Glial cell line-derived neurotrophic factor (GDNF) is required for differentiation of pontine noradrenergic neurons and patterning of central respiratory output.

Genetic mutations affecting signaling by glial cell line-derived neurotrophic factor (GDNF) perturb development of breathing in mice and are associated with congenital central hypoventilation syndrome in humans. However, the role of GDNF in development of brainstem neurons that control breathing is largely unknown. The present study demonstrates that genetic loss of GDNF decreases the number of tyrosine hydroxylase (TH) neurons in the pontine A5 noradrenergic cell group, a major source of inhibitory input to the medullary respiratory pattern generator. This phenotype is associated with a significant increase in the frequency of central respiratory output recorded from the fetal medulla-spinal cord in vitro. In dissociate cultures of the A5 region from rat embryos, GDNF increases TH cell number and neurite growth without affecting total neuronal survival or proliferation of TH neurons. These effects of GDNF are inhibited by function blocking antibodies against endogenous brain-derived neurotrophic factor (BDNF), indicating that GDNF requires BDNF as a cofactor to stimulate differentiation of A5 neurons. Our findings demonstrate that GDNF is required for development of pontine noradrenergic neurons in vivo and indicate that defects in the A5 cell group may contribute to the effects of genetic disruption of GDNF signaling on respiratory control.

Hypotension, autonomic failure, and cardiac hypertrophy in transgenic mice overexpressing the alpha 1B-adrenergic receptor.

alpha(1)-Adrenergic receptors (alpha(1A), alpha(1B), and alpha(1D)) are regulators of systemic arterial blood pressure and blood flow. Whereas vasoconstrictory action of the alpha(1A) and alpha(1D) subtypes is thought to be mainly responsible for this activity, the role of the alpha(1B)-adrenergic receptor (alpha(1B)AR) in this process is controversial. We have generated transgenic mice that overexpress either wild type or constitutively active alpha(1B)ARs. Transgenic expression was under the control of the isogenic promoter, thus assuring appropriate developmental and tissue-specific expression. Cardiovascular phenotypes displayed by transgenic mice included myocardial hypertrophy and hypotension. Indicative of cardiac hypertrophy, transgenic mice displayed an increased heart to body weight ratio, which was confirmed by the echocardiographic finding of an increased thickness of the interventricular septum and posterior wall. Functional deficits included an increased isovolumetric relaxation time, a decreased heart rate, and cardiac output. Transgenic mice were hypotensive and exhibited a decreased pressor response. Vasoconstrictory regulation by alpha(1B)AR was absent as shown by the lack of phenylephrine-induced contractile differences between ex vivo mesenteric artery preparations. Plasma epinephrine, norepinephrine, and cortisol levels were also reduced in transgenic mice, suggesting a loss of sympathetic nerve activity. Reduced catecholamine levels together with basal hypotension, bradycardia, reproductive problems, and weight loss suggest autonomic failure, a phenotype that is consistent with the multiple system atrophy-like neurodegeneration that has been reported previously in these mice. These results also suggest that this receptor subtype is not involved in the classic vasoconstrictory action of alpha(1)ARs that is important in systemic regulation of blood pressure.

End-systolic stress-velocity and pressure-dimension relationships by transthoracic echocardiography in mice.

The purposes of this study were to assess load-independent, end-systolic relationships in mice and compare these relationships to ejection phase indexes in assessing contractility. In 13 mice, ejection phase indexes (shortening fraction and velocity of fiber shortening) and end-systolic relationships [pressure-dimension relationship (ESPDR) and stress-velocity relationship (ESSVR)] were determined using M-mode echocardiography and simultaneous left ventricular pressure. Load was altered with phenylephrine and nitroprusside. Contractility was increased with dobutamine and decreased by induction of hypothyroidism. Ejection phase indexes increased with dobutamine infusion but were not significantly decreased with hypothyroidism. However, end-systolic relationships changed significantly with both dobutamine (gamma-intercepts: ESPDR from 22 to 48 mmHg, ESSVR from 3.7 to 6.6 circ/s, P < 0.05) and hypothyroidism (gamma-intercepts: ESPDR from 22 to 11 mmHg, ESSVR from 3.7 to 3.2 circ/s, P < 0.05). We conclude that end-systolic indexes can be accurately measured in the intact mouse by echocardiography with simultaneous left ventricular pressure recording and appear to be more sensitive to inotropic state than ejection phase indexes.

Cell-cell coupling occurs in dorsal medullary neurons after minimizing anatomical-coupling artifacts.

Dye (Lucifer Yellow) and tracer (Biocytin) coupling, referred to collectively as anatomical coupling, were identified in 20% of the solitary complex neurons tested in medullary tissue slices (120-350 microm) prepared from rat, postnatal day 1-18, using a modified amphotericin B-perforated patch recording technique. Ten per cent of the neurons sampled in nuclei outside the solitary complex were anatomically coupled. Fifty-eight per cent of anatomically coupled neurons exhibited electrotonic postsynaptic potential-like activity, which had peak-to-peak amplitudes of < or = 7 mV, with the same polarity as action potentials; increased and decreased in frequency during depolarizing and hyperpolarizing current injection; was maintained during high Mg2+-low Ca2+ chemical synaptic blockade; and was measured only in anatomically coupled neurons. The high correlation between anatomical coupling and electrotonic postsynaptic potential-like activity suggests that Lucifer Yellow, Biocytin and ionic current used the same pathways of intercellular communication, which were presumed to be gap junctions. Anatomical coupling was attributed solely to the junctional transfer of Lucifer Yellow and Biocytin since potential sources of non-junctional staining were minimized. Specifically, combining 0.26 mM amphotericin B and 0.15-0.5% Lucifer Yellow produced a hydrophobic, viscous solution that did not leak from the pressurized pipette tip < or = 3 microm outer diameter) submerged in artificial cerebral spinal fluid. Moreover, unintentional contact of the pipette tip with adjacent neurons that resulted in accidental staining, another source of non-junctional staining, wits averted by continuously visualizing the tip prior to tight seal formation with infrared video microscopy, used here for the first time with Hoffman modulation contrast optics. During perforated patch recording which typically lasted for 1-3 h. Lucifer Yellow was confined to the pipette, indicating that the amphotericin B patch was intact. However, once the patch was intentionally ruptured at the end of recording, the viscous, lipophilic solution entered the neuron resulting in double labeling. Placing a mixture of amphotericin B, Biocytin and Lucifer Yellow directly into the pipette tip did not compromise tight seal formation with an exposed, cleaned soma, and resulted in immediate (<1 min) steady-state perforation at 22-25 degrees C. This adaptation of conventional perforated patch recording was termed "rapid perforated patch recording". The possible functional implication of cell-cell coupling in the dorsal medulla oblongata in central CO2/H+ chemoreception for the cardiorespiratory control systems is discussed in the second paper of this set [Huang et al. (1997) Neuroscience 80, 41-57].

The effect of nightly nasal CPAP treatment on underlying obstructive sleep apnea and pharyngeal size.

Nasal continuous positive airway pressure (CPAP) is an effective treatment for obstructive sleep apnea (OSA). It is usually prescribed for nightly use; however, recent studies show that patients often do not wear the appliance consistently. Previous studies have also suggested that nasal CPAP may improve a patient's underlying OSA even when the mask is not in place. We investigated 12 men with OSA to see if nasal CPAP used nightly for six weeks would improve their underlying sleep-disordered breathing. We also studied pharyngeal volumes measured using magnetic resonance imaging and a computer-controlled digitizing pad. Patients with more severe OSA had improvement after six weeks; however, they still demonstrated significant OSA. Patients with less severe OSA did not have significant change. We were unable to show a significant difference in any patient's awake pharyngeal volumes. We conclude that patients with OSA should be encouraged to wear their nasal CPAP machines regularly.

Pharyngeal volume in asymptomatic snorers compared with nonsnoring volunteers.

STUDY OBJECTIVE: to determine if asymptomatic snorers have smaller pharyngeal volumes than age- and height-matched nonsnorers. DESIGN: we recruited asymptomatic heavy snorers and nonsnorers for a study. Each snorer was matched by age (+/- 3 years) and height (+/- 2 inches) with a nonsnorer. The nonsnorers were required to be near ideal body weight. All volunteers underwent overnight polysomnography, pulmonary function testing, and magnetic resonance imaging of the pharynx while awake. The volume of the pharynx was determined by a computer with data input from a digitizing instrument. SETTING: Veterans Administration Hospital and University of Florida Teaching Hospital PARTICIPANTS: Nine volunteers were recruited for both the snorer and nonsnorer groups. Each participant was paid $50. There were no interventions. MEASUREMENTS AND RESULTS: There were no differences in sleep variables between the two groups. There was also no significant difference between pharyngeal volumes for the two groups. CONCLUSIONS: The volume of the pharynx in asymptomatic snorers is similar to the volume in age- and height-matched nonsnorers.

Consistency of respiratory measurements from night to night during the sleep of elderly men.

Fourteen healthy elderly men had polysomnography performed on two consecutive nights to assess the consistency of sleep and breathing from night to night. The reported first-night effect was seen on electroencephalographic sleep, leading to lighter or fitful sleep on the first night. Mean values for apneas, hypopneas, and oxygen desaturations did not change from night 1 to night 2. Five of 14 subjects would have changed classification from night to night if a cutoff of five apneas and hypopneas per hour were used to define normality. Because this numerical cutoff may not be valid in the elderly, the clinical significance of such a changing classification is not clear.

Measurement of pharyngeal volume by digitized magnetic resonance imaging. Effect of nasal continuous positive airway pressure.

Pharyngeal size is thought to play an important role in the pathogenesis of snoring and obstructive sleep apnea. It has been hypothesized that nasal continuous positive airway pressure (CPAP) works by enlarging pharyngeal size and splinting the airway open. In this study, we selected 12 heavy snorers and abolished their snoring with nasal CPAP in our sleep laboratory. Using magnetic resonance imaging and a computer program utilizing a digitizing pad, we measured these awake subjects' pharyngeal volumes without and with the nasal CPAP apparatus on at the level used to abolish their snoring. We found an average 27.7% increase in pharyngeal volume with nasal CPAP. We have visually shown an increase in pharyngeal size with the use of nasal CPAP in a cohort of heavy snorers.

Nocturnal oxygen therapy does not improve snorers' intelligence.

STUDY OBJECTIVE: To determine whether nasal oxygen therapy at 2 L/min would (1) reverse nocturnal hypoxemia and (2) improve neuropsychologic function in men who snore heavily. DESIGN: To select heavy-snoring subjects for a treatment protocol, volunteers were screened for one night, breathing air the first half and oxygen the second half of the night. If nocturnal oxygen desaturation occurred in the first half and was improved in the second half of the night, the subject entered a two-month treatment program. In random order, either nocturnal air or nocturnal oxygen was administered for one month each at 2 L/min in a double-blind crossover design. Neuropsychologic testing was done before and after each month. SETTING: Oxygen concentrators were modified to produce either greater than 96 percent oxygen or air at 2 L/min. Machines were delivered to the home of the subject and the machines were used each night, administering inspirate by nasal cannula. PARTICIPANTS: Seventeen asymptomatic men who snored heavily and volunteered for minimal payment of $75. INTERVENTIONS: Air was administered for one month, and oxygen was administered for one month. MEASUREMENTS AND RESULTS: On the screening night, oxygen administration did not improve obstructive sleep apnea, but did improve oxygenation. After one month of oxygen therapy at night, there was no significant benefit to multiple measures of neuropsychologic function. CONCLUSION: In this study, oxygen therapy at night had no effect on neuropsychologic function in men who snore heavily.

Snoring, nocturnal hypoxemia, and the effect of oxygen inhalation.

Men who snore heavily have an increased incidence of hypertension, angina, stroke, and neuropsychologic dysfunction, which may be due to nocturnal oxygen desaturation. Nocturnal oxygen therapy might be beneficial to such individuals by improving oxygenation and relieving tissue hypoxia. Twenty-eight asymptomatic heavy snoring men were recruited for polysomnographic monitoring during sleep. During the first half-night, air was breathed through a nasal cannula, and during the latter half-night, 2 L/min oxygen was administered. Breathing air, 20 subjects demonstrated sleep apneas, hypopneas and nocturnal oxygen desaturation. Eighteen subjects had more than ten apneas plus hypopneas per hour. Thirteen subjects reached low oxygen saturation below 80 percent and eight below 70 percent. Only 13 of the 20 subjects showed improvement with oxygen therapy. Apneas alone were not decreased in frequency and were lengthened with oxygen therapy. Episodes of oxygen desaturation were improved by oxygen therapy and consequently, rates of hypopnea were decreased. Severe sleep apnea, hypopnea and oxygen desaturation are common in asymptomatic male snorers, and oxygen therapy is not always beneficial.

Ingestion of either scotch or vodka induces equal effects on sleep and breathing of asymptomatic subjects.

Polysomnography was performed on 13 asymptomatic men and four women on three consecutive nights in our sleep laboratory. In random order, the subjects ingested either orange juice alone or the equivalent of 1 mL of 100-proof alcoholic beverage (scotch or vodka) per pound of body weight in 1.5 hours or less. All subjects ingested a different beverage on each of the three nights. Blood alcohol level in the subjects before sleep was, for vodka, 73 mg/100 mL, and, for scotch, 74 mg/100 mL. On control nights the subjects showed significantly more time in bed, sleep period time, and total sleep time, and more rapid eye movement sleep. On the scotch and vodka nights, oxygen saturation was significantly lower; there were more episodes of oxygen desaturation in which there was greater than 4% decrease in saturation, more desaturation to levels of less than 90%, and more hypopnea. Comparison of data of scotch with vodka nights showed no significant differences in any variable. Both scotch and vodka ingestion in equal dosage induced sleep-disordered breathing and nocturnal oxygen desaturation in asymptomatic volunteers, and the beverages had equal effects.

Effect of alcohol ingestion on breathing and oxygenation during sleep. Analysis of the influence of age and sex.

Breathing and oxygenation were monitored in 78 asymptomatic volunteers on two successive nights of sleep. Four groups of subjects were recruited: 20 young men, 20 young women, 20 men older than 40 years, and 18 postmenopausal women. In random order, subjects ingested either 2 ml/kg (body weight) of 100-proof vodka in orange juice or a similar amount of water in orange juice before bedtime. Alcohol ingestion shortened sleep in the older men and in the postmenopausal women. No effect of alcohol ingestion on breathing or oxygenation during sleep was seen in any group of women. In men, alcohol ingestion increased the numbers of desaturation episodes and caused more severe oxygen desaturation during sleep. The effect of alcohol ingestion on breathing and oxygenation during the sleep of asymptomatic volunteers appears to be limited to men.

Minimal effect of alcohol ingestion on breathing during the sleep of postmenopausal women.

Eighteen asymptomatic postmenopausal women volunteered to ingest 2 ml of 100-proof vodka per kg of body weight in orange juice on one night and a placebo on another. Overnight sleep monitoring was performed immediately thereafter. Alcohol ingestion caused reduction in total sleep time from 329 to 281 minutes and a decrease in rapid eye movement sleep. There was no difference from placebo in the number of episodes of apnea or hypopnea, or in the frequency, length, or severity of oxygen desaturation. In contrast to the effects of alcohol ingestion in men, the effects on breathing and oxygenation are minimal during the sleep of women if this amount of alcohol is ingested.