Philip G. Wood

Periodic Fluctuations in the Pulmonary Surfactant System in Gould’s Wattled Bat (Chalinolobus gouldii)

Pulmonary surfactant is a mixture of phospholipids, neutral lipids, and proteins that controls the surface tension of the fluid lining the lung. Surfactant amounts and composition are influenced by such physiological parameters as metabolic rate, activity, body temperature, and ventilation. Microchiropteran bats experience fluctuations in these parameters throughout their natural daily cycle of activity and torpor. The activity cycle of the microchiropteran bat Chalinolobus gouldii was studied over a 24‐h period. Bats were maintained in a room at constant ambient temperature (24°C) on an 8L:16D cycle. Diurnal changes in the amount and composition of surfactant were measured at 4‐h intervals throughout a 24‐h period. The C. gouldii were most active at 2 a.m. and were torpid at 2 p.m. Alveolar surfactant increased 1.5‐fold immediately after arousal. The proportion of disaturated phospholipid remained constant, while surfactant cholesterol levels increased 1.5‐fold during torpor. Alveolar cholesterol in C. gouldii was six times lower than in other mammals. Cholesterol appears to function in maintaining surfactant fluidity during torpor in this species of bat.

Control of pulmonary surfactant secretion from type II pneumocytes isolated from the lizard Pogona vitticeps

Pulmonary surfactant, a mixture consisting of lipids and proteins and secreted by type II cells, functions to reduce the surface tension of the fluid lining of the lung, and thereby decreases the work of breathing. In mammals, surfactant secretion appears to be influenced primarily by the sympathetic nervous system and changes in ventilatory pattern. The parasympathetic nervous system is not believed to affect surfactant secretion in mammals. Very little is known about the factors that control surfactant secretion in nonmammalian vertebrates. Here, a new methodology for the isolation and culture of type II pneumocytes from the lizard Pogona vitticeps is presented. We examined the effects of the major autonomic neurotransmitters, epinephrine (Epi) and ACh, on total phospholipid (PL), disaturated PL (DSP), and cholesterol (Chol) secretion. At 37°C, only Epi stimulated secretion of total PL and DSP from primary cultures of lizard type II cells, and secretion was blocked by the β-adrenoreceptor antagonist propranolol. Neither of the agonists affected Chol secretion. At 18°C, Epi and ACh both stimulated DSP and PL secretion but not Chol secretion. The secretion of surfactant Chol does not appear to be under autonomic control. It appears that the secretion of surfactant PL is predominantly controlled by the autonomic nervous system in lizards. The sympathetic nervous system may control surfactant secretion at high temperatures, whereas the parasympathetic nervous system may predominate at lower body temperatures, stimulating surfactant secretion without elevating metabolic rate.

Autonomic control of the pulmonary surfactant system and lung compliance in the lizard.

An increase in body temperature in the bearded dragon, Pogona vitticeps, is accompanied by an increase in the amount of pulmonary surfactant, a mixture of proteins and lipids, with the latter consisting predominantly of phospholipid and cholesterol. This increase may result from a temperature‐induced change in autonomic input to the lungs, as perfusing the isolated lungs of P. vitticeps with either acetylcholine or adrenaline increases surfactant phospholipid release. However, whether acetylcholine acts via intrapulmonary sympathetic ganglia or directly on alveolar Type II cells is unknown. Moreover, the relative importance of circulating catecholamines and pulmonary sympathetic nerves on the control of the surfactant system is also obscure. Here, we describe the mechanism of the modulation of the surfactant system and the effect of this modulation on lung compliance. The role of acetylcholine was determined by perfusing isolated lungs with acetylcholine, acetylcholine and the ganglionic antagonist hexamethonium, or acetylcho‐line, hexamethonium, and the muscarinic antagonist atropine. Perfusing with acetylcholine significantly increased phospho‐lipid release but did not affect cholesterol release. While histo‐logical examination of the lung revealed the presence of a large autonomic ganglion at the apex, blocking sympathetic ganglia with hexamethonium did not prevent the acetylcholine‐medi‐ated increase in phospholipid. However, the increase was inhibited by blocking muscarinic receptors with atropine, which indicates that acetylcholine acts on muscarinic receptors to stimulate phospholipid release. By increasing pulmonary smooth muscle tone, acetylcholine decreased opening pressure and increased static inflation pressures. Plasma levels of nor‐adrenaline and adrenaline increased with increasing temperature and were accompanied by a greater surfactant content in the lungs. While surfactant content was also higher in animals that exercised, plasma levels of adrenaline, noradrenaline, and dopamine were not elevated following exercise. Hence, surfactant release in the lizard lung may increase in response to an increase in plasma catecholamine levels. Acetylcholine, and hence the parasympathetic nervous system, may act to stimulate surfactant release but does not act via pulmonary sympathetic ganglia. We conclude that promoting surfactant secretion via an increase in circulating catecholamines may be inappropriate for a cold lizard with a requirement to conserve energy. As body temperature decreases, release of surfactant via nonadrenergic mechanisms, including cholinergic stimulation, may become increasingly important.

The ontogeny of pulmonary surfactant secretion in the embryonic green sea turtle (Chelonia mydas).

Pulmonary surfactant, consisting predominantly of phosphatidylcholine (PC), is secreted from Type II cells into the lungs of all air‐breathing vertebrates, where it functions to reduce surface tension. In mammals, glucocorticoids and thyroid hormones contribute to the maturation of the surfactant system. It is possible that phylogeny, lung structure, and the environment may influence the development of the surfactant system. Here, we investigate the ontogeny of PC secretion from cocultured Type II cells and fibroblasts in the sea turtle, Chelonia mydas, following 58, 62, and 73 d of incubation and after hatching. The influence of glucocorticoids and thyroid hormones on PC secretion was also examined. Basal PC secretion was lowest at day 58 (3%) and reached a maximal secretion rate of 10% posthatch. Dexamethasone (Dex) alone stimulated PC secretion only at day 58. Triiodothyronine (T3) stimulated PC secretion in cells isolated from days 58 and 73 embryos and from hatchling turtles. A combination of Dex and T3 stimulated PC secretion at all time points.

Surfactant in the gas mantle of the snail Helix aspersa

Surfactant occurs in cyclically inflating and deflating, gas‐holding structures of vertebrates to reduce the surface tension of the inner fluid lining, thereby preventing collapse and decreasing the work of inflation. Here we determined the presence of surfactant in material lavaged from the airspace in the gas mantle of the pulmonate snail Helix aspersa. Surfactant is characterized by the presence of disaturated phospholipid (DSP), especially disaturated phosphatidylcholine (PC), lavaged from the airspace, by the presence of lamellated osmiophilic bodies (LBs) in the airspaces and epithelial tissue, and by the ability of the lavage to reduce surface tension of fluid in a surface balance. Lavage had a DSP/phospholipid (PL) ratio of 0.085, compared to 0.011 in membranes, with the major PL being PC (45.3%). Cholesterol, the primary fluidizer for pulmonary surfactant, was similar in lavage and in lipids extracted from cell homogenates (cholesterol/PL: 0.04 and 0.03, respectively). LBs were found in the tissues and airspaces. The surface activity of the lavage material is defined as the ability to reduce surface tension under compression to values much lower than that of water. In addition, surface‐active lipids will vary surface tension, increasing it upon inspiration as the surface area expands. By these criteria, the surface activity of lavaged material was poor and most similar to that shown by pulmonary lavage of fish and toads. Snail surfactant displays structures, a biochemical PL profile, and biophysical properties similar to surfactant obtained from primitive fish, teleost swim bladders, the lung of the Dipnoan Neoceratodus forsteri, and the amphibian Bufo marinus. However, the cholesterol/PL and cholesterol/DSP ratios are more similar to the amphibian B. marinus than to the fish, and this similarity may indicate a crucial physicochemical relationship for these lipids.

Factors affecting opening and filling pressures in the lungs of the lizard Pogona vitticeps

We have previously reported that levels of pulmonary surfactant in the lungs of the lizard Pogona vitticeps increase with increasing body temperature. Static lung compliance decreases with increasing body temperature, and is only marginally affected by the presence of surfactant. Here, we examined the effects of surfactant, temperature, ventilatory pattern and autonomic neurotransmitters on opening and filling pressures. Isolated lungs were ventilated at either 18 or 37 degrees C at low, intermediate and high ventilatory regimes. The effects of acetylcholine and adrenaline were examined using an isolated perfused lung preparation at 27 degrees C. Changing ventilatory pattern or experimental temperature had no effect on either filling or opening pressures. Removal of surfactant increased both opening and filling pressures. Adrenaline administration reduced opening and filling pressures. Normal variations in surfactant levels, which occur with changes in body temperature, do not affect either opening or filling pressures. A critical amount of surfactant may be necessary to prevent adhesion of epithelial surfaces in the lungs of Pogona vitticeps. The anti-glue function of pulmonary surfactant may be more important at 18 than at 37 degrees C.