Using the Parafilm-assisted Microdissection (PAM) Method to Sample Rodent Nucleus Accumbens

Autor(es)

Pagliusi Jr, Marco ; Brandao, Arthur F. ^[1] ; Zanetti, Gabriel G. ^[1] ; Bonet, Ivan J. M. ^[2] ; Sartori, Cesar R. ^[1] ; Vieira, Andre S. ^[1]

Afiliação do(s) autor(es)

^[1] Pagliusi Jr, Jr., Marco, Univ Estadual Campinas, Dept Struct & Funct Biol, Campinas - Brazil

^[2] Univ Calif San Francisco, Dept Oral & Maxillofacial Surg, San Francisco, CA 94143 - USA

• Tipo de documento: Artigo Científico
• Fonte: BIO-PROTOCOL; v. 10, n. 23 DEC 5 2020.
• Citações Web of Science: 0

Resumo

Microdissection techniques are very important for anatomical and functional studies focused on neuroscience, where it is often necessary microdissect specific brain areas to perform molecular or anatomical analyses. The parafile-assisted microdissection (PAM) was previously described and involves the microdissection of tissue sections mounted on parafilm-covered glass slides. In this work, we describe the use of the PAM method to microdissect rodent nucleus accumbens (NAc). (1) We first describe the best way to perform the mouse euthanasia and how to remove the brain. (2) Next, we describe how to prepare the slides with parafilm (R) that will be used to receive the brain slices. (3) Following, we describe how to handle the brain in the cryostat, how to align the hemispheres and how to identify the NAc antero-posterior limits. (4) We also describe how to perform the staining and dehydration of the slices, a critical step to facilitate the microdissection and preserve macromolecules. (5) In the final step, we describe how to identify the dorso-ventral and latero-medial limits of the NAc and, finally, how to perform the manual microdissection of the area. This is a low-cost technique that allows the researcher to specifically microdissect any brain region, from which intact RNA and proteins can be extracted to perform several molecular analyses (e.g., real-time PCR, Western blot, and RNA-seq). (AU)

• Processo FAPESP: 13/09749-4 - Estudo do efeito do Estresse por Submissão Social sobre o comportamento nociceptivo de camundongos e mecanismos neurobiológicos associados.
• Beneficiário: Marco Oreste Finocchio Pagliusi Junior
• Linha de fomento: Bolsas no Brasil - Mestrado
• Processo FAPESP: 15/26777-7 - Estudo dos efeitos preventivo e terapêutico do exercício físico sobre a hiperalgesia induzida por estresse por subjugação social em camundongos e mecanismos neuroplásticos associados.
• Beneficiário: Cesar Renato Sartori
• Linha de fomento: Auxílio à Pesquisa - Regular