Histomorphometric and immunohistochemical study of the goat omasum during prenatal development.

This work studies the morphological changes taking place in the goat omasum during prenatal development, using scanning electron microscope, light microscopy and immunohistochemical analysis. A total of 140 goat embryos and fetuses were used, from the first stages of prenatal life until birth. Differentiation of the omasum as a separate compartment of the primitive gastric tube was observed at 35 days of prenatal life ([crown-rump length (CRL)] 3 cm, 23% gestation). By 38 days (CRL 4.3 cm, 25% gestation) the omasal wall comprised three layers: an internal epithelial layer, a middle layer of pluripotential blastemic tissue and an external layer or serosa. Omasal laminae appeared in the following order: primary at 38 days (CRL 4.3 cm, 25% gestation), secondary at 50 days (CRL 7.7 cm, 33% gestation), tertiary at 59 days (CRL 12 cm, 39% gestation) and quaternary at 64 days (CRL 13.5 cm, 43% gestation). Neuroendocrine cells were detected by synaptophysin (SYP) at 52 days (CRL 8 cm, 35% gestation), while glial cell markers (glial fibrillary acidic protein - GFAP, and vimentin-VIM) were observed at 64 days (CRL 13.5 cm, 43% gestation) and 38 days (CRL 4.3 cm, 25% gestation), respectively. Sympathetic and parasympathetic nerve fibers and nerve bodies were detected via neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) at 95 days (CRL 20 cm, 63% gestation). In conclusion, prenatal development of the omasum - like that of the rumen - appears to take place somewhat earlier in goats than in sheep or cattle, but at a similar stage to that reported in deer.

Structural, histochemical and immunocytochemical study of the forestomach mucosa in domestic ruminants.

The forestomach plays an important role in the digestion physiology of ruminants. The aim of this study is to clarify the morpho-functional role of the mucosa in each of the three compartments of the forestomach in three domestic ruminants species, viz cattle, buffalo and sheep, by means of structural, histochemical and immunocytochemical methods, including transmission electron microscopy, light microscopy and scanning electron microscopy. These methods were chosen to demonstrate the indirect evidence for the presence of nitric oxide (NO) employing NADPHd and nitric oxide synthase I (NOS I). The various cell layers of the forestomach epithelium are described and illustrated in detail. An intense NADPHd staining was observed in the granulosa, spinosa and basal layers of the epithelium, in particular in the cytoplasm over the nucleus. NOS I immunoreactivity was found in all specimens of the forestomach mucosa. The results of this study might reflect a possible role of NO in delaying the onset of cellular apoptosis in the forestomach mucosa of the domestic ruminants, by playing a role in the production of cell energy.

Functional morphology of unguiculiform papillae of the reticular groove in the ruminant stomach.

The arrangement of the ruminant stomach in four gastric compartments with specialized mucosal papillae along the gastric groove (GG) has been previously described. However, a debate remains about functional implications of these morphological pecularities. This study was aimed to elucidate the relation between the papillar morphology and its putative functions. The GG was obtained from adult bovine stomachs (n=10) and subdivided into (1) proximal, (2) middle, (3) distal portion of the reticular groove (RG) and (4) the area of the reticulo-omasal sphincter (ROS). The specimens were processed for scanning electron microscopy and stereomicroscopy to analyze the density, shape and location of the papillae. Whereas the proximal portion of the RG was characterized by small (1.5mm), conically shaped, smooth papillae, the middle portion exhibited larger papillae (4mm) with sharp borders covered by keratin. Towards the ROS the papillae further increased in size (3-11mm) and showed compound or single processes resembling the shape of arrows, twisted hooks or thorns (unguiculliform papillae). At the ROS the unguiculliform papillae were distributed in clusters groups and along the border of the sphincter. Due to their peculiar morphological features it is suggested that unguiculliform papillae functions as a filter barrier preventing the passage of large-sized food particles into the omasum and avoiding subsequent obstruction of both the RG and the ROS. The data give further evidence that unguiculliform papillae are actively involved in the complex mechanisms of food processing taking place within the ruminant pluricavity stomach.

The surface morphology of the omasum of the African goat.

The microstructure of the omasum of the African goat was investigated macroscopically, as well as by scanning electron microscopy to determine how the omasum is anatomically specialised to enable the goat to graze and browse so successfully in the harsh environments of Africa. The omasum of the African goat was found to be more simple in structure than that of other domesticated ruminants. The omasal laminae were arranged in only 3 orders with the number of laminae varying between 27 and 33, considerably fewer than previously described for other goats, sheep or cattle. The large conical and unguiculiform papillae surrounding the reticulo-omasal orifice, as well as the hooked papillae present on the laminae, were more similar to those of wild ruminants that browsed than domestic grazing species. The papillae on the laminae were also found to be directionally orientated towards the orifices, which may suggest a "sieve mechanism' at the reticulo-omasal orifice and interlaminar spaces. In addition, it may suggest a directional movement of ingesta between the laminae. The stratified squamous epithelium lining the laminae and papillae was lightly keratinised which, along with the microplicae-like surface folds and pits of the superficial cells, indicated that it was functionally structured for absorption. These morphological specialisations of the omasal surfaces of the indigenous African goat demonstrated similarities with that of wild ruminants that are able to graze and browse.

Immunohistochemical localization of chondroitin sulfate in the forestomach of the sheep.

The localization of chondroitin sulfate glycosaminoglycans in the forestomach mucosa was examined by use of monoclonal mouse IgMs against chondroitin sulfate (CS-56 and 2H6). Under the light microscope, immunoreactive sites were observed in subepithelial regions and around blood vessels of the rumen, reticulum and omasum. In the reticular papillae and omasal papillae, large areas of core regions also reacted with the monoclonal antibodies. The cell surfaces of fibroblasts in the core region of the epithelium were intensely stained. Under the electron microscope, heavy staining of collagen fibrils beneath the basement membrane of the mucosal epithelium was observed. In addition, products of immunoreactions were localized at the abluminal surfaces of endothelial cells, in pinocytotic vesicles and on pits of the endothelium, as well as surrounding collagen fibrils. At the core regions of reticular papillae and omasal papillae, cell surfaces, interstitial area of collagen fibrils and elastin fibers were stained. Some of the rough endoplasmic reticulum in fibroblasts was also stained. These results suggest that chondroitin sulfate in the forestomach mucosa might serve as a charge barrier in the transport of charged molecules.

Morphological study of the surface structure of the omasal laminae in cattle, sheep and goats.

The mucosal relief of the omasum in cattle, sheep and goats was examined by the naked eye, stereoscopic microscope and scanning electron microscope. The mucosal relief of the omasum showed wide variations both between and within these species. Omasal papillae on the laminae varied highly in shape, i.e. conical, rounded, claw-like, wart-like, finger-like etc. They decreased gradually in size from the oral to the aboral region of the omasal lamina. Ridge-like structures and linear arrangements of the papillae were also seen on the omasal laminae of cattle and sheep. Many grooves and folds were found on the lateral surface of the omasal papillae in adult cattle and on the inter-papillar surface of the laminae of all three species. The mucosal relief of the omasum in cattle seemed to be more well-developed than those of sheep and goats. Dietary effects on the omasal mucosa were also discussed.

Ultrastructure of the epithelium of the rumen, reticulum and omasum of grey, white and black Karakul lambs.

Mortalities due to digestive disturbances occur in homozygous grey and white lambs after they have reached weaning age. Milk-filled, distended rumens, due to malfunctioning of the oesophageal groove, are found 24 h after birth. Scanning electron microscopical studies revealed that milk caused sloughing of the luminal cells in the forestomachs of the affected lambs, while no sloughing of cells was apparent in control black lambs. The purpose of this study was to compare the ultrastructure of the forestomach mucosa of grey, white and black Karakul lambs; to determine whether the sloughing of luminal cells was evident in sections; and, if possible, to find a reason for the desquamation of the cells. Samples of the forestomach of grey, white and black Karakul lambs were prepared routinely for electron microscopy and studied with a Phillips electron microscope. In all the lambs the mucosa of the forestomach was a stratified squamous epithelium consisting of a stratum basale, stratum spinosum and stratum corneum. In the grey and white lambs the luminal cells of the stratum corneum were electron dense, non-nucleated and vacuolated. Sloughing of luminal cells was observed. In the black lambs no sloughing of cells was evident and the luminal cells were moderately electron-dense, nucleated elements. Desquamation of the luminal cells in the affected lambs revealed the underlying layer with its exposed desmosomal attachment sites. This explained the differences in the appearance of the luminal cells in the three groups of lambs as revealed by the scanning electron microscope.

Histochemical localization of glycosaminoglycans in the omasal papillae of sheep.

Histochemical localization of the glycosaminoglycans in the omasal laminae were examined at light and electron microscopic levels. The core region of the omasal papillae was stained strongly with alcian blue at pH 2.5 and 1.0. The staining was degraded moderately and completely when tissue sections were pretreated with mild and active methylation, respectively. Alcianophilia was moderately decreased with saponification. Furthermore, enzymatic digestion procedures showed that these alcianophilic substances contained a large amount of hyaluronic acid and chondroitin sulphate. Ultrastructurally, a spider web-like structure was widely distributed among the spindle-shaped fibroblast-like cells and fibrous networks of collagen and elastin. These results suggest that the core region of the omasal papillae not only acts as a physical buffer resisting the local pressure from the lumen, but may also influence material transport through the omasal mucosa.

Three-dimensional architecture of the subepithelial connective tissue in the omasal laminae of sheep and cattle.

The three-dimensional architecture of the subepithelial connective tissue in the omasal laminae of sheep and cattle was studied by scanning electron microscopy after treatment with 2 N NaOH solution. The omasal laminae were equipped with highly undulated subepithelial connective tissue showing various projections or ridges. In the sheep, the subepithelial connective tissue in the omasal papillae formed flat papillary projections in the oral and middle regions of the laminae. Ridges were arranged in cellular or rosette-like form in the aboral region. In the interpapillary space, the subepithelial connective tissue formed papillary projections in the oral region, parallel ridges with dentate projections in the middle region and smooth low ridges in the aboral region. In the cattle, the projections of the subepithelial connective tissue were more developed than in the sheep. The reticular or cellular ridges were uniformly arranged. The omasal papillae with horny tips located near the reticulo-omasal orifice showed various shapes of papillary projections. In the interpapillary space, round-edged and complex folded ridges with high undulations and deep grooves were observed. These findings may indicate regional and interspecies diversities in the function of the omasal mucosa.

Scanning electron microscopy of vascular architecture in the mucosa of sheep omasal laminae.

Three dimensional vascular architecture in the omasal laminae of sheep was studied by scanning electron microscopy of corrosion casts. The omasal laminae were equipped with well-developed subepithelial capillary network with various capillary loops and inter-papillar undulations. The capillary loops were more highly developed in the omasal papillae than in the inter-papillar space and more in the oral region than in the aboral. The capillary loops were inconspicuous in the inter-papillar space of the middle and aboral regions and in the omasal papillae of the aboral region. On the other hand, the inter-papillar undulation of the capillary network was more conspicuous in the aboral region than in the other regions of the omasal mucosa. These findings may indicate a regional diversity in the function of the omasal laminae.

Morphometry of mitochondria in ovine forestomach epithelia: high densities of mitochondria correlate with specialised metabolic activity.

The volume density of mitochondria and the surface density of mitochondrial inner membranes were investigated in the stratified squamous epithelia of the oesophagus and forestomachs of sheep, and in the oesophagus and nonglandular part of the ventricle of rats. The higher values found for both parameters in all forestomach regions are thought to be correlated with the special metabolic activity known to take place in these epithelia.

The effect of lumen conditions on oxygen uptake in perfused omasal laminae.

The vascular anatomy of the bovine omasal lamina permitted perfusion of a discrete area of the tissue. As occurs in vivo, oxygen was provided through the vascular system, while the luminal sides of the tissue could be kept in an anaerobic environment, thus allowing study of foregut tissue metabolism under physiologically realistic conditions. O2 consumption of perfused leaves in the presence of anaerobic buffer was 64.9 and 73.5 nmol O2/mg dry weight per h in Expts 1 and 2 respectively, and was elevated (P less than 0.05) when the lumen side of the tissue was exposed to an atmosphere of nitrogen gas. In Expt 1, the rate of O2 consumption was increased (P less than 0.01) by 35% as a result of suspension of a boiled preparation of rumen micro-organisms and particles (less than 1 mm) in the anaerobic lumen buffer. Replacement of the boiled preparation with an unboiled suspension increased O2 consumption further by 11%, but this was not statistically significant (P greater than 0.05). In Expt 2, sequential addition of the following substrates or preparations to the lumen chambers all resulted in stepwise increases (P less than 0.05) in O2 consumption; 8 mM-butyrate, boiled rumen micro-organisms and particles and, finally, unboiled rumen micro-organisms and particles. Identities of the heat-labile and heat-stable components of the microbial and particle suspensions that caused enhancement of O2 removal across the perfused tissue are discussed.