Unit Length Of Dendrite Research Articles

A Kalirin missense mutation enhances dendritic RhoA signaling and leads to regression of cortical dendritic arbors across development

Normally, dendritic size is established prior to adolescence and then remains relatively constant into adulthood due to a homeostatic balance between growth and retraction pathways. However, schizophrenia is characterized by accelerated reductions of cerebral cortex gray matter volume and onset of clinical symptoms during adolescence, with reductions in layer 3 pyramidal neuron dendritic length, complexity, and spine density identified in multiple cortical regions postmortem. Nogo receptor 1 (NGR1) activation of the GTPase RhoA is a major pathway restricting dendritic growth in the cerebral cortex. We show that the NGR1 pathway is stimulated by OMGp and requires the Rho guanine nucleotide exchange factor Kalirin-9 (KAL9). Using a genetically encoded RhoA sensor, we demonstrate that a naturally occurring missense mutation in Kalrn, KAL-PT, that was identified in a schizophrenia cohort, confers enhanced RhoA activitation in neuronal dendrites compared to wild-type KAL. In mice containing this missense mutation at the endogenous locus, there is an adolescent-onset reduction in dendritic length and complexity of layer 3 pyramidal neurons in the primary auditory cortex. Spine density per unit length of dendrite is unaffected. Early adult mice with these structural deficits exhibited impaired detection of short gap durations. These findings provide a neuropsychiatric model of disease capturing how a mild genetic vulnerability may interact with normal developmental processes such that pathology only emerges around adolescence. This interplay between genetic susceptibility and normal adolescent development, both of which possess inherent individual variability, may contribute to heterogeneity seen in phenotypes in human neuropsychiatric disease.

Parsimonious modelling allows generation of the dendrograms of primate striatal medium spiny and pallidal type II neurons using a stochastic algorithm

Data from quantitative three-dimensional analysis of primate striatal medium spiny neurons (MSNs) and pallidal type I and type II neurons were used to search for possible rules underlying the dendritic architecture of these cells. Branching and terminating probabilities per unit length of dendrite were computed from all available measurement points. In the three neuronal groups, terminating probabilities were found to be exponentially increasing functions of the path distance to soma. MSNs and type II branching probabilities could be accurately modelled with decreasing functions of both the metrical (exponential functions) and topological (power functions of the centrifugal branch order) distances to soma. Additionally, type II branching also slightly depended on the distance to the proximal tip of the supporting branches. Type I branching probabilities did not follow these rules accurately. Embedding the modelled probability functions in a stochastic algorithm allowed generation of dendrograms close to those of the real MSNs and pallidal type II neurons, while the algorithm failed to simulate type I dendrites. MSN and pallidal type II neuron branching and terminating probabilities are thus highly dependent on the position in the dendritic arbor. This relationship can be modelled with simple functions and has a strong incidence on the dendrogram structure of the cells concerned. The additional dependence of the branching probability on the within-branch position led us to propose an extension of a previous modelling study by Nowakowski and co-workers which could account for a large range of topological and metrical (length) dendritic tree structures.

P4-394: Increased dendritic spine density in adult gerbil hippocampus following oral UMP and DHA supplementation

Loss of synaptic functions is a major pathological feature of Alzheimer's disease and related neurocognitive disorders. Developing therapeutic strategies to help prevent the degeneration of synapses may be essential to establish an effective remedial program. Several previous studies suggest that dietary supplementation of membrane phospholipid precursors may provide a way for developing such strategies. To explore this possibility, we previously treated normal adult gerbils by orally administering phospholipid precursors, uridine–5'–monophosphate (UMP, a uridine source) and/or docosahexaenoic acid (DHA) daily for 4 weeks. Animals that received the precursors were found to exhibit significant increases in all of the major brain membrane phospholipids, for example, phosphatidylcholine by 40–45% (Wurtman, Ulus, Cansev, et al., ASBMB abstract, San Francisco, CA, 2006). The precursors also increased expression levels of synaptic proteins, such as PSD–95 (38%) and Synapsin–1 (41%). In this study, we investigated the effect of UMP and DHA on neuronal morphology. Normal adult gerbils were given a diet supplemented with UMP (240 mg uridine/kg) and DHA (300 mg/kg, by gavage) daily for up to 2 weeks. Animals were decapitated, and fixed hippocampal slices were stained with DiI at the end of the treatment. Images of hippocampal neurons were obtained by two–photon microscopy. We found that in animals that received the precursors, dendritic spine density (i.e., the number of spines per unit length of dendrite) was increased significantly in CA1 pyramidal neurons in the hippocampus (27% increase, p = .001 vs. control group given a regular, unsupplemented diet). Our findings suggest that administering a uridine source and DHA can enhance neuronal properties associated with synaptic functions. These findings may be explained in part by enhancement of neuronal membrane synthesis with substrate saturation of enzymes that mediate the precursor incorporation into phosphatidylcholine, and in part by activation of UTP receptors, such as R2Y receptors. Testing of possible therapeutic applications of such precursor supplementation is currently underway.

Adult female and male zebra finches show distinct patterns of spine deficits in an auditory area and in the song system when reared without exposure to normal adult song

Male songbirds typically require exposure to normal adult conspecific song during development in order to learn a normal song of their own. Females require exposure to conspecific song during development in order to select high-quality, learned song over the incomplete song produced by males reared in isolation. Altering males' opportunity for song learning during development affects the neuroanatomy of brain regions involved in song production (the song system), but in females the neural effects of song learning are unknown. We raised male and female zebra finches (Taeniopygia guttata) with differing amounts of exposure to singing males during development. At 120 days, we Golgi-stained their brains and measured the frequency of dendritic spines in brain areas used in song perception or production. We found that females reared with little or no exposure to song have 31% fewer dendritic spines per unit length of dendrite in caudomedial nidopallium (NCM), a brain area activated by song perception, compared to control females. The deprived females had small deficits in the frequency of spines in HVC, a region activated by song production in males. Males with limited exposure to song had a 24% lower spine density in HVC than controls but only a 10% lower density in NCM. These data support the hypothesis that NCM is important in auditory learning, while HVC is involved in sensorimotor learning, and that these capacities are differentially emphasized in the two sexes.

Age-dependence in the homeostatic upregulation of hippocampal dendritic spine number during blocked synaptic transmission

Homeostatic regulation of spine number in mature hippocampal neurons results in more dendritic spines when synaptic transmission is blocked, providing a mechanism to compensate for diminished synaptic input. It is unsettled whether blockade of synaptic transmission also elevates spine number during development. To address this question, synaptic transmission was blocked in rat hippocampal slices during critical developmental stages of spine formation at postnatal days (P) 6–P22 and compared to adults. CA1 pyramidal cells were labeled with DiI and maintained for 5 h in one of three conditions, control artificial cerebrospinal fluid (ACSF), block media containing synaptic transmission antagonists in ACSF, or block media containing synaptic transmission antagonists in a nominally calcium-free ACSF with high magnesium. Slices were fixed in mixed aldehydes, sectioned, and the lateral dendrites were imaged in stratum radiatum with confocal microscopy. Dendritic spine density was quantified per unit length of dendrite. At P6–7 there were only a few protrusions emerging from the dendrites, which were predominantly filopodia-like in appearance. At both P11–12 and P15–16 there was a mixture of dendritic spines and filopodia-like structures. By P20–22 dendritic spines predominated and spine density was about 82% of the adult level. Dendritic spine density increased during blocked synaptic transmission at P20–22 as in adults, but was unchanged during blockade at younger ages. When extracellular calcium was nominally zero, dendritic spine density further increased on P20–22 dendrites as in adults. In contrast, spine density decreased along P11–12 dendrites under the nominally zero calcium condition. Under control conditions, dendritic protrusions were longer at P6–7 than at all other ages, which did not differ from one another. When synaptic transmission was blocked, dendritic protrusions further elongated at P6–7 only. Under the nominally zero calcium condition with blocked synaptic transmission, dendritic protrusions shortened at P11–12 only. These findings reveal age-dependent changes in the manifestation of homeostatic control of dendritic spines that could be mediated by maturational changes in mechanisms regulating postsynaptic calcium.

A morphological correlate of synaptic scaling in visual cortex.

We studied the response of dendritic spines in the thalamic-recipient zone of rat visual cortex to simple manipulations of the visual environment. We measured the morphologies of a total of 3824 spines located on the basal dendrites of 60 layer 3 pyramidal cells. As expected from previous studies, we found a significantly lower spine density in dark-reared animals at postnatal day 30 (P30) compared with light-reared controls. Additional analysis revealed that the spines in dark-reared animals were significantly shorter and more bulbous than in light-reared animals. When these two results were combined, we found that the total synaptic area per unit length of dendrite was conserved, compatible with the phenomenon of "synaptic scaling." We also found that the increase in average spine head diameter is reversed by 10 d of light exposure (starting at P20), but surprisingly, the decrease in spine density is not. Thus, not all effects of dark rearing can be reversed by subsequent visual experience, even when the experience occurs during the third postnatal week.

Surface expression and co-localization of NMDA receptor and AMPA receptor on dendritic tree of hippocampal neurons in culture

To investigate the developmental profiles on surface expression and co-localization of NMDA receptor clusters and AMPA receptor clusters on dendrite in cultured hippocampal neurons of rats. Green fluorescent protein tagged GluR2 subunit (GFP-GluR2) and FLAG tagged NR2B subunit (FLAG-NR2B) were transfected into cultured hippocampal neurons at 5 days in vitro (DIV5). FLAG-NR2B containing NMDA receptor clusters and GFP-GluR2 containing AMPA receptor clusters expressed on membrane surface were then labeled in living neurons using anti FLAG mAb/Cy3-conjugated anti-mouse antibody and anti-GFP pAb/Alex488-conjugated anti-rabbit antibody. The numbers of receptor cluster per 100 microm dendrite in the neurons at DIV7 and DIV19 were 39.7+/-5.0 and 64.7+/-6.1 (P<0.01) for NR2B-NMDAR, 59.1+/-3.3 and 99.7+/-6.4 (P<0.01) for GluR2-AMPAR, and 29.9+/-4.5 and 37.5+/-2.5(P<0.05) for the co-localized, respectively. At DIV7 and DIV19, 75.4% and 57.9% NR2B-NMDAR clusters were co-localized with GluR2-AMPAR; and 50.6% and 37.6% GluR2-AMPAR clusters were co-localized with NR2B-NMDAR, respectively. The density of NR2B-NMDAR containing and GluR2-AMPAR containing receptor clusters increases during development of hippocampal neurons in culture. Although the co-localized clusters are increased as well in an unit length of dendrite, the extent to which the two receptor clusters are co-localized decreases. These data imply a possible change in the partnership of AMPA receptor subtype and NMDA receptor subtype at newly formed synapses during development.

Stability in Synapse Number and Size at 2 Hr after Long-Term Potentiation in Hippocampal Area CA1

Long-term potentiation (LTP) is an important model for examining synaptic mechanisms of learning and memory. A key question is whether the enhanced synaptic transmission occurring with LTP involves the addition of new synapses, the enlargement of existing synapses, or a redistribution in synaptic weight among synapses. Two experimental designs were used to address this question. In the first experimental design three conditions were evaluated across hippocampal slices maintained in vitro, including slices with LTP analyzed at 2 hr post-tetanus, slices tetanized in the presence of APV, and control slices receiving test stimulation only. In the second experimental design independent LTP and control (low-frequency stimulation) sites were examined. Synapse density was estimated by an unbiased volume sampling procedure. Synapse size was computed by three-dimensional reconstruction from serial electron microscopy (EM). Serial EM also was used to compute synapse number per unit length of dendrite. In both experimental designs there were no significant effects of LTP on total synapse number, on the distribution of different types of synapses (thin, mushroom, stubby, or branched dendritic spines and macular, perforated, or segmented postsynaptic densities), on the frequency of shaft synapses, nor on the relative proportion of single or multiple synapse axonal boutons. There was also no increase in synapse size. These results suggest that LTP does not cause an overall formation of new synapses nor an enlargement of synapses at 2 hr post-tetanus in hippocampal area CA1, and these results support the hypothesis that LTP could involve a redistribution of synaptic weights among existing synapses.

Dendritic growth of arcuate neuroendocrine neurons following orchidectomy in adult rats

Recent studies have shown that changes in dendritic architecture are an important component of functional plasticity in the adult central nervous system. In the present study, we determined whether gonadectomy induces changes in dendritic architecture in the arcuate nucleus, a target tissue for gonadal hormones. A combination of retrograde labeling with systemically injected Fluoro-Gold and intracellular injection of neurons in a fixed-slice preparation was used to examine the morphology of neuroendocrine neurons in the rat arcuate nucleus. Intracellullary filled arcuate neuroendocrine neurons (8-21 neurons per brain) from intact (n = 5) and orchidectomized (n = 5) animals were reconstructed with the aid of a computer microscope. A quantitative analysis revealed that orchidectomy had no effect on the number and distribution of Fluoro-Gold-labeled neuroendocrine neurons in the rat arcuate nucleus. The morphology of arcuate neuroendocrine neurons in intact animals was relatively simple, with the majority of neurons (79%) having only two primary dendrites and few dendritic spines. Compared with intact controls, arcuate neuroendocrine neurons in the orchidectomized group had significantly larger somatic profile areas and exhibited significant increases in dendrite length, dendrite volume, terminal branch number, and spines per unit length of dendrite. The increase in terminal branch number in orchidectomized animals was due primarily to the appearance of short branches that gave a striking, claw-like appearance to many of the distal dendrites. These results provide evidence for hormonal regulation of dendritic morphology of arcuate neuroendocrine neurons in adult mammals.

Recovery of function is associated with increased spine density in cortical pyramidal cells after frontal lesions and/or noradrenaline depletion in neonatal rats

Rats were given medial frontal lesions at 7 days of age and were tested as adults on tests of forelimb use, forelimb tactile sensitivity, tongue use, hindleg use, and in a spatial navigation task. The brains were processed with a modified Golgi-Cox procedure and dendritic arborization and spine density was measured. The animals showed recovery only on the spatial task and this was associated with an increase in the number of spines per unit length of dendrite. We also reanalyzed Golgi-Cox stained material from an experiment in which animals were depleted of cortical noradrenaline (NA) in infancy and then given frontal lesions on day 7. The NA depletion blocked the recovery from frontal lesions. Analysis of dendritic morphology showed that in otherwise intact rats, NA depletion decreased dendritic arbor but increased spine density to the level of frontal operates. Depleted frontal-operates showed no additional increase in spine density and also showed a decrease in dendritic arborization. These results suggest that recovery from neonatal cortical injury and from neonatal noradrenaline depletion may be supported by changes in both the dendritic arborization and the spine density in the remaining cortex.

Autoradiographic evidence that NMDA receptor-coupled channels are located postsynaptically and not presynaptically in the perforant path-dentate granule cell system of the rat hippocampal formation

In vitro quantitative autoradiography with [ 3H]MK-801 was used to determine K d and B max values for the NMDA receptor-coupled channel in subregions of the rat hippocampal formation. A single form of the channel with an apparent K d in the 15–20 nM range was found for [ 3H]MK-801 binding in the presence of both 1μM glutamate and 1μM glycine. Specific binding was highest in the molecular layer of the dentate gyrus, followed by CA1 stratum radiatum and CA1 stratum oriens. Fewer binding sites were observed in the hilus of the dentate gyrus, cerebral cortex, CA1 stratum pyramidale. CA3 subregion (stratum oriens, stratum pyramidale, stratum radiatum), and thalamus. Selective destruction of dentate granule cells by colchicine microinjections reduced the amount of specific [ 3H]MK-801 binding by half in the molecular layer of the dentate, compared to intact tissue. [ 3H]MK-801 binding did not change in other hippocampal subregions as a consequence of colchicine injection. Electrolytic entorhinal cortical lesions produced no changes in regional MK-801 binding site density in any of the regions under study. To address the tissue shrinkage following entorhinal cortex lesions, detailed analysis of the binding site density per fixed (16 μm) length of granule cell dendrite, and of the aggregate density across the entire molecular layer revealed no change in the number of MK-801 binding sites per unit length of dendrite in the molecular layer of the dentate gyrus. These findings indicate that NMDA receptor-coupled channels are confined to a postsynaptic location in the perforant path-dentate granule cell system of the adult rat.

Distribution of corticocortical and thalamocortical synapses on identified motor cortical neurons in the cat: Golgi, electron microscopic and degeneration study

Details of the terminal connections of corticocortical and thalamocortical fibers on pyramidal and stellate neurons in the cat motor cortex were studied using the electron microscope in combination with the Golgi and axonal degeneration techniques. Corticocortical terminals were examined in 23 identified neurons of which 11 were pyramidal and 12 were stellate. Stellate neurons located in layer III received many degenerating terminals (average8.4 ± 2.2 per unit length of dendrite (ULD) and the majority of these (95%) were found on the proximal dendrites or on the cell bodies. The pyramidal neurons received fewer degenerating terminals (average2.1 ± 0.27/ULD) and these were located on more distal dendritic shafts or on dendritic spines. The majority of these synapses were of the asymmetric type. Thalamocortical terminals were examined in 9 pyramidal and 9 stellate neurons. Pyramidal neurons received many terminals (average6.0 ± 1.23/ULD) and these were found on the basal as well as the apical dendrites and on dendritic spines. Stellate neurons received fewer terminals (average4.2 ± 0.64/ULD) and were located primarily on proximal dendritic shafts. The majority of these synapses were of the asymmetric type. The functional role of these synapses is discussed in relation to the physiological results reported in the preceding paper.