Research capacity building in general practice: A new opportunity in Fremantle, WA

One of the great disappointments of primary care medicine has been the failure to develop a strong research tradition among general practitioners. This has happened despite the great legacy left by William Budd, James MacKenzie, and Will Pickles, and the clear acceptance that such research is necessary to improve patient care

Some observations on the postnatal development of the rat cerebellum following unilateral pedunclotomy

The structure of the cerebellum, its cortex and the cells contained therein have been described. Also, the distribution of both afferent and efferent fibres has been summarised from the literature. The cerebellar development and experiments which have been performed upon it have also been briefly outlined. The technique for surgical pedunclotomy is described and its effect on cerebellar and Purkinje cell development were found. The effect of pedunclotomy on cerebellar growth and the Purkinje cell numbers is differential, a lesion on postnatal day 7 causing the most severe effect. Also, the Purkinje cell size was reduced following pedunclotomy after the age of 3 days. However the cortical histology was otherwise qualitatively normal. The response of the noradrenergic afferente to the cerebellum to pedunclotomy was observed. Qualitatively the density of fibres was equivalent to normal after 4 days, which is in keeping with previous experiments, but there was no hyperinnervation 35 days post pedunclotomy a result not in agreement with the studies using 6-hydroxydopamine. The olivocerebellar projection 35 days after pedunclotomy on days 3 and 7 shows a reinnervation of the deafferented hemicerebellum. However, this response was not seen after pedunclotomy on day 10. The topography of this aberrant path is organised roughly into sagittal bands although not as discretely as that seen in the normal animal. In neonatal rats there was some evidence that this path may already exist in those animals pedunclotomised on days 3 and 7. Also, an additional ipsilateral projection was found following acute pedunclotomy, which entered the cerebellum through the ipsilateral inferior cerebellar peduncle. The results have been discussed in relation to other experiments on cerebellar development and with studies in the literature on plasticity within the central nervous system

Cerebellum and neurodevelopmental disorders: RORα is a unifying force

Errors of cerebellar development are increasingly acknowledged as risk factors for neuro-developmental disorders (NDDs), such as attention deficit hyperactivity disorder (ADHD), autism spectrum disorder (ASD), and schizophrenia. Evidence has been assembled from cerebellar abnormalities in autistic patients, as well as a range of genetic mutations identified in human patients that affect the cerebellar circuit, particularly Purkinje cells, and are associated with deficits of motor function, learning and social behavior; traits that are commonly associated with autism and schizophrenia. However, NDDs, such as ASD and schizophrenia, also include systemic abnormalities, e.g., chronic inflammation, abnormal circadian rhythms etc., which cannot be explained by lesions that only affect the cerebellum. Here we bring together phenotypic, circuit and structural evidence supporting the contribution of cerebellar dysfunction in NDDs and propose that the transcription factor Retinoid-related Orphan Receptor alpha (RORα) provides the missing link underlying both cerebellar and systemic abnormalities observed in NDDs. We present the role of RORα in cerebellar development and how the abnormalities that occur due to RORα deficiency could explain NDD symptoms. We then focus on how RORα is linked to NDDs, particularly ASD and schizophrenia, and how its diverse extra-cerebral actions can explain the systemic components of these diseases. Finally, we discuss how RORα-deficiency is likely a driving force for NDDs through its induction of cerebellar developmental defects, which in turn affect downstream targets, and its regulation of extracerebral systems, such as inflammation, circadian rhythms, and sexual dimorphism

Reinnervation of late postnatal purkinje cells by climbing fibers: neosynaptogenesis without transient multi-innervation.

Synaptic partner selection and refinement of projections are important in the development of precise and functional neuronal connections. We investigated the formation of new synaptic connections in a relatively mature system to test whether developmental events can be recapitulated at later stages (i.e., after the mature synaptic organization has been established), using a model of postlesional reinnervation in the olivo-cerebellar pathway. During the development of this pathway, synaptic connections between climbing fibers (CFs) and Purkinje cells (PCs) are diffuse and redundant before synapse elimination refines the pattern. The regression of CFs during the first 2 postnatal weeks in the rat leads to mono-innervation of each PC. After unilateral transection of the rat olivo-cerebellar pathway and intracerebellar injection of BDNF 24 h after lesion, axons from the remaining inferior olive can sprout into the deafferented hemicerebellum and establish new contacts with denervated PCs at later developmental stages. We found that these contacts are first established on somatic thorns before the CFs translocate to the PC dendrites, recapitulating the morphological steps of normal CF-PC synaptogenesis, but on a relatively mature PC. However, electrophysiology of PC reinnervation by transcommissural CFs in these animals showed that each PC is reinnervated by only one CF. This mono-innervation contrasts with the reinnervation of grafted immature PCs in the same cerebellum. Our results provide evidence that relatively mature PCs do not receive several olivary afferents during late reinnervation, suggesting a critical role of the target cell state in the control of CF-PC synaptogenesis. Thus, synapse exuberance and subsequent elimination are not a prerequisite to reach a mature relationship between synaptic partners

Age-related Purkinje cell death is steroid dependent: RORα haplo-insufficiency impairs plasma and cerebellar steroids and Purkinje cell survival

A major problem of ageing is progressive impairment of neuronal function and ultimately cell death. Since sex steroids are neuroprotective, their decrease with age may underlie age-related neuronal degeneration. To test this, we examined Purkinje cell numbers, plasma sex steroids and cerebellar neurosteroid concentrations during normal ageing (wild-type mice, WT), in our model of precocious ageing (Rora+/sg, heterozygous staggerer mice in which expression of the neuroprotective factor RORα is disrupted) and after long-term hormone insufficiency (WT post-gonadectomy). During normal ageing (WT), circulating sex steroids declined prior to or in parallel with Purkinje cell loss, which began at 18 months of age. Although Purkinje cell death was advanced in WT long-term steroid deficiency, this premature neuronal loss did not begin until 9 months, indicating that vulnerability to sex steroid deficiency is a phenomenon of ageing Purkinje neurons. In precocious ageing (Rora+/sg), circulating sex steroids decreased prematurely, in conjunction with marked Purkinje cell death from 9 months. Although Rora+/sg Purkinje cells are vulnerable through their RORα haplo-insufficiency, it is only as they age (after 9 months) that sex steroid failure becomes critical. Finally, cerebellar neurosteroids did not decrease with age in either genotype or gender; but were profoundly reduced by 3 months in male Rora+/sg cerebella, which may contribute to the fragility of their Purkinje neurons. These data suggest that ageing Purkinje cells are maintained by circulating sex steroids, rather than local neurosteroids, and that in Rora+/sg their age-related death is advanced by premature sex steroid loss induced by RORα haplo-insufficiency

Low-intensity electromagnetic fields induce human cryptochrome to modulate intracellular reactive oxygen species

Exposure to man-made electromagnetic fields (EMFs), which increasingly pollute our environment, have consequences for human health about which there is continuing ignorance and debate. Whereas there is considerable ongoing concern about their harmful effects, magnetic fields are at the same time being applied as therapeutic tools in regenerative medicine, oncology, orthopedics, and neurology. This paradox cannot be resolved until the cellular mechanisms underlying such effects are identified. Here, we show by biochemical and imaging experiments that exposure of mammalian cells to weak pulsed electromagnetic fields (PEMFs) stimulates rapid accumulation of reactive oxygen species (ROS), a potentially toxic metabolite with multiple roles in stress response and cellular ageing. Following exposure to PEMF, cell growth is slowed, and ROS-responsive genes are induced. These effects require the presence of cryptochrome, a putative magnetosensor that synthesizes ROS. We conclude that modulation of intracellular ROS via cryptochromes represents a general response to weak EMFs, which can account for either therapeutic or pathological effects depending on exposure. Clinically, our findings provide a rationale to optimize low field magnetic stimulation for novel therapeutic applications while warning against the possibility of harmful synergistic effects with environmental agents that further increase intracellular ROS

IGF-I induces neonatal climbing-fibre plasticity in the mature rat cerebellum

Following unilateral transection (pedunculotomy) of the neonatal rat olivocerebellar pathway, the remaining inferior olive reinnervates the denervated hemicerebellum with correct topography. The critical period for this transcommissural reinnervation closes between postnatal days 7 and 10 but can be extended by injection of growth factors. Whether growth factor treatment can extend developmental plasticity into a mature, myelinated milieu remains unknown. Rats aged 11-30 days, underwent unilateral pedunculotomy followed 24 h later by injection of insulin-like growth factor 1 (IGF-1) into the denervated cerebellum. In all animals, IGF-1 induced transcommissural olivocerebellar reinnervation, which displayed organisation consistent with normal olivocerebellar topography even following pedunculotomy up to day 20. Thus IGF-1 can reproduce developmental neuroplasticity to promote appropriate target reinnervation in a mature myelinated environment

Role of afferents in the development and cell survival of the vertebrate nervous system

1. During normal development of the vertebrate central nervous system, a considerable number of neurons die. The factors controlling which neurons die and which survive are not fully understood. 2. Target populations are known to maintain their innervating neurons. However, the role of afferents in maintaining their targets is still under review, 3. In the developing nervous system, deafferentation of a neuron population is difficult to achieve because plasticity (structural re-organization) can cause re-innervation of the area. Re-innervation alters, rather than removes, the efferent supply. 4. Afferent input is important for neuronal survival during development because deafferentation increases neuronal death by 20-30% and increasing input diminishes neuronal death. 5. Deafferented neurons die at the normal time for cell death for any given population. This occurs after the arrival of afferent axons but before the completion of connectivity and the onset of function. 6. Neuronal survival is maintained by any input, such as reinnervation by inappropriate fibres, but for optimal survival, morphological maturation and the acquisition of normal physiology tile correct input is required. 7. Afferents maintain their target neurons via a combination of electrical activity and delivery of trophic agents, which adjust intracellular calcium, thereby facilitating protein synthesis, mitochondrial function and suppressing apoptosis. 8. Evidence from animal and in vitro experiments indicates that efferents play an extremely important role in the survival of developing neurons in the immature vertebrate nervous system