Pub Date : 2023-01-01Epub Date: 2023-02-07DOI: 10.1007/978-1-0716-2918-5_5
Ina Ersing, Meghan Rego, Chen Wang, Yijun Zhang, Kate Harten DeMaio, Michelle Tillgren, Alanna Fava, Gabrielle Clouse, Marcella Patrick, Karen Guerin, Melina Fan
Adeno-associated viral vectors (AAV) are frequently used by neuroscientists to deliver tools, such as biosensors and optogenetic and chemogenetic actuators, in vivo. Despite its widespread use, AAV vector characterization and quality control can vary between labs and viral vector cores leading to variable results and irreproducibility. This protocol describes some of the characterization and quality control assays necessary to confirm an AAV vector's titer, genomic identity, serotype and purity.
{"title":"Quality control for Adeno-associated viral vector production.","authors":"Ina Ersing, Meghan Rego, Chen Wang, Yijun Zhang, Kate Harten DeMaio, Michelle Tillgren, Alanna Fava, Gabrielle Clouse, Marcella Patrick, Karen Guerin, Melina Fan","doi":"10.1007/978-1-0716-2918-5_5","DOIUrl":"https://doi.org/10.1007/978-1-0716-2918-5_5","url":null,"abstract":"<p><p>Adeno-associated viral vectors (AAV) are frequently used by neuroscientists to deliver tools, such as biosensors and optogenetic and chemogenetic actuators, <i>in vivo</i>. Despite its widespread use, AAV vector characterization and quality control can vary between labs and viral vector cores leading to variable results and irreproducibility. This protocol describes some of the characterization and quality control assays necessary to confirm an AAV vector's titer, genomic identity, serotype and purity.</p>","PeriodicalId":74285,"journal":{"name":"Neuromethods","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10997381/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140874130","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.1007/978-1-0716-2026-7_11
Yu Liu, Brittany L. Correia, Mia B. Fox, D. Heck
{"title":"Investigating Cerebrocerebellar Neuronal Interactions in Freely Moving Using Multi-electrode, Multi-site","authors":"Yu Liu, Brittany L. Correia, Mia B. Fox, D. Heck","doi":"10.1007/978-1-0716-2026-7_11","DOIUrl":"https://doi.org/10.1007/978-1-0716-2026-7_11","url":null,"abstract":"","PeriodicalId":74285,"journal":{"name":"Neuromethods","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"51711732","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.1007/978-1-0716-2026-7_8
K. Dorgans, B. Kuhn, M. Uusisaari
{"title":"In Vitro Voltage of Subthreshold Activity in Neurons with ANNINE-6plus","authors":"K. Dorgans, B. Kuhn, M. Uusisaari","doi":"10.1007/978-1-0716-2026-7_8","DOIUrl":"https://doi.org/10.1007/978-1-0716-2026-7_8","url":null,"abstract":"","PeriodicalId":74285,"journal":{"name":"Neuromethods","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"51712288","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.1007/978-1-0716-2083-0_1
Mario F. Juruena, A. Cleare, A. H. Young
{"title":"The Role of Hypothalamic-Pituitary-Adrenal Axis in the Pathophysiology of Major Depression: A Translational Research Perspective of the Prednisolone Suppression Test","authors":"Mario F. Juruena, A. Cleare, A. H. Young","doi":"10.1007/978-1-0716-2083-0_1","DOIUrl":"https://doi.org/10.1007/978-1-0716-2083-0_1","url":null,"abstract":"","PeriodicalId":74285,"journal":{"name":"Neuromethods","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"51712525","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.1007/978-1-0716-2022-9_15
Kirupa Suthakar, Lester Torres Cadenas, C. Weisz
{"title":"Strategies for Identification of Medial Olivocochlear Neurons for Patch-Clamp Studies of Synaptic Function Using Electrical Stimulation and Optogenetics","authors":"Kirupa Suthakar, Lester Torres Cadenas, C. Weisz","doi":"10.1007/978-1-0716-2022-9_15","DOIUrl":"https://doi.org/10.1007/978-1-0716-2022-9_15","url":null,"abstract":"","PeriodicalId":74285,"journal":{"name":"Neuromethods","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"51711305","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.1007/978-1-0716-2026-7_15
Robijanto Soetedjo, Y. Kojima
{"title":"Optogenetics in Complex Model Systems (Non-Human Primate)","authors":"Robijanto Soetedjo, Y. Kojima","doi":"10.1007/978-1-0716-2026-7_15","DOIUrl":"https://doi.org/10.1007/978-1-0716-2026-7_15","url":null,"abstract":"","PeriodicalId":74285,"journal":{"name":"Neuromethods","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"51711854","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.1007/978-1-0716-2083-0_10
B. Roy, P. Korla, Yogesh K. Dwivedi
{"title":"The Role of Transcriptional Profiling in Neurobiology and Treatment of Major Depression: A Translational Perspective on RNA Sequencing Platform","authors":"B. Roy, P. Korla, Yogesh K. Dwivedi","doi":"10.1007/978-1-0716-2083-0_10","DOIUrl":"https://doi.org/10.1007/978-1-0716-2083-0_10","url":null,"abstract":"","PeriodicalId":74285,"journal":{"name":"Neuromethods","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"51712729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.1007/978-1-0716-2083-0_18
P. Handschuh, M. Konadu, B. Spurny-Dworak, L. Silberbauer, M. Murgaš, R. Lanzenberger
{"title":"Serotonin Receptors and Antidepressants: Neuroimaging Findings from Preclinical and Clinical Research","authors":"P. Handschuh, M. Konadu, B. Spurny-Dworak, L. Silberbauer, M. Murgaš, R. Lanzenberger","doi":"10.1007/978-1-0716-2083-0_18","DOIUrl":"https://doi.org/10.1007/978-1-0716-2083-0_18","url":null,"abstract":"","PeriodicalId":74285,"journal":{"name":"Neuromethods","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"51713247","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.1007/978-1-0716-2083-0_19
Mu-Hong Chen, T. Su, S. Tsai
{"title":"Ketamine and Other Glutamate Receptor Antagonists As Fast-Actin","authors":"Mu-Hong Chen, T. Su, S. Tsai","doi":"10.1007/978-1-0716-2083-0_19","DOIUrl":"https://doi.org/10.1007/978-1-0716-2083-0_19","url":null,"abstract":"","PeriodicalId":74285,"journal":{"name":"Neuromethods","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"51713308","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.1007/978-1-0716-2525-5_5
Erika P Portero, Leena Pade, Jie Li, Sam B Choi, Peter Nemes
Molecular composition is intricately intertwined with cellular function, and elucidation of this relationship is essential for understanding life processes and developing next-generational therapeutics. Technological innovations in capillary electrophoresis (CE) and liquid chromatography (LC) mass spectrometry (MS) provide previously unavailable insights into cellular biochemistry by allowing for the unbiased detection and quantification of molecules with high specificity. This chapter presents our validated protocols integrating ultrasensitive MS with classical tools of cell, developmental, and neurobiology to assess the biological function of important biomolecules. We use CE and LC MS to measure hundreds of metabolites and thousands of proteins in single cells or limited populations of tissues in chordate embryos and mammalian neurons, revealing molecular heterogeneity between identified cells. By pairing microinjection and optical microscopy, we demonstrate cell lineage tracing and testing the roles the dysregulated molecules play in the formation and maintenance of cell heterogeneity and tissue specification in frog embryos (Xenopus laevis). Electrophysiology extends our workflows to characterizing neuronal activity in sections of mammalian brain tissues. The information obtained from these studies mutually strengthen chemistry and biology and highlight the importance of interdisciplinary research to advance basic knowledge and translational applications forward.
{"title":"Single-Cell Mass Spectrometry of Metabolites and Proteins for Systems and Functional Biology.","authors":"Erika P Portero, Leena Pade, Jie Li, Sam B Choi, Peter Nemes","doi":"10.1007/978-1-0716-2525-5_5","DOIUrl":"https://doi.org/10.1007/978-1-0716-2525-5_5","url":null,"abstract":"<p><p>Molecular composition is intricately intertwined with cellular function, and elucidation of this relationship is essential for understanding life processes and developing next-generational therapeutics. Technological innovations in capillary electrophoresis (CE) and liquid chromatography (LC) mass spectrometry (MS) provide previously unavailable insights into cellular biochemistry by allowing for the unbiased detection and quantification of molecules with high specificity. This chapter presents our validated protocols integrating ultrasensitive MS with classical tools of cell, developmental, and neurobiology to assess the biological function of important biomolecules. We use CE and LC MS to measure hundreds of metabolites and thousands of proteins in single cells or limited populations of tissues in chordate embryos and mammalian neurons, revealing molecular heterogeneity between identified cells. By pairing microinjection and optical microscopy, we demonstrate cell lineage tracing and testing the roles the dysregulated molecules play in the formation and maintenance of cell heterogeneity and tissue specification in frog embryos (<i>Xenopus laevis</i>). Electrophysiology extends our workflows to characterizing neuronal activity in sections of mammalian brain tissues. The information obtained from these studies mutually strengthen chemistry and biology and highlight the importance of interdisciplinary research to advance basic knowledge and translational applications forward.</p>","PeriodicalId":74285,"journal":{"name":"Neuromethods","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9872963/pdf/nihms-1864086.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10030291","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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