As from 1997, the Journal of Anatomy will be published in 8 issues per year instead of the current 6. The purpose is to reduce the time between acceptance and publication. Otherwise there will be no change in editorial policy. The Journal will continue to publish high quality papers on human and comparative anatomy and cell biology, both experimental and descriptive. Articles on experimental pathology are also acceptable if they contribute to the understanding of normal structure and function. Review articles, if topical and authoritative, are welcome. Short reports are published: these should not exceed 5 pages of printed text. Reports on isolated anatomical variations, if of sufficient interest and relevance, are now included in the Correspondence section, to which letters on matters arising out of previously published papers or on questions of general anatomical interest can be submitted.

Several studies have reported the morphological changes induced by unilateral enucleation during early neonatal life on the developing visual system. This study has examined cellular changes in the superior colliculi by removal of a single eye in adult rats. Anaesthetised male hooded rats aged 90 d had their right eyes removed. Groups of nonenucleated control and enucleated rats were killed when aged either 150 or 390 d. The brains were removed and both the right and left superior colliculi dissected out. The volume of the stratum griseum superficiale (SGS) within these colliculi was estimated stereologically by light microscopy, as well as the numerical density and total number of neurons within this cell layer. The volume of the cell layer was reduced by about 40% on the side contralateral to the enucleated eye but not on the ipsilateral side at both survival periods examined. The numerical density of neurons within the SGS was unaffected by the enucleation so that the colliculi contralateral to the enucleated eye showed a substantial loss of neurons within this cell layer. This study demonstrates the importance of the retinal ganglion cell input, even in adult animals, for maintaining the viability of neurons in the SGS layer of the superior colliculus.

Oligodendrocytes, derived from stem cell precursors which arise in subventricular zones of the developing central nervous system, have as their specialist role the synthesis and maintenance of myelin. Astrocytes contribute to the cellular architecture of the central nervous system and act as a source of growth factors and cytokines; microglia are bone-marrow derived macrophages which function as primary immunocompetent cells in the central nervous system. Myelination depends on the establishment of stable relationships between each differentiated oligodendrocyte and short segments of several neighbouring axons. There is growing evidence, especially from studies of glial cell implantation, that oligodendrocyte precursors persist in the adult nervous system and provide a limited capacity for the restoration of structure and function in myelinated pathways damaged by injury or disease.

The thermogenic potential of the interscapular brown fat pad in the naked mole-rat Heterocephalus glaber, that exhibits poikilothermic thermal responses to changing temperatures is reported. Histological and ultrastructural study of the brown fat pad showed that it consists of layers of skeletal muscle interposed between the layers of brown adipose tissue with both unilocular and multilocular adipocytes. Large numbers of mitochondria were present between and around the lipid droplets of these cells. Glyoxylic acid condensation, used to demonstrate catecholaminergic nerves, was evident in low concentrations in the connective tissue between the brown adipocytes. A 3-dimensional computer-aided reconstruction of the fat pad showed the extent and ramification of nerves and blood vessels between the adipocytes. These findings show that although the naked mole-rat is regarded as an endothermic poikilotherm, it possesses anatomical features usually found in homeothermic mammals, which are essential for thermogenesis.

The development of the patella, its associated tendons, and suprapatella of the rabbit knee joint is described from the 17 d fetus to the mature adult. The patellar tendon (ligament) with the patella on its posterior surface is seen in the 17 d fetus and is fully developed by 1 postnatal wk. It is composed of bundles of types I and V collagens separated by endotenons of types III and V collagens. Anteriorly there is an epitenon of types III and V collagens while synovium and a fat pad cover its posterior surface. In the 25 d fetus, the patella is cartilaginous and is separated from the femoral condyles. The cartilage contains type II collagen, but types I, III and V collagens are found along the articular surface. Ossification starts 1 postnatal wk and at 6 wk only the articular cartilage remains. In addition to type II, types III and V collagens are located around the chondrocyte lacunae. The long anterior junction between the patella and its tendon is fibrocartilaginous at 1 wk, but as ossification proceeds this is replaced by bone. Types I and V collagens are found in this region. The suprapatella on the posterior surface of the quadriceps tendon is first seen 1 wk postnatally as an area of irregularly organised fibres and chondrocyte-like cells. Types I, II, III and V collagens are present from 3 wk onwards. It is compared with the fibrocartilage of other tendons that are under compression. The arrangement of the collagens in the patellar tendon is discussed in relation to its use as a replacement for injured anterior cruciate ligaments. It is suggested that the structural differences between the patellar tendon and anterior cruciate ligament preclude the translocated tendon acquiring mechanical strength similar to that of a normal cruciate ligament. The designation ‘patellar ligament’ as opposed to ‘patellar tendon’ is questioned. It is argued that the term patellar tendon reflects its structure more accurately than patellar ligament.

Epidermal growth factor (EGF) has been implicated in the control of embryonic development, but although the receptor is expressed from an early stage, there is little evidence of embryonic expression of EGF. In order to investigate the role of maternally derived EGF during organogenesis, rat embryos were explanted at d 9.5 and cultured in serum depleted of low molecular weight molecules (retenate) which was then supplemented with EGF. Serum depleted of low molecular weight molecules by prolonged filtration loses its capacity to support normal embryonic development, possibly due to the loss of growth promoting factors. The addition of EGF to retenate significantly improved embryonic development with a maximal effect at 8 ng/ml. The addition of an analogue of EGF, long EGF, to retenate also caused a significant increase in development, although at higher concentrations a decrease in its effect was observed, possibly due to down regulation of the EGF receptor. Therefore, embryos may be able to utilise maternally derived EGF during organogenesis. To test the effects of inhibiting the EGF receptor during organogenesis, d 9.5 embryos were cultured in the presence of tyrphostin 47, a specific EGF receptor inhibitor. Tyrphostin 47 caused a significant dose-dependent decrease in the development of embryos which was also observed when tyrphostin 47 was injected into the vitelline circulation at d 11.5 to bypass the effects of the yolk sac. These findings suggest that the EGF receptor is essential for normal organogenesis and may play a role in the control of proliferation and differentiation. Although EGF is not expressed in the rat embryo at this stage, maternally derived EGF may be the ligand for the embryonic EGF receptor.

Skull morphology and histology in the heterozygous offspring of a transgenic founder mouse Del1, harbouring 6 copies of deletion mutation in Col2a1 gene, were compared with those in normal siblings. On visual observation and roentgenocephalometric examination the heads of heterozygous Del1 mice were smaller than normal. Histologically the sizes of cartilaginous structures of the cranial base were reduced. Severe defects were seen in the temporomandibular joint as progressive osteoarthritic lesions. These observations elucidate the relationship between the genotype and phenotype and demonstrate that heterozygous Del1 mice are a useful model for studies on a genetic disturbance where ‘clinical’ manifestations are not evident until adult age.

This Anatomical Society symposium, held at University College, Cork in September 1995 was the first of its kind. The objective was to stimulate and facilitate constructive interaction between experts on topics relating to the various types of glial barrier which form partitions within the nervous system. Some of these barriers are transient and are found only during development, for example, those which define the limits of developing nuclei or fibre bundles. Others are permanent, such as those at the transitional zones which separate the CNS and PNS milieux at nerve root attachments to the neuraxis. Still others, such as glial scars, are seen only following injury when they tend to inhibit neurite regeneration. In experimental circumstances, for example following irradiation or chemical damage, glial barriers may be broken down and even relocated. This may be associated with Schwann cell invasion of the CNS and, experimentally, with Schwann cell and glial cell transplantation into demyelinated areas. Such transplantation studies are in turn related to remyelination and CNS axon regeneration and the factors which facilitate these. Twelve review lectures were given on these topics. Five articles based on these communications are reproduced here. The underlying theme was the relationship between advances in the understanding of fundamental nervous tissue biology, especially as related to glial cells, and potential developments aimed at treating CNS demyelinating diseases and achieving CNS regeneration.

This study sought to investigate the normal muscle fibre size and type distribution of the human erector spinae, both in thoracic and lumbar regions, in a group of 31 young healthy male (n=17) and female (n=14) volunteers. Two percutaneous muscle biopsy samples were obtained under local anaesthesia, from the belly of the left erector spinae, at the levels of the 10th thoracic and 3rd lumbar vertebrae. Samples were prepared for routine histochemistry for the identification of fibre types. Fibre size (cross-sectional area (CSA) and narrow diameter (ND)) was quantified using computerised image analysis. The mean CSA/ND for each fibre type was greater in the thoracic than the lumbar region, but there was no difference between the 2 regions either for percentage type I (i.e. percentage distribution by number), percentage type I area (i.e. relative area of the muscle occupied by type I fibres) or the ratio describing the size of the type I fibre relative to that of the type II. Men had larger fibres than women, for each fibre type and at both sampling sites. In the men, each fibre type was of a similar mean size, whereas in the women the type I fibres were considerably larger than both the type II A and type II B fibres, with no difference between the latter two. In both regions of the erector spinae there was no difference between men and women for the proportion (%) of a given fibre type, but the percentage type I fibre area was significantly higher in the women.

The erector spinae display muscle fibre characteristics which are clearly very different from those of other skeletal muscles, and which, with their predominance of relatively large type I (slow twitch) fibres, befit their function as postural muscles. Differences between thoracic and lumbar fascicles of the muscle, and between the muscles of men and women, may reflect adaptive responses to differences in function. In assessing the degree of any pathological change in the muscle of patients with low back pain, it seems clear that (1) sex cannot be disregarded and (2) ‘atrophied’ (using the criteria from other muscles) type II fibres are not necessarily abnormal for the erector spinae, particularly in women.

An antiserum against secretory vesicles from human seminal fluid (prostasomes) was used to study the localisation and distribution of the respective antigen(s) during prenatal development and pubertal maturation of the human prostate. The crude antiserum stained both secretory and membrane proteins in the adult prostate and other glands, such as pancreas and parotid gland. An immunoaffinity purified fraction from the antiserum selectively reacted with the apical plasma membrane of prostatic epithelium adluminal cells, recognizing a 100 kDa antigen (PMS). Even in the earliest stages of embryonic prostate specimens studied, the adluminal plasma membrane of the epithelial cells from developing glandular anlagen reacted strongly. The occurrence of PMS immunoreactivity in prostatic anlagen was directly correlated with lumen formation. As the antigen is an androgen-independently synthesised membrane protein of the prostate, it may possibly be used as a marker of cell polarity in the normal and pathologically altered prostate.

This review summarises some of the major findings derived from studies using the model of a glia-depleted environment developed and characterised in this laboratory. Glial depletion is achieved by exposure of the immature rodent spinal cord to x-radiation which markedly reduces both astrocyte and oligodendrocyte populations and severely impairs myelination. This glia-depleted, hypomyelinated state presents a unique opportunity to examine aspects of spinal cord maturation in the absence of a normal glial population. An associated sequela within 2–3 wk following irradiation is the appearance of Schwann cells in the dorsal portion of the spinal cord. Characteristics of these intraspinal Schwann cells, their patterns of myelination or ensheathment, and their interrelations with the few remaining central glia have been examined. A later sequela is the development of Schwann cells in the ventral aspect of the spinal cord where they occur predominantly in the grey matter. Characteristics of these ventrally situated intraspinal Schwann cells are compared with those of Schwann cells located dorsally. Recently, injury responses have been defined in the glia-depleted spinal cord subsequent to the lesioning of dorsal spinal nerve roots. In otherwise normal animals, dorsal nerve root injury induces an astrocytic reaction within the spinal segments with which the root(s) is/are associated. Lesioning of the 4th lumbar dorsal root on the right side in irradiated or nonirradiated animals results in markedly different glial responses with little astrocytic scarring in the irradiated animals. Tracing studies reveal that these lesioned dorsal root axons regrow rather robustly into the spinal cord in irradiated but not in nonirradiated animals. To examine role(s) of glial cells in preventing this axonal regrowth, glial cells are now being added back to this glia-depleted environment through transplantation of cultured glia into the irradiated area. Transplanted astrocytes establish barrier-like arrangements within the irradiated cords and prevent axonal regrowth into the cord. Studies using other types of glial cultures (oligodendrocyte or mixed) are ongoing.

The projections of different subpopulations of myenteric neurons in the mouse small and large intestine were examined by combining immunohistological techniques with myotomy and myectomy operations. The myotomies were used to examine the polarity of neurons projecting within the myenteric plexus and showed that neurons containing immunoreactivity for nitric oxide synthase (NOS), vasoactive intestinal peptide (VIP), calbindin and 5-HT projected anally, while neurons with substance P (SP)-immunoreactivity projected orally, in both the small and large intestine. Neurons containing neuropeptide Y (NPY)- and calretinin-immunoreactivity projected locally. In the large intestine, GABA-immunoreactive neurons projected both orally and anally, with more axons tending to project anally. Myectomy operations revealed that circular muscle motor neurons containing NOS/VIP/±NPY and calretinin neurons projected anally both in the small and large intestine, while SP-immunoreactive circular muscle motor neurons projected orally. In the large intestine, GABA-IR circular muscle motor neurons projected both orally and anally. This study showed that although some neurons, such as the NOS/VP inhibitory motor neurons and interneurons, SP excitatory motor neurons and 5-HT interneurons had similar projections to those in other species, the projections of other chemical classes of neurons in the mouse intestine differed from those reported in other species.

MyoD1 alleles distinguish between the Mm musculus and Mm domesticus subspecies of Mus musculus. SJL/J mice, which belong to the Mm musculus subspecies, are able to regenerate skeletal muscle more efficiently than BALB/c mice which possess the Mm domesticus MyoD1 allele. Hence the possibility that regeneration responses are due to species-specific genetic differences including MyoD1 was investigated in this study. Quantitative morphometric analysis following muscle crush injury of 2 Mm musculus strains, MBK and MGL, has indicated that regeneration phenotype is not species-specific and may not be directly related to MyoD1 alleles. These results contrast with prior investigations conducted in SJL/J and BALB/c mice, and indicate that other genes may have a direct influence on skeletal muscle regeneration.

A number of reports suggest that the process of ageing impairs wound repair and that strategies to manipulate the age-related wound healing environment are necessary in order to stimulate repair. The process of cutaneous wound repair is controlled by growth factors in an autocrine and paracrine fashion: it is therefore surprising that the localisation of specific growth factors and their receptors has not been documented in wound healing with respect to chronological age. In this study the temporal profile of growth factor and receptor immunostaining was assessed within acute incisional wounds in an ageing mouse colony. A delay in appearance of platelet derived growth factor (PDGF) A and B isoforms, and PDGF-α and -β receptors was evident with increasing animal age, paralleled by a similar finding for epidermal growth factor (EGF) and EGF receptor. Transforming growth factor (TGF)-7beta;1 and 2 isoforms were increased at all time points in the wounds of younger animals, but the TGF-β3 isoform increased in intensity from d 7 postwounding in the old mice wounds, and basic fibroblast growth factor (bFGF) from d 14. The quantity and distribution patterns of the various growth factors and their receptors may explain the age-related differences in wound healing speed and quality, and possibly suggest new therapeutic targets for manipulating wound healing in the aged.

Retrograde tracing with Fluoro-Gold was used to identify the complete population of knee joint afferents in the lumbar dorsal root ganglia of adult female Wistar rats. There was an average of 581±31 (mean±S.D.) afferents supplying each joint. These were found distributed from L1 to L5 with the great majority localised in the L3 and L4 ganglia. Electron microscopy of the posterior articular nerve of the knee revealed an average of 103±15 (mean±S.D.) myelinated and 513±39 unmyelinated axonal profiles. Since about 50–60% of the unmyelinated profiles would be expected to be sympathetic efferents, these numbers are consistent with the numbers of afferents found by Fluoro-Gold retrograde tracing and suggest that the posterior articular nerve contains about 50% of the total number of knee joint afferents in the rat. Immunohistochemistry revealed that an average of 10% of identified joint afferents expressed substance P-like immunoreactivity and that 33% expressed calcitonin gene-related peptide-like immunoreactivity.

This review covers a number of aspects of the behaviour of oligodendrocyte progenitors following transplantation into the adult CNS. First, an account is given of the ability of transplanted oligodendrocyte progenitors, grown in tissue culture in the presence of PDGF and bFGF, to extensively remyelinate focal areas of persistent demyelination. Secondly, we describe how transplanted clonal cell lines of oligodendrocyte progenitors will differentiate into astrocytes as well oligodendrocytes following transplantation into pathological environments in which both oligodendrocytes and astrocytes are absent, thereby manifesting the bipotentially demonstrable in vitro but not during development. Finally, a series of studies examining the migratory behaviour of transplanted oligodendrocyte progenitors (modelled using the oliodendrocyte progenitor cell line CG4) are described. These show that CG4 cells do not survive (or migrate) when transplanted into the normal adult CNS. However, if they are transplanted into CNS tissue that has previously been exposed to 40 Gy of x-irradiation then transplanted CG4 cells survive, divide and migrate over large distances. Moreover, within an x-irradiated environment, migrating transplanted CG4 cells are able to enter remotely located foci of demyelination and contribute to the remyelination of the demyelinated axons within. These studies demonstrate that although the normal adult CNS does not appear to support survival and migration of the CG4 cell line, it is possible to manipulate the environment in such a way that these nonpermissive properties of the environment can be overcome.

Using antiserum to human parathyroid hormone-related protein (1–16) [PTHrP(1–16)] we have examined tissues of the common frog (Rana temporaria) for the presence of immunoreactive PTHrP (irPTHrP) at the stage of emergence from water to land. irPTHrP was detected in dorsal and ventral stratum granulosum of the skin, in the developing ovary, striated muscle and the choroid plexus epithelium of the brain as well as in the olfactory gland epithelium and olfactory lobe neurons of the brain. In the pituitary and hypothalamus irPTHrP protein could be demonstrated in the median eminence, infundibular stem and principally in the neural lobe and pars distalis of the pituitary with weak reaction in the pars intermedia. In situ hybridisation of the same tissues with an oligonucleotide probe to chicken PTHrP 55–65 clearly showed the presence of mRNA for PTHrP-like molecule in all the tissues containing irPTHrP. There was a major inconsistency in the pituitary in that the highest level of gene expression, assessed by in situ hybridisation, was found in the pars intermedia with only very low expression in the pars distalis and neural lobe and undetectable levels in the infundibular stem and median eminence. These observations suggest that tissues of the frog synthesise a PTHrP-like molecule but that in the pituitary the pars intermedia cells may export the protein to cells in other regions of the pituitary and hypothalamus.

Cell–matrix and matrix–matrix interactions are of critical importance in regulating the development, maintenance and repair of articular cartilage. In this study, we examined the structural colocalisation of type VI collagen and fibronectin in isolated chondrons and long-term agarose cultured chondrocytes extracted from normal adult canine articular cartilage. Using double labelling immunohistochemistry in conjunction with dual channel confocal microscopy and digital image processing we demonstrate that type VI collagen and fibronectin are distributed in a similar staining pattern and are colocalised at the surface of cultured chondrocytes and isolated chondrons. The results suggest that type VI collagen and fibronectin may play a role in both cell–matrix adhesion and matrix–matrix cohesion in the pericellular microenvironment surrounding articular cartilage chondrocytes.

An understanding of migraine must be based on data concerning the anatomy and physiology of the pain-sensitive intracranial structures. Stimulation of the superior sagittal sinus produces changes in brain blood flow and changes in neuropeptide levels similar to those seen in humans during migraine. To better understand the anatomy of the central ramifications of pain-sensitive intracranial structures we have examined the distribution of c-fos immunoreactivity in the monkey when the sinus is stimulated. Six adult Macaca nemestrina monkeys were anaesthetised. The superior sagittal sinus was isolated after a midline craniotomy and a paraffin well created. At 24 h after completion of the surgery the sinus was stimulated electrically for 1 h and the brain subsequently removed and processed for c-fos. In control animals in which the sinus was isolated but not stimulated there was a small amount of c-fos expression in the caudal brainstem and upper cervical spinal cord. Stimulation of the superior sagittal sinus evoked expression of c-fos in the caudal superfical laminae of the trigeminal nucleus and in superficial laminae of the dorsal horn of the C1 level of the upper cervical spinal cord. A lesser amount of c-fos was seen at C2 while no significant labelling above control was observed at C3. These data, while largely confirming the results from the cat concerning the central distribution trigeminovascular afferents, underscore a possibly unique specialisation of trigeminovascular afferents at the C1 level. Given the close evolutionary relationship of the monkey to man it is likely that the cells described in this study represent for primates the nucleus that mediates the pain of migraine.

Schwann cells are excluded from the CNS during development by the glial limiting membrane, an area of astrocytic specialisation present at the nerve root transitional zone, and at blood vessels in the neuropil. This barrier, however, can be disrupted and, with the highly migratory nature of Schwann cells, can result in their invasion and myelination of the CNS in many pathological situations. In this paper we demonstrate that this occurs in a number of myelin mutants, including the myelin deficient (md) and taiep rats and the canine shaking (sh) pup. While it is still relatively uncommon in the rodent mutants, the sh pup shows extensive Schwann cell invasion along the neuraxis. This invasion involves the spinal cord, brain stem, and cerebellum and increases in amount and distribution with age. In situ hybridisation studies using a P0 riboprobe suggest that the likely origin of these cells in the sh pup is the nerve roots, primarily the dorsal roots. Paradoxically, Schwann cell myelination of the CNS increases with time in the sh pup despite a marked, progressive gliosis involving the glia limitans and neuropil. Thus the mechanism by which these cells migrate into the CNS through the gliosed nerve root transitional zone or from vasa nervorum remains unknown. Extensive Schwann cell CNS myelination may have therapeutic significance in human myelin disease.