دورية أكاديمية
أعرض في EDS

Modulating signalling lifetime to optimise a prototypical animal opsin for optogenetic applications.

التفاصيل البيبلوغرافية
العنوان: Modulating signalling lifetime to optimise a prototypical animal opsin for optogenetic applications.
المؤلفون: Rodgers J; Centre for Biological Timing, Division of Neuroscience, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, M13 9PT, UK. jessica.rodgers@manchester.ac.uk., Wright P; Centre for Biological Timing, Division of Neuroscience, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, M13 9PT, UK., Ballister ER; Centre for Biological Timing, Division of Neuroscience, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, M13 9PT, UK.; Department of Biomedical Engineering, Columbia University, New York, NY, 10027, USA.; Department of Microbiology and Immunology, Vagelos College of Physicians and Surgeons of Columbia University, New York, 10032, NY, USA., Hughes RB; Centre for Biological Timing, Division of Neuroscience, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, M13 9PT, UK., Storchi R; Centre for Biological Timing, Division of Neuroscience, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, M13 9PT, UK., Wynne J; Centre for Biological Timing, Division of Neuroscience, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, M13 9PT, UK., Martial FP; Centre for Biological Timing, Division of Neuroscience, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, M13 9PT, UK., Lucas RJ; Centre for Biological Timing, Division of Neuroscience, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, M13 9PT, UK. robert.lucas@manchester.ac.uk.
المصدر: Pflugers Archiv : European journal of physiology [Pflugers Arch] 2023 Dec; Vol. 475 (12), pp. 1387-1407. Date of Electronic Publication: 2023 Dec 01.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Springer Country of Publication: Germany NLM ID: 0154720 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1432-2013 (Electronic) Linking ISSN: 00316768 NLM ISO Abbreviation: Pflugers Arch Subsets: MEDLINE
أسماء مطبوعة: Original Publication: Berlin, New York, Springer.
مواضيع طبية MeSH: Rod Opsins*/genetics , Rod Opsins*/metabolism , Opsins*/genetics , Opsins*/metabolism, Animals ; Mice ; Humans ; Optogenetics/methods ; HEK293 Cells ; Arrestins/genetics ; Arrestins/metabolism
مستخلص: Animal opsins are light activated G-protein-coupled receptors, capable of optogenetic control of G-protein signalling for research or therapeutic applications. Animal opsins offer excellent photosensitivity, but their temporal resolution can be limited by long photoresponse duration when expressed outside their native cellular environment. Here, we explore methods for addressing this limitation for a prototypical animal opsin (human rod opsin) in HEK293T cells. We find that the application of the canonical rhodopsin kinase (GRK1)/visual arrestin signal termination mechanism to this problem is complicated by a generalised suppressive effect of GRK1 expression. This attenuation can be overcome using phosphorylation-independent mutants of arrestin, especially when these are tethered to the opsin protein. We further show that point mutations targeting the Schiff base stability of the opsin can also reduce signalling lifetime. Finally, we apply one such mutation (E122Q) to improve the temporal fidelity of restored visual responses following ectopic opsin expression in the inner retina of a mouse model of retinal degeneration (rd1). Our results reveal that these two strategies (targeting either arrestin binding or Schiff-base hydrolysis) can produce more time-delimited opsin signalling under heterologous expression and establish the potential of this approach to improve optogenetic performance.
(© 2023. The Author(s).)
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معلومات مُعتمدة: United Kingdom WT_ Wellcome Trust
فهرسة مساهمة: Keywords: Arrestin; Deactivation; G-protein; GPCR; Kinetics; Opsin; Optogenetics; Rhodopsin
المشرفين على المادة: 0 (Rod Opsins)
0 (Opsins)
0 (Arrestins)
تواريخ الأحداث: Date Created: 20231130 Date Completed: 20231221 Latest Revision: 20231222
رمز التحديث: 20231222
مُعرف محوري في PubMed: PMC10730688
DOI: 10.1007/s00424-023-02879-9
PMID: 38036775
قاعدة البيانات: MEDLINE
الوصف
تدمد:1432-2013
DOI:10.1007/s00424-023-02879-9