Morphometric analysis of the growth of the normal fetal guinea pig lung

The structural development of the fetal guinea pig lung is described and quantified morphometrically in this report. At 35 days gestation the lung is in the pseudoglandular phase of growth, by 40 days it is in the canalicular phase, and at 50 days the saccular growth phase has begun. At term (67 days), the fetal guinea pig lung appears mature. From the beginning of the canalicular to the end of the saccular phases, the correlation coefficient between lung volume and gestational age is +.98, between internal surface area and gestational age is +.94 and between total number of saccules and gestational age is +.97. Internal surface area (ISA) correlates closely with lung volume (r = +.99) and the correlation coefficient between total number of saccules and lung volume is +.98. At term, lung volume is 4.22 ml. ISA is 0.5 M2, and total number of saccules is 253 million. Parenchymal growth is achieved by increases in both number and size of airspaces in the canalicular phase, primarily by increases in number during the early saccular phase and largely by increases in airspace size near term. The total length of parenchymal elastic tissue increases from 223 M at 45 days gestation to 5,253 M at term. Elastic tissue fibers first appear in the parenchyma of the fetal guinea pig lung during the canalicular phase, when the rate of saccule formation is high. The quantitative increase in elastic tissue correlates closely with the increase in the total number of saccules from day 45 to day 60 of gestation (r = +.99). The rate of elastic tissue growth increases sharply in the late saccular phase, coinciding with the period of greatest saccular expansion. These data suggest an interdependent relationship between saccular growth, i.e., proliferation and expansion, and the development of lung parenchymal elastic tissue.     

Postnatal growth of the sheep lung: a morphometric study

Stereologic methods were used to study lung development in sheep from 2 to 171 days of age. Most growth occurs within the first 2 months when there is a threefold increase in lung volume, but allometric relationships show that this increase does not keep pace with body weight. Alveolar and capillary surface areas increase as lung volume to a power only slightly larger than 1, suggesting a modest increase in complexity; this is confirmed by only a two- to threefold increase in total alveolar number. Allometric power functions are essentially unchanged even during the first 30 days so that throughout development, the surface for gas exchange is added at a fairly constant rate. A slight increase in septal volume during the first 30 days is probably due to relative increase in capillary luminal volume through a progressive distension of capillaries. The results suggest that the active newborn lamb requires an alveolar lung, but in the postnatal period functional needs increase only moderately. In contrast, other mammals, such as the rat, are helpless as newborns and have a primitive saccular lung that must undergo considerable morphogenesis to meet the demands of the active adult.     

Hypophysectomy and porcine fetal lung development

The effect of hypophysectomy on the development of the lung parenchyma and maturation of pulmonary alveolar type II cells was examined in the fetal pig. In fetuses from four different gilts, hypophysectomy was performed on gestational day 69 or 70. Littermates from the same gilts served as controls. Fetuses were delivered by caesarean section at term (113 +/- 1 days of gestation), and the lungs were fixed by intratracheal instillation. Plasma cortisol and thyroxine concentrations in the umbilical artery were markedly lower for hypophysectomized fetuses compared with control fetuses. Body weight was similar for both groups of fetuses. Total lung volume was 53% smaller in hypophysectomized fetuses compared with control fetuses. However, alveolar septal tissue and capillary luminal volumes were similar in both groups. Total lung alveolar surface area was twice as great in control animals compared with hypophysectomized animals. The volumes of epithelium, interstitium, and endothelium of centriacinar alveolar septa per unit surface area of epithelial basal lamina were 3.7, 4.8, and 2.4 times greater in hypophysectomized fetuses compared with control fetuses. Alveolar type II cell composition also differed significantly between groups. The volume fraction of glycogen in type II cells was 51% for hypophysectomized fetuses and 12% for control fetuses, while lamellar body volume fraction was 8% in hypophysectomized fetuses and 23% in control fetuses. The frequency of alveolar type II cell contact with mesenchymal interstitial cells via foot processes was 5 times greater in the lungs of control animals compared with hypophysectomized animals. These findings demonstrate significant effects of hypophysectomy on the morphogenetic and cytodifferentiation activities of all major tissue compartments of the pulmonary gas exchange area during the final trimester of fetal development.     

Cellular changes in the lungs of adrenalectomized rats following left pneumonectomy

The time course and nature of the cellular response to left pneumonectomy, with or without prior adrenalectomy, were evaluated in the right lungs of male Sprague-Dawley rats using morphometric techniques. Animals were studied at days 2, 5, and 14 following pneumonectomy, intervals prior to, during the course of, and following significant compensatory changes in right lung mass. The postoperative increase in right lung mass and volume in pneumonectomized animals involved minimal changes in the ratios of most tissue components, when compared to the lungs of sham-operated controls. A transient disproportionate increase in type II cell volume and epithelial thickness was evident on day 14. Postpneumonectomy changes in the type II epithelium were accentuated in the lungs of adrenalectomized-pneumonectomized animals. Adrenalectomy 5 days prior to pneumonectomy resulted in a substantial increase in the volume of all right lung tissue components, associated with thickening of the alveolar wall and with increases in the volume of both cellular and noncellular interstitium. Effects of adrenalectomy on the endothelium also were evident. In both adrenal-intact and adrenalectomized animals, pneumonectomy increased alveolar number by day 14 but had no effect on the volume of individual alveoli. These results confirm a coordinated pattern of compensatory growth following pneumonectomy in the adrenal-intact rat. The data further suggest that in adrenalectomized animals compensatory lung growth is more poorly synchronized, with pronounced postoperative elevations in volume of the interstitial and type II epithelial compartments leading to increased thickness of the alveolar wall. Adrenal hormones thus appear to be required for coordination and control of compensatory lung growth and for rapid restoration of normal tissue structure.     

Further examination of alveolar septal adaptation to left pneumonectomy in the adult lung

Recent data from our laboratory are presented concerning alveolar septal adaptation following 42-45% lung resection by left pneumonectomy (PNX) in adult foxhounds compared to sham-operated control animals. Results confirm our previous conclusion that compensation in the remaining lung occurs without a net growth of additional alveolar septal tissue. The major ultrastructural responses are (a) alveolar capillary distention, which recruits capillary blood volume and surface area, leading to a 30-50% increase in lung diffusing capacity estimated by morphometry, a magnitude similar to that measured by physiologic methods; (b) a selectively increased volume of type 2 alveolar epithelial cells. These data, taken together with the balanced compensatory growth of alveolar septal cells observed in adult dogs following 55-58% lung resection by right PNX, support a graded alveolar cellular response to chronic mechanical strain with the alveolar epithelial cells being activated first; as strain increases further with greater lung resection other alveolar cells also become activated leading to an overt increase in septal tissue volume. The spatial distribution of lobar mechanical strain and lobar tissue volume assessed by high resolution computed tomography was markedly non-uniform after PNX, suggesting possible non-uniform distribution of alveolar cellular response. The sequential activation of physiologic recruitment and cellular adaptation confer additive functional benefits that optimize long-term exercise performance after PNX.     

Enhanced alveolar growth and remodeling in Guinea pigs raised at high altitude

To examine the effects of chronic high altitude (HA) exposure on lung structure during somatic maturation, we raised male weanling guinea pigs at HA (3800m) for 1, 3, or 6 months, while their respective male littermates were simultaneously raised at low altitude (LA, 1200m). Under anaesthesia, airway pressure was measured at different lung volumes. The right lung was fixed at a constant airway pressure for morphometric analysis under light and electron microscopy. In animals raised at HA for 1 month, lung volume, alveolar surface area and alveolar-capillary blood volume (V(c)) were elevated above LA control values. Following 3-6 months of HA exposure, increases in lung volume and alveolar surface area persisted while the initial increase in V(c) normalized. Additional adaptation occurred, including a higher epithelial cell volume, septal tissue volume and capillary surface area, a lower alveolar duct volume and lower harmonic mean diffusion barrier resulting in higher membrane and lung diffusing capacities. These data demonstrate enhanced alveolar septal growth and progressive acinar remodeling during chronic HA exposure with long-term augmentation of alveolar dimensions as well as functional compensation in lung compliance and diffusive gas transport.     

Localization of surfactant-associated protein C (SP-C) mRNA in fetal rabbit lung tissue by in situ hybridization.

Surfactant is a lipoprotein substance that is synthesized and secreted by alveolar type II epithelial cells and acts to reduce surface tension at the air-alveolar interface. SP-C is a 5,000-D molecular weight, hydrophobic, surfactant-associated protein. In the present study, we used a ribonuclease protection assay to show that SP-C mRNA is induced in rabbit fetal lung tissue early in development, increases in relative concentration as development proceeds, and is present in maximal concentration at term (31 days of gestation). We also used the technique of in situ hybridization to localize SP-C mRNA in fetal, neonatal, and adult rabbit lung tissue. SP-C mRNA was present in all of the epithelial cells of the prealveolar region of day 19 gestational age rabbit fetal lung tissue, i.e., about 7 days before the appearance of differentiated alveolar type II cells in the fetal lung tissue. By day 27 of gestation, SP-C mRNA was restricted to epithelial cells with the morphologic characteristics of alveolar type II cells. SP-C mRNA was not detected in bronchiolar epithelium at any stage of lung development. The intensity of SP-C mRNA hybridization in the prealveolar and alveolar type II epithelial cells increased as a function of gestational age and was maximal at term. The pattern of SP-C mRNA localization in neonatal and adult rabbit lung tissue was consistent with the restriction of SP-C gene expression to differentiated alveolar type II cells. Our data are suggestive that SP-C may serve some as yet unknown function early in lung development because it is present in fetal lung prealveolar epithelial cells much earlier in gestation than are differentiated, surfactant-producing alveolar type II cells.     

Glucocorticoids and hypothyroidism modulate development of fetal lung insulin receptors

We investigated the development of insulin receptors in membranes of fetal rabbit lung during normal ontogeny and the effect of glucocorticoids and hypothyroidism. Specific binding of 125I-insulin to fetal lung membranes increased progressively to a peak at 29 days gestation, declining by 30 days. Scatchard plots were curvilinear and revealed a progressive increase in receptor numbers (X 10(10)/mg protein) from 129 +/- 7 (mean +/- SE) at 22-24 days to 575 +/- 16 at 29 days, declining to 467 +/- 12 at 30 days, term being approximately 31 days. Affinities did not change throughout gestation and were similar to those of adult lung; receptor numbers in adults were significantly lower than in fetuses at 26-30 days. Epinephrine and PGE1 could evoke a doubling of cAMP production in adult and fetal lung membranes until 29 days. Concomitantly with the fall in fetal insulin receptor number at 30 days, cAMP production in response to epinephrine or PGE1 increased fivefold. Induction of fetal hypothyroidism decreased insulin receptor numbers in the lung of the 28-day fetus by 70% from control (P less than 0.001) without a change in receptor affinity. In contrast, betamethasone administration increased fetal lung insulin receptor numbers by 250% (P less than 0.001) but did not alter their affinity; maternal lung insulin receptors were not altered. Thus, normal ontogeny of the fetal lung insulin receptor is characterized by a progressive increase in number followed by decline immediately before parturition associated with a sharp increase of cAMP responsiveness of the membranes. Hypothyroidism and glucocorticoid exposure can modulate the normal development of the fetal lung insulin receptor.     

Postnatal growth of lung parenchyma in the piglet: Morphometry correlated with mechanics

Background: A previous study of piglet lung growth (Mansell et. al. 1989. J. Appl. Physiol., 67:1422–1427) showed transient stiffness to changes in shape and volume immediately after birth. Later, elastic recoil was found to increase as the lung grew in weight and volume. The present study uses morphometry to test possible structural correlates of these two mechanical changes. Methods: Piglet lungs were fixed near full inflation via the airways during the immediate newborn period (6–12 hours, n=3), at 3–5 days (n=6), 25–30 days (n=5), and 80–85 days (n=3). Morphometry comprised arithmetic and harmonic mean thicknesses of alveolar septae and average mean surface curvature. Measurements of curvature and airspace volume were combined to differentiate alveolar expansion from septal proliferation as mechanisms for volumetric growth. Results: The unique mechanical behavior of the newborn lungs was associated with relatively thick alveolar septae. Marked thinning of the septae and resolution of the stiffness to shape and volume change had occurred by 3–5 days. An increase in elastic recoil during the first postnatal month was found to be associated with simple airspace expansion. The second and third months were characterized by septal proliferation and increase in arithmetic mean septal thickness but elastic recoil did not increase further. Harmonic mean septal thickness and airspace volume per gram of lung tissue did not change over the course of the study. Conclusions: 1) A relative stiffness to shape and volume change in freshly newborn piglet lung is associated with relatively thick alveolar septal walls; 2) postnatal development of piglet lung parenchyma involves septal lengthening and thinning followed by septal proliferation; 3) the initial phase of septal lengthening, rather than the later phase of septal proliferation, is associated with increase in parenchymal recoil. © 1995 Wiley-Liss, Inc.     

Development of alveolar septa and cellular maturation within the perinatal lung

To quantitate fetal lung cellularity and regional variation in alveolar maturation, guinea pig lungs were studied at 55, 60, and 65 d of gestation or within 2 h of birth (term = 68 d), and the data were analyzed for intralobar, interlobar, and age-group differences. Nine blocks from each left cranial and caudal lobe were all measured for volume, numerical, and surface densities in tissue (Vv(i,t), Nv(i,t), and Sv(i,t], and total volumes, numbers, and surface areas per lung (V, N, and S) of type I and type II epithelia, presumptive progenitor epithelium (cuboidal cells lacking lamellar bodies [LB]), interstitium, and endothelium. Total fixed lung volume, VL, increased 3-fold from day 55 through birth. At each age, there were no consistent intralobar or interlobar differences in Vv(i,t), Nv(i,t), or Sv(i,t) for any cell type. Within a septal tissue volume of 580 to 670 microliters at all ages, the N and V of type I cells did not vary with age, although their S increased from 1,240 cm2 at day 55 to 3,967 cm2 at birth. The N of morphologic type II cells per lung increased 7-fold from day 55 to day 60 and was constant thereafter, while the N of cuboidal cells decreased proportionally; type II cells contained only 4.8% (vol/vol) of LB at 55 d compared to 18.0% at birth. The N and V of interstitium did not vary by age. While endothelial V was constant over these ages, endothelial S increased from 897 cm2 to 3,398 cm2, and the V of capillary blood and the V of alveolar airspace each increased 4-fold.(ABSTRACT TRUNCATED AT 250 WORDS)     

A novel mechanism of capillary growth in the rat pulmonary microcirculation

Postnatally, the rat lung parenchyma undergoes impressive growth. Within four months of birth, lung volume and alveolar and capillary surface areas increase over 20-fold and capillary volume 35-fold. Investigation of methacrylate casts of the pulmonary microvasculature revealed that, with age, lung capillaries were not only growing in surface and volume but also increasing their network density. We proposed that the capillary bed grows by formation of slender intravascular tissue pillars and termed this type of growth intussusceptive microvascular growth (Caduff et al., Anat. Rec., 216:154-164, 1986). The aim of this investigation was to detect the presence and to analyze the ultrastructure of slender tissue posts (diameter 1-2.5 microns) extending across the capillary lumina in serial electron microscopic sections of rat lung parenchyma (age 44 days). Computer-assisted three-dimensional reconstruction of the capillary lumen confirmed that tissue posts were matching the holes previously observed in casts. Post ultrastructure varied with size from a simple area of interendothelial contact to tissue pillars with a core of interstitial tissue. Based on the changing morphology of the pillars, a hypothesis for their development can be proposed: phase I, creation of a zone of contact between opposite capillary walls (formation of an interendothelial bridge); phase II, reorganization of the intercellular junctions of the endothelium, with central perforation of the capillary layer; phase III, formation of an interstitial post core, with successive invasion by cytoplasmic extensions of myofibroblasts, pericytes, and finally interstitial fibers; and phase IV, growth of the slender pillar to a normal full size capillary mesh. These findings support the new concept of intussusceptive growth of the lung capillary system.     

Ontogeny and nutritional programming of uncoupling protein-2 and glucocorticoid receptor mRNA in the ovine lung

This study investigated the developmental and nutritional programming of uncoupling protein-2 (UCP2), glucocorticoid receptor (GR) and 11β-hydroxysteroid dehydrogenase type 1 (11βHSD1) mRNA in the sheep lung from the time of uterine attachment to 6 months of age. The effect of maternal nutrient restriction on lung development was determined in early to mid gestation (i.e. 28–80 days gestation, period of maximal placental growth, and embryonic and pseudoglandular stages of fetal lung development) and late gestation (i.e. 110–147 days gestation, period of maximal fetal growth, and canalicular and saccular stages of fetal lung development). Fetal lungs were sampled at 80 and 140 days (term ∼148 days) gestation, and sheep lungs at 1, 7, 30 days and 6 months. GR and 11βHSD1 mRNA were maximal at 140 days gestation, whereas UCP2 mRNA peaked at 1 day of age and then declined with postnatal age. Maternal nutrient restriction in both early-to-mid and late gestation had no effect on lung weight, but increased UCP2, GR and 11βHSD1 mRNA abundance at every sampling age. These findings suggest that the developmental ontogeny of UCP2 mRNA in the ovine lung is under local glucocorticoid hormone action and that maternal nutrient restriction has long-term consequences for UCP2 and GR mRNA abundance in the lung irrespective of its timing.     

Ultrastructural morphometric characterization of alveolar type II cells of perinatal sheep: a comparison with adults

Summary The quantitative morphologic changes in alveolar type II cells during the perinatal period were characterized morphometrically in the lungs of fetal lambs at 132, 138, and 147 days gestational age (DGA) and in newborns at 2 days postnatal age (2 DPN). Ultrastructural features were compared with those of type II cells of ewes 365 days old. Lamellar body profile number per type II cell profile was highest at term (147 DGA) and 2 DPN. In adults, the number of lamellar body profiles and volume density of lamellar bodies were equal to those of the 132 DGA fetus. Multivesicular bodies were most common at 138 DGA and in adults. The volume density of cytoplasmic glycogen fell dramatically during the latter part of gestation. The volume density of many cellular organelles increased to the level observed in adults by term (147 DGA). Subcellular composition of type II cells of adult sheep differs from that reported for adult rats chiefly by the volume density of lamellar material within the cytoplasm. Plate-like or globe-like inclusions were present only in the type II cells of adults. Cytoplasmic extensions of the type II cell crossing the basal lamina were most abundant in the 132 and 138 DGA fetal sheep. Cytoplasmic extensions were rare in adults. We conclude that morphologic changes of the alveolar type II cell associated with gestational age follow a species-specific time course. In the sheep, this occurs during the later part of gestation and extends into the neonatal period. Morphologic and morphometric changes appear to correspond with cellular interactions between alveolar type II cells and mesenchymal cells of the interstitium.     

The effects of pregnancy and maternal nutrition on the maternal renin-angiotensin system in sheep

Physiological changes occurring in the mother during pregnancy can determine the outcome of pregnancy in terms of birthweight and neonatal viability. Maternal adaptations include plasma volume expansion linked to enhanced activity of the renin-angiotensin system (RAS). The present study was designed to determine whether these changes occur very early in gestation, and the extent to which maternal nutrient restriction may compromise the maternal RAS. Using sheep, we have investigated the effects of pregnancy per se, maternal nutrient restriction and later restoration of maternal diet on maternal body weight, plasma volume and plasma renin concentration (PRC), and angiotensinogen (Aogen) and arginine vasopressin (AVP) concentration. During the period of placental growth (i.e. 28-80 days gestation) ewes were fed either a nutrient-restricted (NR) diet or were well fed (WF). NR ewes consumed between 3.2 and 3.8 MJ day(-1) of metabolisable energy (ME) which is close to 60 % of requirements taking into account the ME required for both ewe maintenance and growth of the conceptus in order to produce a 4.5 kg lamb at term. WF ewes consumed 150 % of ME requirements. Restoration of maternal diet between 80 and 140 days gestation (i.e. fed to satiety and consuming between 8 and 10.9 MJ day(-1), which is close to 150 % of ME requirements) followed previous nutrient restriction. Between pre-conception and 28 days gestation, plasma volume increased in conjunction with a decline in PRC and Aogen concentration. During the period of nutrient restriction ewe body weight did not increase and plasma volume was lower in NR than WF ewes. During this time there was no effect of maternal nutrition on PRC; however, Aogen concentration was lower in the NR group. From 80 days gestation following the rise in food intake for previously NR ewes, greater increases in ewe body weight, plasma volume and PRC occurred up to term compared with ewes that were well fed throughout gestation. Plasma AVP concentration was not significantly affected by either maternal nutrition or gestational age. In conclusion, the stimulus of moderately severe maternal nutrient restriction evoked smaller rises in maternal weight, plasma volume and Aogen concentration than occurred in ewes that were well fed throughout gestation. Following the restoration of maternal diet after 80 days gestation, PRC gradually rose to peak at term. These adaptations in the maternal RAS during the critical period of placental growth may have long-term effects on fetal development.     

Ultrastructural studies of hepatoblast junctions and liver hematopoiesis of the mouse embryo]

To clarify the morphological changes in hepatoblast connections during the development of fetal liver hematopoiesis, ICR mouse livers of 11 to 19 days of gestation were studied by means of three-dimensional reconstruction, immunohistochemistry, electron microscopy and freeze fracture replica method. Embryonic liver weight showed rapid increase until 19 days of gestation, and an initial steep increase, due to hematopoietic development, was observed at 13 to 15 days of gestation. Hepatoblast volume appeared to be constant until 13 days of gestation, and, thereafter, showed a gradual increase. An 11-day primitive hepatic cord contained a few immature hematopoietic cells among hepatoblasts, and the hepatoblasts made contact with one another by short cytoplasmic projections. The area of the contact surface had a diameter of 4-5 microns, where E-cadherin-mediated adherens junctions were found. At 12-13 days of gestation, hepatoblasts surrounded large ellipsoidal hematopoietic foci, with long cytoplasmic projections. In addition to the adherens junctions, small desmosomes appeared to bind hepatoblasts together, and biliary canaliculi could be recognized between hepatoblasts. At peak stage of liver hematopoiesis at 14 days of gestation, both tight junctions and gap junctions appeared around the biliary canaliculi, and four types of specialized junctions, i.e., adherens junctions, desmosomes, tight junctions and gap junctions, appeared to be fully developed. After 15 days of gestation, hepatocyte volume showed rapid increase, and the surface areas between adjacent hepatocytes were markedly enlarged. As a result, the involuted hematopoietic foci were forced to move from interhepatocytic spaces to perisinusoidal space at the end of the intrauterine life.     

Neonatal hyperoxia alters the pulmonary alveolar and capillary structure of 40-day-old rats

High inspired oxygen concentrations during the neonatal period profoundly inhibit rat lung development, an effect that is partly reversed during recovery in air. Persistent effects of neonatal hyperoxia on the size and number of alveoli or the structure of pulmonary capillaries have not been well defined. Using light and electron microscopic morphometry plus quantitative three-dimensional reconstructions of alveoli, we examined the lungs of 40-day-old rats that were exposed to more than 95% oxygen for the first 7 days after birth. Neonatal hyperoxia administered to rats resulted in abnormally enlarged air spaces at age 40 days. The fraction of the lung consisting of parenchyma was significantly increased and alveolar surface area was 13% lower than controls. There was an abnormal enlargement of alveolar ducts, which reduced by 24% the relative amount of air in the alveoli, compared to that in the alveolar ducts. The number of alveoli per lung and the mean volume of an alveolus were not different between the groups, but alveolar size class distributions were different, with significantly more very small and very large alveoli in 40-day-old rats after neonatal hyperoxia. By scanning electron microscopy, the alveolar surface of the exposed animals had a corrugated appearance, which was especially evident along alveolar ducts. Transmission electron microscopy revealed a greater density of capillaries, particularly in the alveolar regions close to terminal airways. Based on a random sample of the entire parenchymal region, capillary blood volume per cm2 of alveolar basal lamina was 18% greater. The results demonstrate that neonatal exposure to hyperoxia can cause abnormalities in the pulmonary alveolar and capillary structure of 40-day-old rats, and that these changes are similar to some features of broncho-pulmonary dysplasia.     

An immunohistochemical study of the expression of surfactant apoprotein in the hypoplastic lung of rabbit fetuses induced by oligohydramnios.

We shunted amniotic fluid from alternate gestational sacs into the maternal peritoneal cavity between 23 and 30 days gestation in fetal rabbits (full term, 31 days) to investigate the effect of oligohydramnios on surfactant apoprotein A (SP-A) expression by an immunohistochemical morphometric analysis. The amniotic shunt produced a significant decrease in the amniotic fluid volume (P < 0.02), as well as a reduction in the lung weight (P < 0.005) and lung/body weight ratio (P < 0.001), which indicated lung hypoplasia. These fetuses also showed a statistically significant reduction of SP-A expression, ie, SP-A-positive type II cells per unit area (P < 0.05), SP-A-positive type II cells/total cells ratio (P < 0.001), the percentage of SP-A-positive area per unit area (P < 0.005), and the SP-A-positive area/alveolar epithelium area plus the lung interstitium area ratio (P < 0.005). These results suggest that oligohydramnios significantly retards and modifies the structural growth and functional development of alveolar type II cells in SP-A expression. This animal model of hypoplastic lung in fetuses is thus considered to be useful in helping to further develop the treatment for hypoplastic lung.     

Lung growth of the turkey,Meleagris gallopavo: I. Morphologic and morphometric description

To describe lung growth qualitatively and quantitatively from prehatch to adulthood of an unselected line of turkey, a precocial a avian species, 36 male turkeys, three in each age group, were killed at 22 and 25 days of incubation, on hatch day, and at 1, 4, 7, 10, 14, 21, 28, 112, and 420 days of age. Body weight and lung volume were measured. A three-level cascade sampling system was used to prepare lung tissue for morphologic and morphometric observation by light microscopy. Point and intersection counting were used to estimate volume and surface densities of lung compartments relative to lung volume. Absolute volumes and surfaces of lung compartments were calculated. Bilogarithmic regressions provided allometric equations to describe growth of the lung in three phases: (1) Tissue proliferation—explosive growth of lung volume relative to body weight and of the gas-exchange compartment within the lung. At 22 days of incubation there were few air and blood capillaries and a great deal of tissue that looked like mesenchyme between the parabronchin. Within the 6 days prior to hatch, the surface area of air capillaries increased 11-fold and of blood capillaries 27-fold, whereas the volume of interparabronchial tissue decreased 58%. (2) Equilibrated growth—from hatch day to 28 days of age, most lung compartments grew evenly with lung volume. (3) Regulated growth—from 28 days of age to adult, all lung compartments, except large vessels and exchange compartment, grew more slowly than the entire lung. Interatrial septa lengthened and their epithelial covering thinned, infundibula became more apparent, and interparabronchial connective tissue reached a minimal volume density in the adult lung.     

Phagocytosis, chemiluminescence, and cell volume of alveolar macrophages from neonatal and adult pigs

Broncho-alveolar cells lavaged from the lungs of 1- and 7-day-old piglets and adult pigs were examined to determine their alveolar macrophage content, cell size, and ability to phagocytyze emulsified oil droplets and to chemiluminesce in response to opsonized zymosan particles. Alveolar macrophages were identified by specific cytochemical stains and differential cell counts as being 95%, 97%, and 90% of the cell populations, respectively. The cell volume of macrophages from 1-day-old pigs was 383 +/- 40 microns3; while the cell volumes from 7 day and adult pigs were 1,660 +/- 265 and 1,411 +/- 310 microns3. Alveolar macrophage from 1-day-old piglets possessed little ability to engulf oil droplets; however, macrophages from 7-day-old piglets phagocytized these opsonized droplets at a rate equivalent to alveolar macrophages obtained from adult pigs. The phagocytosis of oil droplets by both 7-day and adult alveolar macrophages manifested similar characteristics. Both rates can be described by saturation kinetics, and while temperatures from 37 degrees to 30 degrees C had little effect, the rate declined rapidly between 30 degrees and 16 degrees C. Alveolar macrophages from 1-day-old piglets displayed a lower magnitude of chemiluminescence than cells from 7-day-old and adult pigs and did not always respond to zymosan exposure. On the other hand, 7-day-old and adult pig alveolar macrophages were quite similar in their ability to chemiluminesce. Increasing extracellular glucose from 1 to 20 mM induced a concomitant increase in chemiluminescence. The findings suggest that functionally competent alveolar macrophages emerge in the pig lung approximately 1 week after birth.     

Ontogeny of pulmonary cyclooxygenase-1 (COX-1) and -2 (COX-2)

Prostaglandins synthesized by enzymatic reactions such as cyclooxygenases have been implicated in lung pathophysiology. The goal of this study was to delineate the pulmonary ontogeny of cyclooxygenase enzymes (COX-1 and COX-2) immunohistochemical expression and cellular localization in various microanatomic locations of lungs from pre-term, term, and post-natal lambs. Lung tissues were obtained at 115 and 130 days of gestation from pre-term lambs, 145 days (term; complete gestation), and 15 days post-natally. No significant differences were seen in lung COX-1 expression at various microanatomic locations during pre-term, term, or postnatally. Moderate to strong COX-1 expression was present in macrophages, alveolar septa, bronchial smooth muscle cells, bronchiolar smooth muscle cells, vascular endothelial cells, and vascular smooth muscle cells. Minimal COX-1 expression was present in bronchial and bronchiolar epithelial cells. Most microanatomic locations lacked COX-2 expression with the exception of weak expression that was present in bronchial and bronchiolar epithelial cells at 145 days of full gestation and 15 days post-natally. This work suggests that: (a) COX-1 is constitutively expressed in lungs from pre-term, term, and post-natal lambs in various microanatomic pulmonary locations, (b) there is differential expression of COX-1 and COX-2 in the developing lung, and (c) COX-2 does not appear to play a role in lung fetal development, at least in neonatal lambs.