Development of the pulmonary vasculature in the gray short-tailed opossum (Monodelphis domestica)-3D reconstruction by microcomputed tomography.

In the marsupial gray short-tailed opossum (Monodelphis domestica), the majority of lung development, including the maturation of pulmonary vasculature, takes place in ventilated functioning state during the postnatal period. The current study uses X-ray computed tomography (µCT) to three-dimensionally reconstruct the vascular trees of the pulmonary artery and pulmonary vein in 15 animals from neonate to postnatal day 57. The final 3D reconstructions of the pulmonary artery and pulmonary vein in the neonate and at 21, 35, and 57 dpn were transformed into a centerline model of the vascular trees. Based on the reconstructions, the generation of end-branching vessels, the median and maximum generation, and the number of vessels were calculated for the lungs. The pulmonary vasculature follows the lung anatomy with six pulmonary lobes indicated by the bronchial tree. The pulmonary arteries follow the bronchial tree closely, in contrast to the pulmonary veins, which run between the pulmonary segments. At birth the pulmonary vasculature has a simple branching pattern with a few vessel generations. Compared with the bronchial tree, the pulmonary vasculature appears to be more developed and extends to the large terminal air spaces. The pulmonary vasculature shows a marked gain in volume and a progressive increase in vascular complexity and density. The gray short-tailed opossum resembles the assumed mammalian ancestor and is suitable to inform on the evolution of the mammalian lung. Vascular genesis in the marsupial bears resemblance to developmental patterns described in eutherians. Lung development in general seems to be highly conservative within mammalian evolution.

Development of the terminal air spaces in the gray short-tailed opossum (Monodelphis domestica)- 3D reconstruction by microcomputed tomography.

Marsupials are born with structurally immature lungs when compared to eutherian mammals. The gray short-tailed opossum (Monodelphis domestica) is born at the late canalicular stage of lung development. Despite the high degree of immaturity, the lung is functioning as respiratory organ, however supported by the skin for gas exchange during the first postnatal days. Consequently, the majority of lung development takes place in ventilated functioning state during the postnatal period. Microcomputed tomography (µCT) was used to three-dimensionally reconstruct the terminal air spaces in order to reveal the timeline of lung morphogenesis. In addition, lung and air space volume as well as surface area were determined to assess the functional relevance of the structural changes in the developing lung. The development of the terminal air spaces was examined in 35 animals from embryonic day 13, during the postnatal period (neonate to 57 days) and in adults. At birth, the lung of Monodelphis domestica consists of few large terminal air spaces, which are poorly subdivided and open directly from short lobar bronchioles. During the first postnatal week the number of smaller terminal air spaces increases and numerous septal ridges indicate a process of subdivision, attaining the saccular stage by 7 postnatal days. The 3D reconstructions of the terminal air spaces demonstrated massive increases in air sac number and architectural complexity during the postnatal period. Between 28 and 35 postnatal days alveolarization started. Respiratory bronchioles, alveolar ducts and a typical acinus developed. The volume of the air spaces and the surface area for gas exchange increased markedly with alveolarization. The structural transformation from large terminal sacs to the final alveolar lung in the gray short-tailed opossum follows similar patterns as described in other marsupial and placental mammals. The processes involved in sacculation and alveolarization during lung development seem to be highly conservative within mammalian evolution.

3D reconstruction of the bronchial tree of the Gray short-tailed opossum (Monodelphis domestica) in the postnatal period.

Recent didelphid marsupials resemble the assumed mammalian ancestor and are suitable to inform on the evolution of the mammalian lung. This study uses X-ray computed tomography (µCT) to three-dimensionally reconstruct the bronchial tree of the marsupial Gray short-tailed opossum (Monodelphis domestica) in order to reveal the timeline of morphogenesis during the postnatal period. The development of the bronchial tree was examined in 37 animals from embryonic day 13, during the postnatal period (neonate to 57 days) and in adults. The first appearance and the branching of lobar, segmental and sub-segmental bronchioles in the lungs were documented. Based on the reconstructions, the generation of end-branching airways, the median and maximum generation and the number of branches were calculated for each pulmonary lobe. At birth, the lung of M. domestica has a primitive appearance since it consists of a simple system of branching airways that end in a number of terminal air spaces, lobar bronchioles, and first segmental bronchioles are present. During the postnatal period, the volumes of the lung and bronchial tree steadily increase and development, differentiation, and expansion of the bronchial tree takes place. By 14 days, the fundamental bronchial tree consisting of lobar, segmental, and sub-segmental bronchioles has been established. A mature bronchial tree, including respiratory bronchioles and alveolar ducts is present by day 35. The asymmetry of the right (predominately four lobes) and the left lung (predominately two lobes), as present in M. domestica, can be considered as plesiomorphic for Mammalia. In marsupials, the process of branching morphogenesis, which takes place intrauterine in the placental fetus, is shifted to the postnatal period, but follows similar patterns as described in placentals. Lung maturation in general and the branching morphogenesis in particular seems to be highly conservative within mammalian evolution.

Early postnatal lung development in the eastern quoll (Dasyurus viverrinus).

Early postnatal lung development (1-25 days) in the eastern quoll (Dasyurus viverrinus) was investigated to assess the morphofunctional status of one of the most immature marsupial neonates. Lung volume, surface density, surface area, and parenchymal and nonparenchymal volume proportions were determined using light microscopic morphometry. The lungs of the neonate were at the canalicular stage and consisted of two "balloon-like" airways with few septal ridges. The absolute volume of the lung was only 0.0009 cm3 with an air space surface density of 108.83 cm-1 and a surface area of 0.082 cm2 . The increase in lung volume in the first three postnatal days was mainly due to airspace expansion. The rapid postnatal development of the lung was indicated by an increase in the septal proportion of the parenchyma around day 4, which was reflected by an increase in the airspace surface density and surface area. By day 5, the lung entered the saccular stage of development with a reduction in septal thickness, expansion of the tubules into saccules and development of a double capillary system. The subsequent saccular period was characterized by repetitive septation steps, which increased the number of airway generations. The lungs of the newborn Dasyurus viverrinus must be considered as structurally and quantitatively insufficient to meet the respiratory requirements at birth. Hence, cutaneous gas exchange might be crucial for the first three postnatal days. The lung has to mature rapidly in the early postnatal period to support the increased metabolic requirements of the developing young.

Development of the skin in the eastern quoll (Dasyurus viverrinus) with focus on cutaneous gas exchange in the early postnatal period.

A morphological and morphometric study of the skin development in the eastern quoll (Dasyurus viverrinus) was conducted to follow the transition from cutaneous to pulmonary gas exchange in this extremely immature marsupial species. Additionally, the development of the cardiac and respiratory system was followed, to evaluate the systemic prerequisites allowing for cutaneous respiration. The skin in the newborn D. viverrinus was very thin (36 ± 3 µm) and undifferentiated (no hair follicles, no sebaceous and perspiratory glands). Numerous superficial cutaneous capillaries were encountered, closely associated with the epidermis, allowing for gaseous exchange. The capillary volume density was highest in the neonate (0.33 ± 0.04) and decreased markedly during the first 4 days (0.06 ± 0.01). In the same time period, the skin diffusion barrier increased from 9 ± 1 µm to 44 ± 6 µm. From this age on the skin development was characterized by thickening of the different cutaneous layers, formation of hair follicles (day 55) and the occurrence of subcutaneous fat (day 19). The heart of the neonate D. viverrinus had incomplete interatrial, inter-ventricular, and aortico-pulmonary septa, allowing for the possibility that oxygenated blood from the skin mixes with that of the systemic circulation. The fast-structural changes in the systemic circulations (closing all shunts) in the early postnatal period (3 days) necessitate the transition from cutaneous to pulmonary respiration despite the immaturity of the lungs. At this time, the lung was still at the canalicular stage of lung development, but had to be mature enough to meet the respiratory needs of the growing organism. The morphometric results for the skin development of D. viverrinus suggest that cutaneous respiration is most pronounced in neonates and decreases rapidly during the first 3 days of postnatal life. After this time a functional transition of the skin from cutaneous respiration to insulation and protection of the body takes place.

Skin structure in newborn marsupials with focus on cutaneous gas exchange.

A morphological and morphometric study of the skin of a variety of newborn marsupials (Dasyurus viverrinus, Monodelphis domestica, Trichosurus vulpecula, Isoodon obesulus, Perameles nasuta, Phascolarctos cinereus, Potorous tridactylus, Petrogale penicillata, Thylogale thetidi, Macropus dorsalis) and of a monotreme hatchling (Ornithorhynchus anatinus) was undertaken to assess the possibility of cutaneous gas exchange. Additionally, the lungs of some of these species were investigated to assess its structural degree at birth. The skin in the different newborn marsupials and the monotreme hatchling had a similar structure (no hair follicles and no sebaceous or perspiratory glands) and was in all cases less developed than the skin of altricial eutherians. The thickness of the entire skin (36-186 µm) and its different layers, epidermis (6-29 µm) and dermis (29-171 µm) varied among the marsupial species and reflected the differences in size and developmental degree of the neonates. In the skin of all marsupial neonates and the monotreme hatchling, numerous superficial cutaneous capillaries were encountered, some closely associated with the epidermis, indicating the possibility that the skin participated in gaseous exchange. The skin of the newborn D. viverrinus had the highest capillary volume density and shortest skin diffusion barrier of all marsupial neonates, suggesting that skin gas exchange in the dasyurid neonate might be the most pronounced. A graduation of the skin capillary density among the marsupial neonates inversely followed the respective lung structure and general developmental degree of the neonates.

Comparative anatomy of neonates of the three major mammalian groups (monotremes, marsupials, placentals) and implications for the ancestral mammalian neonate morphotype.

The existing different modes of reproduction in monotremes, marsupials and placentals are the main source for our current understanding of the origin and evolution of the mammalian reproduction. The reproductive strategies and, in particular, the maturity states of the neonates differ remarkably between the three groups. Monotremes, for example, are the only extant mammals that lay eggs and incubate them for the last third of their embryonic development. In contrast, marsupials and placentals are viviparous and rely on intra-uterine development of the neonates via choriovitelline (mainly marsupials) and chorioallantoic (mainly placentals) placentae. The maturity of a newborn is closely linked to the parental care strategy once the neonate is born. The varying developmental degrees of neonates are the main focus of this study. Monotremes and marsupials produce highly altricial and nearly embryonic offspring. Placental mammals always give birth to more developed newborns with the widest range from altricial to precocial. The ability of a newborn to survive and grow in the environment it was born in depends highly on the degree of maturation of vital organs at the time of birth. Here, the anatomy of four neonates of the three major extant mammalian groups is compared. The basis for this study is histological and ultrastructural serial sections of a hatchling of Ornithorhynchus anatinus (Monotremata), and neonates of Monodelphis domestica (Marsupialia), Mesocricetus auratus (altricial Placentalia) and Macroscelides proboscideus (precocial Placentalia). Special attention was given to the developmental stages of the organs skin, lung, liver and kidney, which are considered crucial for the maintenance of vital functions. The state of the organs of newborn monotremes and marsupials are found to be able to support a minimum of vital functions outside the uterus. They are sufficient to survive, but without capacities for additional energetic challenges. The organs of the altricial placental neonate are further developed, able to support the maintenance of vital functions and short-term metabolic increase. The precocial placental newborn shows the most advanced state of organ development, to allow the maintenance of vital functions, stable thermoregulation and high energetic performance. The ancestral condition of a mammalian neonate is interpreted to be similar to the state of organ development found in the newborns of marsupials and monotremes. In comparison, the newborns of altricial and precocial placentals are derived from the ancestral state to a more mature developmental degree associated with advanced organ systems.

The placentation of eulipotyphla-reconstructing a morphotype of the Mammalian placenta.

Placentation determines the developmental status of the neonate, which can be considered as the most vulnerable stage in the mammalian life cycle. In this respect, the different evolutionary and ecological adaptations of marsupial and placental mammals have most likely been associated with the different reproductive strategies of the two therian clades. The morphotypes of marsupial and placental neonates, as well as the placental stem species pattern of Marsupialia, have already been reconstructed. To contribute to a better understanding of the evolution of Placentalia, a histological and ultrastructural investigation of the placenta in three representatives of Eulipotyphla, that is, core insectivores, has been carried out in this study. We studied the Musk shrew (Suncus murinus), the four-toed hedgehog (Atelerix albiventris), and the Iberian mole (Talpa occidentalis). As a result, a eulipotyphlan placental morphotype consisting of a compact and invasive placenta was reconstructed. This supports the widely accepted hypothesis that the stem lineage of Placentalia is characterized by an invasive, either endothelio- or hemochorial placenta. Evolutionary transformations toward a diffuse, noninvasive placenta occurred in the stem lineages of lower primates and cetartiodactyles and were associated with prolonged gestation and the production of few and highly precocial neonates. Compared to the choriovitelline placenta of Marsupialia, the chorioallantoic placenta of Placentalia allows for a more intimate contact and is associated with more advanced neonates.

Evolution and development of fetal membranes and placentation in amniote vertebrates.

We review aspects of fetal membrane evolution and patterns of placentation within amniotes, the most successful land vertebrates. Special reference is given to embryonic gas supply. The evolution of fetal membranes is a prerequisite for reproduction independent from aquatic environments. Starting from a basically similar repertoire of fetal membranes - the amnion, chorion, allantois and yolk sac, which form the cleidoic egg - different structural solutions for embryonic development have evolved. In oviparous amniotes the chorioallantoic membrane is the major site for the exchange of respiratory gases between fetus and outer environment. The richly vascularised yolk sac and allantois in concert with the chorion play an important role in the evolution of placentation in various viviparous amniotes. Highly complex placentas have evolved independently among squamate sauropsids and in marsupial and placental mammals. In conclusion, there seems to be a natural force to improve gas exchange processes in intrauterine environments by reducing the barrier between the blood systems and optimising the exchange areas.

Evolution and development of gas exchange structures in Mammalia: the placenta and the lung.

Appropriate oxygen supply is crucial for organisms. Here we examine the evolution of structures associated with the delivery of oxygen in the pre- and postnatal phases in mammals. There is an enormous structural and functional variability in the placenta that has facilitated the evolution of specialized reproductive strategies, such as precociality. In particular the cell layers separating fetal and maternal blood differ markedly: a non-invasive epitheliochorial placenta, which increases the diffusion distance, represents a derived state in ungulates. Rodents and their relatives have an invasive haemochorial placental type as optimum for the diffusion distance. In contrast, lung development is highly conserved and differences in the lungs of neonates can be explained by different developmental rates. Monotremes and marsupials have altricial stages with lungs at the early saccular phase, whereas newborn eutherians have lungs at the late saccular or alveolar phase. In conclusion, the evolution of exchange structures in the pre- and postnatal periods does not follow similar principles.

Ventilatory response to hypoxia in chicken hatchlings: a developmental window of sensitivity to embryonic hypoxia.

We had reported previously [Szdzuy, K., Mortola, J.P., 2007b. Ventilatory chemosensitivity of the 1-day-old chicken hatchling after embryonic hypoxia. Am. J. Physiol. (Regul. Integr. Comp. Physiol.) 293, R1640-R1649] that hypoxia during incubation blunted ventilatory chemosensitivity in the hatchling. Because the carotid bodies become functional in the last portion of incubation, we asked whether these last days were the critical period for the effects of hypoxia on the development of ventilatory chemosensitivity. White Leghorn chicken eggs were incubated at 38 degrees C either in 21% O(2) (Controls) or in 15% O(2) for the whole 3-week incubation (HxTot) or for only the 1st (Hx1), 2nd (Hx2) or 3rd week of incubation (Hx3). Hatching time had a delay of half a day in HxTot, and was normal in the other groups. Body weight was similar in all hatchlings. Oxygen consumption ( [Formula: see text] ) and pulmonary ventilation (V e) were measured at about 20 h post-hatching. Ventilatory chemosensitivity was evaluated from the degree of hyperpnea (increase in V e) and hyperventilation (increase in [Formula: see text] ) during acute hypoxia (15 and 10% O(2), 20 min each) and acute hypercapnia (2 and 4% CO(2), 20 min each). The responses to hypoxia were similarly decreased in HxTot and in Hx3 compared to controls, and were normal in the other experimental groups; those to hypercapnia were blunted only in HxTot. The results are in agreement with the idea that prenatal hypoxia blunts V e chemosensitivity by interfering with the normal development of the carotid bodies.

Lung development of monotremes: evidence for the mammalian morphotype.

The reproductive strategies and the extent of development of neonates differ markedly between the three extant mammalian groups: the Monotremata, Marsupialia, and Eutheria. Monotremes and marsupials produce highly altricial offspring whereas the neonates of eutherian mammals range from altricial to precocial. The ability of the newborn mammal to leave the environment in which it developed depends highly on the degree of maturation of the cardio-respiratory system at the time of birth. The lung structure is thus a reflection of the metabolic capacity of neonates. The lung development in monotremes (Ornithorhynchus anatinus, Tachyglossus aculeatus), in one marsupial (Monodelphis domestica), and one altricial eutherian (Suncus murinus) species was examined. The results and additional data from the literature were integrated into a morphotype reconstruction of the lung structure of the mammalian neonate. The lung parenchyma of monotremes and marsupials was at the early terminal air sac stage at birth, with large terminal air sacs. The lung developed slowly. In contrast, altricial eutherian neonates had more advanced lungs at the late terminal air sac stage and postnatally, lung maturation proceeded rapidly. The mammalian lung is highly conserved in many respects between monotreme, marsupial, and eutherian species and the structural differences in the neonatal lungs can be explained mainly by different developmental rates. The lung structure of newborn marsupials and monotremes thus resembles the ancestral condition of the mammalian lung at birth, whereas the eutherian newborns have a more mature lung structure.