• Home
  • Search Results
  • Addiction associated N40D mu-opioid receptor variant modulates synaptic function in human neurons.

Addiction associated N40D mu-opioid receptor variant modulates synaptic function in human neurons.

Molecular psychiatry (2019-09-05)
Apoorva Halikere, Dina Popova, Matthew S Scarnati, Aula Hamod, Mavis R Swerdel, Jennifer C Moore, Jay A Tischfield, Ronald P Hart, Zhiping P Pang
ABSTRACT

The OPRM1 A118G single nucleotide polymorphism (SNP rs1799971) gene variant encoding the N40D µ-opioid receptor (MOR) has been associated with dependence on opiates and other drugs of abuse but its mechanism is unknown. The frequency of G-allele carriers is ~40% in Asians, ~16% in Europeans, and ~3% in African-Americans. With opioid abuse-related deaths rising at unprecedented rates, understanding these mechanisms may provide a path to therapy. Here we generated homozygous N40D subject-specific induced inhibitory neuronal cells (iNs) from seven human-induced pluripotent stem (iPS) cell lines from subjects of European descent (both male and female) and probed the impact of N40D MOR regulation on synaptic transmission. We found that D40 iNs exhibit consistently stronger suppression (versus N40) of spontaneous inhibitory postsynaptic currents (sIPSCs) across multiple subjects. To mitigate the confounding effects of background genetic variation on neuronal function, the regulatory effects of MORs on synaptic transmission were recapitulated in two sets of independently engineered isogenic N40D iNs. In addition, we employed biochemical analysis and observed differential N-linked glycosylation of human MOR N40D. This study identifies neurophysiological and molecular differences between human MOR variants that may predict altered opioid responsivity and/or dependence in this subset of individuals.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, clone M2, purified immunoglobulin (Purified IgG1 subclass), buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)
Sigma-Aldrich
Anti-Somatostatin Antibody, clone YC7, culture supernatant, clone YC7, Chemicon®
Sigma-Aldrich
[D-Ala2, N-Me-Phe4, Gly5-ol]-Enkephalin acetate salt, ≥97% (HPLC)
Sigma-Aldrich
Monoclonal Anti-MAP2 (2a+2b) antibody produced in mouse, clone AP-20, ascites fluid
Sigma-Aldrich
Anti-TRA-1-60 Antibody, clone TRA-1-60, clone TRA-1-60, Chemicon®, from mouse
Sigma-Aldrich
Anti-MAP2 Antibody, Chemicon®, from chicken
Sigma-Aldrich
Anti-MAP2 antibody produced in rabbit, ~1 mg/mL, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Anti-Vesicular GABA Transporter (VGAT) Antibody, Chemicon®, from rabbit
Sigma-Aldrich
Anti-Oct-4 Antibody, clone 10H11.2, clone 10H11.2, Chemicon®, from mouse