Synapses In Somatosensory Cortex Research Articles

Single-Synapse Analysis of a Diverse Synapse Population: Proteomic Imaging Methods and Markers

A lack of methods for measuring the protein compositions of individual synapses insitu has so far hindered the exploration and exploitation of synapse molecular diversity. Here, we describe the use of array tomography, a new high-resolution proteomic imaging method, to determine the composition of glutamate and GABA synapses in somatosensory cortex of Line-H-YFP Thy-1 transgenic mice. We find that virtually all synapses are recognized by antibodies to the presynaptic phosphoprotein synapsin I, while antibodies to 16 other synaptic proteins discriminate among 4 subtypes of glutamatergic synapses and GABAergic synapses. Cell-specific YFP expression in the YFP-H mouse line allows synapses to be assigned to specific presynaptic and postsynaptic partners and reveals that a subpopulation of spines on layer 5 pyramidal cells receives both VGluT1-subtype glutamatergic and GABAergic synaptic inputs. These results establish a means for the high-throughput acquisition of proteomic data from individual cortical synapses insitu.

Changes in neural dendrites and synapses in rat somatosensory cortex following neonatal post-hemorrhagic hydrocephalus

Neonatal post-hemorrhagic hydrocephalus is associated with cognitive decline and a serious deterioration in the patient's quality of life. The underlying impairments to neurons are not well understood. Here, we used the method described by Cherian et al. to construct a model of hydrocephalus after intra-ventricular hemorrhage and then observed the subsequent pathological changes in the morphology of neurons labeled by enhanced green fluorescent proteins (EGFP) using the in utero electroporation technique. Injection of venous blood into the lateral ventricles of 7-day-old rats in the operation group caused marked enlargement of the ventricles in 60% (9/15) of the rats after 2 weeks and in 53.3% (8/15) of the rats after 3 weeks. Compared with the control group, the length of the neural dendrites in the somatosensory cortex was shortened and the number of both neuron dendrite branches and synapses was significantly decreased. There was no evidence of cerebral cortical neuron death as shown by positive EGFP cell counting which suggest that neurological dysfunction after intra-ventricular hemorrhage-induced hydrocephalus may be related to the shortening of neural dendrites and the decreased number of synapses in somatosensory cortex and thus provides a possible neurological cause for hydrocephalus-induced cognitive decline and motor dysfunction.

Critical Period Plasticity Is Disrupted in the Barrel Cortex of Fmr1 Knockout Mice

Alterations in sensory processing constitute prominent symptoms of fragile X syndrome; however, little is known about how disrupted synaptic and circuit development in sensory cortex contributes to these deficits. To investigate how the loss of fragile X mental retardation protein (FMRP) impacts the development of cortical synapses, we examined excitatory thalamocortical synapses in somatosensory cortex during the perinatal critical period in Fmr1 knockout mice. FMRP ablation resulted in dysregulation of glutamatergic signaling maturation. The fraction of silent synapses persisting to later developmental times was increased; there was a temporal delay in the window for synaptic plasticity, while other forms of developmental plasticity were not altered in Fmr1 knockout mice. Our results indicate that FMRP is required for the normal developmental progression of synaptic maturation, and loss of this important RNA binding protein impacts the timing of the critical period for layer IV synaptic plasticity.

Dopamine receptor pruning during the peripubertal period is not attenuated by NMDA receptor antagonism in rat

D 1 and D 2 receptors are overproduced and pruned in the mammalian striatum during the periadolescent period. The mechanism that underlies this process in striatum is unknown. However, previous research has shown that the activity-dependent pruning of dendrites and synapses in somatosensory cortex and the visual fields is mediated by glutamatergic actions via N-methyl- d- aspartate (NMDA) receptor and is prevented by pretreatment with the NMDA antagonist MK-801. In order to test the hypothesis that the pruning of dopamine D 1 and D 2 receptors that occurs in the striatum after puberty (which occurs at approximately 40 days of age; P40), male and female rats were treated with saline vehicle or MK-801 (0.3 mg/kg) for 20 or 40 days and sacrificed immediately after the 20 day treatment (P60), 40 day treatment (P80), or 40 day treatment with 40 day recovery (P120). Analyses of the data reveal that none of these three treatment regimens altered striatal D 1 or D 2 receptor density in males or females relative to vehicle controls. At P60, MK-801 treatment failed to alter either D 1 ( F1,16=0.06, P>0.5) or D 2 receptors ( F1,16=0.39, P>0.5) for either sex. Similarly, MK-801 treatment did not affect D 1 or D 2 receptors at P80 ( P>0.3) or at P120 ( P>0.7). These data suggest that the normal 40% reduction in striatal dopamine receptor density that occurs between puberty and adulthood is not dependent on post-pubertal glutamatergic transmission through NMDA receptors.

α7 Nicotinic Acetylcholine Receptors Occur at Postsynaptic Densities of AMPA Receptor-Positive and -Negative Excitatory Synapses in Rat Sensory Cortex

NMDA receptor (NMDAR) activation requires concurrent membrane depolarization, and glutamatergic synapses lacking AMPA receptors (AMPARs) are often considered "silent" in the absence of another source of membrane depolarization. During the second postnatal week, NMDA currents can be enhanced in rat auditory cortex through activation of the alpha7 nicotinic acetylcholine receptor (alpha7nAChR). Electrophysiological results support a mainly presynaptic role for alpha7nAChR at these synapses. However, immunocytochemical evidence that alpha7nAChR is prevalent at postsynaptic sites of glutamatergic synapses in hippocampus and neocortex, along with emerging electrophysiological evidence for postsynaptic nicotinic currents in neocortex and hippocampus, has prompted speculation that alpha7nAChR allows for activation of NMDAR postsynaptically at synapses lacking AMPAR. Here we used dual immunolabeling and electron microscopy to examine the distribution of alpha7nAChR relative to AMPAR (GluR1, GluR2, and GluR3 subunits combined) at excitatory synapses in somatosensory cortex of adult and 1-week-old rats. alpha7nAChR occurred discretely over most of the thick postsynaptic densities in all cortical layers of both age groups. AMPAR immunoreactivity was also detectable at most synapses; its distribution was independent of that of alpha7nAChR. In both age groups, approximately one-quarter of asymmetrical synapses were alpha7nAChR positive and AMPAR negative. The variability of postsynaptic alpha7nAChR labeling density was greater at postnatal day (PD) 7 than in adulthood, and PD 7 neuropil contained a subset of small AMPA receptor-negative synapses with a high density of alpha7nAChR immunoreactivity. These observations support the idea that acetylcholine receptors can aid in activating glutamatergic synapses and work together with AMPA receptors to mediate postsynaptic excitation throughout life.

Developmental disturbance of rat cerebral cortex following prenatal low-dose γ-irradiation: A quantitative study

Pregnant rats were exposed to a single whole-body γ-irradiation on Day 15 of gestation at a dose of 0.27, 0.48, 1.00, or 1.46 Gy. They were allowed to give birth and the offspring were killed at 6 or 12 weeks of age for microscopic and electron microscopic examinations of the cerebrum. Their body weight, brain weight, cortical thickness, and numerical densities of whole cells and synapses in somatosensory cortex were examined. Growth of the dendritic arborization of layer V pyramidal cells was also examined quantitatively with Golgi-Cox specimens. A significant dose-related reduction in brain weight was found in all irradiated groups. Neither gross malformation nor abnormality of cortical architecture was observed in the groups exposed to 0.27 Gy. A significant change was found in thickness of cortex in the groups exposed to 0.48 Gy or more. Cell packing density increased significantly in the group exposed to 1.00 Gy. Significant reduction in the number of intersections of dendrites with the zonal boundaries were found in the groups exposed to 0.27 Gy or more. There was no difference in the numerical density of synapses in layer I between the control and irradiated groups. These results suggested that doses as low as 0.27 Gy could cause a morphologically discernible change in the mammalian cerebrum.