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. 2021 Dec 15;12:7302. doi: 10.1038/s41467-021-27464-5

Fig. 8. Silk-VM organoids are functionally homogeneous.

Fig. 8

a Representative images of functional recordings from the external part using whole-cell patch-clamp technique. Scale bars, 100 µm. b Representative inward sodium- and outward potassium-rectifying current trace of external VM organoid at day 90 triggered by stepwise depolarization. c Whole-cell patch-clamp recordings of external VM organoid cells depicting current-induced APs at day 90 (−85 pA to +165 pA with 20 pA steps). d Spontaneous firings at resting membrane potential indicative of mature DA neuronal physiology in silk-lam VM organoids in the external part at day 90. e Example trace of rebound depolarization after brief membrane depolarization (20 pA) indicative of DA phenotype in externally located cells. f Resting-membrane quantifications between externally (n = 20) and internally localized cells (n = 20) in VM organoids at day 90. Data represent mean ± SD. g Representative images of functional recordings from the internal region of organoid using whole-cell patch-clamp technique. Scale bars, 100 µm. h Representative internal inward sodium- and outward potassium-rectifying current trace of VM organoid at day 90 triggered by stepwise depolarization i, Whole-cell patch-clamp recordings in internal region of VM organoid cells depicting current-induced APs at day 90 (−85 pA to +165 pA with 20 pA steps). j Spontaneous firings at resting-membrane potential indicative of mature DA neuronal physiology in the internal region of silk-lam VM organoids at day 90. k Example trace of rebound depolarization after brief membrane depolarization (20 pA) indicative of DA phenotype in internally located cells. l Quantification of maximum inward sodium current recorded in internal (n = 16 cells) and external (n = 17 cells) regions. Data represent mean ± SD. m Differential fluorescence-intensity profile of intracellular Ca+ levels as a function of time in neurons expressing MAP2–GCamP5 at day 90. n Fluorescence image with marked regions of interest corresponding to recorded cells and three timeframes displaying the change in intracellular fluorescence intensity. Scale bar, 100 µm. o, p Representative analysis of real-time DA release chronoamperometric measurements in conventional and silk-lam VM organoids and q, relative quantification (n = 12).