A small hole was pricked into the cornea of the median ocellus to allow the insertion of a 10 µl-syringe (World Precision Instrument), which was used to inject 200 nl of each drug solution21 (link). Drugs were injected into the brain of immobilized bees along the median ocellar nerve21 (link). This method ensures that drugs migrate through the ocellar tract and distribute within the bee brain in a fast (less than 5 min) and homogenous way76 (link). Thirty min before the experiment, we injected: Epinastine hydrochloride, an OA receptor antagonist46 (link), cis-(Z)-flupentixol dihydrochloride, a DA receptor antagonist47 (link); or PBS (control). Epinastine and flupentixol were obtained from Sigma-Aldrich (Saint-Quentin Fallavier, France). PBS was obtained from EUROMEDEX (Strasbourg, France). Injection time was chosen based on experiments, which showed that the effects of aminergic blockers reach a stable level approximately 30 min after drug application35 (link),77 (link)–79 (link). One mg of each drug was diluted in 1 ml PBS. Final concentrations obtained were 3.5 mM of Epinastine and 1.94 mM of flupentixol. To test for dose-response effects, we prepared for each drug an additional dilution of 1:10000. In all cases, aliquots were kept in −20 °C until use. Each aliquot was used for one whole week and kept during this time in 4 °C.
10 l syringe
Manufactured by World Precision Instruments
The 10 µl-syringe is a precision laboratory instrument designed for accurate and repeatable liquid handling. It features a maximum capacity of 10 microliters (μl) and is constructed with high-quality materials for reliable performance.
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2 protocols using 10 l syringe
1
Intracranial Injection of Aminergic Antagonists in Bees
2
Bicuculline Injection for Post-Stroke Exercise
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Two weeks before MCAO surgery, a burr hole was drilled in the left dorsal striatum in the EX+Vehicle and EX+BICU groups (n=7/group). The cannula was inserted into the dorsal striatum and xed with dental cement. For subsequent pharmaceutical injection, Bicuculline (35 mM, 250 nl, Selleck) or vehicle (14% DMSO + 3% PEG 400 + 3% Tween 80 + 80% ddH 2 O) was infused into a 5 cm-long PE50 catheter connected to a 10 µl syringe (World Precision Instruments) with a glass needle, and then a PE50 catheter was inserted into the cannula in the EX+BICU and EX+Vehicle groups, respectively. Bicuculline was injected at 30 min after exercise training at a ow rate of 25 nL/min. Bicuculline injection was performed starting on the 3 rd and ended on the 14 th day after stroke.
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