In the developing vertebrate retina, ganglion cells fire spontaneous bursts of action potentials long before the eye becomes exposed to sensory experience at birth. These early bursts are synchronised between neighbouring retinal ganglion cells (RGCs), yielding unique spatiotemporal patterns: ‘waves’ of activity sweep across large retinal areas every few minutes. Both at retinal and extraretinal levels, these embryonic retinal waves are believed to guide the wiring of the visual system using hebbian mechanisms of synaptic strengthening.

In the first part of this review, we recapitulate the evidence for a role of these embryonic spontaneous bursts of activity in shaping developing complex receptive field properties of RGCs in the turtle embryonic retina. We also discuss the role of visual experience in establishing RGC visual functions, and how spontaneous activity and visual experience interact to bring developing receptive fields to maturation. We have hypothesised that the physiological changes associated with development reflect modifications in the dendritic arbours of RGCs, the anatomical substrate of their receptive fields. We demonstrate that there is a temporal correlation between the period of receptive field expansion and that of dendritic growth. Moreover, the immature spontaneous activity contributes to dendritic growth in developing RGCs. Intracellular staining of RGCs reveals, however, that immature receptive fields only rarely show direct correlation with the layout of the corresponding dendritic tree. To investigate the possibility that not only the presence of the spontaneous activity, but even the precise spatiotemporal patterns encoded in retinal waves might contribute to the refinement of retinal neural circuitry, first we must clarify the mechanisms mediating the generation and propagation of these waves across development. In the second part of this review, we present evidence that turtle retinal waves, visualised using calcium imaging, exhibit profound changes in their spatiotemporal patterns during development. From fast waves sweeping across large retinal areas and recruiting many cells on their trajectory at early stages, waves become slower and eventually stop propagating towards hatching, when they become stationary patches of neighbouring coactive RGCs. A developmental switch from excitatory to inhibitory GABAA responses appears to mediate the modification in spontaneous activity patterns while the retina develops. Future chronic studies using specific spatiotemporal alterations of the waves will shed a new light on how the wave dynamics help in sculpting retinal receptive fields.

This paper reports 3-dimensional confocal microscopy observations on how nephrogenic aggregates form from the NCAM- and Pax2-positive caps (4–5 cells deep) of condensed metanephric mesenchyme surrounding the duct tips of the mouse kidney. Aggregates of 6–8 cells are first seen at ∼E12.5–12.75 immediately proximal to this cap, closely abutting the duct surface. As the tip advances, NCAM expression is maintained in the cap but is otherwise restricted to aggregates whose cells rapidly epithelialise, forming tubules that invade the duct epithelium. Pax2 expression studies shows how the rind of nephrogenic blastemal cells forms: as duct tips extend towards the kidney surface, the associated Pax2+ cells form patches of cells on the kidney surface. These observations revise our knowledge of the timing and process of nephron initiation.

The primary cilium is a ubiquitous cytoplasmic organelle of unknown function. Ultrastructural evidence of primary cilia in chondrocytes, and their colocalisation with the Golgi apparatus, has led to speculation that these structures are functionally linked. To investigate the relationship between these organelles, we examined the molecular anatomy of the microtubular cytoskeleton in the chondrocytes of chick embryo sterna. Thick cryosections were immunolabelled with antibodies directed against acetylated α-tubulin (C3B9), detyrosinated α-tubulin (ID5) and total α-tubulin (TAT), and imaged at high magnification using confocal laser scanning microscopy. Transmission electron microscopy confirmed the ultrastructure of the chondrocyte primary cilium and its structural relationship to the Golgi apparatus. Detyrosinated and acetylated α-tubulins were concentrated in the centrioles, centrosome and microtubule organising centre adjacent to the nucleus, with total α-tubulin distributed throughout the cytoplasm. ID5 stained the primary cilium at an incidence of 1 per cell, its colocalisation with C3B9 identifying the primary cilium as one of the most stable features of the microtubular cytoskeleton. Primary cilia varied from 1 to 4 μm in length, and 3 patterns of projection into the extracellular matrix were identified; (1) full extension and matrix contact, with minor undulations along the length; (2) partial extension and matrix contact, with a range of bending deflections; (3) cilium reclined against the cell surface with minimal matrix contact. Ultrastructural studies identified direct connections between extracellular collagen fibres and the proteins which decorate ciliary microtubules, suggesting a matrix–cilium–Golgi continuum in hyaline chondrocytes. These results strengthen the hypothesis that the primary cilium acts as a ‘cellular cybernetic probe' capable of transducing environmental information from the extracellular matrix, communicating this information to the centrosome, and regulating the exocytosis of Golgi-derived secretory vesicles.

A total of 112 human embryos (224 upper limbs) between stages 12 and 23 of development were examined. It was observed that formation of the arterial system in the upper limb takes place as a dual process. An initial capillary plexus appears from the dorsal aorta during stage 12 and develops at the same rate as the limb. At stage 13, the capillary plexus begins a maturation process involving the enlargement and differentiation of selected parts. This remodelling process starts in the aorta and continues in a proximal to distal sequence. By stage 15 the differentiation has reached the subclavian and axillary arteries, by stage 17 it has reached the brachial artery as far as the elbow, by stage 18 it has reached the forearm arteries except for the distal part of the radial, and finally by stage 21 the whole arterial pattern is present in its definitive morphology. This differentiation process parallels the development of the skeletal system chronologically. A number of arterial variations were observed, and classified as follows: superficial brachial (7.7%), accessory brachial (0.6%), brachioradial (14%), superficial brachioulnar (4.7%), superficial brachioulnoradial (0.7%), palmar pattern of the median (18.7%) and superficial brachiomedian (0.7%) arteries. They were observed in embryos belonging to stages 17–23 and were not related to a specific stage of development. Statistical comparison with the rates of variations reported in adults did not show significant differences. It is suggested that the variations arise through the persistence, enlargement and differentiation of parts of the initial network which would normally remain as capillaries or even regress.

Programmed cell death or apoptosis occurred in anuran amphibian larval pancreas as a remodelling agent, and was responsible for the reduction of the gland volume during metamorphosis. Apoptotic cells were recognisable by their morphological characteristics and could be immunocytochemically detected by means of the TUNEL reaction, which evidenced nuclear DNA fragmentation. During the last stages of prometamorphosis, that is in the period of hindlimb differentiation, only a few TUNEL positive cells occurred, whereas they increased at the beginning of metamorphic climax, that is at forelimb emergence and during tail regression. Under the electron microscope, the typical morphological characteristics of apoptosis were observed: decrease in size, and the presence of wide intercellular spaces and nuclei with dense chromatin masses arranged in crescents. The fragmentation of these cells produced the so-called ‘apoptotic bodies’: portions of cytoplasm lined by a membrane, containing nuclear fragments and cytoplasmic organites. Dead cell elimination is hypothesised to occur by phagocytic ingestion.

The purpose of this study was to document and compare the architectural parameters (fibre bundle length, angle of pennation) of human skeletal muscle in cadaveric specimens and live subjects. The medial (MG) and lateral (LG) gastrocnemius, and posterior (PS) and anterior (AS) soleus were examined bilaterally in 5 cadavers (mean age 72.6, range 65–83 y) and 9 live subjects (mean age 76.3, range 70–92 y). Data were obtained from direct measurement of cadaveric specimens and from ultrasonographic scans of the live subjects. In cadaveric muscle, fibre bundles were isolated; their length was measured in millimetres and pennation angles were recorded in degrees. In live muscle, similar measurements were taken from ultrasonographic scans of relaxed and contracted muscle. For the scans of relaxed muscle, subjects were positioned prone with the foot at a 90° angle to the leg, and for scans of contracted muscle, subjects were asked to sustain full plantarflexion during the scanning process. Fibre bundle length and angle of pennation were compared at matched locations in both groups. It was found that the relationship between cadaveric and in vivo values for fibre length and angle of pennation varied between muscle parts. The cadaveric architectural parameters did not tend to lie consistently towards either extreme of relaxation or contraction. Rather, within MG, PS and AS, cadaveric fibre bundle lengths lay between those for relaxed and contracted in vivo muscle. Similarly both the anterior and posterior cadaveric fibre angles of pennation lay between the in vivo values within LG and PS. In summary, architectural characteristics of cadaveric muscle differ from both relaxed and contracted in vivo muscle. Therefore, when developing models of skeletal muscle based on cadaveric studies, the architectural differences between live and cadaveric tissue should be taken into consideration.

This study was undertaken to investigate the morphological characteristics and lectin affinity of the testicular lamina propria in healthy boars and in unilateral and bilateral abdominal cryptorchid boars. The lamina propria of scrotal testes from healthy boars and unilateral cryptorchid boars was constituted by an innermost noncellular layer, the basal lamina, and by 2 layers of peritubular cells, each separated by a fibrous layer. The noncellular layers contained collagen fibres and glycoconjugates with abundant N-acetylgalactosamine, galactose, fucose, N-acetylglucosamine and neuraminic acid residues. The inner peritubular cell layer was composed of myoid cells, the outer layer of fibroblasts. In the abdominal testes of unilateral and bilateral cryptorchid boars, the lamina propria of nondegenerating and degenerating seminiferous tubules appeared thickened due to an increased content of collagen fibres and glycoconjugates. Glycoconjugates showed decreased amounts of fucose, neuraminic acid and galactose, and increased amounts of N-acetylglucosamine residues. The basal lamina formed infoldings toward the seminiferous epithelium and contained small cells. Both inner and outer peritubular cells were fibroblasts of immature appearance. In degenerated seminiferous tubules of bilateral cryptorchid boars, the lamina propria was composed of a thickened and collagenised basal lamina, without peritubular cells and with a low content of glycoconjugates. In scrotal testes, therefore, the lamina propria was implicated in tubular contractility and in mediating the communication and the substrate diffusion between seminiferous tubules and interstitial tissue. Cryptorchidism induced morphological and histochemical alterations in the lamina propria of abdominal testes, which may be linked to evidence from other studies of lack of tubular contractility and defective cell–cell communication and substrate diffusion. The severity of these anomalies correlated with the severity of Sertoli cell alterations.

Aponeurotic deformation measurements have traditionally been taken by loading dissected muscles; thus the values obtained may not reflect in vivo function. In the present study, we estimated dimensional changes in the central aponeurosis of the intact human tibialis anterior muscle upon loading induced by muscle contraction. Measurements were taken in seven males, and involved real-time ultrasound scanning of the tibialis anterior muscle at 30° of plantarflexion at rest and during isometric dorsiflexion maximum voluntary contraction (MVC). At each contraction state, the length of the aponeurosis, the width along its length, and its area were estimated from sagittal-plane and axial-plane sonographs. In the transition from rest to MVC, the length of the aponeurosis increased by 7% (P < 0.05), its width increased by up to 21% (P < 0.05), and its area increased by 17% (P < 0.05). These results indicate that the in vivo tibialis anterior aponeurosis behaves as a compliant material upon active muscle shortening. The methodology employed allows cross-sectional and longitudinal design investigations, circumventing the problems associated with epimysial removal under in vitro experimental conditions.

The iliolumbar ligament has been described as the most important ligament for restraining movement at the lumbosacral junction. In addition, it may play an important role in restraining movement in the sacroiliac joints. To help understand its presumed restraining effect, the anatomy of the ligament and its orientation with respect to the sacroiliac joints were studied in 17 cadavers. Specific dissection showed the existence of several distinct parts of the iliolumbar ligament, among which is a sacroiliac part. This sacroiliac part originates on the sacrum and blends with the interosseous sacroiliac ligaments. Together with the ventral part of the iliolumbar ligament it inserts on the medial part of the iliac crest, separate from the interosseous sacroiliac ligaments. Its existence is verified by magnetic resonance imaging and by cryosectioning of the pelvis in the coronal and transverse plane. Fibre direction, length, width, thickness and orientation of the sacroiliac part of the iliolumbar ligament are described. It is mainly oriented in the coronal plane, perpendicular to the sacroiliac joint. The existence of this sacroiliac part of the iliolumbar ligament supports the assumption that the iliolumbar ligament has a direct restraining effect on movement in the sacroiliac joints.

Expression of androgen receptor (AR), estrogen receptor alpha (ER-α) and estrogen receptor beta (ER-β) in the testis of the marbled newt (Triturus marmoratus marmoratus) was investigated, with special attention to changes during the annual testicular cycle, using light microscopy immunohistochemistry and Western blot analysis. Primordial germ cells, primary and secondary spermatogonia and spermatocytes showed a positive reaction to the 3 receptor antibodies during the annual reproductive cycle. Follicular cells were positive to AR, ER-α and ER-β during the spermiogenesis and quiescence periods in the glandular tissue. Interstitial cells showed reactivity to AR, ER-α and ER-β in the spermiogenesis and the quiescence periods, and presented no labelling to these receptors in the proliferative period. These findings suggest that, as in mammals, there is an androgen-estrogen regulation of the function and development of the newt testis.

This paper reports on the origin and distribution of the coronary arteries in normal mice and in mice of the iv/iv strain, which show situs inversus and heterotaxia. The coronary arteries were studied by direct observation of the aortic sinuses with the scanning electron microscope, and by examination of vascular corrosion casts. In the normal mouse, the left and right coronaries (LC, RC) arise from the respective Valsalva sinus and course along the ventricular borders to reach the heart apex. Along this course the coronary arteries give off small branches at perpendicular or acute angles to supply the ventricles. The ventricular septum is supplied by the septal artery, which arises as a main branch from the right coronary. Conus arteries arise from the main coronary trunks, from the septal artery and/or directly from the Valsalva sinus. The vascular casts demonstrate the presence of intercoronary anastomoses. The origin of the coronary arteries was found to be abnormal in 84% of the iv/iv mice. These anomalies included double origin, high take-off, slit-like openings and the presence of a single coronary orifice. These anomalies occurred singly or in any combination, and were independent of heart situs. The septal artery originated from RC in most cases of situs solitus but originated predominantly from LC in situs inversus hearts. Except for this anomalous origin no statistical correlation was found between the coronary anomalies and heart situs or a particular mode of heterotaxia. The coronary anomalies observed in the iv/iv mice are similar to those found in human hearts. Most coronary anomalies appear to be due to defective connections between the aortic root and the developing coronaries. iv/iv mice may therefore constitute a good model to study the development of similar anomalies in the human heart.

Cartilage fails by fibrillation and wearing away. This study was designed to identify the microscopic failure processes in the collagen network of bovine cartilage using scanning electron microscopy. Cartilage samples from fibrillated cartilage from the bovine patella were removed from the bone, fixed, digested to remove proteoglycans, freeze-fractured, and processed for SEM. The architecture of the collagen network in the normal cartilage was first defined, and then the failure processes were identified by examining sites of fibrillation and at crack tips. The bovine patellar cartilage was organised with a superficial layer composed of 3–5 lamina, attached to a sub-superficial tissue by angled bridging fibrils. Collagen in the sub-superficial tissue was organised in lamina oriented in the radial direction up to the transition zone. Failure of the system occurred by cracks forming in superficial layer and lamina, creating flaps of lamina that rolled up into the larger ‘fronds’. Larger cracks not following the laminar planes occurred in the transition, mid, and deep zones. Failure at the crack tips in the sub-superficial tissue appeared to be by peeling of collagen fibrils, as opposed to breaking of collagen fibrils, suggesting a ‘glue’ bonding the collagen fibrils in a parallel fashion. Cracks propagated by breaking these bonds. This bond could be a site of disease action, since weakening of the bond would accelerate crack propagation.

Mammalian Langerhans cells (LC) are epidermal dendritic cells which originate in bone marrow and migrate toward the T cell area of lymph nodes, where they act as professional antigen-presenting cells. A variety of cell surface markers, such as the ectoenzyme adenosine triphosphatase (ATPase), Ia and CD1a antigens, have been used extensively to identify LC. Ultrastructural identification of this cell type in the mammalian epidermis is made by the demonstration of a typical and unique cytoplasmic organelle, the Birbeck granule (BG). Although we had earlier demonstrated the coexpression of ATPase and Ia antigens on epidermal dendritic cells of the chicken epidermis, the presence of the BG has not previously been documented. The aim of the present study was to investigate whether chicken epidermal LC-like cells possess an organelle similar to the BG, and thus to complete their identification. Our findings are the first demonstration of characteristic rod-shaped, racket-shaped and disc-shaped intracytoplasmic organelles, morphologically similar to the mammalian BG, in avian LC.

According to the usual description in most anatomy texts, the median nerve in the forearm passes between the 2 heads of pronator teres. It continues distally between flexor digitorum superficialis and profundus almost to the retinaculum. Muscular branches leave the nerve near the elbow and supply all superficial muscles of the anterior part of the forearm except flexor carpi ulnaris. Many variations of the median nerve in the forearm have been reported (Urban & Krosman, 1992). The palmaris profundus is also a rare anomaly of the forearm (Dyreby & Engber, 1982). It originates from the radial side of the common flexor tendon in the proximal forearm and inserts into the undersurface of the palmar aponeurosis. The origin of palmaris profundus may be close to the median nerve and its branches, and may be involved in compressive neuropathy of the anterior interosseous nerve. Its tendon crossing through the carpal canal has been implicated in the carpal tunnel syndrome (reviewed by Lahey & Aulicino, 1986). In some cases, palmaris profundus was found enclosed in a common fascial sheath with the median nerve (Stark, 1992; Sahinoglu et al. 1994). To indicate its close association with the median nerve, the palmaris profundus was also named ‘musculus comitans nervi mediani’ (Sahinoglu et al. 1994). This article reports an unusual loop of the median nerve encircling an anomalous palmaris profundus in the forearm, which, to the best of our knowledge, has not been previously described.