Peripheral changes above and below injury level lead to prolonged vascular responses following high spinal cord injury.

Autonomic dysreflexia (AD) is a debilitating disorder producing episodes of extreme hypertension in patients with high-level spinal cord injury (SCI). Factors leading to AD include loss of vasomotor baroreflex control to regions below injury level, changes in spinal circuitry, and peripheral changes. The present study tested for peripheral changes below and above injury level 6 wk after a transection at the fourth thoracic spinal level. Changes in vascular conductance were recorded in the femoral, renal, brachial, and carotid arteries in response to intravenous injections of two alpha-adrenergic agonists, phenylephrine (PE; 0.03-100 microg/kg) and methoxamine (Meth; 1-300 microg/kg). Unlike PE, Meth is not subject to neuronal reuptake. Ganglionic blockade (0.6 mg/kg chlorisondamine) was used to eliminate the central component of the cardiovascular response. After ganglionic blockade, SCI animals exhibited prolonged vasoconstriction in response to PE in all blood vessels measured compared with those in intact animals (all, P < 0.035). However, the PE dose-response curves obtained after ganglionic blockade revealed no significant difference in the potency between the two groups (all, P > 0.06), indicating that the prolonged vasoconstriction was not due to supersensitivity to PE. In contrast to PE, vascular responses to Meth did not vary between intact and SCI groups (all P > 0.108). These results show the development of a widespread peripheral change producing prolonged vasoconstriction in response to PE, but not Meth, possibly due to reduced neuronal reuptake of PE after SCI. This is the first study to report such a change in blood vessels not only below but also above injury level. Interventions to correct this reduced reuptake may help limit the development of AD.

Mild neuritic changes are increased in the brains of fatally injured older motor vehicle drivers.

Given the expected increase in the older population and driving in this age group, concerns have been raised about the safety of older drivers. People over 65 years are over-represented in motor vehicle fatalities when calculated by distance driven. They are also at risk of neurodegenerative diseases, such as Alzheimer's disease, that affect cognitive function. We have examined the brains of older drivers (15M:12F) who died as a result of a motor vehicle accident (MVA) to determine the extent of Alzheimer's disease-related neurofibrillary changes (neuritic plaques and neurofibrillary tangles), Lewy body pathology and cerebrovascular disease and compared them to a control group of older licenced drivers (23M:5F) who died of other causes. The prevalence of moderate or severe neuritic plaque pathology was less than expected for the general population of this age and there was no difference between the groups. However, mild neuritic plaque pathology was increased for MVA deaths compared to controls. There was no evidence of vascular dementia or dementia with Lewy bodies. The current mandatory age-related re-licencing procedures in NSW may contribute to the low percentage of drivers with severe pathology. Further research into the role of mild pathology in cognitive impairment and older drivers is warranted.

Cardiovascular and temperature changes in spinal cord injured rats at rest and during autonomic dysreflexia.

In patients with high spinal cord injuries autonomic dysfunction can be dangerous, leading to medical complications such as postural hypotension, autonomic dysreflexia and temperature disturbance. While animal models have been developed to study autonomic dysreflexia, associated temperature changes have not been documented. Our aim here was to use radiotelemetry and infrared thermography in rodents to record the development of cardiovascular and skin temperature changes following complete T4 transection. In adult male Wistar rats (n=5), responses were assessed prior to spinal cord injury (intact) and for 6 weeks following injury. Statistical analysis by a repeated-measure ANOVA revealed that following spinal cord injury (SCI), rats exhibited decreased mean arterial pressure (MAP, average decrease of 26 mmHg; P<0.035) and elevated heart rate (HR, average increase of 65 bpm, P<0.035) at rest. The basal core body temperature following SCI was also significantly lower than intact levels (-0.9 degrees C; P<0.0035). Associated with this decreased basal core temperature following SCI was an increased skin temperature of the mid-tail and hindpaw (+5.6 and +4.0 degrees C, respectively; P<0.0003) consistent with decreased cutaneous vasoconstrictor tone. Autonomic dysreflexia, in response to a 1 min colorectal distension (25 mmHg), was fully developed by 4 weeks after spinal cord transection, producing increases in MAP greater than 25 mmHg (P<0.0003). In contrast to the tachycardia seen in intact animals in response to colorectal distension, SCI animals exhibited bradycardia (P<0.0023). During episodes of autonomic dysreflexia mid-tail surface temperature decreased (approximately -1.7 degrees C, P<0.012), consistent with cutaneous vasoconstriction. This is the first study to compare cardiovascular dysfunction with temperature changes following spinal cord transection in rats.

Increased neurofibrillary tangles in the brains of older pedestrians killed in traffic accidents.

BACKGROUND/AIMS: Older people are over-represented in pedestrian fatalities, and it has been suggested that the presence of cognitive impairment or dementia in these individuals may contribute to their accidents. Using neuropathological methods, we aimed to compare the prevalence of dementia pathology in fatally injured older pedestrians with similarly aged ambulatory subjects who died from other causes. METHODS: The brains of 52 pedestrians (65-93 years) and 52 controls (65-92 years) were assessed for neurofibrillary tangles (NFT), neuritic plaques, Lewy bodies and vascular lesions using established neuropathological criteria. RESULTS: The examination for Alzheimer's disease (AD) pathology showed that 43% of the pedestrians had NFT scores of III-VI using Braak and Braak staging, compared with 23% of the controls (p < 0.05, Fisher's exact test), indicating incipient, possible or probable AD. There were no differences in the prevalence of pathology for vascular dementia or dementia with Lewy bodies. CONCLUSION: These results suggest that cognitive decline associated with AD, even in the earliest stages of the disease, may be a factor in fatal traffic accidents for older pedestrians. Special measures for pedestrian safety are necessary in areas with high densities of older citizens and especially for those diagnosed as having a mild cognitive impairment or AD.

Developmental onset of functional activity in the wallaby whisker cortex in response to stimulation of the infraorbital nerve.

This study used the extrauterine development of a marsupial wallaby to investigate the onset of functional activity in the somatosensory pathway from the whiskers. In vivo recordings were made from the somatosensory cortex from postnatal day (P) 55 to P138, in response to electrical stimulation of the infraorbital nerve supplying the mystacial whiskers. Current source density analysis was used to localize the responses within the cortical depth. This was correlated with development of cortical lamination and the onset of whisker-related patches, as revealed by cytochrome oxidase. The earliest evoked activity occurred at P61, when layers 5 and 6 are present, but layer 4 has not yet developed. This activity showed no polarity reversal with depth, suggesting activity in thalamocortical afferents. By P72 synaptic responses were detected in developing layer 4 and cytochrome oxidase showed the first hint of segregation into whisker-related patches. These patches were clear by P86. The evoked response at this age showed synaptic activity first in layer 4 and then in deep layer 5/upper layer 6. With maturity, responses became longer lasting with a complex sequence of synaptic activity at different cortical depths. The onset of functional activity is coincident with development of layer 4 and the onset of whisker-related pattern formation. A similar coincidence is seen in the rat, despite the markedly different chronological timetable, suggesting similar developmental mechanisms may operate in both species.

Cholinergic depletion by IgG192-saporin retards development of rat barrel cortex.

This study examines the role of cholinergic projections from the basal forebrain on development of the rodent barrel cortex. Pups were administered the immunotoxin IgG192-saporin (0.1 microg) intraventricularly at postnatal day (P) 0 and sacrificed at P1-P7. One ventricle was injected with saporin while the other side received saline, allowing comparison between the two sides of the same animal, as well as with controls receiving saline only. Compared to control animals, neuronal loss in the basal forebrain was present on both sides of saporin-treated pups but was significantly greater on the toxin-treated side, in all age groups and regions sampled. Depletion of acetylcholine did not prevent the formation of the barrel pattern, however it delayed its emergence by approximately 1 day. At P4, the thickness of layer IV barrel cortex was also significantly reduced; this reduction was undetectable by P7. From P3 to P5, the ratios of intensity of staining for acetylcholinesterase between the barrel centres and septa on the toxin-treated side were significantly lower than those on the saline side, although normal densities were present by P7. Thus, the depletion of cholinergic innervation at birth causes a transient delay in the development of the barrel pattern during the first postnatal week.

Extent and distribution of vascular brain injury in pediatric road fatalities.

This study used a multidisciplinary approach to examine the brains of pediatric road trauma fatalities in the Sydney area over a 3-year period. The brains of 32 children (0-16 years) were examined: 20 pedestrians, nine passengers, and three cyclists. The extent and distribution of brain injury was assessed, peak linear head acceleration determined, and the severity of brain damage was compared to that previously reported for adults using the same scoring method. Skull fractures (20/32) and subarachnoid haemorrhage (22/32) were the commonest head injuries. In general, the neuropathology was similar to that seen in adults, with a high percentage of damage in the corpus callosum and gliding contusions within the subcortical white matter. Intracerebral hemorrhage was relatively rare. For frontal and occipital head impacts, the corpus callosum was the most injured part of the brain, followed by the deep central structures and the temporal lobes, whereas for lateral impacts, the injuries were more evenly distributed. Comparison of the current data for children with the vascular injury sector scores reported for adults suggests that the brains of children are more severely damaged for the same peak linear head acceleration.

Transplantation of nasal olfactory tissue promotes partial recovery in paraplegic adult rats.

Recent reports have highlighted the potential therapeutic role of olfactory ensheathing cells for repair of spinal cord injuries. Previously ensheathing cells collected from the olfactory bulbs within the skull were used. In humans a source of these cells for autologous therapy lies in the nasal mucosa where they accompany the axons of the olfactory neurons. The aim of the present study was to test the therapeutic potential of nasal olfactory ensheathing cells for spinal cord repair. Olfactory ensheathing cells cultured from the olfactory lamina propria or pieces of lamina propria from the olfactory mucosa were transplanted into the transected spinal cord. Three to ten weeks later these animals partially recovered movement of their hind limbs and joints which was abolished by a second spinal cord transection. Control rats, receiving collagen matrix, respiratory lamina propria or culture medium, did not recover hind limb movement. Recovery of movement was associated with recovery of spinal reflex circuitry, assessed using the rate-sensitive depression of the H-reflex from an interosseous muscle. Histological analysis of spinal cords grafted with olfactory tissue demonstrated nerve fibres passing through the transection site, serotonin-positive fibres in the spinal cord distal to the transection site, and retrograde labelling of brainstem raphe and gigantocellularis neurons from injections into the distal cord, indicating regeneration of descending pathways. Thus, olfactory lamina propria transplantation promoted partial restoration of function after relatively short recovery periods. This study is particularly significance because it suggests an accessible source of tissue for autologous grafting in human paraplegia.

Afferent ingrowth and onset of activity in the rat trigeminal nucleus.

A novel in vitro preparation, consisting of the rat brainstem with the trigeminal ganglion attached, has been used to study the anatomical and functional development of the trigeminal nucleus from embryonic day (E)13 to postnatal day (P)6. Neurobiotin injections into the trigeminal ganglion showed that primary afferents had reached the trigeminal tract by E13 and had grown simple, mainly unbranched, collaterals into all levels of the nucleus by E15. By E17, these collaterals were extensively branched, with occasional boutons present. Patches of intense neurobiotin-labelled terminals, corresponding to whisker-related patterns, were first seen at E20 and became clearer over the next few days. Terminal arbours at this stage were relatively localized and densely branched, with many boutons. Responses from the trigeminal nucleus were recorded with suction electrodes, following stimulation of the trigeminal ganglion. Recordings from the main sensory nucleus showed a postsynaptic response was first present at E15. At E16, bath application of AP5 and DNQX showed that the response contained both NMDA and AMPA components, with NMDA predominating (75%). The NMDA : AMPA ratio remained high until P1, then gradually declined to 50% by P6. The postsynaptic response was also reduced by bath application of bicuculline, indicating the presence of a GABAA-mediated excitatory component. GABAergic excitation was present at all ages but was maximal from E20 to P1, the age at which whisker-related patterns are developing. It is hypothesized that both GABAergic excitation and NMDA receptor activation play a role in the consolidation of trigeminal connections, and are thus important in the development of whisker-related patterns.

Fatal head injury in children: a new approach to scoring axonal and vascular damage.

As part of a multidisciplinary study of brain damage in children fatally injured in motor vehicle accidents, a simple method to quantify and visualise the distribution and extent of injury has been developed. Vascular and axonal injury were assessed using coronal brain sections stained for haematoxylin and eosin, or reacted immunohistochemically for beta-amyloid precursor protein. Subsequent analysis was carried out using NIH Image software, and the resulting information is displayed in schematic diagrams. These summary diagrams simply and clearly show the distribution of injury in both the coronal and horizontal planes. This technique offers an advantage over previous scoring methods in that it provides both a quantitative and a visual summary of the distribution and extent of brain injury. This information can then be used to compare the injury distribution and severity with estimated impact points and acceleration data.

Serotonergic neurons in the brainstem of the wallaby, Macropus eugenii.

The organisation and cytoarchitecture of the serotonergic neurons in a diprotodont marsupial were examined by using serial sections of the brainstem processed for serotonin immunohistochemistry and routine histology. The topographic distribution of serotonergic neurons in the brainstem of the adult wallaby (Macropus eugenii) was similar to that of eutherian mammals. Serotonergic neurons were divided into rostral and caudal groups, separated by an oblique boundary through the pontomedullary junction. Approximately 52% of the serotonergic neurons in the wallaby brainstem were located in the rostral midline nuclei (caudal linear nucleus, dorsal, median, and pontine raphe nuclei and the interpeduncular nucleus), whereas 21% were found in the caudal midline region (nuclei raphe magnus, obscurus, and pallidus). The remaining serotonergic neurons (27%) were located in more lateral regions such as the pedunculopontine tegmental nuclei, the supralemniscal nuclei (B9 group), and the ventrolateral medulla. The largest serotonergic group, the dorsal raphe, contained one-third of the brainstem serotonergic neurons and showed five subdivisions, similar to that described in other species. In contrast, the median raphe did not show clear subdivisions. The internal complexity of the raphe nuclei and the degree of lateralisation of serotonergic neurons suggest that the wallaby serotonergic system is similar in organisation to that described for the cat and rabbit. This study supports the suggestion that the serotonergic system is evolutionally well conserved and provides baseline data for a quantitative study of serotonergic innervation of the developing cortex in the wallaby.

Spontaneous activity in the perinatal trigeminal nucleus of the rat.

The role of electrical activity in establishing appropriate connections in the trigeminal pathway remains controversial. We report here a novel observation of spontaneous activity in the perinatal trigeminal sensory nucleus between embryonic day (E) 16 to postnatal day (P) 8. Most of these spontaneous bursts had the same polarity and were of comparable amplitude to the trigeminal nerve-evoked response. This synchronized activity was abolished by bicuculline or kynurenic acid. Recording from the teased trigeminal root during the spontaneous bursts also showed a corresponding afferent depolarization. We speculate that the GABAergic depolarization of pre- and postsynaptic cells may contribute to the generation of the spontaneous activity and its propagation throughout the trigeminal sensory pathway, even in the absence of activity initiated from peripheral sensory receptors.

Development of whisker-related patterns in marsupials: factors controlling timing.

In mature rodents, whisker-related patterns are known to be present in three levels of the brain: the brainstem trigeminal nuclei, the ventrobasal thalamus and the somatosensory cortex. These patterns have been demonstrated using neuroanatomical tracing techniques, histological and histochemical staining methods and electrophysiological recordings. The development and topography of these patterns are dependent on an intact periphery. But what governs when patterns form at the three levels? Possibilities include a controlling signal from the periphery or local mechanisms at each site, such as the arrival of afferent inputs or the maturation of target tissue. In this review, we report on the maturation of the whisker pathway in a marsupial, the wallaby, where the slow tempo of development is a feature. At each level, afferent fibres grow into the region of termination many weeks before the whisker-related pattern emerges. The results suggest that the maturity of the target tissue as well as signals from the periphery combine to trigger pattern formation at each level of the pathway.

Cholinergic depletion reduces plasticity of barrel field cortex.

This experiment examines the impact of the cholinergic input from the basal forebrain on the plasticity of the vibrissa-related somatosensory cortex. Newborn rat pups received intraventricular injections of the cholinergic immunotoxin IgG192-saporin, after bilateral removal of the C-line whisker follicles. Compared with saline-injected control animals, unilateral injections of 0.1 microg IgG192-saporin decreased the number of cholinergic neurons on the toxin injected side by 78% in the basal nucleus of Meynert and the vertical limb of the diagonal band of Broca, 80% in the magnocellular preoptic nucleus and the horizontal limb of the diagonal band of Broca, and 54% in the medial septal nucleus. Neuronal loss contralateral to the toxin was approximately half that on the ipsilateral side. The size of the C and D row barrels were compared from tangential sections through the barrel field. In control animals, D row barrels expanded into C row territory, giving a ratio of areas for D/C barrels of 2.03. Depletion of the cholinergic neurons reduced the expansion of D row barrels and hence decreased the D/C ratio, with a greater reduction on the toxin-treated side (1.43, P <0.005) compared with the contralateral side (1.64, P <0.05). This study implicates the basal forebrain cholinergic projection in somatosensory cortical plasticity.

Bcl-x and Bcl-2 immunoreactivity in postnatal mouse barrel fields.

BcL-xL and Bcl-2 proteins were identified by paraffin section immunohistochemistry in the neuropil core of the barrelettes of the caudalis spinal trigeminal nucleus 6 h after birth. They were subsequently identified at progressively more rostral levels of the trigeminal pathway, peaking in barreloid neuropil cores in the ventral posterior thalamic nucleus at postnatal day (P)4.5, and in cortical barrel cores at P7. Labelling was confined to the cores of barrel-like structures surrounded by immunonegative shells and became progressively less distinct at all levels from P8. These protein products, which are usually considered to control the onset of apoptosis, may serve other functions in the axon terminal fields of the trigeminal pathway.

Development of commissural neurons in the wallaby (Macropus eugenii).

We have examined the development of the laminar and areal distribution of cortical commissural neurons in a marsupial mammal, the wallaby Macropus eugenii. In this species, commissural axons approach the major cerebral commissure, the anterior commissure, via either the internal capsule or the external capsule and first cross the midline at postnatal day 14 (P14). By retrogradely labelling these axons with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine (DiI) at P15, we show here that the cell bodies of these neurons are restricted to a region of cortex adjacent to the rhinal fissure. Most of these labelled neurons are located in the compact cell zone of the cortical plate, with only a few labelled cells found in the zone of loosely packed cells deep to this layer. Over the subsequent 66 days, commissural neurons are found progressively more dorsally, rostrally, and caudally, so that, by P80, they are present throughout the extent of the neocortex. At this age, they are mainly pyramidal in morphology and form a single band within the deeper part of layer 5 of the developing cortex. From P80 to adulthood, the distribution of commissural neurons has been assessed in the visual cortex by using retrograde transport of horseradish peroxidase. At P80, labelled neurons with immature pyramidal morphology are present throughout the occipital cortex; as in DiI material, somata are located in deep layer 5. At P165, previously shown to be the age when commissural axon numbers peak, widespread labelling is present in the occipital region, with labelled cells now found in two bands corresponding to layers 3 and 5. After this age, neurons become more restricted in distribution, so that, by adulthood, commissural neurons are no longer apparent throughout area 17 but are restricted to a localised region around the area 17/18 boundary. Within this region, labelling is still present in layers 3 and 5 but is more dense in layer 3. The gradual restriction of commissural fields seen here in the wallaby is similar to that reported in the neocortex in many eutherians. These findings also support studies in eutheria, suggesting that subplate neurons do not appear to play a major role in commissural development.

Timecourse of development of the wallaby trigeminal pathway: III. Thalamocortical and corticothalamic projections.

The development of trigeminal projections between the thalamus and cortex has been investigated in the marsupial mammal, the wallaby, by using a carbocyanine dye, horseradish peroxidase conjugated to wheat germ agglutinin (WGA-HRP), Neurobiotin, and biocytin as pathway tracers. The appearance of whisker-related patterns in the cortex in relation to their appearance in the brainstem and thalamus was examined, as was the presence or absence of a waiting period for thalamocortical afferents and the identity of the first cortical cells to project to the thalamus. Thalamic afferents first reached the cortex at postnatal day (P) 15 and were distributed up to the deep edge of the compact cell zone in the superficial cortical plate throughout development, in both dye and WGA-HRP labelled material, with no evidence of a waiting period. The initial corticothalamic projection, detected by retrograde transport of WGA-HRP from the thalamus, occurred at P60 from layer 5 cells. This was confirmed by labelling of corticothalamic axons after cortical injections of Neurobiotin and biocytin. Scattered, labelled cells seen before P60 after dye labelling from the thalamus presumably resulted from transcellular labelling via thalamic afferents. Clustering of afferents in layer 4 and cell bodies and their dendrites in layers 5 and 6 first occurred simultaneously at P76. There is a sequential onset of pattern formation, first in brainstem, then in thalamus, and finally in cortex, with a long delay between afferent arrival and pattern formation at each level. Independent regulation at each level, likely depending on target maturation, is suggested.

Effect of neonatal capsaicin and infraorbital nerve section on whisker-related patterns in the rat trigeminal nucleus.

In the present study, we investigated the effect of neonatally administered capsaicin on whisker-related pattern formation in the rat trigeminal complex. Both normal whisker-related patterns of barrelettes and the modified patterns seen after neonatal section of the infraorbital nerve were assessed. Capsaicin caused no change in the pattern or size of cytochrome oxidase (CO) barrelettes in the principal trigeminal nucleus (Vp) or trigeminal nucleus interpolaris (Vi) or caudalis (Vc). Injections of horseradish peroxidase (HRP) or wheatgerm agglutinin conjugated to HRP (WGA-HRP) into the posteroorbital (PO) whisker follicle in vehicle-treated animals showed that WGA labelled a larger number of trigeminal ganglion cells than HRP (203 +/- 23; cf. 158 +/- 19), with an increased labelling of small-diameter neurons (HRP: 25.9 +/- 7.7 microm; WGA: 23.2 +/- 7.2 pm). Capsaicin caused a loss of smaller diameter cells but had no effect on the location, cross-sectional area, or rostrocaudal extent of the transganglionically labelled HRP terminations in Vp, Vi, Vc, and cervical dorsal horn. WGA-HRP labelling revealed similar, but less dense, central terminal areas as HRP and an additional area of superficial terminals in the caudal medulla; these were also unaffected by capsaicin treatment. After infraorbital nerve section, CO patches and transganglionically labelled afferent terminations, corresponding to innervated nonmystacial whiskers, were approximately doubled in size. Capsaicin had no effect on the increased size of these spared whisker patches or their afferent terminal areas. These results suggest that barrelette formation is not dependent on unmyelinated afferents and that the changes in response properties seen after capsaicin, such as increased receptive fields, reflect functional changes rather than anatomical expansion of afferent terminal areas.

Peripheral and central predictors of whisker afferent morphology in the rat brainstem.

Prior studies suggest that whisker afferents have but one central projection pattern, despite their association with differing peripheral receptors that predict central morphology in other systems. Target factors in barrelettes are thought to dictate afferent projection patterns; yet, barrelettes differ in their size, shape and development. We tested the hypothesis that whisker afferents have differing morphologies that are predicted by peripheral and central factors. Branching patterns and collaterals of 78 Neurobiotin-stained afferents were compared in rats. Fibers from one whisker had precisely somatotopic projections but highly varied morphologies. For the entire sample, analysis of variance revealed significant intrafiber variance in collateral number and arbor shape that was attributed to the target subnucleus. Significant interfiber variance did not reflect response adaptation rate, direction sensitivity, whisker row origin or parent fiber bifurcation in the trigeminal root. Instead, we found the following. 1) Mandibular fibers had more elongated arbors than maxillary axons. In subnuclei interpolaris and principalis, mandibular fibers had larger arbors with more boutons/collateral than maxillary axons; in oralis and interpolaris, mandibular fibers had fewer collaterals than those of the maxillary division. 2) Upper lip whisker axons had more boutons than those from the B-D row in all subnuclei. 3) Rostral whisker are afferents had larger arbors and more boutons than those from middle or caudal arcs due to significant arc effects in interpolaris and oralis. Thus, whisker afferents are not structurally uniform, and some morphological features are predictable. Intrafiber variance is attributed to the central target; interfiber variance reflects maxillary versus mandibular origin, upper lip origin and whisker rostrocaudal arc.

Anterior commissure of the wallaby (Macropus eugenii): adult morphology and development.

In metatheria, all neo- and paleo-cortical commissural connections are made by the anterior commissure. We have examined the adult morphology of this commissure and its development in a diprotodontid metatherian, the wallaby (Macropus eugenii), at both the light and electron microscope level. The total number of axons in the adult anterior commissure was 21.7 million, of which 55-62% were myelinated. The dorsal two thirds of the commissure, containing neocortical commissural axons, showed a higher percentage of larger, myelinated axons than the ventral one third, which contains paleocortical commissural axons. The commissure also showed a topographical gradient, with cells in the dorsal cortex projecting through the dorsal region of the commissure, the fasciculus aberrans. In the rostrocaudal axis, axons from the frontal cortex tended to pass more anteriorly through the commissure and those from the occipital more posteriorly, but there was extensive overlap of projections from different areas. The gestation length of this wallaby is 28.3 days, and all commissural development occurs postnatally. The anterior commissure first appeared at P (postnatal day) 14, at which time commissural fibres were apparently derived from the external capsule exclusively. Commissural fibres passing through the internal capsule, and joining the anterior commissure via the fasciculus aberrans, were first noted at P18. By that age there were 94,000 to 161,000 axons. Peak axon counts of 50 to 63 million occurred between P100 and P150. The number of growth cones in a single midline section peaked at approximately P114 (480,000) and dropped to 0 by P170. The distribution of growth cones was analysed during the early stages of anterior commissure development (P18, P30, P82). At P18, growth cones were concentrated in the dorsal parts of the commissural bundle, suggesting a ventrodorsal sequence of addition of axons. There was no apparent preferential association of growth cones with the periphery of the commissure or glial structures at any of the three ages examined. The results show that axonal overproduction and regression in cortical commissural connections are features of development in diprotodontid metatheria, as in eutheria.