Morphological Changes Of Synapses Research Articles

Administration of miR-195 Inhibitor Enhances Memory Function Through Improving Synaptic Degradation and Mitochondrial Dysfunction of the Hippocampal Neurons in SAMP8 Mice.

Mitochondrial dysfunction is an early feature of Alzheimer's disease (AD) and miR-195 is involved in mitochondrial disorder through targeting MFN-2 protein in hippocampal neurons of AD. To clarify if administration of miR-195 inhibitor could enhance the memory deficits through improving hippocampal neuron mitochondrial dysfunction in SAMP8 mice. The expression of miR-195 was detected by RT-qPCR in primary hippocampal neurons and HT-22 cells treated with Aβ1-42. Morris water maze (MWM) was used to assess the learning and memory function in SAMP8 mice administrated with antagomir-195. Transmission electron microscopy was employed to determine the morphological changes of synapses and mitochondria of hippocampus in SAMP8 mice. Mitochondrial respiration was measured using a high-resolution oxygraph. The expression of miR-195 were upregulated in the primary hippocampal neurons and HT-22 cells induced by Aβ1-42. Inhibition of miR-195 ameliorated the mitochondrial dysfunction in HT-22 cells induced by Aβ1-42, including mitochondrial morphologic damages, mitochondrial membrane potential, respiration function, and ATP production. Administration of antagomir-195 by the third ventricle injection markedly ameliorated the cognitive function, postsynaptic density thickness, length of synaptic active area, mitochondrial aspect ratio, and area in hippocampus of SAMP8 mice. Finally, antagomir-195 was able to promote an increase in the activity of respiratory chain complex CI and II in SAMP8 mice. This study demonstrated that miR-195 inhibitor ameliorated the cognitive impairment of AD mice by improving mitochondrial structure damages and dysfunction in the hippocampal neurons, which provide an experimental basis for further exploring the treatment strategy of AD.

The stress sensitization of adult rat Increased by stress in childhood and the neural mechanism

Objective To observe the influence of stress in childhood on adult rat's stress sensitization and the neural mechanism. Methods Sixty male Sprague-Dawley rats were randomly divided into stress group (thirty) and non-stress group(thirty). The rats in stress group were given 14 repeated daily forced swimming. The emotional behaviors were tested at the beginning of the experiment and when stress of forced swimming ended 8 weeks later. When the appraisal of behavior ended,the pyramidal neuron in the left hippocampal CA3 regions were counted under the light microscope, the pictures of synapses in the left hippocampal CA3 regions were got under the electromicroscope,and the morphological changes of synapses were analyzed with the automatic image analyzing system. Results (1) The score (1.20±0.60) of emotional behavior of stress group increased more than non-stress group(-0.13±0.83) (P<0.05). (2) The number of pyramidal neuron in the left hippocampal CA3 re-gions of stress group rats was small than that of non-stress group rats. (3) The number density [(1.01±0.15) N/ μm3 ], prosop density (0.20±0.04 )μm2/μm3], the average area of synaptic linkage [(0.05±0.02) μm2], the length of active areas [(314.78±46.32) nm], the curvatures (1.02±0.04), the thickness of postsynaptic dense material [(46.35±6.81) nm] in the left hippocampal CA3 regions of the stress group were small than those of non-stress group (P<0.05). Conclusion The stress in childhood could increase the stress sensitization of adult rats,and the decrease of synaptic plasticity might be the mechanism. Key words: Forced swimming test; Stress sensitization; Synapse; Pyramidal neuron; Rats

Aprendizado e memória

Memory is broadly divided into declarative and nondeclarative forms of memory. The hippocampus is required for the formation of declarative memories, while a number of other brain regions including the striatum, amygdala and nucleus accumbens are involved in the formation of nondeclarative memories. The formation of all memories require morphological changes of synapses: new ones must be formed or old ones strengthened. These changes are thought to reflect the underlying cellular basis for persistent memories. Considerable advances have occurred over the last decade in our understanding of the molecular bases of how these memories are formed. A key regulator of synaptic plasticity is a signaling pathway that includes the mitogen activated protein (MAP) kinase. As this pathway is required for normal memory and learning, it is not surprising that mutations in members of this pathway lead to disruptions in learning. Neurofibromatosis, Coffin-Lowry syndrome and Rubinstein-Taybi syndrome are three examples of developmental disorders that have mutations in key components of the MAP kinase signaling pathway.

Remodeling of Synaptic Membranes after Induction of Long-Term Potentiation

Several morphological changes of synapses have been reported to be associated with the induction of long-term potentiation (LTP) in the CA1 hippocampus, including an transient increase in the proportion of synapses with perforated postsynaptic densities (PSDs) and a later occurrence of multiple spine boutons (MSBs) in which the two spines arise from the same dendrite. To investigate the functional significance of these modifications, we analyzed single sections and reconstructed 134 synapses labeled via activity using a calcium precipitation approach. Analyses of labeled spine profiles showed changes of the spine head area, PSD length, and proportion of spine profiles containing a coated vesicle that reflected variations in the relative proportion of different types of synapses. Three-dimensional reconstruction indicated that the increase of perforated spine profiles observed 30 min after LTP induction essentially resulted from synapses exhibiting segmented, completely partitioned PSDs. These synapses had spine head and PSD areas approximately three times larger than those of simple synapses. They contained coated vesicles in a much higher proportion than that of any other type of synapse and exhibited large spinules associated with the PSD. Also the MSBs with two spines arising from the same dendrite that were observed 1-2 hr after LTP induction included a spine that was smaller and a PSD that was smaller than those of simple synapses. These results support the idea that LTP induction is associated with an enhanced recycling of synaptic membrane and that this process could underlie the formation of synapses with segmented PSDs and eventually result in the formation of a new, immature spine.

Morphological changes of synapses induced by urinary stimulation in the hamster accessory olfactory bulb.

Urinary pheromones are considered to regulate reproductive functions in various rodent species. The effects of urinary stimuli on synaptic plasticity in the accessory olfactory bulb (AOB), which is the primary nucleus of the vomeronasal system, were studied. Adult male hamsters were divided into four groups and each group was exposed to one of the following four materials: distilled water, female hamster urine, female rat urine, and male hamster urine. After 15 days, the sizes of synapses in the glomerular and the mitral/tufted (MT) cell layers of the AOB were measured. The glomerular synapses, located between the axons of sensory cells and the dendrites of MT cells, were larger in the groups exposed to either the female hamster or the female rat urine compared with those for the distilled water and male hamster urine groups. In the MT cell layer, the synapses are of two types: asymmetrical excitatory synapses and symmetrical inhibitory ones. Exposure of adult male hamsters to female hamster urine induced a reduction in the size of asymmetrical synapses, while on exposure to other kinds of urine there was no synaptic change. The sizes of the symmetrical synapses were not changed by any urinary stimulus. The present study revealed that morphological changes of synapses in the AOB were induced by urinary stimuli. Different urines induced different morphological responses. It is suggested that this synaptic plasticity is responsible for regulation of the output of pheromonal information from the AOB to the higher centers of the vomeronasal system.

Synaptic vesicle ultrastructural changes in the rat hippocampus induced by a combination of alpha-linolenate deficiency and a learning task.

Rats fed either a safflower oil (alpha-linolenate-deficient) or a perilla oil (alpha-linolenate-sufficient) diet through two generations (F1) showed significant differences in the brightness-discrimination learning task. In this task, correct responses were lever-pressing responses, which were reinforced with dietary pellets, and incorrect responses were those with no reinforcement. The inferior learning performance in the safflower oil group was caused mainly by the inferior ability to rectify the incorrect responses through the learning sessions. In the safflower oil group after the learning task, the average densities of synaptic vesicles in the terminals of the hippocampus CA1 region were decreased by nearly 30% as compared with those in the perilla oil group, and it is notable that this difference was not detected without the learning task. These results suggest that dietary oil-induced morphological changes in synapses in the hippocampus of rats are related to the differential learning performance and that the turnover rate of synaptic vesicles in the hippocampus may be an important factor affecting learning performance.

1416 Morphological changes of synapses in the mice accessory olfactory bulb associated with olfactory memory

1416 Morphological changes of synapses in the mice accessory olfactory bulb associated with olfactory memory

The isolation of novel intracellular factors by differential screening methods

Synaptic plasticity, a physiological basis of learning and memory, is mainly classified into two categories: 1) relatively short-term changes in electrical activities and 2) more long-lasting morphological changes in synapses. Studies on neuronal differentiation have provided detailed clarification of many of the morphological changes in synapses. Although it has been demonstrated that neuronal differentiation is induced by a variety of stimuli, the mechanism of neuronal differentiation has never been sequentially understood. Since there must be unknown factors relevant to these complicated processes, it is important to find and identify the novel intracellular factors that are able to induce the differentiation of neurons. Differential screening is useful cloning method to identify molecules without any information about their structures. Genes expressed in a distinct pattern among two or more groups, eg. different drug-treated cells, tissues and so on, can be isolated. To identify novel neuronal differentiation factors, we differentially screened approximately 500,000 primary clones from the cDNA library of NG108-15 cells treated with TPA and diBu-cAMP for 72 hr. Using two single strand cDNA probes, which were reverse-transcribed from poly(A)+ RNA, TA-20 was isolated from cells treated with TPA and diBu-cAMP (probe TA) or from cells treated with diBu-cAMP alone (probe A) for 72 hr. Clones that hybridized preferentially to the probe TA were further investigated by Southern and Northern blots. Thus the identified clone TA20 is a novel gene and plays functional roles as a neuronal differentiation factor.

Morphological changes in cultures of hippocampus following prenatal irradiation in the rat

The effect of prenatal irradiation was studied in organotypic cultures of hippocampus, prepared from newborn rats that had been exposed to whole-body irradiation of 1 Gy from a 60Co-source at day 13 of pregnancy. Light and electron microscopic observations showed remarkable damage to neuronal mitochondria accompanied by extensive swelling, vacuolation of the Golgi complex, and formation of multilamellar bodies and vesicles of the lysosomal type. In contrast to neuronal alterations, no delay in synaptogenesis or onset of myelination was observed based upon the absence of significant morphological changes in synapses and myelin sheaths. Using this tissue culture model it could be confirmed that prenatal exposure to irradiation, even at low doses, induces specific morphological changes in the brain.

Rapid introduction of long-lasting synaptic changes at crustacean neuromuscular junctions.

In this review we present recent evidence implicating second-messenger systems in two forms of long-lasting synaptic change seen at crustacean neuromuscular junctions. Crustacean motor axons are endowed with numerous terminals, each possessing many individual synapses. Some synapses appear to be quiescent or impotent, but can be recruited in response to imposed functional demands. Supernormal impulse activity leads to long-term facilitation (LTF) which persists for many hours. During the persistent phase, additional synapses are physiologically effective, and morphological changes in synapses are seen at the ultrastructural level. Pulsatile application of serotonin, a neuromodulator, also enhances synaptic transmission, but this enhancement declines more rapidly than LTF. Elevation of intraterminal Ca2+ is neither necessary nor sufficient for long-lasting enhancement of transmission, but activation of A-kinase is necessary. LTF is set in motion by an unknown depolarization-dependent mechanism leading to A-kinase activation, whereas serotonin facilitation depends for its initiation on the phosphatidylinositol system. The initial phase of serotonin facilitation may be accounted for by production of inositol triphosphate, whereas the secondary long-lasting phase appears to require participation of both C kinase and A kinase. Neither LTF nor serotonin facilitation requires an intact neuron; both are presynaptic phenomena expressed by the nerve terminals. Brief comparison is made with long-lasting synaptic changes in other systems.

Glial contribution to amino acid content and metabolism of the deafferented dentate gyrus.

The time course of tissue content and evoked release of endogenous amino acids was analyzed in the partially deafferented dentate gyrus of the rat hippocampus 2-24 days following unilateral lesion of the perforant path. Amino acids in tissue extracts and perfusates were determined after precolumn derivatization and hplc separation. The astrocytic glial cell reaction was monitored with immunohistochemistry of S-100. The tissue content of glutamate decreased significantly on the lesioned side, whereas only a moderate reduction in taurine, aspartate, and alanine occurred. Glutamine was significantly elevated at 7 days. The evoked efflux of glutamate was reduced at 2 and 7 days, whereas no change was seen at longer survival periods. The evoked release of GABA and aspartate increased on the denervated side after 12 and 24 days. The rate of carbon utilization into amino acid pools was followed with 14C-glucose and 14C-acetate. The incorporation of acetate showed a peak 2-9 days following lesion, which paralleled in time the hypertrophic glial cells. The incorporation of glucose decreased during this period. The metabolic events are discussed in relation to the morphological changes in synapses and glial cells.

Use of aldehyde fixatives to determine the rate of synaptic transmitter release.

Aldehyde fixation continues to be useful to prepare synapses for freeze-fracture, but it may increase the rate of transmitter release. The effects of different aldehyde fixatives on spontaneous quantal release (m.e.p.p.s), and on the corresponding synaptic vesicle exocytosis at frog nerve-muscle synapses were investigated with the hope of finding a way to minimize side effects of fixation. Increases in m.e.p.p.s of up to 50 S-1 occurred during fixation, despite the species of aldehyde used in the fixative, and this fixative effect decreased only slightly as aldehyde concentration was increased. Increases in m.e.p.p frequency were not blocked by tetrodotoxin, by lowering external calcium and raising external magnesium concentration, or by lowering the total osmotic strength of the fixative. The smallest increase in m.e.p.p. frequency was in 3% glutaraldehyde and corresponded to the lowest level of synaptic vesicle exocytosis seen by freeze-fracture, 0.15 per micrometers of active zone. The effects of aldehyde fixation on m.e.p.p. frequency and synaptic vesicle exocytosis could not be avoided, but this study suggests how its effect on morphological changes in synapses might be minimized.