Development of the Retina in the Porcine Fetus A Light Microscopic Study

Morphogenesis of the porcine retina was studied using light microscopy from 4 weeks of gestation until birth (18 to 310 mm crown-rump length), and compared with the adult stage (6 months). Tissue samples were examined from the posterior and peripheral parts of the retina. At 18 mm the retina consists of an inner marginal layer and an outer layer of neuroblastic cells. At 18-40 mm the latter layer is divided into an inner and an outer neuroblastic layer by the transient layer of Chievitz. Subsequently, the development of the different retinal layers begins at the inner retinal border and moves progressively outwards; it also spreads from the posterior to the peripheral part of the neural retina. Many cells of the inner neuroblastic layer are prospective ganglionic cells which migrate inwards, thus forming the ganglion cell layer and the inner plexiform layer at 90 mm. At 120 mm, primitive horizontal cells appear within the outer neuroblastic layer. Separation of this layer into the inner nuclear, outer plexiform and outer nuclear layers is first evident at 180 mm. At this stage all retinal layers are present, except the layer of the photoreceptor cells which is not widespread until at 220 mm. The inner and outer segments of the photoreceptor cells lengthen considerably during the last month of gestation. During the late fetal stage the nerve fiber layer, the inner and outer plexiform layers and the layer of rods and cones all continue to increase in thickness. Concurrently, the ganglion cell layer and the inner and outer nuclear layers have reached their maximal thickness and become thinner. After the total thickness of the neural retina amounts to approximately 180 microns at two to three weeks before birth, it then thins to approximately 160 microns in the adult stage.     

Abnormal prion accumulation associated with retinal pathology in experimentally inoculated scrapie-affected sheep

The purpose of this study was to characterize the patterns of PrP(Sc) immunoreactivity in the retinae of scrapie-affected sheep and to determine the extent of retinal pathology as indicated by glial fibrillary acidic protein immunoreactivity (GFAP-IR) of Müller glia. Sections from the retina of 13 experimentally inoculated scrapie-affected and 2 negative control sheep were examined with immunohistochemical staining for PrP(Sc), GFAP, and PrP(Sc)/GFAP double staining. GFAP-IR of Müller glia is suggestive of retinal pathology in the absence of morphologic abnormality detected by light microscopy. Sheep with the least amount of PrP(Sc) in the retina have multifocal punctate aggregates of prion staining in the outer half of the inner plexiform layer and rarely in the outer plexiform layer. In these retinae, GFAP-IR is not localized with prion accumulation, but rather is present in moderate numbers of Müller glia throughout the sections of retina examined. The majority of sheep with retinal accumulation of PrP(Sc) have intense, diffuse PrP(Sc) staining in both plexiform layers, with immunoreactivity in the cytoplasm of multiple ganglion cells and lesser amounts in the optic fiber layer and between nuclei in nuclear layers. This intense PrP(Sc) immunoreactivity is associated with diffuse, intense GFAP-IR that extends from the inner limiting membrane to the outer limiting membrane. This is the first report of a prion disease in a natural host that describes the accumulation of PrP(Sc) in retina associated with retinal pathology in the absence of overt morphologic changes indicative of retinal degeneration.     

Cytologic appearance of retinal cells included in a fine‐needle aspirate of a meningioma around the optic nerve of a dog

A 6‐year‐old Wirehair Dachshund had a meningioma around the optic nerve that caused exophthalmos. A benign mesenchymal tumor was suspected based on the cytologic pattern of a fine‐needle aspirate, and a meningioma was diagnosed by histopathologic examination. In addition to the meningioma cells, the cytologic smears included groups of cells from apparently 4 layers of normal retina. In particular, uniform rod‐shaped structures in the cytologic sample could suggest rod‐shaped bacteria, but these structures were identified as cylindrical outer segments of photoreceptor rod cells. Other retinal structures recognized included pigmented epithelial layer cells with their uniquely formed pigment granules, the characteristic bi‐lobed, cleaved nuclei from the outer nuclear layer, and nerve tissue likely from the outer plexiform layer of the retina.     

Immunohistochemical study of caveolin-1 and -2 in the rat retina

The expression of caveolin-1 and -2 in the retina was examined; Western blot analysis showed that both were present. Immunohistochemistry indicated that caveolin-1 was expressed in the majority of retinal layers, including the ganglion cell layer, inner plexiform layer, outer plexiform layer, and in the vascular endothelial cells of the retina. Caveolin-2 was primarily immunostained in the vessels, but in a few other elements as well. This is the first demonstration of caveolin differential expression in the retina of rats, and suggests that caveolin plays an important role in signal transduction in glial cells and neuronal cells.     

Outer retinal degeneration in two closely related Goeldi's monkeys (Callimico goeldii)

This case report comprises studies of four Goeldi's monkeys (Callimico goeldii) from the same enclosure. Globe samples from two related C goeldii (the female C goeldii and her male offspring) were available for a histopathological evaluation. Both cases presented histopathologically evident outer retinal degeneration with differences in severity. There was marked outer retinal atrophy characterized by loss of the outer and inner photoreceptor segments, and depletion of the outer retinal nuclear layer. Furthermore, we report a reduction in the thickness of the outer retinal plexiform, inner retinal nuclear layer, and inner retinal plexiform layer in these C goeldii monkeys. To the authors’ knowledge, these findings have not yet been reported in wild‐ or captive‐bred population of C goeldii.     

Early Embryonic Development of the Camel Lumbar Spinal Cord Segment

The lumbar spinal cord segment of the camel embryo at CVRL 2.4 to 28 cm was examined. Major changes are occurring in the organization of the lumbar spinal cord segments during this early developmental period. At the CVRL 2.4, 2.7 and 3.6 cm the three primary layers, ependymal cells layer, mantle cells layer, marginal cells layer in the developing lumber spinal cord segment were demonstrated. The mantle layer is the first to show striking differentiation, while the marginal layer is represented by thin outer rim. Proliferation and differentiation of the neuroepithelial cells in the developing spinal cord produce the thick lateral walls, thin roof and floor plates. The spinal ganglion and dorsal root of the spinal nerve are differentiated. At 2.7 cm CVRL differential thickening of the lateral walls produces a shallow longitudinal groove called sulcus limitans , which separates the dorsal part (alar plate) from ventral part (basal plate). The ventral root of the spinal nerve, the spinal cord and ganglion are embedded in loose mesenchyme, which tends to differentiate into spinal meninges. At 3.6 cm CVRL the basal plate, which is the future ventral gray horn, seem to be quite voluminous and the dorsal and ventral roots unite to form the beginning of the spinal nerve. At 5.5 cm CVRL the alar plates enlarge forming the dorsal septum. At 8.4 cm to 10.5 cm CVRL the basal plates enlarge, and bulge ventrally on each side of the midline producing the future ventral medium fissure, and the white and gray matters can be recognized. At 28 cm CVRL the lumen of the spinal cord is differentiated into the central canal bounded dorsally and ventrally by dorsal and ventral gray commissures, and therefore the gray matter takes the appearance of a butterfly.The lumber spinal nerve and their roots are well distinguished.     

Ocular lesions of 6-aminonicotinamide toxicosis in rabbits

6-Aminonicotinamide, an antimetabolite of nicotinamide, given by intraperitoneal injection produced diarrhea, ascending paresis/paralysis, death, and bilateral ocular alterations in both sexes of New Zealand white and Dutch belted rabbits. Ocular vascular lesions consisted of iridal congestion with iridal hemorrhage and associated acute iritis and aqueous flare in some rabbits. Cytoplasmic swelling, vacuolation, and loss of staining affinity that represented hydropic change were present in both layers of ciliary epithelium and the inner layer of iridal epithelium. Cells in the outer layer of the iridal processes and the ciliary ridge, were most severely affected. Vacuoles were also present in the retinal pigment epithelium and scattered throughout the outer plexiform layer of the retina with a few in the inner and outer nuclear layers. Ocular alterations were prevented by simultaneous administration of nicotinamide and their development appeared related to nicotinamide deficiency. No ocular alterations were caused by nicotinamide administration alone.     

Early Morphological Differentiation of the Bipolar Neurons in the Chick Retina A Golgi Analysis

We report a histogenetic study of the bipolar cells of the chick embryo retina between days 5 and 9, using the Golgi technique. On day 8, at the level of the developing outer plexiform layer, small delicate spines appear on the outer processes of the bipolar cells, which represents the commencement of their dendritic ramification. At a later staige in development che detachment of Landolt's club from the outer limiting membrane is observed in some cells The inner ramifications of the bipolar cell, at the inner plexiform layer, appear later than those of the outer process.     

Prenatal Development of the Eye Tunics in the Dromedary Camel (Camelus dromedarius)

Studies of ocular development in the dromedary camel (Camelus dromedarius) have not been reported previously. The aim of the present investigation was therefore to document the major landmarks and the time course in the prenatal development of the eye tunics in dromedary camel and its accommodation with the surrounding hard environment of the desert. Serial histological sections of dromedary camel embryos and foetuses were used. Age estimation was made on the basis of gestational size, crown vertebral‐rump length (CVRL), which ranged 1.2–110 cm. The eye of the dromedary camel developed in a similar manner to that of the human and domestic animals eyes; the principal differences were in the time of occurrence of certain developmental events, pigmented peripheral cornea near the limbus, a remarkably thickened Descemet's membrane and pigmentation in the corneo‐scleral junction, which represent an adaptive modification in relation to a severe environment.     

羊驼视网膜组织学结构

用光镜观察了羊驼视网膜的组织学结构,以及3级神经元的细胞胞核层数及胞核特点。结果表明：羊驼视网膜结构与昼行性的哺乳动物的视网膜结构相似,但米勒细胞较明显、水平细胞为一层、内网层细胞很厚等形成了羊驼视网膜结构的特性,显示了羊驼视网膜结构和机能与其行为学特性的一致性。     

Seven-week asynchronous twins in a buffalo heifer (Bubalus bubalis) associated with superfetation – A case report

A female buffalo (Bubalus bubalis) of the Bulgarian Murrah breed aged 1,090 days was observed to give birth to a second newborn (normally developed male) after she had calved (normal female) 49 days earlier. This phenomenon is highly associated with her melatonin treatment within a trial for induction of puberty, the last ear implants being placed approximately 50 days before the assumed date of first mating, to which point the level of progesterone had increased dramatically. Despite none of the matings of the dam was visually witnessed to prove ovulation over an existing gestation, we take the liberty to qualify this phenomenon as superfetation, ruling out the other possible phenomena, namely embryonic diapause as it is highly unlikely to occur in any livestock species, and differentiated development of twin foetuses as it is associated with foetal malformation, which was not observed in this case.     

Ultrastructural ocular lesions of 6-aminonicotinamide toxicosis in rabbits

6-Aminonicotinamide, given by intraperitoneal injection to male and female Dutch belted rabbits, produced swelling and vacuolation of ciliary and iridal epithelium plus vacuolation of the retinal pigment epithelial and outer plexiform layers of the retina. By transmission electron microscopy, inner and outer ciliary epithelial cells and inner iridal epithelial cells contained numerous coalescing, membrane-bound vacuoles of the cytocavitary network. These vacuoles were viewed as numerous interconnecting, intracytoplasmic cavities in scanning electron micrographs. Swelling of vacuolated epithelial cells and the presence of fibrin and proteinaceous fluid in the ciliary stroma resulted in thickening of the anterior ciliary processes with the formation of surface alterations detectable by scanning electron microscopy. In transmission electron micrographs the vacuoles in the retinal pigment epithelium were large, electron-lucent spaces and the vacuoles in the outer plexiform layer of the retina appeared to be intracytoplasmic spaces in axons of photoreceptor cells. Distention of cytocavitary structures has been reported in glial cells of animals given 6-aminonicotinamide and this change was apparently due to alterations in ion and water movement across cellular membranes that resulted in intracellular edema.     

Studies on the development of omasum in west african dwarf goats (<Emphasis Type="Italic">Capra hircus</Emphasis>)

This work studied the sequential morphological changes of omasum in foetuses, neonates and adult West African Dwarf (WAD) goats by gross and light microscopic methods. The mean omasal volume was 1.37 ± 0.36, 4.2 ± 0.4, 8.7 ± 6.9 and 60.1 ± 8.6 ml for gestation day 87 and 146 foetuses, neonates and adults respectively. Grossly the mucosa exhibited longitudinally oriented primary, secondary, tertiary and quartnery laminae with smooth surface in foetuses and numerous papillae in the adults. Microscopically it was lined by stratified squamous epithelium which was divided into larger lighter luminal and smaller darker basal zones in gestation day 60 foetuses. The core of the laminae contained extension of the inner muscular tunic. The basal zone developed lateral evaginations (corial papillae) into the lighter zone of the laminae in gestation day 106 foetuses. By term these corial papillae approached the luminal surface. The papillae emerged above the luminal surface in 4 week old neonates. The papillae were fully developed in adult goats. The mean papillary height and width were -205.1 ± 34.0, 67.0 ± 9.6; 235.0 ± 86.5, 185.0 ± 42.3 and 570.3 ± 60.0, 290.1 ± 66.3 μm for foetuses, neonates and adult goats. The inner circular layer was thicker than the outer longitudinal layer of the muscular tunic. The central muscular layer of the laminae originated from the inner circular layer with attachment to the outer muscular layer in adults. This attachment could be adaptation for better anchorage and muscular contraction of the laminae in WAD goats.     

Histo-ontogenetic study of the parotid salivary gland of Indian buffalo

The present study was aimed at elucidating the histogenesis of parotid gland of buffalo. The study was carried out on buffalo foetuses (n = 36), during different stages of prenatal life. The foetuses were categorised into three groups based on their curved crown rump length (CVRL). The primordial anlage of parotid salivary gland was evident at 40th day of development whereas the primary ducts, in the form of cords, were first observed at 81st day of prenatal life. The capsule formation as well as the lobulation of the gland was initiated at 127th day. At 141st day, the duct system of gland was completed. The terminal tubules attained the structure of acini at 167th day. The myoepithelial cells first appeared as flattened basal cells initially around the developing acinar cells at 167th day. The typical compound tubulo-acinar nature of the gland was first observed at 185th day. Purely serous acinar cells were seen from 185th day onwards. The micrometrical studies revealed that the mean diameter of acinar cells, intercalated ducts, striated ducts and large ducts increased with the advancement of age. The serous acinar cells were devoid of acidic as well as neutral mucopolysaccharides in prenatal age groups; however, large ducts with goblet cells exhibited positive reaction. Combined PAS-AB method revealed mixed reaction in acinar cells as well as in large ducts. Fine lipid droplets were observed in intralobular as well as interlobular connective tissue; however, phospholipids were observed in the cell membrane of secretory cells and ducts.     

Distribution of red-spotted grouper nervous necrosis virus (RGNNV) antigens in nervous and non-nervous organs of European seabass (Dicentrarchus labrax) during the course of an experimental challenge

The distribution of red-spotted grouper nervous necrosis virus (RGNNV) antigens was examined by immunohistochemistry in the nervous and non-nervous organs of juvenile European seabass (Dicentrarchus labrax) during the course of an intramuscular infection. Histological changes resulting from the infection were evaluated from 3 days to 2 months post-infection. The specific antibody response was also studied 2 months post-challenge. Viral proteins were present throughout the experimental period in the retina (inner nuclear layer, ganglion layer, outer limiting membrane, and outer plexiform layer), brain (cerebellum and tectum opticum), and liver (hepatocytes and endothelial cells). These proteins were also observed in the renal tubular cells, white pulp of spleen, and in fibroblasts and cartilage of caudal fin. This is the first report of RGNNV proteins appearing in these organs, where the immunostaining was only detected at certain sampling times after the onset of mortality. Brain and retina of virus-exposed fish showed high levels of vacuolation, while accumulation of fat vacuoles was observed in the liver. RGNNV infection also induced a specific antibody response as measured by an ELISA. In summary, this is the first study demonstrating the presence of viral proteins in cells of caudal fin, kidney and spleen of European seabass.     

Intestinal development of bovine foetuses during gestation is affected by foetal sex and maternal nutrition

We aimed to evaluate the effects of maternal nutrition (MN) and foetal sex on the intestinal development of bovine foetuses throughout different days of gestation (DG). Forty‐four multiparous, dry Holstein × Gyr cows with average initial body weight of 480 ± 10 kg were fed the same diet of either restricted feeding at 1.15% of body weight (CO, n = 24) or fed ad libitum (overnourished, ON, n = 20). Six cows from CO group and five cows from ON group were slaughtered at 139, 199, 241 and 268 DG, and foetuses were necropsied to evaluate the intestinal development. The mass, length and density of foetal intestines were not affected by MN (p ≥ 0.260). An interaction between MN and DG was observed for the villi length of jejunum (p = 0.006) and ileum (p < 0.001). Villi length of jejunum and ileum was higher (p < 0.10) in foetuses from ON‐fed cows than in foetuses from CO‐fed cows at 139 DG. However, at 199 DG, the villi length of jejunum and ileum of foetuses from CO‐fed cows was higher than in foetuses from ON‐fed cows. Despite these differences, MN did not affect the villi length of jejunum and ileum at 268 DG (p > 0.10). Female foetuses had greater small intestine mass (p = 0.093), large intestine mass (p = 0.022), small intestine mass in proportion to body mass (p = 0.017) and large intestine mass in proportion to body mass (p < 0.001) than male foetuses. Female foetuses had also longer small intestine (p = 0.077) and greater small intestine density (p = 0.021) and villi length of jejunum (p = 0.001) and ileum (p = 0.010) than males. We conclude that MN affects the pathway for the development of foetal villi length throughout the gestation in bovine foetuses without changing the final villi length. Female foetuses had higher intestinal mass, density and villi length than males during the foetal phase in bovines.     

Photoreceptor cell dysplasia in two Tippler pigeons

Two 12-week-old Tippler pigeons were evaluated for ocular abnormalities associated with congenital blindness. The pigeons were emaciated and blind. Biomicroscopy and direct and indirect ophthalmoscopy findings of the Tippler pigeons were normal with the exception of partially dilated pupils at rest. Scotopic (blue stimuli) and photopic monocular electroretinograms were extinguished in the blind Tippler pigeons. Histological and electron microscopy studies revealed reduced numbers of rods and cones, and an absence of the double cone complex. The photoreceptor cells' outer segments were absent, and the inner segments were short and broad. The number of cell nuclei in the outer and inner nuclear layers was decreased, and the internal and external plexiform layers were reduced in width. Photoreceptor cell endfeet with developing synaptic ribbons were present in the external plexiform layer. Inflammatory cell and subretinal debris was not seen. The electroretinographic, histopathological, and ultrastructural findings of the blind Tippler pigeons support the diagnosis of a photoreceptor cell dysplasia.     

Experimental Neospora Caninum Infection in Pregnant Dairy Heifers Raises Concentrations of Pregnancy‐Associated Glycoproteins 1 and 2 in Foetal Fluids

Plasma concentrations of PAG‐1 are used for pregnancy diagnosis and as a marker of placental/foetal well‐being, while those of PAG‐2 may be an indicator of abortion risk in Neospora caninum‐infected cows. Studies have shown that N. caninum infection modifies PAG‐1 and PAG‐2 patterns in maternal blood plasma. However, no prior work has examined the effects of N. caninum infection on concentrations of PAGs in foetal fluids. In this study, PAG‐1, PAG‐2 and pH levels were determined in the amniotic and allantoic fluids of foetuses collected at 152 days of gestation from control uninfected dams and from dams experimentally infected with N. caninum on Day 110 of gestation. Foetal fluids from infected foetuses had significantly higher PAG‐2 concentrations (p = 0.026) and pH values (p = 0.02) than fluids from non‐infected foetuses. In infected foetuses, significantly higher concentrations of PAG‐1 (p < 0.001) and PAG‐2 (p < 0.001) were detected in fluid samples showing antibodies against N. caninum than those without antibodies. Moreover, pH values were significantly higher (p = 0.011) in foetal fluid samples with antibodies than in samples from non‐infected foetuses. In conclusion, this is the first report on the effect of N. caninum infection on PAG levels in foetal fluids. Our results indicate that following the experimental infection of dams with N. caninum on Day 110 of gestation, foetal fluids collected from the infected foetuses of these dams featured higher PAG‐1 and PAG‐2 levels and pH values than fluids from non‐infected controls, provided that the samples tested showed the presence of antibodies. The clinical implications of these findings are that following infection with N. caninum, most cows will experience some level of placental damage and that this injury correlates with foetal fluid PAG levels and pH.     

Comparative studies on the distribution of glutamate transporters in the retinae of the Mongolian gerbil and the rat

Glutamate is the major excitatory amino acid transmitter in vertebrate retinae. Glutamate transporters therefore play an important role in the precise control of glutamate concentration in the synaptic cleft by regulating extracellular glutamate concentration. In the present study, we performed an analysis of the expressions of three glutamate transporters in gerbil retina using immunohistochemistry. In the gerbil retina, excitatory amino acid carrier 1 and glutamate transporter 1 immunoreactivity was predominant in the ganglion cells but not amacrine or bipolar cells. Glutamate/aspartate transporter (GLAST) immunoreactivity was observed in the radial gliocytes of which the dense network of fine processes was localized in the inner and outer plexiform layers. GLAST immunoreactivity was also detected in astrocytes in the nerve fibre layer. These results demonstrate that three glutamate transporters show specific distributions in the gerbil retina and suggest that the glutamate re-uptake system in the gerbil retina may be different from that of the rat.