analyses. More detailed information about its specific features and intended applications is not available at this time.
Automatically generated - may contain errors
4 protocols using gr127935
1
Detailed ACSF compositions for in vitro experiments
2
Pharmacological Dissection of Dural Stimulation-Evoked Neuronal Responses
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The effects of CSD induction on dural stimulation-evoked neuronal responses in
the TCC were dissected pharmacologically. All drugs for intravenous injection
were dissolved in saline and dosed at a volume 1 ml/kg. The selective
5-HT1B/1D receptor antagonist, GR127935 (21 (link)) (Tocris; Ellisville, MO) was
injected at a dose of 3 mg kg−1. Naloxone hydrochloride, an opioid
receptor antagonist (Tocris; Ellisville, MO), was injected at a dose of
1.5 mg kg−1.
the TCC were dissected pharmacologically. All drugs for intravenous injection
were dissolved in saline and dosed at a volume 1 ml/kg. The selective
5-HT1B/1D receptor antagonist, GR127935 (21 (link)) (Tocris; Ellisville, MO) was
injected at a dose of 3 mg kg−1. Naloxone hydrochloride, an opioid
receptor antagonist (Tocris; Ellisville, MO), was injected at a dose of
1.5 mg kg−1.
3
Detailed ACSF compositions for in vitro experiments
4
Antagonist GR127935 and ZJW Preparation
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The 5-HTR1D antagonist GR127935 (GR) (TOCRIS, Ellisville, USA) was dissolved in phosphate buffer saline (PBS) and then diluted into different concentrations. ZJW was prepared using a formula of Chinese herb Rhizoma Coptidis (60 g) and Evodia rutaecarpa (10 g), at a ratio of 6:1. All the herbs were purchased from Longhua Hospital herbal pharmacy department. Briefly, the mixture (70 g) was extracted twice for 1 h each time by refluxing in ethanol (1: 8, v/v). The filtrates were concentrated and dried in vacuum at 60 °C. Its preparations were standardized, regulated, and quality controlled according to the guidelines defined by China Food and Drug Administration (CFDA). High-performance liquid chromatography (HPLC) was used to identify the components of ZJW extract, and confirm the final concentration of ZJW extract to ensure the quality and stability. Detailed procedures were followed with the published protocol [6 (link)]. ZJW was dissolved in PBS and diluted into three different concentrations, 25, 50 and 100 μg/mL, corresponding to low, medium and high dose of ZJW (ZJW-L, ZJW-M and ZJW-H).
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