Cadherin-13 is a critical regulator of GABAergic modulation in human stem-cell-derived neuronal networks

Britt Mossink,Jason M Keller,Shan Wang,Sophie Jansen,Ilse Van Der Werf,Maria Rosaria Vitale ,Brooke Latour ,Dirk W Schubert ,Anouk H.a Verboven ,Johanna Eva Maria Zöller ,Katrin Linda,Monica Frega,Hans Van Bokhoven,Klaus‐Peter Lesch ,Moritz Negwer,Teun Klein Gunnewiek ,Alessia Anania,Astrid R Oudakker ,Martijn Selten,Jon Ruben Van Rhijn ,Chantal Schoenmaker,Jitske Bak,Eline J.h Van Hugte ,Nael Nadif Kasri

doi:10.1038/s41380-021-01117-x

Abstract

Activity in the healthy brain relies on a concerted interplay of excitation (E) and inhibition (I) via balanced synaptic communication between glutamatergic and GABAergic neurons. A growing number of studies imply that disruption of this E/I balance is a commonality in many brain disorders; however, obtaining mechanistic insight into these disruptions, with translational value for the patient, has typically been hampered by methodological limitations. Cadherin-13 (CDH13) has been associated with autism and attention-deficit/hyperactivity disorder. CDH13 localizes at inhibitory presynapses, specifically of parvalbumin (PV) and somatostatin (SST) expressing GABAergic neurons. However, the mechanism by which CDH13 regulates the function of inhibitory synapses in human neurons remains unknown. Starting from human-induced pluripotent stem cells, we established a robust method to generate a homogenous population of SST and MEF2C (PV-precursor marker protein) expressing GABAergic neurons (iGABA) in vitro, and co-cultured these with glutamatergic neurons at defined E/I ratios on micro-electrode arrays. We identified functional network parameters that are most reliably affected by GABAergic modulation as such, and through alterations of E/I balance by reduced expression of CDH13 in iGABAs. We found that CDH13 deficiency in iGABAs decreased E/I balance by means of increased inhibition. Moreover, CDH13 interacts with Integrin-β1 and Integrin-β3, which play opposite roles in the regulation of inhibitory synaptic strength via this interaction. Taken together, this model allows for standardized investigation of the E/I balance in a human neuronal background and can be deployed to dissect the cell-type-specific contribution of disease genes to the E/I balance.

Highlights

Neuronal network activity is controlled by a tightly regulated interplay between excitation (E) and inhibition (I)
A growing number of studies imply that the E/I balance is disrupted in many neurodevelopmental disorders (NDDs) [3, 4], including monogenic disorders, where the causative mutations are typically related to altered neuronal excitability and/or synaptic communication [5,6,7], as well as polygenic disorders, such as autism spectrum disorders (ASD) and attention-deficit hyperactivity disorder (ADHD) [4, 8]
RNAseq analysis at days in vitro (DIV) 49 further confirmed that E/I networks highly express SST, MEF2C, and genes expressed in mature fast-spiking neurons (FGF13 [39], LGL2 [39], PVALB), as well as genes coding for Glutamate and GABA transporters (SLC17A6/7, GAD1/2) and GABAergic neuron development (DLX1-6, LHX6, ZEB2, SOX6, Fig. 1f, and Supplementary Table 1)

Summary

Introduction

Neuronal network activity is controlled by a tightly regulated interplay between excitation (E) and inhibition (I). This interplay maintains a certain E/I ratio via balanced synaptic communication between glutamatergic and GABAergic neurons [1, 2], resulting in the so called. We recently showed that in the hippocampus, CDH13 is located to the presynaptic compartment of inhibitory GABAergic neurons, of parvalbumin (PV+) and somatostatin (SST+) expressing neurons, and that Cdh knockout (KO) mice (Cdh13−/−) show an increased inhibitory, but not excitatory synaptic input onto hippocampal CA1 pyramidal neurons [9] These mice display deficits in learning and memory [9]. The mechanism via which CDH13 regulates GABAergic synapses remains unknown

Methods

Results

Conclusion