The presence of a cholinergic innervation of arterioles within the gut wall is suggested by pharmacological studies of nerve mediated vasodilatation, but attempts to identify nerve cells that give rise to cholinergic vasodilator fibres have yielded discrepant results. In the present work, antibodies to the vesicular acetylcholine transporter protein (VAChT) were used to investigate the relationships of immunoreactive nerve fibres to submucosal arterioles. Comparison was made with cerebral arteries, which are known to be cholinergically innervated. Double labelling immunohistochemical techniques revealed separate VAChT and tyrosine hydroxylase (TH) immunoreactive (IR) fibres innervating all sizes of arteries of the submucosa of the stomach, ileum, proximal colon, distal colon and rectum as well as the cerebral arteries. Arterioles of all digestive tract regions had greater densities of TH-IR innervation than VAChT-IR innervation. In the ileum, double labelling for VAChT-IR and VIP-IR or calretinin-IR showed more VAChT-IR than either VIP-IR or calretinin-IR fibres. Calretinin-IR and VAChT-IR were colocalised in a majority of calretinin-IR axons, but VIP-IR and VAChT-IR were not colocalised. All calretinin-IR nerve cells in submucous ganglia were immunoreactive for choline acetyltransferase, but only 1–2% of VIP-IR nerve cells were immunoreactive. Extrinsic denervation of the ileum did not alter the distribution of VAChT-IR fibres, but it eliminated TH-IR fibres. Removal of myenteric ganglia (myectomy) did not alter the distribution of fibres with VAChT or TH-IR. This work thus provides evidence for cholinergic innervation of intrinsic arterioles throughout the digestive tract and indicates that the fibres in the small intestine originate from submucosal nerve cells.

Connexin 43 (Cx43) has been reported to be expressed in vascular smooth muscle cells and endothelial cells. Evidence for possible variations in Cx43 distribution within different parts of the vascular system is limited. We have therefore investigated the expression of Cx43 in the endothelia and media of 11 vessels of different size and function in the rat, using immunofluorescence and confocal laser scanning microscopy. The results showed that punctate Cx43 staining was abundant in the endothelia and media of all of the 5 elastic arteries examined. In the media, the amount of Cx43 staining decreased as the size of the elastic arteries became smaller. In the 6 muscular arteries examined, 2 different patterns of Cx43 staining were observed. In the first type, Cx43 expression was high in the endothelium but virtually absent from the media. Mesenteric resistance, hepatic and tail arteries were examples. In the second type, Cx43 staining was absent from both the media and the endothelia. The coronary, basilar, and middle cerebral arteries showed this appearance. The results suggest that expression of Cx43 is largely restricted to elastic arteries in the arterial system of the rat. The lack of immunodetectable Cx43 from the media of all muscular arteries examined, and from the endothelia of some of these arteries, raises the possibility of significant differences in the form of expression of Cx43 in these vessels or the presence of other connexins.

Serial transverse histological sections of the human craniovertebral junction (CVJ) of 4 normal human embryos (aged 45 to 58 d) and of a fetus (77 d) were used to create 3-dimensional computer models of the CVJ. The main components modelled included the chondrified basioccipital, atlas and axis, notochord, the vertebrobasilar complex and the spinal cord. Chondrification of the component parts of CVJ had already begun at 45 d (Stage 18). The odontoid process appeared to develop from a short eminence of the axis forming a third occipital condyle with the caudal end of the basioccipital. The cartilaginous anterior arch of C1 appeared at 50–53 d (Stages 20–21). Neural arches of C1 and C2 showed gradual closure, but there was still a wide posterior spina bifida in the oldest reconstructed specimen (77 d fetus). The position of the notochord was constant throughout. The normal course of the vertebral arteries was already established and the chondrified vertebral foramina showed progressive closure. The findings confirm that the odontoid process is not derived solely from the centrum of C1 and that there is a ‘natural basilar invagination’ of C2 during normal embryonic development. On the basis of the observed shape and developmental pattern of structures of the cartilaginous human CVJ, we suggest that certain pathologies are likely to originate during the chondrification phase of development.

The mammalian neuromuscular junction has been extensively studied by different methods to understand better the biological aspects of its normal development, ageing and pathological conditions, such as disorders of neuromuscular transmission. In the present report, a new technique is described that combines confocal microscopy with the use of a vital nerve terminal dye (4-Di-2-ASP) and rhodamine-alpha-bungarotoxin to stain postsynaptic acetylcholine receptors in the same endplate. Nerve terminals in the sternomastoid muscles of living adult mice were stained with 4-Di-2-ASP, which labels intracellular compartments of the nerve terminal containing mitochondria. Slides of these muscles were viewed by confocal microscopy and images were stored on magnetic optical discs. This procedure was compatible with subsequent acetylcholine receptor staining with rhodamine-α-bungarotoxin and observation under the confocal microscope. Classical features of the adult neuromuscular junction were displayed, such as the branched-pattern distribution of the nerve terminals and receptors and their complete colocalisation. In addition, nerve fibres from intramuscular nerve branches with their neighbouring cells, nuclei and muscle fibre striations could also be visualised. We conclude that the present technique can complement existing methods of investigation of nerve terminal anatomy and pathology, particularly where preservation of 3-dimensional relationships is required and intracellular disturbances involving mitochondrial organisation, such as ageing or other degenerative disorders, may be present.

The 3-dimensional ultrastructure of mouse renal glomeruli under normal haemodynamic conditions was studied by scanning electron microscopy using an in vivo cryotechnique followed by freeze-substitution, and compared with glomeruli prepared by conventional fixation methods. Mouse kidneys were frozen with a cryoknife apparatus and a liquid isopentane-propane mixture (−193°C). Surface areas of the frozen tissues were freeze-fractured with a scalpel in liquid nitrogen. The specimens were routinely freeze-substituted, freeze-dried, ion-sputtered, and then observed in a scanning electron microscope at an accelerating voltage of 5 kV. Renal glomeruli showed good ultrastructural preservation of the surface tissues. Podocytes with interdigitating foot processes covering capillary loops exhibited smooth surface contours and their cell surfaces were arranged more tightly than those seen by the conventional fixation method. Filtration slits between foot processes were found to be narrow. The internal structure of the glomerular tuft was seen in the freeze-fracture faces. The capillary lumen with variously shaped erythrocytes was kept open in frozen glomeruli under normal blood circulation conditions. The ultrastructure of renal glomeruli, as revealed by the in vivo cryotechnique with freeze-substitution, appears to be closer to that of the living state.

Data on femoral neck-shaft angles were collected for 30 modern, historic and prehistoric human population samples, and analysed with respect to sexual dimorphism, bilateral asymmetry, geographical patterning and general economic level. Although some samples had modest sexual dimorphism in this angle, and some individuals exhibited clear asymmetry, there were no consistent patterns of sexual or side differences across human populations. Similarly, there was no evidence for geographic differences, since broad regional groups lacked significant differences and mean angles were not correlated with latitude. However, there is a significant increase in mean neck-shaft angles across populations with an increasingly sedentary existence and with mechanisation. The last reflects the developmental plasticity of this feature with respect to habitual load levels during ontogeny of the hip region.

This report deals with the as yet undetermined issue of whether cell-surface associated microtubules in certain cochlear epithelial cells are centrosomally nucleated and subsequently migrate to microtubule-capturing sites located at the surface regions in question. Alternatively, the cells may possess additional nucleating sites which are noncentrosomal and surface-associated. These alternative possibilities have been investigated for highly polarised epithelial cells called supporting cells in the mouse and guinea pig organ of Corti using antibodies to pericentrin and γ-tubulin. There is substantial evidence that both proteins are essential components of microtubule-nucleating sites in cells generally. Each mature supporting cell possesses a large microtubule array that is remotely located with respect to its centrosome (more than 10 μm away). The antibodies bind to a cell's centrosome. No binding has been detected at 2 other microtubule-organising centres that are associated with the ends of the centrosomally-remote microtubule array while it is being constructed. Such arrays include thousands of microtubules in some of the cell types that have been examined. If all a cell's microtubules are nucleated by its centrosome then the findings reported above imply that microtubules escape from the centrosomal nucleating site and migrate to a new location. Furthermore, capture of the plus and minus ends of the errant microtubules is taking place because both ends of a centrosomally-remote microtubule array are attached to sites that are precisely positioned at certain cell surface locations. Minus ends are locating targets with an exactitude comparable to that which has been demonstrated for plus ends in certain cell types. These cells apparently operate a single control centre strategy for microtubule nucleation that is complemented by precise positioning of plus and minus end-capturing sites at the cell surface.

Segmental duplications (also termed ‘fenestrations’) of the fetal anterior cerebral artery are a relatively rare anomaly. Their occurrence was studied in 200 fetuses at 20–40 wk gestational age. Duplications were found in 21 of the 200 fetuses, i.e. 10.5%. In 2 fetuses, i.e. 1%, the duplications were bilateral. In 1 case, i.e. 0.5%, 2 duplications were found on the left anterior cerebral artery. There were 15 segmental duplications on the left anterior cerebral artery, i.e. 62.5% of all duplications, and 9 on the right, i.e. 37.5%. The duplications were located in the distal part of the A1 segment in 45.7%, at the level of the anterior communicating artery (A1–A2) in 37.7%, and in the initial part of the A2 segment in 16.6%. The duplications had 4 different forms namely oviform, fissured, triangular and punctate.

In tonsils of the soft palate of pigs high endothelial venules (HEV) occur throughout the parafollicular region. In some regions of the venules the endothelial lining is uniformly high; in other areas, mainly the nuclear region of the endothelial cell protrudes into the lumen and the cytoplasmic region is attenuated. In this portion of the venule endothelial cells may traverse the lumen of the vessel to form intravascular bridging processes. The endothelial cells are characterised by pinocytotic vesicles, many surface microvilli, cytoplasmic processes and local cell-cell contacts with adjacent endothelial cells. These specialised HEV may play a role in the unusual pattern of lymphocytes from tonsillar tissue into blood. In the tonsils of the soft palate of pigs, HEV may mediate both lymphocyte migration to and from the lymphoid tissue. Further investigation is required to determine the importance of these 2 migration pathways.

Remodelling of bone occurs in response to physical stress. Habitual squatting is associated with modifications of the neck of the talus (squatting facets) and its trochlear/malleolar surfaces (trochlear extensions), and individual populations exhibit different incidences of these modifications that reflect their lifestyle. The occurrence of talar modifications was therefore investigated in a population of late Byzantine (13th century AD) adult male skeletons. Lateral squatting facets occurred most frequently (37.7%), but medial (0.6%), combined (0.6%) and continuous (gutter-like) facets (0.6%) were also observed. Lateral (8.0%), medial (10.9%) and continuous (lateral/central/medial) extensions (4.6%) of the trochlear surface were all present in the late Byzantine population. There was no evidence of side dimorphism. The occurrence of lateral squatting facets in the late Byzantine population was greater than that reported for modern Europeans, but similar to that reported for some populations of modern Indians. The frequency of occurrence of trochlear extensions in the late Byzantine population was substantially less than in modern Indian populations, but similar to modern Europeans. Therefore, it is unlikely that precisely the same factors determine the expression of squatting facets and trochlear extensions.

The most commonly performed procedure for treating coronary artery stenosis is percutaneous transluminal coronary angioplasty (PTCA) and, where the vessel lumen is severely narrowed, coronary artery bypass grafting (CABG). In PTCA, regions of atherosclerotic plaques are disrupted, and the vessel lumen increased by inflating a balloon catheter. In CABG an autologous saphenous vein into coronary artery interposition graft is performed in order to bypass occluded regions of epicardial coronary arteries. Both interventions cause varying degrees of vascular damage and the long-term efficacy of these procedures is limited by a high incidence of neointimal formation and subsequent vascular restenosis (Bach et al. 1994; Bryan & Angelini, 1994).

The endothelium-derived constrictor peptide, endothelin-1 (ET-1) (Yanagisawa et al. 1988), also possesses mitogenic activity on vascular smooth muscle cells (Hirata et al. 1989) and has been suggested as playing a role in atherosclerosis (Dashwood et al. 1993; Zeiher et al. 1994) and intimal hyperplasia (Dashwood et al. 1993; Douglas et al. 1994).

Traumatic avulsions of ligament or tendon insertions rarely occur at the actual interface with bone, which suggests that this attachment is strong or otherwise protected from injury by the structure of the insertion complex. In this study we describe the terminal extent of quadriceps tendon fibres where they insert into the patellae of adult rabbits, humans, dogs and sheep. Specimens were examined by scanning electron microscopy (SEM) and light microscopy (LM). To facilitate tracing of tendon fibres the specimens were decalcified for SEM, and polarised light microscopy (PLM) was used in the LM segment of the study. By SEM it was possible to identify mature bone by the presence of osteocytes and a lamellar organisation. PLM and SEM showed that, unlike tendon fibres elsewhere, those in the calcified fibrocartilage were not crimped. No specific cement line was identified by SEM. Tendon fibres interdigitated among separate bone lamellar systems, (osteons or marrow spaces), but did not merge with the collagen systems of individual lamellae. The interdigitation was more extensive and the margin between tendon and bone was less distinct in the anterior third of the insertion. The segment of calcified tendon which interdigitated with bone stained less intensely blue and was less cellular than the more proximal calcified fibrocartilage zone adjacent to the tidemark. Lamellar collagen fibres of the bony trabeculae in the anterior patella were unusually parallel and longitudinal in orientation, making distinction of interposed tendon fibres difficult on LM and PLM sections. LM, SEM and transmission electron microscopy of rabbit patellae at birth revealed that anterior quadriceps tendon fibres extended over the patella in a fibrous cellular layer. By 2 wk of age, this layer had acquired chondroid features (i.e. cell lacunae and metachromasia) and contained vessels extending from patellar marrow. At 6 wk of age, part of this fibrocartilaginous layer was replaced by mature bone and osteoid. In the young adult animal, the quadriceps tension interdigitates extensively with the patellar bone. This segment of the insertion is perhaps the remnant of calcified fibrocartilage which has been remodelled by bone formation.

The term superficial ulnar artery (SUA) is applied to an artery which arises from the axillary, brachial or superficial brachial arteries and courses over the origins of the superficial forearm muscles to join at the midlevel of the forearm with the ulnar artery, sometimes replacing it (Wankoff, 1962; Lippert & Pabst, 1985; Nakatani et al. 1996, 1997). Its incidence ranges from 0.7% (Adachi, 1928) to 3.3% (Müller, 1903). The SUA has been reported with different terminologies; arteria antebrachialis superficialis ulnaris (Gruber, 1867; Müller, 1903; Adachi, 1928), high origin of the ulnar artery (Rodriguez-Baeza et al. 1995; Aharinejad et al. 1997) and SUA with a high origin (Fuss, 1988).

The present observation adds several aspects to the anatomical variations of the superficial ulnar arteries already described, most notably its anastomosis with the anterior interosseous artery. This anastomosis has previously been reported twice (Quain, 1844; Gruber, 1867) but without explanation as to the course taken by the anastomotic branch. The SUA was found in the right arm of a 66-y-old male during routine dissections in the Unit of Anatomy and Embryology at the Autonomous University of Barcelona.

In order to clarify the fundamental structure of the host defence mechanism in chicken caeca, a detailed analysis of the distribution of lymphoid nodules (LNs) was carried out on longitudinal sections of both the mesenteric (side of the ileocaecal ligament) and the antimesenteric mucosa. An overwhelming majority of solitary or aggregated LNs were located in the mesenteric mucosa, although a few were also found in the antimesenteric mucosa. Of the total LNs, 45.7% were detected at the proximal 7.8% section in the caecal tonsil. LNs (21.4%) were also concentrated in the distal 22.0% section corresponding to the apex. A moderate concentration of LNs (13.1%) was found at the transitional 20.0% region between the base and body. Approximately 80.2% of total LNs were found at the above 3 regions in the mesenteric mucosa. In many cases, the frequency of LNs in the caecal tonsils was opposite to that at the apices. Aggregated LNs were mainly found in the caecal tonsils, transitional region and apex. Almost all aggregated LNs consisted of fundamental nodular units possessing M cells in their follicle associated epithelia. The aggregated LNs in the above 3 regions therefore could provide immunological surveillance against caecal luminal contents. In particular, the cooperative function between LNs of the caecal tonsil and apex might be highly important in maintaining the caecal microenvironment.

The Summer meeting of the Anatomical Society of Great Britain and Ireland was held at the Anatomy Department, Edinburgh University from 15 to 17 July 1997. It included a symposium on ‘Computer Modelling for Anatomists and Clinicians’ on Wednesday 16 July. The following are abstracts of communications and demonstrations presented at the meeting.

A study was carried out on the discomalleolar ligament by dissection of adult human cadavers. The ligament corresponds to the most internal portion of the superior lamina of the temporomandibular joint capsule. It extends from the posterointernal portion of the temporomandibular joint disc, penetrates the petrotympanic fissure and reaches the malleus of the middle ear. Because of its morphology and anatomical arrangement the discomalleolar ligament should be considered as an intrinsic ligament of the temporomandibular joint and distinguished from the tympanic portion of the sphenomandibular ligament (anterior ligament of the malleus).

The desired properties required for successful embalming of cadavers for gross anatomy teaching include: (1) good long-term structural preservation of organs and tissues with minimal shrinkage or distortion; (2) prevention of over-hardening, while maintaining flexibility and suppleness of internal organs; (3) prevention of desiccation; (4) prevention of fungal or bacterial growth and spread within a specific cadaver and to other cadavers in the dissection room; (5) reduction of potential biohazards (spread of infection to dissection personnel and students); (6) reduction of environmental chemical hazards (especially from formaldehyde and phenol) in order to comply with increasingly severe health and safety regulations and a new awareness of possible dangers of these chemicals in the workplace; and (7) retention of colour of tissues and organs while minimising oxidation effects that result in ‘browning’.

The recent exchange on cartilage ultrastructure, the localisation of keratan sulphate (KS) therein and the influence of oxygen tension on both (Stockwell & Scott, 1997; Archer et al. 1997; Caterson, 1997) raised important issues which need clarification and extension. We wish to correct certain statements and to examine more rigorously principles which will establish a foundation for future discussion.