To investigate the claim that the primate paranasal sinuses possess not a functional but a structural role associated with the skull architecture (Blaney, 1990), the relationship between the maxillary sinus and the skull architecture was studied ontogenetically in 30 skulls of male and female Japanese macaques (Macaca fuscata). Coronal CT scan series and computerised 3-dimensional images served to evaluate the maxillary sinus. The definitive hemispherical shape of the sinus was already achieved after the completion of the primary dentition. Sinus volume increased with a trend indicating positive allometry. When compared with an ontogenetic data set of orang-utan (Koppe et al. 1995), however, the growth rate of the maxillary sinus of M. fuscata was significantly less. The maxillary sinus both of male and female macaques enlarged according to a common growth pattern. However, no sexual dimorphism could be established for the maxillary sinus size. Although the volume of the right maxillary sinus was normally bigger than that of the left side, the results suggested that asymmetry in maxillary sinus volume is related neither to skull size nor sex. Whereas a correlation analysis showed close relationships between the maxillary sinus volume and external cranial dimensions, the partial correlation coefficients revealed that these relationships were highly influenced by skull size. Although it cannot be ruled out that the paranasal sinuses are to some extent linked to the skull architecture, this study does not support a solely structural role for these air cavities.

Neurons cannot negotiate an elongation across the peripheral (PNS)–central nervous system (CNS) transitional zone and grow into or out of the spinal cord in the mature mammal. The astrocytic rich CNS part of the spinal nerve root is most effective in preventing regeneration even of nerve fibres from transplanted embryonic ganglion cells. Regeneration of severed nerve fibres into the spinal cord occurs when the transition zone is absent as in the immature animal. Before the establishment of a transition zone there is also new growth of neuronal processes from dorsal horn neurons distally to the injured dorsal root. Thus the experimental strategy to reestablish spinal cord to peripheral nerve connectivity has been to delete the transitional region and implant severed ventral or dorsal roots into the spinal cord. Dorsal root implantation resulted in reestablished afferent connectivity by new neuronal processes from secondary sensory neurons in the dorsal horn of the spinal cord extending into the PNS. The ability for plasticity in these cells allowed for a concurrent retention of their original rostral projection. Ventral root implantation into the spinal cord corrected deficit motor function. In a long series of experiments performed in different species, the functional restitution was demonstrated to depend on an initial regrowth of motor neuron axons through spinal cord tissue (CNS). These findings have led to the design of a new surgical strategy in cases of traumatic spinal nerve root injuries.

Timing and pattern of expression of alkaline phosphatase was examined during early differentiation of the 1st arch skeleton in inbred C57BL/6 mice. Embryos were recovered between 10 and 18 d of gestation and staged using a detailed staging table of craniofacial development prior to histochemical examination. Expression of alkaline phosphatase is initiated at stage 20.2 in the plasma membrane of mesenchymal cells in the distal region of the first arch. Expression is strongest in osteoid (unmineralised bone matrix) and presumptive periosteum at stage 21.32. Mineralisation begins at stage E23. Expression is present in the mineralised bone matrix. Secondary cartilages form in the condylar and angular processes by stage M24. The cartilaginous cells and surrounding cells in the processes are all alkaline phosphatase-positive and surrounded by the common periosteum, suggesting that progenitor cells of the processes, dentary ramus and secondary cartilages all originate from a common pool. Nonhypertrophied chondrocytes of Meckel's cartilage express alkaline phosphatase at stage M23. Expression in these chondrocytes is preceded by the expression in their adjacent perichondrium. This is true of chondrocytes in all other cranial cartilages examined. 3-D reconstruction of expression in Meckel's cartilage also revealed that the chondrocytes of Meckel's cartilage which express alkaline phosphatase and the matrix of which undergoes mineralisation are those surrounded by the alkaline phosphatase-positive dentary ramus. By stage 25, coincident with mineralisation in the distal section of Meckel's cartilage, most chondrocytes are strongly positive. The perichondria of malleus and incus cartilages express alkaline phosphatase at stage M24. Nonhypertrophied chondrocytes along these perichondria also express alkaline phosphatase. Superficial and deep cells in the dental laminae of incisor and 1st molar teeth become alkaline phosphatase-positive at the bud stage, stages 21.16 and 21.32, respectively. Dental papillae are negative until stage M24 when alkaline phosphatase expression begins in the dental papillae and follicles of the incisor teeth and the dental follicles of the 1st molar teeth. The dental papillae of the 1st molar teeth express alkaline phosphatase at stage 25. Expression in the dental papillae and follicles appears to coincide with cellular differentiation of follicle from papilla. The presumptive squamosal, ectotympanic and gonial membrane bones, lingual oral epithelial cells connected to the dental laminae of the incisor teeth, hair follicle papillae and sheath and surrounding dermis all express alkaline phosphatase in a stage-specific manner.

Fourteen steps of spermatid development in the tammar wallaby (Macropus eugenii), from the newly formed spermatid to the release of the spermatozoon into the lumen of the seminiferous tubules, were recognised at the ultrastructural level using transmission and scanning electron microscopy. This study confirmed that although the main events are generally similar, the process of the differentiation of the spermatid in marsupials is notably different and relatively more complex than that in most studied eutherian mammals and birds. For example, the sperm head rotated twice in the late stage of spermiogenesis: the shape of the spermatid changed from a T-shape at step 10 into a streamlined shape in step 14, and then back to T-shape in the testicular spermatozoa. Some unique figures occurring during the spermiogenesis in other marsupial species, such as the presence of Sertoli cell spurs, the nuclear ring and the subacrosomal space, were also found in the tammar wallaby. However, an important new finding of this study was the development of the postacrosome complex (PAC), a special structure that was first evident as a line of electron dense material on the nuclear membrane of the step 7 spermatid. Subsequently it became a discontinuous line of electron particles, and migrated from the ventral side of the nucleus to the area just behind the posterior end of the acrosome, which was closely located to the sperm–egg fusion site proposed for Monodelphis domestica (Taggart et al. 1993). The PAC and its possible role in both American and Australian marsupials requires detailed examination. Distinct immature features were discovered in the wallaby testicular spermatozoa. A scoop shape of the acrosome was found on the testicular spermatozoa of the tammar wallaby, which was completely different to the compact button shape of acrosome in ejaculated spermatozoa. The fibre network found beneath the cytoplasm membrane of the midpiece of the ejaculated sperm also did not occur in the testicular spermatozoa, although the structure of the principal piece was fully formed and had no obvious morphological difference from that of the epididymal and ejaculated spermatozoa. The time frame of the formation of morphologically mature spermatozoa in the epididymis of the tammar wallaby needs to be determined by further studies.

A quick-freezing and deep-etching method in combination with erythrocyte splitting was used to examine the cytoplasmic aspect of whole-mount human erythrocyte membranes. Various external forces induced alterations in membrane skeletal organisation during the splitting procedure. The initial change was elongation in the peripheral part of the membrane skeleton, examined by immunostaining with a monoclonal antispectrin antibody. Under severe stretching conditions, a linear rearrangement of filamentous components was evident; these were disposed parallel to the rim of the erythrocyte, while the central part of the concavity exhibited a more compacted structure. These changes resulted in a different distribution of membrane skeletal components between central rigid and peripheral flexible areas in biconcave erythrocytes. It is suggested that the reversible membrane skeletal changes in the flexible areas which resist the external forces are important for maintaining the normal framework of biconcave human erythrocytes.

Adult muscle is highly vascularised, with blood vessels being essential for adequate oxygenation of the tissue and for supporting increased metabolic demands. Whether this is the case during muscle development has not been examined. Resin histology was used to map the muscle splitting process and conventional transmission electron microscopy to examine early muscle differentiation at the midlimb level or later at the mid radius/ulna level in the chick wing bud from stages 24 (4.5 d) to 36 (10 d) (Hamburger & Hamilton, 1951). Microinjection of India Ink into the extra-embryonic vasculature was used to visualise the patent muscle microcirculation. The results showed that the premuscle masses are present at stage 24 and initial splitting of the muscle masses commences at stage 28. The final muscle pattern is not established until stage 36. At stage 26 the cells within the premuscle masses exhibited a mesenchymal morphology, but at stage 28 overt muscle differentiation was evident with myofibrils present within myoblasts. Undifferentiated mononucleated cells were interspersed with the differentiating myoblasts. The ratio of mononucleated cells[ratio ]myoblasts decreased and the myoblasts became plumper and increasingly packed with myofibrils with age. There was no evidence of secondary myotube formation at any of the stages examined. Vascular invasion of the limb occurred at stage 35 just prior to the establishment of the final muscle pattern. This was surprising as it was assumed that myogenic differentiation would be both oxygen and nutrient dependent. The results of this study provide descriptions of the splitting of the premuscle masses through to the establishment of the final muscle pattern at the midlimb or mid radius/ulna level of the chick wing bud together with the differentiation of the myogenic cells within the developing muscles. However, the relationship between muscle patterning at the tissue level and muscle differentiation at the cellular level with vascularisation remains unclear. It is hoped that the results of the study may provide the basis for future investigations into mechanisms involved in muscle patterning and the signalling mechanisms for vascular invasion.

The management of peripheral nerve injury remains a major clinical problem. Progress in this field will almost certainly depend upon manipulating the pathophysiological processes which are triggered by traumatic injuries. One of the most important determinants of functional outcome after the reconstruction of a transected peripheral nerve is the length of the gap between proximal and distal nerve stumps. Long defects (> 2 cm) must be bridged by a suitable conduit in order to support axonal regrowth. This review examines the cellular and acellular elements which facilitate axonal regrowth and the use of acellular muscle grafts in the repair of injuries in the peripheral nervous system.

In the course of an extensive comparative, structural and developmental study of the cranial and postcranial dermal skeleton (teeth and scales) in osteichthyan fishes, we have undertaken investigations on scale development in zebrafish (Danio (Brachydanio) rerio) using alizarin red staining, and light and transmission electron microscopy. The main goal was to know whether zebrafish scales can be used as a model for further research on the processes controlling the development of the dermal skeleton in general, especially epithelial–mesenchymal interactions. Growth series of laboratory bred specimens were used to study in detail: (1) the relationship of scale appearance with size and age; (2) the squamation pattern; and (3) the events taking place in the epidermis and in the dermis, before and during scale initiation and formation, with the aim of searching for morphological indications of epithelial-mesenchymal interactions. Scales form late in ontogeny, generally when zebrafish are more than 8.0 mm in standard length. Within a population of zebrafish of the same age scale appearance is related to standard length, but when comparing populations of different age the size of the fish at scale appearance is also related to age. Scales always appear first in the posterior region of the body and the squamation then extends anteriorly. Scales develop in the dermis but closely apposed to the epidermal–dermal boundary. Cellular modifications occurring in the basal layer of the epidermis and in the dermis before scale formation clearly indicate that the basal epidermal cells differentiate first, before any evidence of differentiation of the progenitors of the scale-forming cells in the dermis. This strongly suggests that scale differentiation could be initiated by the epidermal basal layer cells which probably produce a molecular signal towards the dermis below. Subsequently dermal cells accumulate close to the epidermis, and differentiate to form scale papillae. The late formation of the scales during ontogeny is due to a late colonisation of the dermis by the progenitors of the scale-forming cells. Because of their late formation during ontogeny and of their regular pattern of development, scales in zebrafish represent a good model for further investigations on the general mechanisms of epithelial-mesenchymal interactions during dermal skeleton development, and in particular for the study of the gene expression patterns.

Nuclear condensation during spermiogenesis in the ostrich follows the basic pattern established in other vertebrates. The fine granular nuclear substance of early spermatids is gradually replaced by numbers of coarse dense granules which appear to arise by aggregation of smaller dispersed elements of the chromatin. The granules increase in size and eventually coalesce to form the compact homogenous mass of chromatin typical of the mature sperm. In ostrich spermatids, however, the aggregation of the nuclear material produces large numbers of longitudinally oriented rod-shaped structures in addition to some granular material. Although fibrillar chromatin has been observed during spermiogenesis in a number of vertebrate species, the hollow nature of the rod-shaped chromatin granules in ostrich spermatids is a unique phenomenon. The spiralisation of the chromatin material observed in ostrich spermatids and in some other nonpasserine birds is possibly related to the reduction in nuclear length demonstrated during spermiogenesis in these species. In common with other nonpasserine birds, spermiogenesis in the ostrich is characterised by the appearance both of a circular and a longitudinal manchette. The circular manchette consists of a single row of microtubules reinforced by additional peripherally arranged microtubules. Links between adjacent microtubules, and between the nucleolemma and some of the microtubules, are evident. The longitudinal manchette consists of arrays of interconnected microtubules arranged in approximately 4–6 staggered, ill defined rows. This structure seems to originate as a result of the rearrangement of the microtubules of the circular manchette and is only formed once the process of chromatin condensation is well advanced. Based on the sequence of morphological events observed during spermiogenesis in the ostrich, it is concluded that the circular manchette is responsible for the initial transformation in shape of the spermatid nucleus. Thereafter, the chromatin condenses independently within the confines of the nucleolemma with the circular manchette merely acting to maintain the shape of the nucleus while this process is underway, to compress the nuclear membrane, and possibly to orientate the subunits of the condensing chromatin. The longitudinal manchette appears to assist in the translocation of material during spermatid elongation. There are indications that the developing acrosome is instrumental in effecting nuclear shaping of the apical (subacrosomal) head region of the ostrich spermatid.

Some 30 lectins in combination with glycosidase digestion and immunohistochemistry with 5 antibodies directed against antigens of the ABO and Lewis blood group systems were used to analyse the distribution and synthesis of glycoconjugates in the epithelium of the large airways in man. Both mucous gland cells and goblet cells were labelled by 12 of 30 lectins and by the antibodies, dependent on the ABO, Lewis, and secretor status. The corresponding binding patterns of the serous gland cells differed markedly from those of goblet and mucous gland cells and in general were not dependent on the ABO, Lewis, and secretor status. After digestion with neuraminidase and fucosidase, binding of soy bean agglutinin and peanut agglutinin to goblet and mucous gland cells was increased. Binding of peanut agglutinin to serous gland cells was stronger only after the digestion with neuraminidase. Digestion with O-glycosidase after the use of neuraminidase or fucosidase resulted in a decrease of peanut agglutinin binding to goblet and mucous gland cells. The present results show that the secretory products of goblet and mucous gland cells on the one hand and those of serous cells on the other differ considerably with respect to their terminal glycosylation. The glycosyltransferases coded by genes of the ABO and Lewis blood group and secretor systems are active only in goblet and mucous gland cells, resulting in the presence of the corresponding antigens. Precursor substances of blood group antigens types 1, 2, and 3 are found only in these cell types. In serous gland cells, blood group systems do not influence the glycosylation of glycoproteins. The results of the digestion with O-glycosidase indicates the presence of O-glycosylation in mucous gland and goblet cells, but not in serous gland cells.

Tammar wallaby spermatozoa undergo maturation during transit through the epididymis. This maturation differs from that seen in eutherian mammals because in addition to biochemical and functional maturation there are also major changes in morphology, in particular formation of the condensed acrosome and reorientation of the sperm head and tail. Of spermatozoa released from the testes, 83% had a large immature acrosome. By the time spermatozoa reached the proximal cauda epididymis 100% of sperm had condensed acrosomes. Similarly 86% of testicular spermatozoa had immature thumb tack or T shape head-tail orientation while only 2% retained this immature morphology in the corpus epididymis. This maturation is very similar to that reported for the common brush tail possum, Trichosurus vulpecula. However, morphological maturation occurred earlier in epididymal transit in the tammar wallaby. By the time spermatozoa had reached the proximal cauda epididymis no spermatozoa had an immature acrosome and thumbtack orientation. Associated with acrosomal maturation was an increase in acrosomal thiols and the formation of disulphides which presumably account for the unusual stability of the wallaby sperm acrosome. The development of motility and progressive motility of tammar wallaby spermatozoa is similar to that of other marsupials and eutherian mammals. Spermatozoa are immotile in the testes and the percentage of motile spermatozoa and the strength of their motility increases during epididymal transit. During passage through the caput and corpus epididymis, spermatozoa first became weakly motile in the proximal caput and then increasingly progressively motile through the corpus epididymis. Tammar wallaby spermatozoa collected from the proximal cauda epididymis had motility not different from ejaculated spermatozoa. Ultrastructural studies indicated that acrosomal condensation involved a complex infolding of the immature acrosome. At spermiation the acrosome of tammar wallaby spermatozoa was a relatively large flat or concave disc which projected laterally and anteriorly beyond the limits of the nucleus. During transit of the epididymal caput and proximal corpus the lateral projections folded inwards to form a cup like structure the sides of which eventually met and fused. The cavity produced by this fusion was lost as the acrosome condensed to its mature form as a small button-like structure contained within the depression on the anterior end of the nucleus. During this process the dorsal surface of the immature acrosome and its outer acrosomal membrane and overlying plasma membrane were engulfed into the acrosomal matrix. This means that the dorsal surface of the acrosomal region of the testicular tammar wallaby sperm head is a transient structure. The dorsal acrosomal surface of the mature spermatozoon appears ultrastructurally to be the relocated ventral surface of the acrosomal projections which previously extended out beyond the acrosomal depression on the dorsal surface of the nucleus of the immature spermatozoon.

Changes in the ovine mammary gland epithelium during initiated involution were studied by light and electron microscopy. Apoptosis of the duct and alveolar epithelial cells was first identified at 2 d after weaning, reached a peak at 4 d and then progressed gradually thereafter. Apoptotic cells were phagocytosed by intraepithelial macrophages and alveolar epithelial cells. Occasional apoptotic epithelial cells were observed in the alveolar and duct lumina. The highly vacuolated cells in the alveolar and duct lumina were confirmed to be macrophages as they were CD45+, MHC class II+. Changes in myoepithelial cells involved shrinkage and extension of cytoplasmic processes into the underlying stroma and no apoptosis was observed. Regression of the blood capillaries was also by apoptosis. The resulting apoptotic bodies were either taken up by adjacent endothelial cells or were shed into the capillary lumen to be phagocytosed later by mural endothelial cells or blood monocytes. The mammary glands were completely involuted by 30 d after weaning. It was concluded that the mammary gland involutes by apoptosis, a process which allows deletion of cells without the loss of the basic architecture and the integrity of the epithelial lining of the gland.

The organisation of the stromal cell compartment in the mouse lymph node was studied by light and electron microscopy after tissue impregnation by the zinc iodide-osmium (ZIO) method. Fibroblastic reticular cells (FRCs) represented the main stromal cell population. These cells were located both in the cortical region and in the medulla and exhibited various configurations. In the cortex, FRCs were fusiform in shape and came into close proximity with the floor of the subcapsular sinus. In the medulla, the FRCs were shaped like irregular dendritic cells which formed a complex 3-dimensional network. The FRCs surrounded vascular structures such as capillaries and/or high endothelial venules; in these instances they were organised in a discontinuous sheath-like fashion around the vessel wall. By light and electron microscopy, FRCs have been observed to come in close spatial relationship with a number of cells in the lymph node, including sinus endothelial cells, the endothelium of high endothelial venules and capillaries, various types of lymphocytes, follicular dendritic cells and interdigitating cells. These microanatomical features are consistent with the proposal that FRCs may be involved in the communicative networks between the different lymph node compartments. In particular, the FRCs may be involved in the transport of molecules from the sinus compartment to the high endothelial venules or to the distinct cell populations in the lymphoid parenchyma.

Exposure of male Albino Swiss rats to the nonsteroidal antiandrogen flutamide during the period from gestational day (d) 10 to birth resulted in feminisation of the external genitalia and the suppression of growth of the male reproductive tract. In adulthood, testes were found to be located in diverse positions. True cryptorchidism occurred in 10% of cases, whereas 50% of testes descended to the scrotum and 40% were located in a suprainguinal ectopic region. Varying degrees of tubule abnormality were seen in the testes of flutamide-treated animals, ranging from completely normal tubules with full spermatogenesis (and the expected frequency of the stages of spermatogenesis) to severely abnormal tubules lined with Sertoli cells only. For each individual testis, the overall severity of tubule damage was strongly correlated with its adult location, with intra-abdominal testes worst affected and scrotally-located testes least; only the latter contained normal tubules. Similarly, intra-abdominal testes were the smallest in weight and contained the least testosterone. By contrast, postnatal treatment of male rats with flutamide from birth to postnatal d 14 did not impair development of the external genitalia, the process of testicular descent or adult spermatogenesis. These findings confirm that androgen blockade during embryonic development interferes with testicular descent but also demonstrate that (1) prenatal flutamide treatment per se has a detrimental effect on adult testis morphology but (2) the degree of abnormality of the testes is strongly influenced by location.

The crush model of injury in skeletal muscle is widely used in the investigation of tissue degeneration and regeneration. Previously, such trauma has been induced by using forceps to crush the muscle, commonly applying sufficient pressure to bring the mid-arms of the forceps together. This report introduces a reliable electromechanical device designed to generate reproducible focal lesions in skeletal muscle of mice. The tibialis anterior was crushed in 17 young adult mice. Two days after injury, the muscles were examined microscopically. By morphometric analysis, it was determined that the volumes of the lesions produced were similar (mean 0.499 mm3±0.098, range 0.278−0.601 mm3), and that the full extent of the damaged muscle was easily distinguished and readily quantifiable. This will allow a more precise comparison in future investigations into regenerative differences between age groups, satellite cell activation and the inflammatory response.

Wheat germ agglutinin–horseradish peroxidase conjugate (WGA–HRP) was injected into the dorsal root ganglia (L5–S1) of the cat and used as an anterograde tracer substance for intra-axonal labelling of peripheral nerve endings in joint capsule and cranial (anterior) cruciate ligament (CCL). We believed that the high specificity of WGA–HRP for neural tissue along with the high visibility of its reaction product could help resolve controversies concerning the sensory innervation of the cruciate ligaments. Substantial amounts of WGA–HRP were transported in tibial nerve axons to the level of the knee. However, using standard HRP histochemistry we found that the capsular tissue and ligament synovia disintegrated during the incubation reaction. This problem was avoided by air drying the tissue slices on glass slides prior to reaction. Abundant labelling occurred in the posterior capsule with dense filling of axons and terminal endings. Sensory endings displayed features consistent with Ruffini endings and pacinian corpuscles. Sensory endings were located throughout the CCL in its sagittal plane, in the subsynovial layers and between collagen fascicles. In each CCL we observed 5–17 ovoid and elongated endings with dense terminal arborisations. These endings were between 100 and 150 μm long, were encapsulated, and gave rise to 1 or 2 axons. Large (up to 1.5 mm in maximum extent) elongated regions of dense, inhomogeneous labelling were found in the body of several CCLs. These resembled Golgi tendon-like endings, with the exception of their large size. We conclude that anterograde transport of HRP to the knee is a useful technique for labelling mechanoreceptors and axons in knee tissue. However, recently developed immunohistochemical analysis of peripheral tissue using protein gene product 9.5 appears to be the method of choice and should be employed for further study of human and animal cruciate ligament innervation.

The Trembler-J (TrJ) mouse has a point mutation in the gene coding for peripheral myelin protein 22 (PMP22). Disturbances in PMP22 are associated with abnormal myelination in a range of inherited peripheral neuropathies both in mice and humans. PMP22 is produced mainly by Schwann cells in the peripheral nervous system where it is localised to compact myelin. The function of PMP22 is unclear but its low abundance (∼5% of total myelin protein) means that it is unlikely to play a structural role. Its inclusion in a recently discovered family of proteins suggests a function in cell proliferation/differentiation and possibly in adhesion. Nerves from TrJ and the allelic Trembler (Tr) mouse are characterised by abnormally thin myelin for the size of the axon and an increased number of Schwann cells. We report ultrastructural evidence of abnormal Schwann cell-axon interactions. Schwann cell nuclei have been found adjacent to the nodes of Ranvier whereas in normal animals they are located near the centre of the internodes. In some fibres the terminal myelin loops faced outwards into the extracellular space instead of turning inwards and terminating on the axon. In severely affected nerves many axons were only partially surrounded by Schwann cell cytoplasm. All these features suggest a failure of Schwann cell–axon recognition or interaction. In addition to abnormalities related to abnormal myelination there was significant axonal loss in the dorsal roots.

Important functions of the oviduct during reproduction include the provision of an optimal environment for gametes and zygotes and nutrition of the early embryo. These functions are ensured by the secretion of an oviductal fluid which is known to contain organ-specific glycoproteins. Glycoconjugates of the apical glycocalyx are considered to play a major role in cell recognition and interaction processes. In the present investigation, binding patterns of Con A, HPA, LTA, RCA I, UEA I, and WGA were studied in defined segments of the oviduct (infundibulum, ampulla, isthmus) and in the uterus during the oestrous cycle. The carbohydrate distribution showed regional as well as time dependent differences. LTA, HPA and WGA reacted strongly with Golgi regions and secretory granules in the oviduct epithelium during the follicular phase, indicating high secretory activity. LTA, HPA, and UEA I also revealed a varying carbohydrate composition of the glycocalyx during the oestrous cycle. Prominent regional differences in glycoconjugate expression were shown in oviductal segments by LTA and HPA binding: during the follicular phase, LTA binding sites were only present on epithelial cells of the isthmic segment, the ampulla and infundibulum being unreactive. D-N-acetyl-galactosamine residues were demonstrated on ciliated epithelial cells of the ampulla and infundibulum exclusively during oestrus. The glycocalyx of uterine epithelial cells was clearly defined by HPA, WGA, LTA, RCA I and UEA I; LTA binding was restricted to the secretory phase. The observation of regional and time dependent variability in glycoconjugate distribution strongly indicates their specific physiological functions in reproductive processes.

An immunocytochemical and cytochemical study has been made on the ultrastructural localisation of type III (endothelial) nitric oxide synthase, endothelin-1 and the binding sites of lectin from Bandeirea simplicifolia to the endothelium surface-associated glycoproteins in the rat left common carotid artery at 1 and 28 d after Fogarty embolectomy balloon catheter-induced injury. Controls were carotid arteries from sham operated rats. In the controls, the immunoreactivity to nitric oxide synthase-III and endothelin-1 was localised in different proportions in vascular endothelial cells (36·9%±4·3 and 7·6%±2·7, respectively); immunoreactivity was confined to the cytoplasm and the membranes of intracellular organelles and structures. In contrast, staining with lectin was localised on the luminal surface of all endothelial cells. 1 d after injury, platelets were adherent to the endothelium-denuded intima. Some of the platelets displayed immunoreactivity to nitric oxide synthase-III and endothelin-1 and were stained with lectin. 28 d after injury, a neointimal thickening of substantial size was present. Subpopulations of the regrown endothelial cells covering the luminal surface of the neointima showed positive immunoreactivity to nitric oxide synthase-III and endothelin-1 but there was a significant decrease in the proportion of nitric oxide synthase-III-containing endothelial cells (17·2%±1·9; P < 0·001) and a significant increase in the proportion of endothelin-1-containing endothelial cells (36·9%±4·7; P < 0·001) compared with the controls. Staining with lectin was associated with the cell membrane of all endothelial cells and in addition with cells located ‘deeper’ in the neointima which showed lectin-positive plasmalemma, Golgi complex and multivesicular bodies/lysosomes. In conclusion, regenerated endothelial cells of the neointima showed reduced population (2–fold) of nitric oxide synthase-III- and increased population (5–fold) endothelin-1-positive cells. The subendothelial location of some lectin-stained cells after balloon catheter injury indicates the heterogeneity of the neointima and suggests that some of these cells are involved in early angiogenesis. 24 h and 28 d after injury some platelets showed positive immunoreactivity for nitric oxide synthase-III and endothelin-1.

Morphological features of the cap enameloid and dental epithelial cells were investigated by light and transmission electron microscopy during the various stages of enameloid mineralisation and early maturation in the tilapia. The pattern of mineralisation along collagen fibrils in the enameloid differed from that in the dentine. Many matrix vesicles were found in the predentine and in the enameloid, suggesting that they may be involved in the initial mineralisation in both regions. Most of the organic matrix disappeared from the cap enameloid during mineralisation and maturation. The disappearance of the organic matrix could be divided into 2 stages. Initially a fine network-like matrix, which probably consisted of glycosaminoglycans and extended between collagen fibrils, began to disappear. At the same time, fine crystallites and electron-dense, fine granular material covered the collagen fibrils as mineralisation of the enameloid began. In the second stage, the maturation of the enameloid, the collagen fibrils degenerated completely and disappeared from the cap enameloid, being replaced by large numbers of large crystals. At the mineralisation stage, the numbers of lysosomal bodies tended to increase in the inner dental epithelial (IDE) cells, which contained a well developed Golgi apparatus and rough endoplasmic reticulum (rER). At the early stage of maturation, a ruffled border was noted at the distal ends of the IDE cells, which contained many mitochondria and lysosomal bodies, but less rER. These features suggest that the cells actively absorb the organic matrix, which includes collagen fibrils, in the cap enameloid. The outer dental epithelial (ODE) cells were translucent cells that contained well developed labyrinthine canalicular spaces from the onset of the mineralisation stage to the middle stage of maturation. The IDE and ODE cells were clearly involved in the mineralisation of the cap enameloid at the mineralisation and maturation stages.