Fine Structure of the Pecten Oculi of the Red-Tailed Hawk (Buteo jamaicensis)

The pecten oculi of the red-tailed hawk (Buteo jamaicensis) has been examined by light and electron microscopy. In this species the pecten is very large and of the pleated type. It consists of 17-18 accordion folds which are joined apically by a heavily pigmented bridge of tissue which holds the pecten in a fan-like shape, widest at its base. It is situated over the optic nerve head and extends into the vitreous. Within each fold are numerous capillaries, larger supply and drainage vessels and melanocytes. The capillaries are specialized and display extensive microfolds on both the luminal and abluminal borders. The endothelial cell bodies are thin with most organelles present in a paranuclear location. The capillaries are surrounded by thick fibrillar basal laminae which are probably structurally important and which often enclose pericytes. The melanocytes which are most plentiful in the bridge region and peripherally in the pecten, form an incomplete sheath around the capillaries and other blood vessels. These melanocytes are also felt to be fulfilling a structural role within the pecten. The morphology of the pecten of the red-tailed hawk is indicative of a heavy involvement in the transport of materials to the avascular avian retina.     

Light and electron microscopic studies on the pecten oculi showing blood–retina barrier properties in Turkey’s native Gerze chicken

The pecten oculi is a highly vascularized and pigmented organ that projects from the optic disc into the vitreous body in the avian eye. In this study, the pecten oculi of Turkey's native Gerze chicken was examined by light and scanning electron microscopy. Furthermore, the localization of some adherens junction components (E-cadherin and pan-cadherin) in intact vessels of the blood–retina barrier was investigated by immunohistochemistry. In the Gerze chicken, the pecten oculi was a thin structure, which was located over the head of the discus nervi optici and projected from the retina into the corpus vitreum. The pecten oculi consisted of 18–21 highly vascularized pleats, joined apically by a bridge and resembled an accordion in appearance. Hyalocytes and melanocytes were observed around the small and large vessels. The morphometric data of the pecten oculi showed that there were no statistical differences in terms of sex. The immunohistochemical analysis of the pecten oculi, which is used as a model for the investigation of the formation and maturation of the barrier properties in the central nervous system, revealed cytoplasmic E-cadherin and pan-cadherin immunoreactivity in the endothelial cells of the small, large and capillary vessels. These observations suggest that while the morphological and histological structure of the Gerze chicken's pecten oculi was generally similar to that of other diurnal domestic birds, the pecten oculi, a model system for vascular differentiation and the blood–retina barrier, expressed different cadherins.     

Fine Structure of the Pecten Oculi in the American Crow (Corvus brachyrhynchos)

The pecten oculi of the American crow (Corvus brachyrhynchos) has been examined by both light and electron microscopy. In this species the pecten is very large and of the pleated type. It consists of 22 - 25 accordion folds that are joined apically by a bridge of tissue which holds the pecten in a fan-like shape widest at its base. Within each fold are numerous capillaries, larger supply and drainage vessels and many melanocytes. The capillaries are extremely specialized for transport functions and display extensive microfolds on both their luminal and abluminal borders. Except for the nuclear region which contains most of the organelles, the endothelial cell bodies are extremely thin. These capillaries are surrounded by thick fibrillar basal laminae which are conjectured to be structurally important. Pericytes are a common feature of these capillaries. The numerous pleomorphic melanocytes interspersed between the capillaries are also felt to be important in structural support of the pecten. The pecten is considered to be comparable to the falciform process of some teleosts, the conus papillaris of reptiles, the supraretinal vessels of amphibians and teleosts and the intraretinal vessels of mammals which are all felt to be alternative methods of bringing nutrients to the inner retina.     

Macroanatomic,light and scanning electron microscopic structure of the pecten oculi in northern bald ibis (Geronticus eremita)

Eyes are the most primarily required sensory organs during the migration of migratory birds and Northern Bald Ibises (Geronticus eremita) are known to make long migrations. This study examined for the first time the structure of pecten oculi in northern bald ibises by using macroscopic anatomy as well as light and electron microscopic methods. In the study 20 eye globes from 10 adult bald ibises were used. The pecten was of pleated type. As in most bird species, it was located on the optic nerve head and projects into the vitreous from the optic nerve head. The wider basal part was observed to attached to retina and its free apical part was found in camera vitrea bulbi embedded in corpus vitreum. The pecten had 13–14 accordion like pleats lying between the basal and apical parts. In addition to arterial and venous vessels, numerous capillary vessels as well as melanocytes were observed within each pleat. The bridge binding the pleats at the apical part showed a stronger pigmentation compared to other parts of the pecten. The results of the study indicated that the general morphology of pecten oculi in northern bald ibises which is a migratory bird species were similar to that in other diurnal bird species.     

The development of the pecten oculi in the chick

The development of the pecten oculi, a structure peculiar to the avian eye, was studied by scanning electron microscopy (SEM) correlated with light microscopy (LM) in embryonic and adult chickens. The development of the chick pecten was divided into 4 phases: (1) formation of the primordial pecten (Hamburger-Hamilton's stages 27 to 29), (2) formation of the plate-like pecten (stages 30 to 34), (3) pleat formation and pigmentation (stages 35 to 37), and (4) bridge formation and high-vascularization (stage 38 to adult). The primordial pecten is formed entirely from the ectoderm by fusion of the inwardly-projecting edges of the optic fissure. The primordial pecten grows into a tall, thin plate rising from a broad base. The pecten begins to fold slightly at stage 35. The number of pleats increases rapidly, from 7 at stage 35, to 16 at stage 36, 18 at stage 37, and 19 to 20 at stage 40. The bridge begins to form at stage 38 by a swelling of the apical edge of the pecten and completes its development by the twentieth post-hatch day. Blood vessels appear first in the broad base of the plate-like pecten, then become more numerous and gradually extend into the pleats. The pecten becomes more vascular than cellular at stage 43, and it is highly vascularized in the adult. The pleat surface becomes conspicuously irregular with increased vascularization. The peripectinate cells, located on the pecten, are already present at stage 27.     

Angiogenesis and Vascular Regression in the Corpus Luteum Periodicum

Introduction and Aim: The corpus luteum is one of the most intensely vascularized tissues. Luteal angiogenesis is strictly controlled and blood vessels regress completely within a short period of time. The aim of this study was to investigate vascular dynamics in relation to cellular and molecular mechanisms of luteal angiogenesis and anti‐angiogenesis. Material and Methods: Endothelial cells of blood vessels in paraffin sections of bovine corpora lutea from different stages were examined by labelling with the lectin Bandeiraea simplicifolia agglutinin I. Angiogenesis was studied by morphometry of the capillaries, and immunolocalization of the angiogenic factor VEGF and VEGF‐receptor 2. Presence of apoptotic luteal and endothelial cells was investigated using the TUNEL test and transmission electron microscopy. Results: During development of the corpus luteum (day 3–8 of the oestrous cycle) a dense capillary network (8–12% area ratio) is established and maintained until day 17. Early regression (day 18–24) is characterized by a remarkable decrease of capillaries (1% area ratio). In the regressing corpus luteum the number of apoptotic luteal cells is closely correlated (r = 0.9) to the number of apoptotic endothelial cells. VEGF is immunolocalized in luteal cells (day 3–17), smooth muscle cells and endothelial cells of arterioles of the regressing corpus luteum. During late luteal regression, a moderate increase of capillaries (2.5% area ratio) is obvious. Conclusions: The dynamic changes of the capillarity during development and regression of the cyclic corpus luteum correlate with VEGF and VEGF‐R2 activities. In contrary to expectations the late stage of luteal regression is accompanied by angiogenesis. One reason for this phenomenon may be an increase in metabolic activity resulting in re‐organization of blood vessels already regressed.     

Clinicopathological Features of Hemangioma in Two Young Horses

The clinicopathological and immunohistochemical features of two vascular tumors in two young horses are described in the present work. These animals were referred to the Veterinary Teaching Hospital of the University of Córdoba because of the presence of hyperpigmented plaques located in the medial aspect of the left leg and also around the hock (case 1) and in the right front leg (case 2). Some of the lesions showed deep ulceration and severe protrusion with abundant bleeding. The histopathological study revealed that lesions were composed of nonencapsulated, proliferated, closely packed small blood vessels, some of which showed irregular shape, whereas others were similar to capillaries, arterioles, and venules. Neoplastic cells expressed vimentin and factor VIIIar, suggesting their endothelial nature, whereas in the wall of some proliferated vessels, some cells expressed vimentin, α-smooth muscle actin and desmin, an immunophenotype consistent with pericytes, and small muscle cells. These features agree with those reported in human juvenile hemangioma rather than with hemangioma in adult horses.     

Morphology and Aquaporin Immunohistochemistry of the Uterine Tube of Saanen Goats (Capra hircus): Comparison Throughout the Reproductive Cycle

The expression of six different aquaporins (AQP1, 2, 3, 4, 5 and 9), integral membrane water channels that facilitate bi‐directional passive movement of water, was investigated by immunohistochemistry in the uterine tube of pre‐pubertal and adult Saanen goats (Capra hircus), comparing the different phases of the oestrous cycle. Regional morphology and secretory processes were markedly different during the goat oestrous cycle. The tested AQP molecules showed different expression patterns in comparison with already studied species. AQP1‐immunoreactivity was evidenced at the endothelium of blood vessels and in nerve fibres, regardless of the tubal tract and cycle period. AQP4‐immunoreactivity was shown on the lateral plasmalemma in the basal third of the epithelial cells at infundibulum and ampulla level in the cycling goats, more evidently during follicular than during luteal phase. No AQP4‐immunoreactivity was noticed at the level of the isthmus region, regardless of the cycle phase. AQP5‐immunoreactivity, localized at the apical surface of epithelial cells, increased from pre‐puberty to adulthood. Thereafter, AQP5‐immunoreactivity was prominent during the follicular phase, when it strongly decorated the apical plasmalemma of all epithelial cells at ampullary level. During luteal phase, immunoreactivity was discontinuous, being weak to strong at the apex of the secretory cells protruding into the lumen. In the isthmus region, the strongest AQP5‐immunoreactivity was seen during follicular phase, with a clear localization in the apical plasmalemma of all the epithelial cells and also on the lateral plasmalemma. AQP2, 3 and 9 were undetectable all along the goat uterine tube. Likely, a collaboration of different AQP molecules sustains the fluid production in the goat uterine tube. AQP1‐mediated transudation from the blood capillaries, together with permeation of the epithelium by AQP4 in the basal rim of the epithelial cells and final intervening of apical AQP5, could be involved in fluid production as well as in secretory processes.     

Concentrations of retinol, 3,4-didehydroretinol, and retinyl esters in plasma of free-ranging birds of prey

This study investigated vitamin A compounds in the plasma of healthy free‐ranging Central European raptors with different feeding strategies. Plasma samples of nestlings of white‐tailed sea eagle [white‐tailed sea eagle (WTSE), Haliaeetus albicilla) (n = 32), osprey (Pandion haliaetus) (n = 39), northern goshawk (Accipiter gentilis) (n = 25), common buzzard (Buteo buteo) (n = 31), and honey buzzard (Pernis apivorus) (n = 18) and adults of WTSE (n = 10), osprey (n = 31), and northern goshawk (n = 45) were investigated with reversed‐phase‐high‐performance liquid chromatography (RP‐HPLC). In WTSE, northern goshawks and common buzzards retinol were the main plasma component of vitamin A, whilst in ospreys and honey buzzards, 3,4‐didehydroretinol predominated. The median of the retinol plasma concentration in the nestlings group ranged from 0.12 to 3.80 μm and in the adult group from 0.15 to 6.13 μm . Median plasma concentrations of 3,4‐didehydroretinol in nestlings ranged from 0.06 to 3.55 μm . In adults, northern goshawks had the lowest plasma concentration of 3,4‐didehydroretinol followed by WTSE and ospreys. The plasma of all investigated species contained retinyl esters (palmitate, oleate, and stearate). The results show considerable species‐specific differences in the vitamin A plasma concentrations that might be caused by different nutrition strategies.     

In vitro measurement of post‐natal changes in proliferating satellite cell frequency during rat muscle growth

Satellite cells, resident myogenic stem cells found in postnatal skeletal muscle, are most abundant during early postnatal development and sharply decline in frequency thereafter to adult levels in mice and rats. Therefore, postnatal changes in satellite cell mitotic activities are important aspects for further understanding a muscle growth strategy. In large meat‐production animals, however, the traditional in vivo proliferation assay may be less realistic because it requires intra‐peritoneal (ip) injection of huge dosage of mutagenic nucleosides, ³H‐labeled thymidine or bromodeoxyuridine (BrdU), at each age‐time of sacrifice. We report in the present pilot study using rats that in vivo proliferation activity of satellite cells can be evaluated by an in vitro BrdU‐incorporation assay in early cultures. Briefly, satellite cells were prepared from upper hind‐limb and back muscles and maintained for 24 h with imposing by BrdU addition for the last 2 h, followed by the regular immunocytochemistry for determining BrdU‐incorporated cell percentage. This in vitro assay demonstrated a rapid decrease in proliferating satellite cell frequency to the adult level during about 3‐month period after birth, and yielded a high correlation to the measurements by the in vivo BrdU ip‐injection method during the postnatal period examined from day‐2 to month‐11. The in vitro proliferation assay may be further adaptable for large domestic animals by the combination with a muscle biopsy technique that enables age‐interval sampling from the same growing animals.     

Age‐ and Gender‐Dependent Changes of Bovine Myocardium Architecture

Growth, gravidity and lactation put high demands on the performance of the myocardium. The aim of this study, which was performed in 40 female and 20 male bovines ranging from 1 to 4.5 years old, was to determine gross and microscopic morphometric data of bovine myocardium to establish a comparative measure of myocardial growth during juvenile development. During the developmental stage of young adulthood, age‐related increases in female myocardial characteristics included cardiac mass, left and right ventricular mass and the ratio of cardiac mass to loose connective tissue. Age‐related decreases were observed in the number of myocyte nuclei per mm² and the thickness of the right ventricular wall. Sex differences in these parameters were found between 2‐year‐old bulls (N = 20) and 2‐year‐old heifers (N = 10), with males having heavier hearts, thicker ventricular walls, less myocytes in the left ventricle and less connective tissue in both ventricles. Age and sex had no influence on the ratio of capillaries to myocytes, estimated at 0.98 in the adult bovine. Capillary density does not change during juvenile development, but cross‐sectional capillary area does adapt to myocyte cross‐sectional area, accounting for this relatively constant ratio.     

New insights in vascular development: vasculogenesis and endothelial progenitor cells

In the course of new blood vessel formation, two different processes – vasculogenesis and angiogenesis – have to be distinguished. The term vasculogenesis describes the de novo emergence of a vascular network by endothelial progenitors, whereas angiogenesis corresponds to the generation of vessels by sprouting from pre-existing capillaries. Until recently, it was thought that vasculogenesis is restricted to the prenatal period. During the last decade, one of the most fascinating innovations in the field of vascular biology was the discovery of endothelial progenitor cells and vasculogenesis in the adult. This review aims at introducing the concept of adult vasculogenesis and discusses the efforts to identify and characterize adult endothelial progenitors. The different sources of adult endothelial progenitors like haematopoietic stem cells, myeloid cells, multipotent progenitors of the bone marrow, side population cells and tissue-residing pluripotent stem cells are considered. Moreover, a survey of cellular and molecular control mechanisms of vasculogenesis is presented. Recent advances in research on endothelial progenitors exert a strong impact on many different disciplines and provide the knowledge for functional concepts in basic fields like anatomy, histology as well as embryology.     

Labelling of rat endothelial cells with antibodies to vWF, RECA-1, PECAM-1, ICAM-1, OX-43 and ZO-1

Labelling with endothelium specific monoclonal antibodies, von Willebrand Factor (vWF), rat endothelial cell antigen-1 (RECA-1), platelet-endothelial cell adhesion molecule-1 (PECAM-1), intercellular adhesion molecule-1 (ICAM-1), OX-43 and zonula occludentes-1 (ZO-1), was investigated in cryostat sections of vessels from rats of different ages using a confocal microscope. The results showed that labelling of the vWF was positive in endothelial cells from adult, fetal and different ages of embryonic rat. Labelling with RECA-1 was weakly positive in adult rat aorta and lung endothelial cells but not in embryonic yolk sac endothelial cells. Labelling using PECAM-1, ICAM-1 and OX-43 was negative in both adult and embryonic endothelial cells. ZO-1 showed positive but very weak reactivity in embryonic yolk sac endothelial cells. The expression of vWF on vessels from adult and 19.5-day fetal tissues was strongly positive. However, the expression of vWF in embryonic endothelial cells was dependent on the gestational age. While the 11.5-day yolk sac vessels stained weakly, staining gradually increased in 13.5-, 15.5- and 17.5-day-old yolk sac vessels. The results suggest that vWF is a reliable endothelial cell marker in rat vascular endothelial cells, including both fetal and embryonic stages.     

Angiogenesis and Interstitial Pressure in a Rat Tumour Model

Introduction and Aim:  The corpus luteum is one of the most intensely vascularized tissues. Luteal angiogenesis is strictly controlled and blood vessels regress completely within a short period of time. The aim of this study was to investigate vascular dynamics in relation to cellular and molecular mechanisms of luteal angiogenesis and anti-angiogenesis.
Material and Methods:  Endothelial cells of blood vessels in paraffin sections of bovine corpora lutea from different stages were examined by labelling with the lectin Bandeiraea simplicifolia agglutinin I. Angiogenesis was studied by morphometry of the capillaries, and immunolocalization of the angiogenic factor VEGF and VEGF-receptor 2. Presence of apoptotic luteal and endothelial cells was investigated using the TUNEL test and transmission electron microscopy.
Results:  During development of the corpus luteum (day 3–8 of the oestrous cycle) a dense capillary network (8–12% area ratio) is established and maintained until day 17. Early regression (day 18–24) is characterized by a remarkable decrease of capillaries (1% area ratio). In the regressing corpus luteum the number of apoptotic luteal cells is closely correlated ( r  = 0.9) to the number of apoptotic endothelial cells. VEGF is immunolocalized in luteal cells (day 3–17), smooth muscle cells and endothelial cells of arterioles of the regressing corpus luteum. During late luteal regression, a moderate increase of capillaries (2.5% area ratio) is obvious.
Conclusions:  The dynamic changes of the capillarity during development and regression of the cyclic corpus luteum correlate with VEGF and VEGF-R2 activities. In contrary to expectations the late stage of luteal regression is accompanied by angiogenesis. One reason for this phenomenon may be an increase in metabolic activity resulting in re-organization of blood vessels already regressed.     

Bacillary angiomatosis in an immunosuppressed dog

A dog being treated with immunosuppressive doses of prednisone and azathioprine for pancytopenia of unknown origin, developed, over a 2‐week period, multiple erythematous nodular lesions in the skin including footpads. Skin samples revealed lesions identical to those of human bacillary angiomatosis (BA). The nodules were composed of multifocal proliferations of capillaries, each lined by protuberant endothelial cells. The capillary clusters were separated by an oedematous connective tissue, lightly infiltrated with degenerate inflammatory cells, including neutrophils and macrophages. Tissue sections stained with Warthin–Starry silver stain revealed large numbers of positively stained bacilli in the stromal tissue, most heavily concentrated around the proliferating capillaries. Lesions of vascular degeneration and inflammation were evident. Bartonella vinsonii subsp. berkhoffii genotype 1 was independently amplified and sequenced from the blood and the skin tissue. The pathognomonic nature of the histological lesions, demonstration of compatible silver‐stained bacilli in the tissue, and identification of B. vinsonii subsp. berkhoffii in the blood and tissue indicates that this is most likely the aetiologic agent responsible for the lesions. Antibiotic therapy was successful in resolving the nodules. It would appear that B. vinsonii subsp berkhoffii, like Bartonella henselae and Bartonella quintana, has the rare ability to induce angioproliferative lesions, most likely in association with immunosuppression. The demonstration of lesions identical to those of human BA in this dog is further evidence that the full range of clinical manifestations of human Bartonella infection occurs also in canines.     

Post‐hatch Changes in the Immunoexpression of Desmin,Smooth Muscle Actin and Vimentin in the Testicular Capsule and Interstitial Tissue of the Japanese quail (Coturnix coturnix japonica)

The post‐hatch development of immunoreactivity to desmin, smooth muscle actin (SMA) and vimentin in the testicular capsule and interstitial tissue of day‐old to adult quails was described in this study. The tunica albuginea of the testicular capsule was composed mainly of myoid cells. A zonal arrangement of desmin and SMA immunostaining was observed in myoid cells of the tunica albuginea in 1‐ to 24‐day‐old quails. Immunostaining for SMA and desmin was uniform in the tunica albuginea of adult birds. Vimentin immunostaining in the testicular capsule was demonstrated in mesothelial cells, endothelial cells and fibroblasts. The interstitial tissue contained mesenchymal cells, peritubular myoid cells, Leydig cells and fibroblasts. Desmin‐immunopositive mesenchymal cells were present in the interstitial tissue of 1‐ to 17‐day‐old quails. Peritubular myoid cells expressed strong desmin immunostaining in all developmental stages, while the intensity of SMA immunostaining increased with testicular maturation. Vimentin was demonstrated in Leydig cells and fibroblasts, while the peritubular myoid cells displayed strong vimentin immunostaining only in adult birds. Strong vimentin immunostaining was demonstrated in the endothelial cells of capsular and interstitial blood vessels. The tunica media of these blood vessels displayed desmin and SMA immunostaining. The results of the study have established that variability exists in the distribution and intensity of desmin, SMA and vimentin immunostaining in the testicular capsule and interstitial tissue of the post‐hatch Japanese quail.     

Retinal vascular patterns in domestic animals

In this paper a morphological study of the retinal vascular patterns in various species of domestic animals is reported. A classification of these patterns into four well-defined groups is described. In the domestic ruminants, pigs and carnivores the retina contains a compact plexus of blood vessels located in the major part of the light-sensitive portion of the retina (euangiotic or holangiotic pattern). In other domestic animals blood vessels are present only in a smaller part of the retina. In the rabbit, vessels are confined to a broad horizontal band coincident with the area of dispersion of the myelinated nerve fibres. The larger of these vessels are readily visible macroscopically (merangiotic pattern). In the horse and the guinea pig the retinal blood vessels are minute and restricted to the direct neighbourhood of the optic disc (paurangiotic pattern). The avian retina is completely avascular (anangiotic pattern), but a densely vascularised pecten oculi is attached to the linear optic nerve head and protrudes far into the inferior part of the vitreous body.     

Immunolocalization of angiogenic growth factors in the ovine uterus during the oestrus cycle and in response to Steroids

The vascular changes associated with endometrial maturation in preparation for embryo implantation depend on numerous growth factors, known to regulate key angiogenic events. Primarily, the vascular endothelial growth factor (VEGF) family promotes vascular growth, whilst the angiopoietins maintain blood vessel integrity. The aim was to analyse protein levels of VEGFA ligand and receptors, Angiopoietin‐1 and 2 (ANG1/2) and endothelial cell receptor tyrosine kinase (TIE‐2) in the ovine endometrium in the follicular and luteal phases of the oestrus cycle and in response to ovarian steroids. VEGFA and its receptors were localized in both vascular cells and non‐vascular epithelium (glandular and luminal epithelium) and stroma cells. VEGFA and VEGFR2 proteins were elevated in vascular cells in follicular phase endometrium, compared to luteal phase, most significantly in response to oestradiol. VEGFR1 was expressed by epithelial cells and endothelial cells and was stimulated in response to oestradiol. In contrast, Ang‐1 and Ang‐2 proteins were elevated in luteal phase endometrium compared to follicular phase, and in response to progesterone, evident in vascular smooth muscle cells and glands which surround TIE‐2‐expressing blood vessels. Our findings indicate that VEGFA is stimulated by oestradiol, most predominantly in follicular phase endometrium, and Ang‐1 and 2 are stimulated by progesterone and were increased during the luteal phase of the oestrus cycle, during the time of vascular maturation.