Margaret B. Lowrie

Dependence of postnatal motoneurones on their targets: review and hypothesis

Motoneurones are known to die (1) during embryonic development (naturally occurring cell death), (2) early in postnatal development after axonal injury, and (3) as a consequence of disease, such as spinal muscular atrophy or (in later life) amyotrophic lateral sclerosis. Naturally occurring motoneurone death has been extensively investigated, and interaction with the target muscle has emerged as an important factor for survival of embryonic motoneurones. Evidence that this target dependence of motoneurones continues postnatally is discussed in this review, as is the possible nature of the retrograde signal from the muscle. An explanation for the role of the muscle in motoneurone survival is also proposed, which may be applicable in situations where motoneurone death occurs postnatally. This proposal takes into account the changing functional demands imposed on motoneurones as a result of the gradual maturation of the CNS, and suggests that during development the muscle induces the motoneurones to become competent to carry out these requirements.

Evidence That Spinal Interneurons Undergo Programmed Cell Death Postnatally in the Rat

Programmed cell death has been demonstrated in several specific neuronal populations as a mechanism for modulating the population size following differentiation, but its applicability to all neuronal types is unclear. Evidence for programmed cell death in some populations such as the numerous spinal interneurons has been lacking. We have studied the incidence of apoptosis in the rat spinal cord with three different methods and found a previously undocumented wave of apoptosis occurring in spinal grey matter shortly after birth. The apoptotic morphology was confirmed ultrastructurally. Dying cells were identified as neurons by immunocytochemical labelling for neuronal markers and had an anatomical distribution which indicated that most of the apoptotic cells were interneurons not motoneurons. This wave of apoptosis has the characteristics of a discrete developmental process and occurs later than that of either ventral horn motoneurons or dorsal root ganglion cells, to which most spinal interneurons are connected. These findings indicate that interneurons do undergo programmed cell death, and we suggest that this occurs in response to the earlier reduction in size of their main synaptic targets.

Cell death of spinal interneurones

The occurrence of neuronal death during development is well documented for some neuronal populations, such as motoneurones and dorsal root ganglion cells, whose connecting pathways are clearly defined. Cell survival is thought to be regulated largely by target and input connections, a process that serves to match the size of synaptically linked neuronal populations. Far less is known about interneurones. It is assumed that most interneurone populations are excluded from this process because their connections are more diffuse. Recent studies on the rat spinal cord have indicated that interneurone death does occur, both naturally during development and induced following peripheral nerve injury. Here the evidence for spinal interneurone death is reviewed and the factors influencing it are discussed. There are many functional types of interneurones in the spinal cord that may differ in vulnerability to cell death, but it is concluded that for most spinal interneurones the traditional view of target regulation is unlikely. Instead it is proposed that developmental interneurone death in the spinal cord forms part of a plastic response to altered sensory activation rather than a size-matching exercise. There is also emerging evidence that interneurone death may play a more direct role in some neurodegenerative diseases than hitherto considered.

Cadherin-10 is a novel blood–brain barrier adhesion molecule in human and mouse

Maintenance of the specialised environment of the central nervous system requires barriers provided by the endothelium of brain microvessels (the blood-brain barrier (BBB)) or the epithelium lining the ventricles (CSF-brain barrier) or the choroid plexus (blood-CSF barrier). Inter-endothelial junctions are more extensive in the BBB than in other tissues, with elaborate tight junctions. However, few differences in the molecular composition of these junctions have been described. Here, we show, in both human and mouse brain, that the type II classical cadherin, cadherin-10, is expressed in BBB and retinal endothelia, but not in the leaky microvessels of brain circumventricular organs (CVO), or in those of non-CNS tissues. This expression pattern is distinct from, and reciprocal to, VE-cadherin, which is reduced or absent in tight cortical microvessels, but present in leaky CVO vessels. In CVO, the barrier function is switched from the microvasculature to the adjacent ventricular epithelium, which we also find to express cadherin-10. In the vessels of gliobastoma multiforme tumours, where BBB is lost, cadherin-10 is not detected. This demonstration of a distinctive expression pattern of cadherin-10 suggests that it has a pivotal role in the development and maintenance of brain barriers.

Time Course of Motoneurone Death after Neonatal Sciatic Nerve Crush in the Rat

Sciatic motoneurones were retrogradely labelled with long-lasting fluorescent dyes prior to unilateral nerve crush in either 3-day-old or adult rats. The number of surviving labelled motoneurones at intervals after nerve injury were compared to the number in the contralateral control ventral horn and in unoperated animals. Following adult nerve crush there was no significant reduction in the number of labelled motoneurones, but after neonatal nerve crush the count was reduced to about 35%. Most of the cell death occurred during the first 6 days after nerve injury, mainly from the lower half of the motor column, but about one third died between 6 and 12 days, mainly from the upper part. These results suggest that less mature motoneurones tend to die earliest, before the muscle is reinnervated. Those in the upper, more mature part of the motor pool survive longer but may still die during reinnervation. At least two types of glial cell were secondarily labelled by this method, distinguished by their response to nerve injury.

Nerve injury in adult rats causes abnormalities in the motoneuron dendritic field that differ from those seen following neonatal nerve injury

Disruption of neuromuscular contact by nerve-crush during the early postnatal period causes increased activity and abnormal reflex responses in affected motoneurons, but such changes are not found after nerve-crush in adult animals. We found previously that neonatally lesioned cells develop an abnormal dendritic field, which may explain the functional changes. Here we have studied the dendritic morphology of the same motoneuron pool after nerve-crush at maturity in order to correlate the observed alterations in morphology with physiological findings. One to two months after sciatic nerve-crush in adult animals, motoneurons supplying the extensor hallucis longus muscles of the rat were retrogradely labelled with cholera toxin subunit-B conjugated to horseradish peroxidase. The dendritic tree of labelled cells was then analysed. Following adult nerve-crush, the dendritic tree of the motoneurons was smaller but did not display the localised increase in dendritic density seen after neonatal nerve-crush. These findings support the view that such specific morphological changes contribute to the physiological abnormalities seen only after neonatal nerve injury.

Motoneurones That Innervate the Rat Soleus Muscle Mature Later than Those to the Tibialis anterior and Extensor digitorum longus Muscles

The response of motoneurones that innervate either the soleus or tibialis anterior (TA) and extensor digitorum longus (EDL) muscles to increased locomotor activity or to nerve injury at different stages after birth was examined. Inceased locomotor activity of rat pups was induced by daily treatment with L-dopa during the first 12 days after birth, and the number of surviving motoneurones to the soleus or TA/EDL muscles was established by retrograde labelling. Treatment with L-dopa resulted in the loss of a significant number of motoneurones within the soleus motor pool but had no effect on the survival of those motoneurones innervating the TA/EDL. Furthermore, following nerve injury during the first few days postnatally, more motoneurones within the soleus motor pool die than in the TA/EDL pool. These results indicate that motoneurones to the soleus muscle mature later than those to the TA/EDL muscles.

Dendritic Development in Normal Lumbar Motoneurons and Following Neonatal Nerve Crush in the Rat

Motoneurons from the rat were retrogradely labelled with cholera toxin-horseradish peroxidase at intervals during normal postnatal development and following nerve crush at birth. Normal cells displayed a relatively steady increase in total visible dendritic density which was largely confined to the dorsomedial direction. After nerve crush at birth, dorsomedially orientated dendrites failed to achieve normal density, resulting in a significantly smaller dendritic tree by adulthood. There was also a transient, abnormal extension of dendrites in the medioventral direction which had regressed to normal levels by maturity. The predominance of changes in the dorsally directed region of the dendritic tree suggests that dendritic development of motoneurons is influenced by synaptic inputs in the dorsal horn.

N-Methyl-d-aspartate receptors in motoneurones after unilateral axotomy in the neonatal rat

Quantitative autoradiography was used to characterise the binding of the N-methyl-D-aspartate receptor antagonist [3H]dizocilpine maleate in the ventral horns of the lumbar spinal cord of normal, sham-operated, and axotomized neonatal rats. Specific binding sites were revealed on the cell membranes of the motoneurones. In the normal neonate both the Bmax and the Kd values for the binding declined over the first 14 days of life. At 14 days the Kd value was similar to that in adult rats. Unilateral sciatic nerve section was performed in neonates on the day of birth. Axotomy caused the death of approximately 53% of motoneurones. The Bmax and Kd values for [3H]dizocilpine maleate binding declined in the first 2 weeks on operated and contralateral sides in both axotomized and sham-operated animals. However, the specific binding per motoneurone was significantly higher on the sectioned side in axotomized animals at all times examined after the day of birth. The results are consistent with two populations of NMDA receptors with different binding affinities being present in motoneurones at birth, and the lower affinity receptor gradually disappearing over the first few days. The lower affinity receptor may be responsible for the plasticity of motoneurones during embryonic and neonatal life, and for determining which motoneurones die after axotomy.

Both Afferent and Efferent Connections Influence Postnatal Growth of Motoneuron Dendrites in the Rat

Spinal motoneurons from mature rats, which had received one of 5 different surgical procedures neonatally, were retrogradely labelled with a cholera toxin-horseradish peroxidase conjugate and their dendritic morphology was analysed. The motoneurons studied were those innervating extensor digitorum longus and the procedures disrupted their motor and sensory connections to varying degrees. Disruption of motor contact with the target muscle retarded dendritic growth in the transverse plane, particularly in the dorso-medial direction. Disruption of sensory as well as motor contact resulted additionally in an increase in dendritic density in the longitudinal plane, largely along the rostral-caudal axis. The findings suggest that dendritic development of motoneurons is influenced by both afferent and efferent target contacts and that these effects can be differentiated.