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

Autor:	Pagliusi M Jr; Department of Structural and Functional Biology, University of Campinas, Campinas, Brazil., Brandão AF; Department of Structural and Functional Biology, University of Campinas, Campinas, Brazil., Zanetti GG; Department of Structural and Functional Biology, University of Campinas, Campinas, Brazil., Bonet IJM; Department of Structural and Functional Biology, University of Campinas, Campinas, Brazil., Sartori CR; Department of Structural and Functional Biology, University of Campinas, Campinas, Brazil., Vieira AS; Department of Structural and Functional Biology, University of Campinas, Campinas, Brazil.
Jazyk:	angličtina
Zdroj:	Bio-protocol [Bio Protoc] 2020 Dec 05; Vol. 10 (23), pp. e3836. Date of Electronic Publication: 2020 Dec 05 (Print Publication: 2020).
DOI:	10.21769/BioProtoc.3836
Abstrakt:	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 parafilm ® -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 ® 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). Competing Interests: Competing interestsWe declare no competing interests. (Copyright © 2020 The Authors; exclusive licensee Bio-protocol LLC.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu