Developmental toxicity in rat fetuses exposed to the benzimidazole netobimin.

Netobimin (NTB) is a prodrug of albendazole (ABZ) and is used as a broad-spectrum anthelmintic both in human and veterinary medicine. Pregnant Sprague-Dawley rats were treated po with 50, 59.5 and 70.7 mg/kg of NTB on Gestational Day (GD) 10. The results, observed on GD 20, demonstrated that NTB induced a significant increase of resorptions. Moreover, decreased fetal body weight and an increase in skeletal malformations were observed in treated groups. We report the first study in which vascular malformations are described in rats after the administration of a benzimidazole compound. An interesting relationship between intercostal vessel and rib malformations was found.

The lack of genital ridge vascularization in the early chick embryo: implications in the migration of the primordial germ cells.

It is known that chick primordial germ cells (PGCs) in early embryonic development migrate via the blood vascular system to colonize the gonadal anlagen. Classically, two factors have been involved in the extravasation of PGCs from the blood stream: chemotactic and mechanical factors. An accurate knowledge of the vascular system of the genital ridge is therefore necessary. However, development of gonadal vascularization in bird embryos has been scarcely studied. Our previous studies have shown that the gonadal arteries develop from the mesonephric arteries. The purpose of this work was to study the implications of the development of the vascular system of the chick genital ridge on PGCs colonization. We selected the Hamburger and Hamilton (H-H) stage 18, since the genital ridge is well developed and PGCs actively extravasate. Forty chick embryos of this stage were processed for scanning electron microscopy of vascular corrosion casts and of critical point-dried specimens as well as light microscopy. Our results are conclusive. We could not find any vessel or capillary network supplying the genital ridge; the dorsal aorta and the primordia of the mesonephric arteries were the closest vessels. However, numerous interendothelial spaces were found in the dorsal aorta at the level of the genital ridge. It is suggested that the interendothelial gaps may be very important in the exchange of substances between the avascular genital ridge and the aortic endothelium at this developmental stage. Two different routes are thought to be involved in PGC migration to the gonadal anlage at this stage: the aortic endothelium and the mesonephric arteries. Whereas mechanical factors may be important for extravasation of PGCs in the mesonephric arteries, no reasons have been found from the morphological point of view to support a slowness of the blood flow in the dorsal aorta at the level of the genital ridge facilitating the extravasation.

Injection technique and scanning electron microscopic study of the arterial pattern of the 20 gestation days (G20) rat fetus.

A technique to obtain microvascular corrosion casts of the G20 rat fetus and the normal pattern of the main arteries of the G20 rat fetus are described. The casts were studied by means of scanning electron microscopy (SEM). The arterial pattern is similar to that described in the adult; however, several variations have been found. It is concluded that the use of vascular corrosion casts studied by SEM may be particularly helpful to observe the extremely small arteries of rat fetuses. Moreover, we suggest that this technique may be useful in practical teratological studies.

Study of the distribution of albendazole-sulphoxide (ABZ-SO) in fertilized egg compartments.

In order to use the chicken embryo in teratogenic studies, it is necessary to know the internal volume in which a xenobiotic distributes. The inoculation of a xenobiotic in one of the compartments of the fertilized egg is the usual technique used in these studies. Neither the concentration nor the moment in which the xenobiotic comes into contact with the chicken embryo have been considered. Predicting the internal volume of distribution in the egg from some of the external parameters that do not interfere with the normal development is necessary. A simple method to calibrate these external parameters and their correlation with the different compartments of the fertilized eggs as well as the different distribution of the xenobiotic in these compartments has been successfully demonstrated. After injection of ABZ-SO, the maximum concentration in the embryo is reached by 36 h. The mean AUC for the albumen (sharp and obtuse end), yolk, and embryo were 78.4, 40.7, 79.2, and 10.8 micrograms.h/ml respectively. The results obtained about the kinetics of the diffusion of ABZ-SO indicate that this compound does not have a homogeneous distribution in all the compartments of the fertilized egg. These results highlight that whenever fertilized eggs are used as a screening for the possible toxicity of a drug or other substances, the dose of the xenobiotic to be injected has to be precisely determined in accordance with the total volume and the stage of embryonic development selected to be affected, starting from the previous knowledge of when and how much substance accedes to the embryo.

Angiogenesis in the gonadal capillary network of the chick embryo.

Seventy-one chick embryos of both sexes at the 35 Hamburger and Hamilton (H-H) developmental stage were processed for scanning electron microscopy of vascular corrosion casts and of critical point dried specimens, as well as transmission electron- and light microscopy, in order to study the angiogenic structures. The gonadal subepithelial capillary network was located at the level of the tunica albuginea under the covering epithelium. The casts showed a densely-meshed capillary network and numerous sprouting (nodular protrusions or capillary sprouts) and non-sprouting (enlarged vessels and angiogenic holes) angiogenic structures that were randomly distributed and mixed. Four types of angiogenic holes were encountered in the casts: primary (diameter < 2.5 microns), secondary (diameter > 2.5 microns), tertiary (variable diameter and circular narrowings on one side), and open angiogenic holes. We suggest that the different morphologies reflect evolution of these holes. Furthermore, the open angiogenic hole would probably either form nodular protrusions at its open ends, which tend to join with other nodular protrusions and neighboring capillaries and form new vessels; or there would be fusion with two or more neighboring open holes. Correlative critical point dried sections showed fenestrations in the capillary walls and transcapillary pillars that corresponded to the angiogenic holes found in the casts. Ultrathin sections of the vessels presented typical characteristics of growing endothelium: large nuclei with loosely textured chromatin, abundant cytoplasm rich in cell organelles and intraluminal endothelial processes.

Development and degeneration of the arterial system in the mesonephros and metanephros of chicken embryos.

BACKGROUND: The general morphology of the mesonephric and metanephric arteries in chicken embryos has already been described previously. Moreover, the general basis of glomerulogenesis has also been established. However, the degeneration of the mesonephric vascular system, and especially glomerular degeneration, have not been well established yet. Also the morphology of the metanephric angiogenic buds has not been studied yet. METHODS: Scanning electron microscopy of vascular corrosion casts and critical point dried specimens as well as light microscopy of serially sectioned material has been used in this study. Mesonephric degeneration coincides in time with metanephros growth and maturation in the developmental stages of chicken embryos chosen for this investigation (7.5, 9, 11, and 14 days of incubation). RESULTS: The arterial system of the mesonephros in embryonic chicken is similar to that of the anuran kidney, as described in the literature. The morphology of the degenerating mesonephric glomeruli shows that the glomerular capillaries are more thick, tortuous, and numerous than those in normal glomeruli. The podocytes also show degeneration. The arterial system of the metanephros grows directly from the aorta and from the mesonephric arterial system. During these stages of rapid growth, the metanephros shows angiogenic buds. These angiogenic buds can be either pointed or round blind endings. CONCLUSIONS: The distribution and topography of the mesonephric and metanephric arteries is in general accordance with the literature. The process of glomerular degeneration in the mesonephros seems similar to glomerular senescence in man but is different from that of the aged rat glomeruli. The round angiogenic buds observed in the metanephros resemble tumoral angiogenic buds in some aspects. However, both angiogenesis and the degenerative phenomena are part of the normal developmental process. Consequently, the involved mechanisms are probably under sole genetic control. The system studied here offers therefore the possibility to study vascular growth and degeneration on the same model in physiological conditions without application of vasoactive or pathological agents.

Vascular architecture of the lactating and non-lactating teat of the bitch: a scanning electron- and light microscope study.

Tissues from fourteen mammary glands of eight bitches were prepared for scanning electron microscopy of vascular corrosion casts and for histology to study the vasculature of the lactating and non-lactating teats. The densely-meshed mammary dermal capillary network formed ridges and troughs. The teat ducts were vascularized by a relatively densely-meshed capillary network which drained into veins longitudinally oriented to the ducts. Between eight and fifteen teat duct openings were seen on the tip of the teat, that were sometimes divided by a septum. The inner vascularization of the teat showed that the main papillary arteries divided into undulating secondary papillary arteries which presented numerous semi-constrictions and loops. Their structure may help during erection of the teat. Arteriovenous anastomoses found at different points may participate in blood flow maintenance during suckling, heat regulation and teat erection. Veins freely anastomosed and ran longitudinally to the axis of the teat. They exhibited numerous bicuspid valves. In non-lactating teats, vessels showed the same main architecture and characteristics mentioned above, although these were considerably less marked. The structure of the vascular elements in the teat of the bitch could favor blood flow during suckling and suggest that vessels adapt to the physiological situation.