Neuronal NO modulates spontaneous and ANG II-stimulated fetal swallowing behavior in the near-term ovine fetus

Abstract

Spontaneous fetal swallowing occurs at a markedly higher rate compared with spontaneous adult drinking activity. This high rate of fetal swallowing is critical for amniotic fluid volume regulation. Central NO is critical for maintaining the normal rate of fetal swallowing, as nonselective inhibition of NO (with central N G -nitro-l-arginine methyl ester) suppresses spontaneous and angiotensin II (ANG II)-stimulated swallowing. We sought to differentiate the contributions of central endothelial vs. neuronal NO in the regulation of spontaneous and stimulated fetal swallowing, using a selective neuronal NO synthase (nNOS) inhibitor. Six time-dated pregnant ewes and fetuses were chronically prepared with fetal vascular and intracerebroventricular (icv) catheters and electrocorticogram (ECoG) and esophageal electromyogram electrodes and studied at 130 ± 1 days of gestation. After an initial 2-h baseline period (0–2 h), the selective nNOS inhibitor N-propyl-l-arginine (NPLA) was injected icv (2–4 h). At 4 h, the dose of NPLA was repeated, together with ANG II, and fetal swallowing was monitored for a final 2 h. Four fetuses also received an identical control study (on an alternate day) in which NPLA was replaced with artificial cerebrospinal fluid (aCSF). Suppression of nNOS by icv NPLA significantly reduced mean (± SE) spontaneous fetal swallowing (1.35 ± 0.12 to 0.50 ± 0.07 swallows/min; P< 0.001). Injection of ANG II in the presence of NPLA had no dipsogenic effect on fetal swallowing (0.68 ± 0.09 swallows/min). In the aCSF study, icv aCSF did not change fetal swallowing (0.93 ± 0.10 vs. 0.95 ± 0.09 swallows/min), whereas icv ANG II resulted in a significant increase in the rate of fetal swallowing (2.0 ± 0.04 swallows/min; P = 0.001). We speculate that the suppressive dipsogenic effects of central NPLA indicate that spontaneous and ANG II- stimulated fetal swallowing is dependent on central nNOS activity.