title: Evaluation of voltage-sensitive fluorescence dyes for monitoring neuronal activity in the embryonic central nervous system.
abstract: Using an optical imaging technique with voltage-sensitive dyes (VSDs), we investigated the functional organization and architecture of the central nervous system (CNS) during embryogenesis. In the embryonic nervous system, a merocyanine-rhodanine dye, NK2761, has proved to be the most useful absorption dye for detecting neuronal activity because of its high signal-to-noise ratio (S/N), low toxicity and small dye bleaching. In the present study, we evaluated the suitability of fluorescence VSDs for optical recording in the embryonic CNS. We screened eight styryl (hemicyanine) dyes in isolated brainstem-spinal cord preparations from 7-day-old chick embryos. Measurements of voltage-related optical signals were made using a multiple-site optical recording system. The signal size, S/N, photobleaching, effects of perfusion and recovery of neural responses after staining were compared. We also evaluated optical responses with various magnifications. Although the S/N was lower than with the absorption dye, clear optical responses were detected with several fluorescence dyes, including di-2-ANEPEQ, di-4-ANEPPS, di-3-ANEPPDHQ, di-4-AN(F)EPPTEA, di-2-AN(F)EPPTEA and di-2-ANEPPTEA. Di-2-ANEPEQ showed the largest S/N, whereas its photobleaching was faster and the recovery of neural responses after staining was slower. Di-4-ANEPPS and di-3-ANEPPDHQ also exhibited a large S/N but required a relatively long time for recovery of neural activity. Di-4-AN(F)EPPTEA, di-2-AN(F)EPPTEA and di-2-ANEPPTEA showed smaller S/Ns than di-2-ANEPEQ, di-4-ANEPPS and di-3-ANEPPDHQ; but the recovery of neural responses after staining was faster. This study demonstrates the potential utility of these styryl dyes in optical monitoring of voltage changes in the embryonic CNS.
authors: Department of Health and Nutrition Sciences, Faculty of Human Health, Komazawa Women's University, 238 Sakahama, Inagi-shi, Tokyo, 206-8511, Japan.
authors: Saad
authors: S
authors: Habib-E-Rasul Mullah
authors: Ryo
authors: R
authors: Komuro
authors: Ping
authors: P
authors: Yan
authors: Shihori
authors: S
authors: Hayashi
authors: Motoki
authors: M
authors: Inaji
authors: Yoko
authors: Y
authors: Momose-Sato
authors: Leslie M
authors: LM
authors: Loew
authors: Katsushige
authors: K
authors: Sato
mesh_terms: D000042:Absorption; D000818:Animals; D001578:Benzopyrans / Q000737:chemistry / Q000378:metabolism; D002642:Chick Embryo; D004558:Electric Stimulation; D004563:Electrochemistry; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D007211:Indoles / Q000737:chemistry / Q000378:metabolism; D008564:Membrane Potentials; D061848:Optical Imaging; D011726:Pyridinium Compounds / Q000737:chemistry / Q000378:metabolism; D013343:Styrenes / Q000737:chemistry / Q000378:metabolism
chemical_list: D001578:Benzopyrans; D005456:Fluorescent Dyes; D007211:Indoles; D011726:Pyridinium Compounds; C096453:RH 795; D013343:Styrenes; C054158:RH 414
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