The Development of the Vascular System in Quail Embryos: a Combination of Microvascular Corrosion Casts and Immunohistochemical Identification
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Scanning Microscopy Volume 5 Number 4 Article 17 10-16-1991 The Development of the Vascular System in Quail Embryos: A Combination of Microvascular Corrosion Casts and Immunohistochemical Identification M. C. DeRuiter University of Leiden B. Hogers University of Leiden R. E. Poelmann University of Leiden L. Vanlperen University of Leiden A. C. Gittenberger-de Groot University of Leiden Follow this and additional works at: https://digitalcommons.usu.edu/microscopy Part of the Biology Commons Recommended Citation DeRuiter, M. C.; Hogers, B.; Poelmann, R. E.; Vanlperen, L.; and Gittenberger-de Groot, A. C. (1991) "The Development of the Vascular System in Quail Embryos: A Combination of Microvascular Corrosion Casts and Immunohistochemical Identification," Scanning Microscopy: Vol. 5 : No. 4 , Article 17. Available at: https://digitalcommons.usu.edu/microscopy/vol5/iss4/17 This Article is brought to you for free and open access by the Western Dairy Center at DigitalCommons@USU. It has been accepted for inclusion in Scanning Microscopy by an authorized administrator of DigitalCommons@USU. For more information, please contact [email protected]. Scanning Microscopy, Vol. 5, No. 4, 1991 (Pages 1081-1090) 0891- 7035/91$3.00+. 00 Scanning Microscopy International, Chicago (AMF O'Hare), IL 60666 USA THE DEVELOPMENT OF THE VASCULAR SYSTEM IN QUAIL EMBRYOS : ' A COMBINATION OF MICROVASCULAR CORROSION CASTS AND IMMUNOHISTOCHEMICAL IDENTIFICATION M.C.DeRuiter , B.Hogers , R.E.Poelmann L.Vanlperen, A.C .Gittenberger-de Groot* Dept. of Anatomy and Embryology, University of Leiden. The Netherlands. (Received for publication June 11, 1991, and in revised form October 16, 1991) Abstract Introduction Although vascular casts, obtained by injection with methacrylates, are frequently used The last decade has seen more frequent application of the microcorrosion casting to investigate the adult vascu_lar system, l1ttl_e data are available for embryonic stages technique in combination with scanning electron . In this microscopy to investigate the vascular system. paper we use MercoxR in quail _embryos in the period of 2 to 7 days after Most of the investigators studied the vascu lar incubation. The system in adult or postnatal microvascular corrosion casts were evaluated In tissues , e.g. the vascularization of the vertebral column the scanning electron microscope (SEM) with (Konerding special attention and Blank 1987) , lung (Caduff et al. to the developme_nt and 1986· remodelling of the Schraufnagel 1989; Schraufnagel and large arteries and veins. Our Schmid 1988), esophagus (Aharinejad et al. results show that the remodelling of the large 1989) stomach (Imada et al. arteries and veins together 1987), uterus with their developing (Kardon et al. 1982) and testis (Murakami et tributary vessels can be visualized from very _al. early embryonic stages 1989). Only a small number of studies onwards . However , concerned the development of the vascular complete replication of a developing vascular system depends on diameter and system during the embryonic and fetal period. regularity of Burton and Palmer (1989) investigated the the lumen. In the stages investigated, the chorioallantoic membrane of chick embryos, 6 to vascular lumen, even of the largest vessels, is 1 days after incubation. The development of the still very irregular o . Detailed cellular ch_aracteristics renal vascular system in chick embryos has like nuclear impressions of endothelial cells, as been described by often seen in adult material, were seldom found Ditrich and Splechtna (1989). Sumida (1988) studied light in the embryos. microscopically To examine whether blind methacrylate casts ot abnormal aortic arches of ending sprouts are completely or incompletely 12 to 16-day-old rat embryos, which were replicated in a developing vascular system, additional series treated with bisdiamine. Bockman and coworkers of quail embryos were stained (1989, 1990), studying the abnormal immunohistochemically with a monoclonal anti development of the pharyngeal arch arteries body (MB1) specific for endothelial and_ hemo after ablation of the neural crest, showed some poietic cells. It seems that a plexus consisting of endothelial precursors (endothelial cells lacking a micrographs of vascular casts of 3 to 5-day-old chick embryos lumen) is present in the developing organ before . _ _ _ In their the formation of a discussion with the reviewers, DItnch lumen and assembly into and Splechtna vessels , which are connected to an adjacent (1989) remarked that it is almost impossible to fill artery or vein . Expa nsion of the the arterial blood vessels of vascular system chick embryos younger than 6 days. may in part be due to IncorporatIon of these Because of our interest in the embryonic development and endothelial precursors in the wall of existing vessels. changes of the vascular system (DeRuiter et a~ 1989, 1990A, 1990B ), we have used Mercox Key words: Angiogenesis , antibodies , embryos, with a modified injection method in avian, endothelium, quail, scanning electron specifically qua il embryos. Special attention was microscopy, vascu logene sis. paid to casts of budding vessels. To study developing vessels with the mIcrocorrosIon • Address for correspondence: casting technique , i t is necessary to examine A.C. Gittenberger -de Groot whether the casts represent complete or Department of Anatomy and Embryology, incomplete replications of the vascular system. University of Leiden, P.O.Box 9602 , Therefore additional series of quail embryos 2300 RC Leiden . The Netherlands . were incubated with MB1 , a species-specific Phone No: ( .. 31 )71276691 monoclonal antibody aqainst endothelial 1081 MC DeRuiter, B Hogers, RE Poelmann, et al. precursors and endothelial and hemopoietic cells were incubated with MB1 monoclonal antibody , (Peault et al . 1983). Moreover, the combination which detects quail endothelial cells (Peault et of these two techniques has the advantage to al. 1983) , diluted in PBS with 0 .05% Tween-20 study the relation between the formation of a and 0.1 % BSA. The overnight incubation at room PBS lumenized vascular system from endothelial temperature was followed by washing in precursors and the possibility of blood flow in a with 0 .05% Tween-20. Hereafter the slides were developing organ. incubated for 2 hrs with the second antibody Rabbit anti-Mouse conjugated to Horse radish peroxidase . After washing in PBS , the staining Materials and Methods reaction was performed with 0.04 % Diamino benzidine tetrahydrochloride in 0.05M Tris-Maleic Vascular casts and SEM acid (pH 7 .6) with 0 .07% lmidazole and 0.06%% Quail embryos ( Coturnix coturnix japonica) Hydrogen peroxide for 10 min , followed by of 2 to 7 days ' incubation were used in this washing in buffer . Negative controls omitting corrosion casting study . The quail embryos were MB1 and /or the second antibody were also removed from the yolk sac with iridectomy included in the staining reaction. Lastly, the scissors and transferred to phosphate buffered sections were briefly counterstained with Mayer 's saline (PBS), pH 7.2. The serosa and amnion hematoxylin . were removed . The intra-embryonic vascular resistance of the venous system is lower than that of the Results arterial system. To prevent the lack of arterial filling, the Mercox (Mercox CL-28 with catalyst SEM of the vascular casts MA; Fa Okenshoji Co., Ltd, Chou-ku, Tokyo 104) In vascular casts of 2-day-old embryos , two was injected into the outflow tract of the heart. dorsal aortae are present. At the cardiac level, The low venous pressure of embryos older than the first local fusion as small left-right connec 3 days was increased by ligating the vitelline tions between the paired arteries , are present veins. In embryos younger than 3 days , damage (Figs.1 A and 1 B). In the caudal part of the of yolk sac vessels should be avoided. To embryo, both dorsal aortae diverge laterally to prevent overpressure , leakage was induced in blend into a plexus of small vessels, which will the tail vessels. Glass needles (tip diameter: 4 to give rise to the omphalomesenteric arteries. The 20 µm) were mounted to disposable needles omphalomesenteric arteries communicate with (21G), which in turn were connected to a the vitelline vessels of the yolk sac , which drain disposable 1 ml syringe. Because the fragile , into the omphalomesenteric veins . In none of the immature vessels could easily be ruptured by 2-day-old embryos could the dorsal interseg high pressure , the filling had to be controlled mental arteries be completely injected , but for a under the dissecting microscope. A small small part at their origin from the dorsal aortae . amount , 100 to 200 µI Mercox, was mixed with The proximal part of the posterior cardinal veins , 1% catalyst , which is recommended by studies draining this intersegmental system , is filled on adult material . retrogradely (Fig .18) . In an early 2-day-old The site of injection varied with the stage of embryo the posterior and anterior cardinal veins development. Embryos with a looped heart (2-3 drain through the common cardinal vein in the days) and those without a complete ventricular omphalomesenteric vein (Fig .1 B), but at the end septum (3.5 to 4 days) were injected into the of the second day the common cardinal vein and outflow tract of the ventricle , while older embryos the omphalomesenteric vein drain separately into (quail 5 to 7 days) were injected in one of the the sinus venosus (Fig .1 C). ventricles. At each age about 7 successful