Figure 6.
AcD neurons have less inhibitory synapses at the AIS. (A) Representative images of DIV14 hippocampal neurons stained with GAD1 for inhibitory interneurons (top row), and pCaMKII (bottom row) for excitatory neurons. Neuronal morphology is highlighted by MAP2. Red arrowhead indicates GAD1 positive neurons (top row) and pCaMKII negative neurons (bottom row). Scale bar is 50 µm. (B) Quantification of GAD1 positive (GAD1+) and pCaMKII negative (pCaMKII−) neuron percentage in DIV14 dissociated hippocampal cultures. Mean ± SEM, GAD1: n = 3 independent preparations, pCaMKII: n = 6 independent preparations. (C) Top row: Representative images of nonAcD (left) and AcD (right) neurons co-immunostained with markers for pre- and postsynaptic compartments of an inhibitory synapse (VGAT and gephyrin, respectively), for AIS (TRIM46), and for somato-dendritic compartment (MAP2). Scale bar is 10 µm. White dashed rectangle indicates the AIS. Bottom row: corresponding zoom-ins of white dashed rectangular in upper row. Scale bar is 5 µm. White arrowheads indicate co-localized gephyrin and VGAT clusters. Related to Video 10. (D) Number of single gephyrin clusters per 5 μm at the AIS of nonAcD and AcD neurons. Mean ± SEM, seven independent cultures, nonAcD: n = 111 cells, AcD: n = 97 cells. Grey dot indicates the value of individual cell. Orange and cyan triangle indicate mean of each independent culture. (E) Number of single VGAT clusters per 5 μm at the AIS of nonAcD and AcD neurons. Mean ± SEM, seven independent cultures, nonAcD: n = 102 cells, AcD: n = 104 cells. Grey dot indicates value of individual cell. Orange and cyan triangle indicate mean of each independent culture. (F) Number of VGAT clusters co-localize with gephyrin per 5 μm at the AIS of nonAcD and AcD neurons. Mean ± SEM, three independent cultures, nonAcD: n = 34 cells, AcD: n = 32 cells. Grey dot indicates value of individual cell. Orange and cyan triangle indicate mean of each independent culture. Mann–Whitney test: not significant (two-sided): not significant (n.s.) P > 0.05, *P < 0.05, ***P < 0.0001. More detailed statistical information see Data S1.

AcD neurons have less inhibitory synapses at the AIS. (A) Representative images of DIV14 hippocampal neurons stained with GAD1 for inhibitory interneurons (top row), and pCaMKII (bottom row) for excitatory neurons. Neuronal morphology is highlighted by MAP2. Red arrowhead indicates GAD1 positive neurons (top row) and pCaMKII negative neurons (bottom row). Scale bar is 50 µm. (B) Quantification of GAD1 positive (GAD1+) and pCaMKII negative (pCaMKII−) neuron percentage in DIV14 dissociated hippocampal cultures. Mean ± SEM, GAD1: n = 3 independent preparations, pCaMKII: n = 6 independent preparations. (C) Top row: Representative images of nonAcD (left) and AcD (right) neurons co-immunostained with markers for pre- and postsynaptic compartments of an inhibitory synapse (VGAT and gephyrin, respectively), for AIS (TRIM46), and for somato-dendritic compartment (MAP2). Scale bar is 10 µm. White dashed rectangle indicates the AIS. Bottom row: corresponding zoom-ins of white dashed rectangular in upper row. Scale bar is 5 µm. White arrowheads indicate co-localized gephyrin and VGAT clusters. Related to Video 10. (D) Number of single gephyrin clusters per 5 μm at the AIS of nonAcD and AcD neurons. Mean ± SEM, seven independent cultures, nonAcD: n = 111 cells, AcD: n = 97 cells. Grey dot indicates the value of individual cell. Orange and cyan triangle indicate mean of each independent culture. (E) Number of single VGAT clusters per 5 μm at the AIS of nonAcD and AcD neurons. Mean ± SEM, seven independent cultures, nonAcD: n = 102 cells, AcD: n = 104 cells. Grey dot indicates value of individual cell. Orange and cyan triangle indicate mean of each independent culture. (F) Number of VGAT clusters co-localize with gephyrin per 5 μm at the AIS of nonAcD and AcD neurons. Mean ± SEM, three independent cultures, nonAcD: n = 34 cells, AcD: n = 32 cells. Grey dot indicates value of individual cell. Orange and cyan triangle indicate mean of each independent culture. Mann–Whitney test: not significant (two-sided): not significant (n.s.) P > 0.05, *P < 0.05, ***P < 0.0001. More detailed statistical information see Data S1.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal