A novel rare variant R292H in RTN4R affects growth cone formation and possibly contributes to schizophrenia susceptibility.

Reticulon 4 receptor (RTN4R) plays an essential role in regulating axonal regeneration and plasticity in the central nervous system through the activation of rho kinase, and is located within chromosome 22q11.2, a region that is known to be a hotspot for schizophrenia (SCZ) and autism spectrum disorder (ASD). Recently, rare variants such as copy-number variants and single-nucleotide variants have been a focus of research because of their large effect size associated with increased susceptibility to SCZ and ASD and the possibility of elucidating the pathophysiology of mental disorder through functional analysis of the discovered rare variants. To discover rare variants with large effect size and to evaluate their role in the etiopathophysiology of SCZ and ASD, we sequenced the RTN4R coding exons with a sample comprising 370 SCZ and 192 ASD patients, and association analysis using a large number of unrelated individuals (1716 SCZ, 382 ASD and 4009 controls). Through this mutation screening, we discovered four rare (minor allele frequency <1%) missense mutations (R68H, D259N, R292H and V363M) of RTN4R. Among these discovered rare mutations, R292H was found to be significantly associated with SCZ (P=0.048). Furthermore, in vitro functional assays showed that the R292H mutation affected the formation of growth cones. This study strengthens the evidence for association between rare variants within RTN4R and SCZ, and may shed light on the molecular mechanisms underlying the neurodevelopmental disorder.

Contractile effect of acetylcholine on isolated ampullar segment of Fallopian tubes.

The Fallopian tubes are scarcely innervated with cholinergic nerve fibers. Acetylcholine released from these nerves contracts smooth muscles of the tubes. The aim of our study was to investigate the effect of acetylcholine on the Fallopian tubes using selective antagonists in different hormonal settings. We have investigated effects of acetylcholine on isolated ampulla of Fallopian tubes taken from 83 patients during abdominal hysterectomy with adnexectomy. Twenty-eight patients were in follicular, 36 were in luteal phase of menstrual cycle, and 19 patients were in menopause. Selective and non-selective muscarinic and nicotinic receptor blockers were used for investigation of the effects. Acetylcholine (from 1.8 to 658.6 microM/l) produced concentration-dependent tonic contraction of ampulla taken from the patients in follicular phase, luteal phase and menopause. The nicotinic receptor blocker mecamylamine (6.5 microM/l) and local anesthetic lidocaine (230.8 microM/l) did not affect the effect of acetylcholine. While M(1)- and M(2)-selective muscarinic receptor blockers pirenzepine (1.6 microM/l) and methoctramine (0.9 microM/l) did not show specific effect, atropine (0.01 microM/l) and selective M(3)-receptor blocker p-fluoro-hexahydro-sila-difenidol (pFHHSiD, 0.2 microM/l) effectively blocked contractions caused by acetylcholine (maximal pA(2) values 9.74 and 7.54, respectively). The affinity of pFHHSiD for muscarinic receptors was highest in the follicular phase. The results of our study suggest existence of functional M(3) muscarinic receptors in ampulla of Fallopian tubes, located on the smooth muscle cells.