For sessions 1–8, rats were trained and tested in the punishment task. Punishment sessions were identical to acquisition sessions, except that a designated lever was also punished with a 0.5 sec, 0.5 mA footshock on an FR-10 schedule. The same lever (left or right) was designated as “punished” throughout the experiment for each rat (whether left or right was punished was counterbalanced between rats). Immediately before the first 2 d of punishment, rats received bilateral infusions of 0.9% phosphate-buffered saline or of the GABA agonists baclofen and muscimol (BM; 1 mM baclofen, 0.1 mM muscimol; Sigma–Aldrich) to assess the role of the target region in the acquisition of punishment. For microinjections, a 33-gauge microinjection cannula (Plastics One) was inserted into the guide cannula and connected to a 10-µL glass syringe (Hamilton Company) operated by an infusion pump (World Precision Instruments). The microinjection cannula projected a further 1 mm ventral to the tip of the guide cannula. Drugs were infused at a rate of 0.25 µL/min over 2 min, and the microinjection cannula was left in place for a further 1 min to permit diffusion of the injectate. Rats also received bilateral infusions of either saline or BM on days 6 and 7 (counterbalanced within subject) to test for the effect of PFC region inactivation on expression of punishment.
10 l glass syringe
Manufactured by Hamilton Company
Sourced in United States
The 10 µL glass syringe is a precision laboratory instrument designed for accurate volumetric measurements and fluid handling. It features a glass barrel and a plunger, allowing for precise control and delivery of small liquid volumes.
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4 protocols using 10 l glass syringe
1
Punishment Task in Rats: GABA Modulation
2
Induction of Ocular Inflammation in Mice
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Local, intravitreal injection of 1 µg LPS (Salmonella typhimurium; Sigma) was used to induce ocular inflammation as previously described.22 (link),36 (link) Mice were anesthetized via intraperitoneal injection of ketamine (100 mg/kg)/xylazine (10 mg/kg). Each eye was then injected with either 1 µL LPS diluted in sterile PBS vehicle or PBS alone using a 10 µL glass syringe (Hamilton Company, Reno, NV, USA).
3
Punishment Task Acquisition and Expression
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On Days 1–8, rats were trained and tested in the punishment task. Punishment sessions were identical to acquisition sessions, except that a designated lever was also punished with a 0.5 sec, 0.5-mA footshock on an FR-10 schedule. The same lever (left or right) was designated as “punished” throughout the experiment for each rat but which lever was designated as punished was counterbalanced between rats. Immediately before the first 2 d of Phase I, rats received bilateral infusions of 0.9% phosphate-buffered saline or of the GABA agonists baclofen and muscimol (BM; 1 mM baclofen, 0.1 mM muscimol; Sigma-Aldrich) to assess the role of BLA in the acquisition of punishment. For microinjections, a 33-gauge microinjection cannula (Plastics One) was inserted into the guide cannula and connected to a 10 µL glass syringe (Hamilton Company) operated by an infusion pump (World Precision Instruments). The microinjection cannula projected a further 1 mm ventral to the tip of the guide cannula. Drugs were infused at a rate of 0.25 µL/min over 2 min, and the microinjection cannula was left in place for a further 1 min to permit diffusion of the injectate. Rats also received bilateral infusions of either saline or BM on Days 6 and 7 (counterbalanced, within subject) to test for the effect of BLA inactivation on expression of punishment.
4
Footshock Punishment in Rats: LHb Role
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On Days 1–5, rats were trained and tested in the punishment task. Punishment sessions were identical to acquisition sessions, except that a designated lever was also punished with a 0.5 s, 0.5 mA footshock on an FR-10 schedule. The same lever (left or right) was designated as “punished” throughout the experiment for each rat but which lever was designated as punished was counterbalanced between rats. Immediately before the first 2 days of punishment, rats received bilateral infusions of 0.9% phosphate-buffered saline or of the AMPA antagonist NBQX (1 µg/µl; Sigma-Aldrich, Sydney, Australia) to assess the role of LHb in the acquisition of punishment. This dose of NBQX is sufficient to prevent aversion-related signals exciting the LHb [16] (link) and has also yielded behaviourally significant yet specific effects in previous experiments [17] (link). For microinjections, a 33-gauge microinjection cannula (Plastics One, Roanoke, VA, USA) was inserted into the guide cannula and connected to a 10 µl glass syringe (Hamilton Company, NV, USA) operated by an infusion pump (World Precision Instruments, FL, USA). The microinjection cannula projected a further 1 mm ventral to the tip of the guide cannula. Drugs were infused at a rate of 0.25 µl/min over 2 min, and the microinjection cannula was left in place for a further 1 min to permit diffusion of the injectate.
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