Secreted amyloid-β precursor protein functions as a GABABR1a ligand to modulate synaptic transmission

A physiological function for sAPP? Although the pathological role of the amyloid-β precursor protein (APP) in Alzheimer's disease is well studied, the physiological role of this protein has remained elusive. Rice et al. found that the secreted ectodomain of APP (sAPP) binds to GABABR1a, the metabotropic receptor for the inhibitory neurotransmitter γ-aminobutyric acid (GABA) (see the Perspective by Korte). Binding suppressed synaptic vesicle release and modulated synaptic transmission and plasticity in mice. A short, 17–amino acid peptide in APP bound to GABABR1a's sushi 1 domain, conferring structure to this unstructured domain. Therapeutics targeting this interaction could potentially benefit a range of neurological disorders in which GABA signaling is implicated. Science, this issue p. eaao4827; see also p. 123 Amyloid-β precursor protein suppresses vesicle release from presynaptic boutons by binding to the GABAB1a receptor. INTRODUCTION More than 30 years have passed since the amyloid-β precursor protein (APP) was identified. Although the role of APP in Alzheimer’s disease has been studied widely, its normal physiological function in the brain has remained elusive. APP undergoes ectodomain shedding by α-, β-, or η-secretase to release secreted APP (sAPPα, sAPPβ, or sAPPη, respectively). sAPPα affects synaptic transmission and plasticity and is sufficient to rescue synaptic defects in App knockout mice. This has led to speculation of a yet unidentified cell-surface receptor for sAPPα. RATIONALE To elucidate the physiological function of APP, we sought to identify the cell-surface receptor mediating its effects on synaptic function. To identify candidate synaptic interactors for sAPPα, we performed affinity-purification experiments using recombinant sAPPα to pull down interacting proteins from synaptosome extracts, followed by mass spectrometric analysis of bound proteins. We identified the γ-aminobutyric acid type B receptor (GABABR), the metabotropic receptor for the inhibitory neurotransmitter γ-aminobutyric acid (GABA), as the leading candidate for a synaptic, cell-surface receptor for sAPPα. We then performed a combination of cell-surface binding assays and in vitro biophysical techniques to determine the interacting domains and structural consequences of binding. We investigated whether sAPPα can modulate GABABR function by assessing miniature excitatory and inhibitory postsynaptic currents (mEPSCs and mIPSCs, respectively) and synaptic vesicle recycling in mouse hippocampal neuron cultures, short-term plasticity in acute hippocampal slices from mice, and in vivo neuronal activity in the hippocampus of anesthetized mice. RESULTS Recombinant sAPPα selectively bound to GABABR subunit 1a (GABABR1a)–expressing cells. Binding was mediated by the flexible, partially structured extension domain in the linker region of sAPP and the natively unstructured sushi 1 domain specific to GABABR1a. sAPPβ and sAPPη, which both contain the extension domain, also bound to GABABR1a-expressing cells. Conversely, APP family members APP-like proteins 1 and 2, which lack a conserved extension domain, failed to bind GABABR1a-expressing cells. Acute application of sAPPα reduced the frequency of mEPSCs and mIPSCs and decreased synaptic vesicle recycling in cultured mouse hippocampal neurons. In addition, sAPPα enhanced short-term facilitation in acute hippocampal slices from mice. Together, these findings demonstrate that sAPP reduces the release probability of synaptic vesicles. These effects were dependent on the presence of the extension domain in sAPP and were occluded by a GABABR antagonist. A short APP peptide corresponding to the GABABR1a binding region within APP stabilized the natively unstructured sushi 1 domain of GABABR1a, allowing determination of its solution structure using nuclear magnetic resonance spectroscopy and the generation of a structural model of the APP–sushi 1 complex. Application of a 17–amino acid APP peptide mimicked the effects of sAPPα on GABABR1a-dependent inhibition of synaptic vesicle release and reversibly suppressed spontaneous neuronal activity in vivo. CONCLUSION We identified GABABR1a as a synaptic receptor for sAPP and revealed a physiological role for sAPP in regulating GABABR1a function to modulate synaptic transmission and plasticity. Our findings provide a potential target for the development of GABABR1a isoform–specific therapeutics, which is relevant to a number of neurological disorders in which GABABR signaling is implicated. sAPP is a functional GABABR1a-specific ligand. In the presence of sAPP (right), the extension domain (ExD) of sAPP binds the sushi 1 domain specific to GABABR1a. Binding induces a conformational change in the sushi 1 domain and leads to increased short-term facilitation and decreased neuronal activity via inhibition of neurotransmitter release. N, amino terminus; C, carboxyl terminus; α, β, and γ, G protein subunits coupled to GABABR subunit 2 (GABABR2); E1 and E2, sAPP domains. ILLUSTRATION: SOMERSAULT18:24 Amyloid-β precursor protein (APP) is central to the pathogenesis of Alzheimer’s disease, yet its physiological function remains unresolved. Accumulating evidence suggests that APP has a synaptic function mediated by an unidentified receptor for secreted APP (sAPP). Here we show that the sAPP extension domain directly bound the sushi 1 domain specific to the γ-aminobutyric acid type B receptor subunit 1a (GABABR1a). sAPP-GABABR1a binding suppressed synaptic transmission and enhanced short-term facilitation in mouse hippocampal synapses via inhibition of synaptic vesicle release. A 17–amino acid peptide corresponding to the GABABR1a binding region within APP suppressed in vivo spontaneous neuronal activity in the hippocampus of anesthetized Thy1-GCaMP6s mice. Our findings identify GABABR1a as a synaptic receptor for sAPP and reveal a physiological role for sAPP in regulating GABABR1a function to modulate synaptic transmission.