For measurement of brain dopamine turnover, rats were sacrificed either directly after winning an aggressive interaction in the intruder-paired compartment or after exposure to the intruder-unpaired compartment (Fig. 2A ). Winning was defined as the combination of an attack by the test subject and the submissive posture of the intruder rat after an aggressive interaction of at least five minutes. Rats were lightly anesthetized with isoflurane (Baxter AB, Sweden), and decapitated using a guillotine. Upon collection, brains were flash frozen in dry-ice cooled isopentane. Brains were sectioned into 60 µm coronal slices using a cryostat (Leica 3050 S; Leica Biosystems, Germany). Nucleus accumbens were identified using a brain atlas (Paxinos & Watson) (Fig. 2G ) and dissected bilaterally using disposable biopsy punches with plungers (INTEGRA, USA). All tissues were stored at −80 °C until further processing. Dopamine and its metabolites 3-methoxytyramine (3-MT), 3,4-dihydroxyphenylacetic acid (DOPAC), and homovanilic acid (HVA) were analyzed using high performance liquid chromatography (HPLC) as described previously [35 (link), 36 (link)].
Disposable biopsy punches with plungers
Manufactured by Integra LifeSciences
Sourced in United States
Disposable biopsy punches with plungers are single-use medical devices used to obtain tissue samples during diagnostic procedures. The core function of these punches is to excise a small, cylindrical section of tissue from the patient's skin or other accessible body part. The plunger mechanism assists in the controlled extraction of the tissue sample.
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2 protocols using disposable biopsy punches with plungers
1
Measurement of Brain Dopamine Turnover
2
GLP-1R Expression in Rat Brain Regions
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Brains were collected from 17-week-old rats, frozen, and placed in the cryostat. SuM, LH, nucleus accumbens (encompassing both the shell and the core regions), and NTS were collected using disposable biopsy punches with plungers (INTEGRA, USA) to assess GLP-1R expression using quantitative real-time PCR (qPCR). RNA was isolated using the RNeasy Lipid Tissue Mini Kit (Quiagen, Germany) and GLP-1R levels were quantified using Taqman gene expression kits from Life Technologies using beta actin as the housekeeping gene (Primer information: GLP-1R: Rn00562406_m1; beta actin: RN00667869_m1). Comparative threshold cycle method [31] (link) was used to quantify relative mRNA expression. For both sexes SuM GLP-1R expression relative to beta actin was set as 1 to visualize the expression in SuM relative to other brain regions.
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