Morphological characteristics of boar efferent ductules and epididymal duct.

The aim of the present study was to provide a comprehensive morphological analysis of the porcine epididymis in view of the specific functions being performed in different regions of this organ. Blood supply and microvasculature of efferent ductules and epididymal duct were investigated by means of corrosion casts which were analysed macroscopically and by scanning electron microscopy. This revealed blood supply to the testis and epididymis to be closely related. The capillary pattern was typical for the efferent ductules, the caput, corpus, and distal cauda epididymidis, respectively. Corrosion casts were also used to visualize the course of the efferent ductules themselves. Tissue samples from different regions of the efferent ductules and epididymal duct were examined by light microscopy and both scanning and transmission electron microscopy, with special attention being payed to transitional areas. Morphological criteria allowed the distinction of three segments within the efferent ductules and of the initial segment, proximal caput, distal caput, corpus, proximal cauda, and distal cauda regions of the epididymal duct. Components of the endocytic apparatus of efferent ductule principal cells were identified by ferritin uptake. Ultrastructural evidence of absorption in the epididymal duct was particularly prominent in proximal and distal caput. Extensive cisternae of rough endoplasmic reticulum and a well-developed Golgi apparatus were indicative of active protein synthesis and secretion especially in the distal caput and corpus regions. However, assignment of various organelles in principal cells of the epididymal duct to either absorptive or secretory pathways still remains tentative.

Uterine glands of the pig during pregnancy. An ultrastructural and cytochemical study.

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 placenta of the pig. I. Finestructural changes of the placental barrier during pregnancy.

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.

The placenta of the pig. II. The ultrastructure of the areolae.

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.

[Enzyme histochemical studies of the swine placenta. Histoptics of enzymes in interareolar placental epithelia]. / Enzymhistochemische Untersuchungen an der Schweineplacenta. I. Histotopik von Enzymen in den interareolären Placenta-Epithelien.

In porcine interareolar placental epithelia, the following enzymes were demonstrated by histochemical methods after 30, 58, 80, 100, and 110 d of pregnancy, respectively: beta-N-acetylhexosaminidase, beta-galactosidase, beta-glucuronidase, alpha-mannosidase, acid phosphatase, alkaline phosphatase, nonspecific esterases, cytochrome oxidase, 5-nucleotidase, leucine aminopeptidase, adenosine triphosphatase, diaphorases (NADH, NADPH), glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, succinate dehydrogenase, isocitrate dehydrogenase (NAD, NADP), beta-hydroxybutyrate dehydrogenase, glycero-3-phosphate dehydrogenase, NAD-glycero-3-phosphate dehydrogenase, glutamate dehydrogenase (NAD, NADP), lactate dehydrogenase. The results show that most of the enzyme activities remained almost unchanged during the period of investigation. Only G-6-PDH and 6-PGDH activities increased within the uterine epithelium and nonspecific esterase activity within uterine as well as chorionic epithelia during the 2nd half of pregnancy. Within chorionic and uterine epithelia, hydrolases but not dehydrogenases demonstrated a higher activity at the bases of chorionic villi as compared to the apices and flanks of the latter. The action and influence of the demonstrated enzymes on metabolism, energy transfer, secretory, and resorptive activities of chorionic and uterine epithelia are discussed.

[Enzyme histochemistry of the pig placenta. III. Histotopics of enzymes in the uterine epithelium]. / Enzymhistochemische Untersuchungen an der Schweineplacenta. III. Histotopik von Enzymen im Uterindrüsenepithel.

Within the uterine glands, the following enzymes were demonstrated by histochemical methods after 30, 58, 80, 100, and 110 d of pregnancy, respectively: beta-N-acetyl-hexosaminidase, beta-galactosidase, beta-glucuronidase, alpha-mannosidase, acid phosphatase, alkaline phosphatase, esterases, cytochrome oxidase, 5-nucleotidase, leucine aminopeptidase, adenosine triphosphatase, diaphorases (NADH, NADPH), glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, succinate dehydrogenase, isocitrate dehydrogenase (NAD, NADP), beta-hydroxybutyrate dehydrogenase, glycero-3-phosphate dehydrogenase, NAD-glycero-3-phosphate dehydrogenase, glutamate dehydrogenase (NAD, NADP), lactate dehydrogenase. The results show that the activities of G-6-PDH, 6-PGDH, and cytochrome oxidase increase within secreting cells during the 2nd half of pregnancy. The activities of the other enzymes remained almost unchanged during the period of investigation. The description of our results distinguishes between gland neck, middle, and distal part of the secretory unit, respectively. In general, the enzyme activities are similar within the middle and distal gland segments, but lower in the epithelia of the neck region. The activity of dehydrogenases was medium to intensive within the middle and distal gland segments, but only low to medium within the neck portion. Of the hydrolases, the acid phosphatase, ATPase, leucine aminopeptidase, and beta-galactosidase demonstrated an intensive activity within activity secreting cells. The enzyme activities of the gland epithelia are compared with these of the uterine surface epithelia and the histochemical results are discussed in context with their significance in histiotrophic nutrition.

[Enzyme-histochemical studies of the pig placenta. II. Histotopics of enzymes in the areolar placenta epithelium]. / Enzymhistochemische Untersuchungen an der Schweineplacenta. II. Histotopik von Enzymen in den areolären Placentaepithelien.

In porcine areolar placental epithelia, the following enzymes were demonstrated by histochemical methods after 30, 58, 80, 100, and 110 d of pregnancy, respectively: beta-N-acetyl-hexosaminidase, beta-galactosidase, beta-glucuronidase, alpha-mannosidase, acid phosphatase, alkaline phosphatase, nonspecific esterases, cytochrome oxidase, 5-nucleotidase, leucine aminopeptidase, adenosine triphosphatase, diaphorases (NADH, NADPH), glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, succinate dehydrogenase, isocitrate dehydrogenase (NAD, NADP), beta-hydroxybutyrate dehydrogenase, glycero-3-phosphate dehydrogenase, NAD-glycero-3-phosphate dehydrogenase, glutamate dehydrogenase (NAD, NADP), lactate dehydrogenase. The results show that the enzyme activities remained almost unchanged during the period of investigation. Of the dehydrogenases, the diaphorases as well as succinate and lactate dehydrogenase demonstrated generally an intensive activity within the epithelia. The activity of the other dehydrogenases was only low. The activity of unspecific esterase was very intensive within the uterine epithelia but remarkably low within chorionic epithelia. Contrarily, the reaction of adenosine triphosphatase was more intensive within chorionic than uterine epithelia. All investigated glucosidases reacted distinctly positive within chorionic epithelia, but only beta-N-acetyl-hexosaminidase and beta-galactosidase in uterine epithelia. The high activity of acid phosphatase, especially within the chorionic epithelium, seems to be connected with uteroferrin, an iron-binding protein. The histochemical results are discussed in context with the function of the areolae in histiotrophic nutrition and iron transport.

[The glandular compartments of the canine placenta--a scanning electron microscopic study]. / Die Drüsenkammern der Hundeplazenta--eine rasterelektronenmikroskopische Untersuchung.

The glandular chambers in the spongy zone grow out of the superficial endometrial glands during the implantation of the embryo. To our knowledge, no scanning electron microscopic investigations of the spongy layer in the placenta of the bitch have been published so far. We examined the placenta at the 44. day of pregnancy by scanning electron microscopy. The glandular epithelium in the spongy zone forms extensive folds, which fill up most of the lumen of the glandular chambers. They are lined by a simple columnar epithelium with high apical protrusions. The luminal surface of the cells is bordered by short microvilli. Numerous vesicles are stacked in the cytoplasm. The tips of the chorionic tufts project into the lumen of the glandular chambers. The lumen is filled with masses of mucus separating the trophoblast from the glandular epithelium. With respect to its morphology, the trophoblast lining the tips of the chorionic tufts is very different from the trophoblast in the other parts of the placenta. The cells are swollen and club-shaped. The cytoplasm is completely supplanted by big vacuoles. We feel that the trophoblast in the glandular chambers is unable to perform any resorptive activity. The significance of the mucus in the glandular chambers in stopping the invasive growth of the trophoblast is discussed.

[The vascularization of the epididymis of the boar with special reference to perfusion fixation]. / Die Vaskularisation des Ebernebenhodens unter besonderer Berücksichtigung der Perfusionsfixation.

Vasculature of the epididymis was investigated by means of corrosion casts. In the boar, epididymal arteries form a complex network around their stem vessel, the testicular artery. Proper perfusion fixation or complete casting therefore require direct injection into one of these branches. To reach the distal cauda, cannulation of the deferential artery is further needed. Connections between all of these feeding vessels occur at the level of the vascular cone. A prominent anastomosis between an epididymal branch and the testicular artery is regularly observed under the caput epididymis. Epididymal veins drain to a large extent into the pampiniform plexus. Unlike the situation in other species, vascularization of testis and epididymis are closely associated in the boar.

Exposure of rainbow trout (Oncorhynchus mykiss) to nonylphenol is associated with an increased chloride cell fractional surface area.

Nonylphenol is a biodegradation product of a widely used group of non-ionic detergents. Because of its ubiquitous distribution and persistence, nonylphenol is present in surface waters as a pollutant. Little is known about its biological effects at environmentally relevant concentrations other than its action as a xenoestrogen. The goal of the present paper was to study the trout gill surface epithelium as the major interface between fish and water in view of possible morphological alterations due to exposure to nonylphenol. Rainbow trout were intermittently exposed to 10 micrograms/l nonylphenol and gill samples from experimental and control animals were investigated by scanning electron microscopy. Gill surface epithelium was scrutinised for changes in chloride cell density and their status regarding cell surface modifications. In addition, chloride cell fractional surface area (CCFA) was determined by morphometrical methods. Statistical analysis revealed a highly significant increase of CCFA in animals exposed to nonylphenol as compared to control animals (P = 0.0001). Semi-quantitative assessment of the other parameters suggested a higher chloride cell density and a larger proportion of chloride cells bearing microvilli. Taken together, these results provide evidence that exposure of trout to nonylphenol is associated with a substantial increase in the active interface of chloride cells with water. We interpret these findings as being a means to further the fish's capacity for calcium exchange.

Immunohistochemical demonstration of lumbar intervertebral disc innervation in the dog.

Low back pain is a common ailment in dogs, particularly in specific breeds such as the German shepherd dog. A number of structures such as facet joint capsules, ligaments, dorsal root ganglia, periosteum, vertebral endplates and meninges have been associated with this condition. Yet, in spite of all diagnostic efforts, the origin of pain remains obscure in a substantial proportion of all cases. A further structure often being involved in vertebral column disorders is the intervertebral disc. The presence of nerves, however, is a precondition for pain sensation and, consequently, structures lacking innervation can be left out of consideration as a cause for low back pain. Nerve fibres have been demonstrated at the periphery of the intervertebral disc in man, rabbit and rat. With regard to the dog, however, the extent of intervertebral disc innervation is still being disputed. The goal of the present study, therefore, was to substantiate and expand current knowledge of intervertebral disc innervation. Protein gene product (PGP) 9.5 was used for immunohistochemical examination of serial transversal and sagittal paraffin sections of lumbar discs from adult dogs. This general marker revealed nerve fibres to be confined to the periphery of the intervertebral discs. These results indicate that even limited pathological processes affecting the outer layers of the intervertebral disc are prone to cause low back pain.

Macrophage-lymphocyte cluster formation in the medullary sinus of lymph node after immunization with sheep red blood cells (SRBC).

Normally the lymphatic sinuses of the lymph node are loosely packed with lymphocytes and free macrophages as well as with macrophages adhering to the fibrocellular trabeculae. After immunization with SRBC cluster formation occurs in the medullary sinuses of rats between a central macrophage and peripherally located lymphocytes. These rosette-like clusters are nearly identical with the clusters found during primary and secondary immune response against SRBC in vitro and seem to be the in vivo equivalent for the same immune response.

[Electron microscope localization of thiamine pyrophosphatase and nucleoside diphosphatase in lymphocytes]. / Elektronenmikroskopische Lokalisation von Thiaminpyrophosphatase und Nucleosiddiphosphatase in Lymphozyten

In ductus thoracicus lymphocytes of the rat the localization of specific phosphatases was studied by means of ultrahistochemistry. Thiaminepyrophosphatase was found in the outer lamellae of the Golgi-field as well as in lysosomes, whereas nucleosiddiphosphatases could be localized on the plasma membrane at low pH. These findings are correlated with the phaenotype of B- and T-lymphocytes. Their functional significance is discussed.

The junctions between efferent ductules and epididymal duct in the boar.

Sperm transit through the male excurrent duct system is dependent on complete luminal patency. Since male excurrent ducts are derived embryologically from originally separate structures, the junctions between mesonephric tubules and the mesonephric duct are of major interest with respect to obstructive disorders. Therefore, we investigated the junctions between efferent ductules and epididymal duct in adult boars by means of serial semithin and ultrathin sections. Based on the anastomosing pattern and on the site of transition from low columnar, ciliated epithelium to high columnar, non-ciliated epithelium, three different types of anastomoses were identified, one of which was associated with luminal stenosis and sperm accumulation. The occurrence of end-to-side junctions with the epithelial transition being localized within the lumen of the tributary may thus impede normal sperm transport.

Con A- and WGA-binding sites on bovine epididymal spermatozoa: TEM of specimens in toto.

Binding sites for Con A and WGA were detected on bovine spermatozoa during epididymal maturation. We used colloidal gold as an EM-marker. The spermatozoa were treated according to a two-step method for lectin and colloidal gold, then adsorbed to lysine-coated nickel grids and subsequently examined by TEM in toto. Using this method we rapidly got information about the topographic distribution of lectin-binding sites. Major differences exist for WGA between caput and cauda spermatozoa. Conceding that cell-thickness poses some limitation, we consider this method to be practical and especially useful in studies concerning topographic distribution of cell surface components in single cell systems.

A scanning electron-microscopic analysis of the morphology of equine lower lip sinus hair.

Sinus hairs, which are tactile organs of most mammals, are differentiated into cavernous and sinus types. The horse has the cavernous type. Horse lower lips were perfused in diluted Karnovsky's fixative and sinus hair processed for scanning electron microscopy. Anastomosing trabeculae of different thickness and shape originate from the internal connective tissue layer and extend to the external layer, thus forming a dense meshwork suspending the hair root like a net. However, many of the distal trabeculae do not reach the external wall of the blood sinus but end in a finger-like knob within the sinus, thus sharing some features of the sinus type. The sinus wall and trabeculae are completely covered by polygonal endothelial cells showing characteristically protruding nuclei, sparse microvilli and distinct cell borders. Endothelial cells on the trabeculae are more elongated. Openings of small blood vessels were found exclusively on the internal wall, indicating that blood exchange occurs via the internal wall only and that turnover of blood may be minimal. The supposed significance and function of the finger-like trabeculae in the distal part of the blood sinus lying close to the majority of nerve terminals is discussed.