Vibeke Dantzer

Electron microscopy of the initial stages of placentation in the pig

SummaryTo elucidate the morphology of the initial stages of epitheliochorial placentation in the pig, material from 10 sows of the Danish Landrace and from one Göttinger minipig gilt from day 13 to day 26 of gestation was processed for scanning and transmission electron microscopy. The observed foetomaternal interaction from day 19 1/2 minipig placenta corresponded well to the observations on the Danish Landrace placenta. From the results and the discussion it was concluded that the following structures were implicated in the initial phases of placentation in the pig: (1) Protruding epithelial proliferations of the uterine epithelium enclosed by chorionic caps serving to immobilize the blastocyst (days 13 and 14). (2) A thick glycocalyx on the maternal and a thin one on the foetal epithelium before contact. (3) Close apposition between the apical plasma membranes from trophoblastic and uterine epithelium (day 14). (4) Development of interdigitating microvilli (days 15–16). (5) Formation of apical domes on the uterine epithelium closely related to the trophoblast provided with long cytoplasmic extensions into a luminal space between the apical domes, apparently representing a transition from histiotropic to haemotrophic nutrition (days 15–20). (6) Placentation, development of interdigitating microvilli between foetal and maternal epithelium, was extended but not terminated in the peripheral zone at day 26.

Placental vascular corrosion cast studies: A comparison between ruminants and humans

The microvasculature of both the ruminant placentomes of cattle, sheep, and goats and the human placenta were compared, using corrosion casts of blood vessels and scanning electron microscopy. The fetal vascular trees of ruminant and human placenta differ in form and size, which correlates with the degree of ramification; however, their architecture of stem, intermediate, and terminal villi is similar. In the human, the system of serially linked capillary convolutions of terminal villi is longer than that in ruminants. Therefore, in guaranteeing blood flow against flow resistance, the human vessels particularly need a straight course, anastomoses, and sinusoidal dilations. Specifically in the ruminants studied, the venous vessels outweigh the arterial ones by volume and by number. They are suggested to be absorptive for substances metabolized in the zone of the capillary complex. The most extreme interspecies difference relates to the maternal vasculature, which, in contrast to the fetal system, is a closed system in the ruminant septas and an open lacunal intervillous space in the human. Converging and differing morphological vascular phenomena of ruminants and human placenta are discussed in terms of maternofetal exchange related to placental efficiency. In summary, the ruminant placenta, concerning the fetal vascular tree, in many aspects is workable as a model for the human. Microsc. Res. Tech. 38:76–87, 1997.

A low-salt diet facilitates Cl secretion in hen lower intestine

SummaryThe regulation of sodium and chloride transport in hen coprodeum by mineralocorticoids was investigated with isolated epithelia under short-circuit conditions. Unidirectional fluxes of Na and Cl were measured by isotopes and modulated by amiloride, theophylline and bumetanide. Hens were maintained either on low-NaCl diet (LS) or on high-NaCl diet (HS). Plasma aldosterone (PA) levels of these groups were measured with radioimmunoassay. A group of HS hens received injections of aldosterone on a 6-hr schedule before experiments. Another group of LS hens was resalinated, and experiments carried out on a 24-hr interval.Salt deprivation stimulated PA levels ninefold, compared to HS hens. Na absorption was stimulated according to previous reports. Electrogenic Cl secretion was elicited by theophylline and partially inhibited by bumetanide. Modulation of PA levels by diet, resalination or aldosterone injection changed the magnitude of electrogenic Cl secretion in parallel between 0.5 μeq/cmau2 · hr (HS) and 4 μeq/cm2 · hr (LS), with pronounced alteration in tissue resistance.The results demonstrate a new action of aldosterone which besides stimulating Na absorption also directly or indirectly elicits Cl secretion. Evidence is presented for a hormonal adaptation of chloride transport in this epithelium. There was a morphological change of the apical plasma membrane and further experiments will have to clarify the exact cellular nature of this process.

Structural and functional aspects of porcine placental microvasculature

SummaryThe microvascular architecture of the pig placenta was studied by serial semithin histological sections for light microscopy, which were compared with scanning electron microscopy of artificially exposed materno-fetal contact surfaces as well as of vessel casts prepared from the maternal, fetal, and combined maternal and fetal sides.The superficial reliefs from the exposed surfaces as well as from the casts are almost identical with the complementary maternal and fetal sides. In order to meet the physiological needs of materno-fetal exchange for the rapidly growing fetuses, these reliefs develop from a simple to a more complex system during pregnancy and can be described as follows: (1) The degree of interlocking increases between the fetal ridges or bulbous protrusions and maternal ridges of different orders separated by maternal troughs of variable depth, most clearly seen on vessel casts. It creates a three-dimensional nontact area. (2) The structure of precapillary vessels as well as of the meshwork, and the diameter of capillaries of the maternal and fetal sides, adapt during gestation giving a good distribution of oxygenated blood into the maternal capillaries; these, with the development of large prevenous connecting capillaries on the fetal side, favour a high arterio-venous difference of fetal blood O2 pressure. (3) The vascular architecture of endometrial and fetal ridges and troughs develop into a crosscurrent to countercurrent materno-fetal blood interrelationship.Our demonstration of the materno-fetal capillary interrelationship in the porcine placenta thus shows that the latter is a much more efficient organ for exchange than hitherto assumed.

Fetal villosity and microvasculature of the bovine placentome in the second half of gestation

The architecture of the fetal villous tree and its vasculature in the bovine placentome were studied in the second half of gestation using both conventional histology and histology of ink‐filled blood vessels. These were compared with corrosion casts of plastic fillings of the vasculature, prepared for scanning electron microscopy. This combination of morphological methods allows perception of the villous tree throughout gestation from broad‐conical to tall‐conical form where branch ramification occurs mainly at right angles to the stem. The stem villus typically contains a single central artery and several peripheral veins arranged in parallel. The proximal branches to the stem, the intermediate villi, contain a central arteriole and accompanying venules. The distal branches, the terminal villi, enclose capillary convolutions which consist of an afferent arterial capillary limb, capillary loops and efferent venous capillary limbs. Vascular interconnections exist within the terminal villi, as capillaries or venules between the capillary convolutions, serially bridging them in up to 5 places, and as capillary anastomoses between the capillary loops. Coiling and sinusoidal dilatations of these loops develop near the end of gestation. The intraplacentomal rearrangement of villous trees with progressive gestation and their morphological vascular adaptations are discussed in relation to placental function, including the ever increasing need for transplacental substance exchange. This adaptation allows the blood to traverse the shortest possible arterioarteriolar route to the periphery of the trees where exchange takes place. The need for an increasing blood flow stimulates capillary growth and at the same time optimises the blood flow reaching the placental barrier represented by the vessel cast surface. The capillaries also carry the blood back into the very voluminous system of venules and veins where back diffusion may occur. The total volume of terminal villi of bovine placentome, the ‘working part’ of villous trees, hence distinctly increases with respect to the stem and intermediate villi, the ‘supplying part’ of the villous tree. In morphological terms the efficiency of the bovine transplacental diffusional exchange is higher than in the closely related ‘co‐ruminants’ sheep and goats and distinctly higher when compared with the human placenta.

Scanning Electron Microscopy of Exposed Surfaces of the Porcine Placenta

The three-dimensional development of the separated parts of the porcine placenta from 9 Danish Landrace sows at gestational stages from 20 to 100 days was studied. After cautious separation of the allantochorion and the endometrium in a 1 mM buffered ethylenediaminetetraacetic acid solution, the separated parts were processed for scanning electron microscopy by routine methods. The macroscopic enlargement resulted from primary and secondary circular folds or plicae, which were permanent on the maternal side, whereas they were mainly non-permanent on the fetal side. The areolar placenta and interareolar placenta with formation of permanent microscopic folding on both sides were described. The observations of the separated parts yielded new information on the development of surface enlargement during gestation and revealed a hitherto unknown regular architecture of the endometrium by the formation of parallel primary ridges or rugae with secondary ridges or rugae at their sides subdividing the maternal troughs or fossae. This configuration on the maternal side explains the transformation of the regular chorionic ridges from the 63-day stage into bulbous protrusions at the 100-day stage. Based on these observations the precise terminology used above was proposed.

The organisation of the enteric nervous system in the submucous and mucous layers of the small intestine of the pig studied by VIP and neurofilament protein immunohistochemistry

The arrangement of the enteric ganglia and nerve fibre plexuses was examined in the submucous and mucous layers and around Peyers patches of the porcine small intestine to clarify their organisation. Immunohistochemistry of vasoactive intestinal peptide (VIP) and neurofilament proteins in wholemounts, chopped or paraffin sections was used to locate the neural elements. The ganglia of the internal and external submucous plexuses were situated at 2 different topographic locations, being clearly demarcated by the submucosal vascular arcades and differing in neuronal composition. The internal submucous plexus was the only contributor to the plexus surrounding the follicles of Peyers patches as a continuous mesh of 3 ganglionated nerve subplexuses. VIP‐immunoreactive fibres from this mesh innervated the dome. The mucosal plexus, which was subdivided into 4 subunits — the outer proprial, inner proprial, pericryptal and villous plexuses — contained a few solitary neuronal perikarya. Labelling for neurofilament proteins revealed Dogiel types II, IV and VI neurons. The observations reveal several new features in the enteric nervous system of the pig and clarify its nomenclature.

Glycosylation in the near-term epitheliochorial placenta of the horse, donkey and camel: A comparative study of interbreeding and non-interbreeding species

Studies from this laboratory have shown great diversity in the glycosylation of tissues comprising the interhaemal barrier of species with different placental types. This diversity may be one of the factors preventing interbreeding between species. Glycan expression within the uterine epithelium and trophoblast of the interhaemal barrier was examined to test this proposition in three species with similar diffuse, microcotyledonary, epitheliochorial allantochorionic types of placenta: the horse (Equus caballus) and donkey (Equus asinus), which can interbreed with each other, and the camel (Camelus dromedarius), which cannot interbreed with either of the other two species. A panel of 14 lectins was used and it was found that glycosylation patterns were generally similar between placental tissues of the horse and donkey, except for the expression of non-bisected complex N-glycan and some sialic acids, whereas those of the camel showed striking differences in the binding of lectins to many structures carrying terminal residues of fucose, N-acetyl galactosamine and beta-galactose, as well as to complex N-glycans and sialic acids. These results are consistent with the proposition that interbreeding species carry similar glycans in tissues forming the interhaemal barrier whereas glycodiversity is one of the factors preventing implantation and subsequent placental development in interspecific hybrids.

Comparative Morphological Aspects of Placental Vascularization

The placental vascular architecture is interesting not only from the morphological point of view, but also as a basis for functional interpretation. The materno-fetal vascular arrangement in the placental exchange areas is one important criterion for the effectiveness of transplacental transport.

THE DEVELOPMENT OF THE ENDOTHELIOCHORIAL MINK PLACENTA : LIGHT MICROSCOPY AND SCANNING ELECTRON MICROSCOPICAL MORPHOMETRY OF MATERNAL VASCULAR CASTS

 The development of the mink endotheliochorial placenta has been studied by means of light microscopy and scanning electron microscopy of maternal vascular corrosion casts. The placental blood vessels of three groups of mink, representing early, intermediate and near-term gestational ages were either perfusion fixed for histology, or instilled with liquid plastic in order to prepare vascular casts, which were examined qualitatively and/or quantitatively. The maternal component of the placental vascular system evolves from preimplantation blood vessels between the endometrial glands, into which the initial feto-maternal contact is made. The influence of highly invasive syncytiotrophoblast provokes a transition of the maternal capillaries into extensively anastomosing sinusoids with a subsequent modification of their endothelial cells into large cells with luminal protrusions. Three-dimensionally, the sinusoids are arranged as vascular crypts. This implies a villous-crypt type of interdigitation for the mink, but since the fetal capillaries surround the maternal sinusoids as a dense network a labyrinth is formed. The vascular crypts are supplied by very short arterioles, branching from maternal stem arteries, which arise from branches of the uterine artery and move straight to the surface of the endometrium. Venous outlets of the sinusoids converge onto venules and large stem veins in the deepest portion of the endometrium. This architectural pattern persists until term. Morphometry was used to confirm the qualitative observations in vascular casts. The diameter of materal vascular crypts significantly increased from 137.3±21.9 µm in early gestation up to 217.8±80.9 µm in the intermediate stage and 431.8±119.5 µm near-term, when compared to the paraplacental zone in early gestation (82.2±19.5 µm). The capillary or sinusoidal diameter also increased significantly from intermediate stage (42.9±11.8 µm) to near term (60.1±16.7 µm), whereas the difference in the paraplacental zone (7.3±2.1 µm) and early gestation (13.0±3.2 µm) was not statistically significant.