Max R. Bennett

The concept of long term potentiation of transmission at synapses

The phenomenon of long term potentiation (LTP) of synaptic transmission, whereby a series of conditioning trains of impulses potentiate the size of synaptic potentials for periods in excess of hours, was discovered in the mammalian hippocampus by Lomo [1966, Acta Physiol. Scand. 68(Suppl. 277), 128] and subsequently characterized by Bliss and Lomo (1970, J. Physiol. 207, 61P). The search for the underlying mechanisms of LTP makes for fascinating reading. The induction of associative LTP was shown by Collingridge et al. (1982, J. Physiol. 334, 33-46) to be dependent on the presence of N-methyl-D-aspartate receptors, following the discovery of these receptors by Watkins and Evans (1981, A. Rev. Pharmac. Toxic. 21, 165-204). There has, however, been continuing controversy as to whether the maintenance phase of LTP over periods of hours may be attributed to an increase in the amount of transmitter released or to an increase in the number of glutamate receptors or both. There is more agreement on the important role or protein kinases in the maintenance phase of LTP. The role of LTP in memory is just now being elucidated.

The formation of topographical maps in developing rat gastrocnemius muscle during synapse elimination.

1. The rat lateral gastrocnemius muscle (LG) is a complex of four muscle compartments, each defined in terms of its unique innervation by a single primary nerve branch of the muscle nerve. A study has been made of the topographical distribution of motor units in the medial compartment of the LG (LGM) both before and after the loss of polyneuronal innervation that accompanies development. 2. Glycogen depletion methods showed that the distribution of single motor units depended on the rostro‐caudal origins of their axons in the spinal cord: rostral axons possessed motor units almost exclusively confined to the medial half of the LGM; intermediate axons possessed motor units primarily in the intermediate and lateral part of the LGM; caudal axons possessed motor units that were not restricted to any particular part of the LGM. 3. Myosin ATPase staining showed that about 80% of the LGM consists of type II A fibres, whilst the remainder are type II B. Physiological determination of the contractile properties of motor units indicated two classes of units: those that were relatively fatigue resistant and did not show a sag property (like fast‐twitch, fatigue‐resistant fibres or FR) and those that were relatively fatigable and did show a sag property (like fast‐twitch, fatigable fibres or FF). 4. Glycogen depletion was also used to determine the distribution of motor units in the LGM at 7 days post‐natal, when most fibres still receive a polyneuronal innervation. The LGM primary nerve branch innervated a confined sub‐volume of muscle fibres which is similar to the mature pattern. However, rostral axons possessed motor units that extended into the lateral half of the LGM, a position from which they are excluded in the adult. 5. These observations suggest that the axons of rostral and intermediate units form a topographical map within adult FR motor units (type II A fibres) in the LGM. The results suggest that competition between axon terminals for synaptic sites plays a role in the elimination of inappropriately positioned terminals and subsequent emergence of the topographical map.

Comparative study on the distribution patterns of P2X1–P2X6 receptor immunoreactivity in the brainstem of the rat and the common marmoset (Callithrix jacchus): Association with catecholamine cell groups

The present study investigated the topographical distribution of P2X1–P2X6 receptor subtypes in the rat and common marmoset hindbrain by immunohistochemistry. In addition, double‐labeling immunofluorescence was used to determine the extent of colocalization between catecholamine cell groups and the various P2X receptors. The data demonstrate a widespread distribution pattern for all six P2X receptors throughout both the rat hindbrain and the marmoset hindbrain, although distinctions between species, brain nuclei, and P2X receptor subtypes exist. In rat, dense staining for the P2X receptors was found in the nucleus of the solitary tract (NTS), medial vestibular nucleus, and medial and lateral parabrachial nuclei. Moderate staining was observed in the hypoglossal nucleus, cuneate nucleus, inferior olive, prepositus hypoglossi, rostral ventrolateral medulla (RVLM), and locus coeruleus. Staining was also observed in the gracile nucleus, the mesencephalic trigeminal nucleus, and the central pontine gray. In marmoset, prominent P2X receptor‐like immunoreactivity occurred in the NTS, medial cuneate nucleus, prepositus hypoglossi, and medial vestibular nucleus. Moderate staining was observed in the area postrema, dorsal motor nucleus of the vagus, lateral cuneate, lateral reticular, spinal trigeminal nucleus, RVLM, and inferior olive. Immunofluorescent double labeling of tyrosine hydroxylase (TH)‐containing cells revealed that all subtypes of P2X receptors show some degree of colocalization with TH. The highest proportion of TH and P2X receptor double labeling was in the A5 region (with the P2X2 subunit), whereas the lowest proportion of double‐labeled cells occurred in the C2 region of the NTS for the P2X5 subunit. These findings support a role for extracellular adenosine 5′‐triphosphate in fast synaptic neurotransmission within the brainstem. J. Comp. Neurol. 427:485–507, 2000.

A systematic review and meta-analysis of magnetic resonance imaging measurement of structural volumes in posttraumatic stress disorder

Posttraumatic stress disorder (PTSD) is a debilitating condition associated with mild to moderate cognitive impairment and with a prevalence rate of up to 22% in veterans. This systematic review and quantitative meta-analysis explore volumetric differences of three key structural brain regions (hippocampus, amygdala and anterior cingulate cortex (ACC)), all of which have been implicated in dysfunction of both salience network (SN) and default mode network (DMN) in PTSD sufferers. A literature search was conducted in Embase, Medline, PubMed and PsycINFO in May 2013. Fifty-nine volumetric analyses from 44 articles were examined and included (36 hippocampus, 14 amygdala and nine ACC) with n=846 PTSD participants, n=520 healthy controls (HCs) and n=624 traumatised controls (TCs). Nine statistical tests were performed for each of the three regions of interest (ROIs), measuring volume differences in PTSD subjects, healthy and traumatised controls. Hippocampal volume was reduced in subjects with PTSD, with a greater reduction in the left hippocampus. A medium effect size reduction was found in bilateral amygdala volume when compared with findings in healthy controls; however, no significant differences in amygdala volume between PTSD subjects and trauma-exposed controls were found. Significant volume reductions were found bilaterally in the ACC. While often well matched with their respective control groups, the samples of PTSD subjects composed from the source studies used in the meta-analyses are limited in their homogeneity. The current findings of reduced hippocampal volume in subjects with PTSD are consistent with the existing literature. Amygdala volumes did not show significant reductions in PTSD subjects when compared with volumes in trauma-exposed controls-congruous with reported symptoms of hypervigilance and increased propensity in acquisition of conditioned fear memories-but a significant reduction was found in the combined left and right hemisphere volume analysis when compared with healthy controls. Bilateral volume reductions in the ACC may underpin the attentional deficits and inabilities to modulate emotions that are characteristically associated with PTSD patients.

Stress and trauma: BDNF control of dendritic-spine formation and regression

Chronic restraint stress leads to increases in brain derived neurotrophic factor (BDNF) mRNA and protein in some regions of the brain, e.g. the basal lateral amygdala (BLA) but decreases in other regions such as the CA3 region of the hippocampus and dendritic spine density increases or decreases in line with these changes in BDNF. Given the powerful influence that BDNF has on dendritic spine growth, these observations suggest that the fundamental reason for the direction and extent of changes in dendritic spine density in a particular region of the brain under stress is due to the changes in BDNF there. The most likely cause of these changes is provided by the stress initiated release of steroids, which readily enter neurons and alter gene expression, for example that of BDNF. Of particular interest is how glucocorticoids and mineralocorticoids tend to have opposite effects on BDNF gene expression offering the possibility that differences in the distribution of their receptors and of their downstream effects might provide a basis for the differential transcription of the BDNF genes. Alternatively, differences in the extent of methylation and acetylation in the epigenetic control of BDNF transcription are possible in different parts of the brain following stress. Although present evidence points to changes in BDNF transcription being the major causal agent for the changes in spine density in different parts of the brain following stress, steroids have significant effects on downstream pathways from the TrkB receptor once it is acted upon by BDNF, including those that modulate the density of dendritic spines. Finally, although glucocorticoids play a canonical role in determining BDNF modulation of dendritic spines, recent studies have shown a role for corticotrophin releasing factor (CRF) in this regard. There is considerable improvement in the extent of changes in spine size and density in rodents with forebrain specific knockout of CRF receptor 1 (CRFR1) even when the glucocorticoid pathways are left intact. It seems then that CRF does have a role to play in determining BDNF control of dendritic spines.

A Retinal Ganglion Cell Neurotrophic Factor Purified from the Superior Colliculus

Abstract: Dissociated neonatal rat retinal ganglion cells can be maintained by the addition of an extract from the neonatal superior colliculus. This extract can support 95% of ganglion cells over 24 h in culture; in addition it promotes the expression of neurites from these cells. This report describes the purification of a neurotrophic factor from the superior colliculus which supports the survival of 80% of retinal ganglion cells over 24 h in vitro. The purification procedure involves a combination of dye‐ligand, anion‐exchange, and molecular sieve chromatography. The purified neurotrophic factor has a Stokes radius of approximately 200 Å using molecular sieve chromatography in the presence of a chaotropic agent. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis of the purified factor indicates that it is a glycoprotein that migrates with a molecular mass >400 kDa. Further characterization of this high‐molecular‐mass glycoprotein by enzymatic digestion demonstrated that it is a chondroitin sulfate pro‐teoglycan. This factor is clearly distinguishable from other neurotrophic factors that have an effect on retinal ganglion cells such as brain‐derived neurotrophic factor and fibroblast growth factor. The chondroitin sulfate proteoglycan from the neonatal superior colliculus is the first proteoglycan to be identified as a neurotrophic factor.

P2X1 receptor membrane redistribution and down-regulation visualized by using receptor-coupled green fluorescent protein chimeras

The P2X(1) purinergic receptor subtype occurs on smooth muscle cells of the vas deferens and urinary bladder where it is localized in two different size receptor clusters, with the larger beneath autonomic nerve terminal varicosities. We have sought to determine whether these synaptic-size clusters only form in the presence of varicosities and whether they are labile when exposed to agonists. P2X(1) and a chimera of P2X(1) and green fluorescent protein (GFP) were delivered into cells using microinjection, transient transfection or infection with a replication-deficient adenovirus. The P2X(1)-GFP chimera was used to study the time course of P2X(1) receptor clustering in plasma membranes and the internalization of the receptor following prolonged exposure to ATP. Both P2X(1) and P2X(1)-GFP clustered in the plasma membranes of Xenopus oocytes, forming patches 4-6 microm in diameter. Human embryonic kidney 293 (HEK293) cells, infected with the adenovirus, possessed P2X(1) antibody-labeled regions in the membrane colocalized with GFP fluorescence. The ED(50) for the binding of alpha,beta-methylene adenosine triphosphate (alpha,beta-meATP) to the P2X(1)-GFP chimera was similar to native P2X(1) receptors. ATP-generated whole-cell currents in oocytes or HEK293 cells expressing either P2X(1) or P2X(1)-GFP were similar. Exposure of HEK293 cells to alpha, beta-meATP for 10-20 min in the presence of 5 microM monensin led to the disappearance of P2X(1)-GFP fluorescence from the surface of the cells. These observations using the P2X(1)-GFP chimera demonstrate that P2X(1) receptors spontaneously form synaptic-size clusters in the plasma membrane that are internalized on exposure to agonists.

Schizophrenia: susceptibility genes, dendritic-spine pathology and gray matter loss.

Gray matter loss in the cortex is extensive in schizophrenia, especially in the prefrontal-temporal-network (PTN). Several molecules such as neuregulin-1 (NRG1) and its ErbB4 receptor are encoded by candidate susceptibility genes for schizophrenia. The question arises as to how these genes might contribute to the observed changes in gray matter. It is suggested that one pathway involves molecules such as NRG1/ErbB4 determining the efficacy of N-methyl-D-aspartate receptors (NMDARs) found on dendritic spines at synapses in the PTN. The growth of dendritic spines is modulated by NRG1/ErbB4 through NMDARs as these activate small Rho-GTPases, such as kalirin, which control the actin cytoskeleton in the spines responsible for their growth. Another pathway involves NRG1/ErbB determining the proliferation and differentiation of oligodendrocytes in the white matter as well as their capacity for myelination, the integrity of which determines the stability of nerve terminals on dendritic spines. A causal chain is established between failure of the products of susceptibility genes for schizophrenia, the decrease of dendritic spines and synaptic terminals, and the loss of gray matter. It is suggested than an important focus for future research in schizophrenia is to identify interventions that prevent the loss of dendritic spines and synapses during the prodromal period or earlier during development as well as to re-establish dendritic spines and synapses lost subsequent to this period. This will help reestablish neural networks in the PTN and so the loss of gray matter in the PTN.

Embryonic chick retinal ganglion cells identified “in vitro”

SummaryWhen HRP is injected into the optic tecta of embryonic or newly hatched chicks, the ganglion cells in the contralateral retina can be successfully dissociated into culture and identified at any time by appropriate histochemical staining. Histological examination of whole mounts of retinae both ipsilateral and contralateral to an injection site indicated that no HRP diffused out of an injected tectum, and that the only reaction product that could be visualized was restricted to the ganglion cell layer of the contralateral eye. Because retinal ganglion cells are the only retinal neurons to project to the optic tectum, the intraxonal retrograde transport of HRP to these cells allows their unequivocal identification from amongst the heterogeneous population of retinal neurons present after dispersal into single cells in monolayer culture. The presence of HRP in the cell bodies did not appear to impair their ability to survive, grow or express neurites. Counts of labelled cells from progressively aged birds confirmed that the peak number of generated ganglion cells occurs on embryonic day 10, and that there is a 40% decline in the number of these neurons over the following 3 days. However, when labelled ganglion cells from 10 day embryos were grown in culture with optic tectum, all the ganglion ceils survived over the following 4 days, including those destined to die in vivo. This trophic effect cannot be induced by cerebellum, but is partly induced by media first conditioned over tectal cells. The trophic effect exerted by optic tectum appears therefore to be specific and chemically mediated.We suggest that the death of retinal ganglion cells in vivo may be a consequence of the inability of some cells to establish adequate supplies of a growth factor from the optic tectum.

Glutamate induces directed chemotaxis of microglia

Microglia in the brain possess dynamic processes that continually sample the surrounding parenchyma and respond to local insults by rapidly converging on the site of an injury. One of the chemotaxic agents responsible for this response is ATP. Here we show that the transmitter glutamate is another such chemotaxic agent. Microglia exposed to glutamate increase their cell membrane ruffling and migrate to a source of glutamate in cell culture and in spinal cord slices. This chemotaxis is meditated by α‐amino‐3‐hydroxy‐5‐methylisoxazole‐4‐propionic acid (AMPA) and metabotropic glutamate receptors on the microglia. Chemotaxis is dependent on redistribution of actin filaments in the cells and on tubulin following receptor activation. Thus glutamate, which is released at synapses as well as from damaged cells, can mediate rapid chemotaxic responses from microglial cells.