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. Author manuscript; available in PMC: 2022 Aug 19.
Published in final edited form as: Cell. 2021 Jul 26;184(17):4547–4563.e17. doi: 10.1016/j.cell.2021.07.003

Figure 7. Tau-V337M susceptibility to glutamate excitotoxicity is reversed by antagonists of excitatory receptors and PIKFYVE inhibition.

Figure 7.

(A) Longitudinal imaging method for tracking neuronal survival in cerebral organoids.

(B) Survival of SYN1::GFP+ neurons in 4-month tau-V337M and isogenic V337V organoids without glutamate treatment. n = 80 neurons from 5 organoids per group. Log rank test. ns, not significant

(C) Images of 4-month tau-V337M and isogenic V337V organoids treated with 5 mM glutamate. Neurons labeled with a lentivirus encoding SYN1::GFP. Scale bars, 50 μm.

(D) Survival of SYN1::GFP+ neurons in tau-V337M and isogenic V337V 4-month organoids with glutamate treatment. n = 100 neurons from 5 individual organoids per group. Log rank test. ****p < 0.0001.

(E) Percentage of surviving neurons following 7 days of glutamate treatment from 3- to 4-month organoids. Each point represents an independent experiment among three isogenic pairs. Two-tailed unpaired t test, *p = 0.0436.

(F) Schematic of the proposed mode of action for apilimod and effect on neuronal vulnerability to excitotoxic stress.

(G) Survival of SYN1::GFP+ neurons in tau-V337M organoids with glutamate treatment and DMSO. 3i, 10 μM CNQX + 10 μM MK-801 + 2 mM nimodipine or 10 μM apilimod. n = 120 neurons from 5 individual organoids per group. Log rank test, **p < 0.01 ****p < 0.0001.

(H) Images of 4-month tau-V337M organoids treated with 5 mM glutamate and DMSO. 3i, 10 μM CNQX + 10 μM MK-801 + 2 μM nimodipine or 10 μM apilimod. Neurons were labeled with SYN1::GFP. Scale bars, 50 μm.

(I) Relative PIKFYVE expression in ND03231 organoids by qRT-PCR following treatment with a PIKFYVE ASO, normalized to 18S expression. Bars represent mean expression ± SEM with n = 6 organoids per group. Two-tailed unpaired t test, **p < 0.01.

(J and K) Survival of SYN1::GFP+ neurons in 4-month tau-V337M (J) and tau-V337V (K) organoids with glutamate and 10 μM negative control (NC) or PIKFYVE ASO treatment. n = 120 neurons from 5 individual organoids per group. Log rank test, ***p = 0.001, ****p < 0.0001.

See also Figure S7.