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The purpose of this study was to isolate endothelial cells from different organs of porcine fetuses and to examine the binding of endothelial markers including lectins. Endothelial cells were isolated from the aorta, cerebral cortex, myocardium, ovary and testis. Binding of the antibodies to von Willebrand factor (vWF) and angiotensin converting enzyme (ACE), the presence of Weibel Palade bodies (WPB), uptake of acetylated low density lipoprotein (acLDL), and labelling with the lectins Bandeiraea simplicifolia agglutinin I (BS I), Peanut agglutinin (PNA), Dolichos biflorus agglutinin (DBA) and Ulex europaeus agglutinin I (UEA I) were examined. Cell preparations displayed cobblestone-like morphology with the exception of testicular endothelium, which formed arcuate structures. Endothelium isolated from the brain labelled more strongly than any other cell line with the lectin PNA, but it did not express ACE. In contrast to other cell preparations, myocardial endothelium showed very low binding of anti-vWF. Ovarian endothelium was able to perform in vitro angiogenesis. Moreover, these endothelial cells possessed the largest number of WPB. Testicular endothelium displayed highest binding of vWF. Endothelium isolated from the aorta, in contrast to all other endothelial cells, did not take up acLDL. These results demonstrate that organ- and tissue-specific heterogeneity is already expressed in fetal endothelium.

The present study was undertaken to define the temporal pattern and distribution of cells positive for chromogranin A (CgA) and tyrosine hydroxylase (TH) in various developmental stages of fetal bovine adrenal gland. CgA is an acidic protein, co-stored and co-released with amines and a variety of peptide hormones and neurotransmitters in dense core vesicles of neural and endocrine cells and can be used as a marker for these cells and their malignant counterparts. TH is the rate-limiting enzyme in catecholamine biosynthesis and reflects noradrenergic differentiation. The expression of CgA and TH was examined by immunohistochemistry. CgA immunoreactivity appears first in 35-day-old bovine fetuses. By the end of the second month, CgA-labelled cells are scattered throughout the entire primordium of the adrenal gland, and at a fetal age of 85-91 days most of these cells concentrate in the developing adrenal medulla. From this stage onwards, immunoreactive cells of the marginal zone of the medulla exhibit significantly stronger CgA immunoreaction than the central area. TH immunoreactivity appeared in the adrenal primordium for the first time at the end of the second month of gestation. The distribution pattern of TH-positive cells was similar to that described for CgA, and no significant differences in topographical arrangement between TH- and CgA-positive cells can be detected. The results show that bovine adrenal chromaffin cells express CgA already during their earliest stages of development and prior to TH. The stronger immunoreaction of marginal adrenal medullary cells suggests an adrenalcortical effect of glucocorticoids on the expression of CgA.

The ultrastructure of the areolae in the porcine placenta is described. The areolae occur on day 30 of pregnancy as dome-shaped formation over the openings of the uterine glands. The lumen of the areolae is filled with the secretions of the uterine glands, the so-called histiotroph. The areolae lining epithelium is high collumnar, possessing long microvilli, a well-developed apical tubular system and numerous coated vesicles. This indicates that the epithelium has a high absorptive capacity. Our histochemical investigations reveal a high content of glycoproteins within the areolar lumen. The importance of one of the glycoprotein components of the histiotroph, uteroferrin, is discussed in connection with iron transfer from mother to the fetus.

The ultrastructure of the porcine uterine glands is described from material taken from 11 pregnant pigs at exactly known stages of gestation (day 30; 58; 80; 100; 110). Fixation was performed by perfusion via a branch of the uterine artery and the tissue was routinely processed for electron microscopy. Additionally, cytochemical studies (phosphotungstic acid reaction for glycoproteins, according to Rambourg 1967; acid phosphatase reaction; ultrastructural localization of cellular iron, according to Parmley et al. 1978) were performed. On day 30 of pregnancy the uterine glands are coiled, simple tubular glands with a narrow lumen. The epithelial lining is simple columnar and consists basically of two cell types, ciliated cells and secretory cells. The secretory activity of the glandular epithelium is low; only a few secretory granules are present in the supranuclear cytoplasm. At midpregnancy the ultrastructure of the glands has significantly changed and the cells now show all the characteristics of high secretory activity: numerous parallel cisternae of rough endoplasmic reticulum, an extensively developed Golgi apparatus and many secretory granules which give a positive reaction for acid phosphatase and glycoproteins. The lumina of the glands are significantly enlarged and filled with a great amount of a granular, acid phosphatase-positive material. In the last third of pregnancy, only minor changes in the ultrastructure of the uterine glands are observed. The secretory activity is still high. The amount of rough endoplasmic reticulum has further increased and parallel arrays of cisternae occupy a considerable part of the supranuclear cytoplasm. The importance of the uterine secretion for embryonic nutrition and development is only partly understood. One of the secreted glycoproteins, uteroferrin, is believed to play an important role in the iron transfer from mother to fetus. From midpregnancy onward, a special cell type, the "granule laden cell" is found scattered between normal secretory cells of the uterine glands. Contrary to the opinion of Perry and Cromby (1982), we could demonstrate that these cells frequently extend to the lumen of the gland; hence the term "basal cell" seems inappropriate for this cell type.

The fine structure of bovine Sertoli cells was studied from the 4th to the 40th week post natum in order to correlate the progressive acquisition of normal adult morphology with functional development. The considerable increase in tubular size during the first 20 weeks is due to the proliferation of both presumptive Sertoli and germ cells. Aside from this, the presumptive Sertoli cells are seen to expand radially and lengthen considerably. From then on however, the observed increase in tubular diameter during the later period of postnatal development is solely due to the great increase in the number of germ cells. Presumptive Sertoli cells undergo morphological differentiation to mature Sertoli cells during the first 28 weeks of proliferative development. The maturation process includes distinct changes in cell shape, nucleus and cellular organelles, as well as an increase in and differentiation of Sertoli cell surface specializations. At 24 weeks the development of inter-Sertoli cell junctions has reached a point of differentiation where, in our opinion, a functional blood-testis barrier can be expected. During the first 8 weeks an extensive development of rough endoplasmic reticulum and a well-developed Golgi apparatus can be observed, which suggests a high secretory activity in the presumptive Sertoli cells at this time. We speculate that these secretory activities may play a role in the formation of the basal lamina which is extremely well developed during early postnatal life. The subsequent reduction of the basal lamina correlates well with diminished secretory activity in the Sertoli cells.

The finestructural changes of the interareolar porcine placenta during pregnancy are described. After perfusion fixation of the placenta the change in the thickness of the placental barrier from day 30 to day 110 of gestation is much more evident than after immerson fixation as has been used by all former authors. The alterations are due to the indentation of both the trophoblast and uterine epithelium by their corresponding capillary-network. This indentation is limited to the lateral wall and the summit of the chorionic ridges, while at the base the trophoblast as well as the uterine epithelium remains high columnar. This indicates that in the interareolar porcine placenta, which is represented by the chorionic ridges and the corresponding endometrial folds, at least two different areas with different structure and function may be discerned. 1) The lateral side and the top of the chorionic ridges seem to be predestinated for gaseous exchange. The placental barrier in this area is often less than 2 micrometers. 2) The transport of blood-borne nutrients takes place at the base of the chorionic ridges. This transport seems to be facilitated by an intercellular channel system between the uterine epithelial cells.