title: A new and reliable guide for studies of neuronal loss based on focal lesions and combinations of in vivo and in vitro approaches.
abstract: In this study, we describe a simple and reliable method to study neuroprotective effects in living and organized neural tissue. This method, which was based on retinal explants for in vivo focal lesions, was conceived as a collection of modular procedures, which can be customized for particular demands. With this model, it is possible to combine immunohistochemistry with image data analysis to track the two- or three-dimensional redistribution of proteins as a time/space function of primary cell loss. At the same time, it is possible to finely control the exposure of the tissue to specific drugs and molecules. In order to illustrate the use of the proposed method, we tested the effects of two different nanotube compounds on retinal explant viability. Transcriptome analyses can be separately performed in the lesion focus and penumbra with laser capture microdissection followed by polymerase chain reaction analyses. In addition, other common experimental drawbacks, such as high individual variance, are eliminated. With intraocular injections, treatments can be verified in vivo, with one eye serving as the experimental tissue and the other serving as the control tissue. In summary, we describe a flexible and easy method, which can be useful in combination with a broad variety of recently developed neuroprotective strategies, to study neurodegeneration.
authors.affiliation: Núcleo de Cognição e Sistemas Complexos, Centro de Matemática, Computação e Cognição, Universidade Federal do ABC, Santo André, São Paulo, Brasil.
authors.forename: Vera
authors.initials: V
authors.lastname: Paschon
authors.forename: Guilherme Shigueto Vilar
authors.initials: GS
authors.lastname: Higa
authors.forename: Lais Takata
authors.initials: LT
authors.lastname: Walter
authors.forename: Erica
authors.initials: E
authors.lastname: Sousa
authors.forename: Erica
authors.initials: E
authors.lastname: de Sousa
authors.forename: Fausto Colla Cortesão
authors.initials: FC
authors.lastname: Zuzarte
authors.forename: Vivian Roca Schwendler
authors.initials: VR
authors.lastname: Weber
authors.forename: Rodrigo Ribeiro
authors.initials: RR
authors.lastname: Resende
authors.forename: Alexandre Hiroaki
authors.initials: AH
authors.lastname: Kihara
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000494:pharmacology; D052158:Aptamers, Peptide / Q000494:pharmacology; D002645:Chickens; D005136:Eye Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D020869:Gene Expression Profiling; D056965:Injections, Intraocular; D008297:Male; D057054:Molecular Imaging; D043942:Nanotubes; D018696:Neuroprotective Agents / Q000494:pharmacology*; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D012160:Retina / Q000166:cytology* / Q000187:drug effects / Q000293:injuries / Q000378:metabolism; D055351:Retinal Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D059010:Single-Cell Analysis; D046509:Tissue Culture Techniques
chemical_list: D052157:Aptamers, Nucleotide; D052158:Aptamers, Peptide; D005136:Eye Proteins; D018696:Neuroprotective Agents; D034741:RNA, Small Interfering