Postsynaptically silent synapses in single neuron cultures

We have used the synapses that isolated hippocampal cells in culture form onto themselves (autapses) to determine if some synapses lack functional AMPA receptors (AMPARs). A comparison of the synaptic variability of the AMPAR- and NMDAR-mediated evoked responses, as well as of miniature synaptic responses, indicates that a population of events exists that only contains an NMDAR component. Spillover of glutamate from adjacent synapses cannot explain these results because in single cell cultures all synaptic events mediated by AMPARs should be detected. Immunocytochemical analysis of these cultures clearly reveals a population of synapses with puncta for NR1 (NMDAR) but not for GluR1 (AMPAR). These results provide strong anatomical and physiological evidence for the existence of postsynaptically silent synapses.     

Inhibition as binding controller at the single neuron level

Natural stimulus for a neuron is a sum of large number of unitary excitatory postsynaptic potentials (EPSP) slightly dispersed in time. We analyze, based on numerical solution of the Hodgkin and Huxley equations, how does the triggering ability of the compound stimulus depend on the relative timing of the EPSPs it comprises. The dependences found suggest that a neuron stimulated from many synaptic inputs can be treated as performing elementary binding function and that inhibition serves as a controller of that kind of binding. The transient process characterized by EPSP operates in this context as a short-term memory mechanism inherent to a single neuron.     

Computer analysis of single neuron activity during conditioned feeding task

Human recombinant tumor necrosis factors alpha and beta (TNF-alpha and TNF-beta), at and above 1 ng/ml (approximately equal to 70 pM), caused a dose- and time-dependent enhancement of 45Ca release from neonatal mouse calvarial bones in vitro. In addition, TNF-alpha and TNF-beta (3-100 ng/ml) caused a dose-dependent stimulation of prostaglandin E2 (PGE2) formation in the calvarial bones. TNF-alpha also enhanced the biosynthesis of PGI2, as assessed by analysis of the stable breakdown product 6-keto-PGF1 alpha. The stimulatory actions of TNF-alpha and TNF-beta on PGE2 formation was maximal at 12 h. Indomethacin, flurbiprofen, and meclofenamic acid, three structurally unrelated nonsteroidal antiinflammatory drugs, abolished PGE2 biosynthesis induced by TNF-alpha and TNF-beta (100 ng/ml). The 45Ca release stimulated by TNF-alpha and TNF-beta (100 ng/ml), however, was only slightly reduced by indomethacin, flurbiprofen, and meclofenamic acid. The partial inhibitory effect of indomethacin on 45Ca release was seen over a wide range of TNF-alpha concentrations, without affecting the concentration producing half-maximal stimulatory response. TNF-alpha and TNF-beta (100 ng/ml) stimulated bone matrix breakdown, as assessed by analysis of the release of 3H from bone prelabeled with [3H]proline. Also, the stimulatory effect of TNF-alpha and TNF-beta on bone matrix degradation was partially reduced by indomethacin. Hydrocortisone (1 microM) and dexamethasone (0.1 microM) abolished TNF-alpha- and TNF-beta-induced production of PGE2. In contrast to the cyclooxygenase inhibitors, the corticosteroids did not affect the stimulatory action by the cytokines on 45Ca release.(ABSTRACT TRUNCATED AT 250 WORDS)     

Synapse-specific control of synaptic efficacy at the terminals of a single neuron

The regulation of synaptic efficacy is essential for the proper functioning of neural circuits. If synaptic gain is set too high or too low, cells are either activated inappropriately or remain silent. There is extra complexity because synapses are not static, but form, retract, expand, strengthen, and weaken throughout life. Homeostatic regulatory mechanisms that control synaptic efficacy presumably exist to ensure that neurons remain functional within a meaningful physiological range. One of the best defined systems for analysis of the mechanisms that regulate synaptic efficacy is the neuromuscular junction. It has been shown, in organisms ranging from insects to humans, that changes in synaptic efficacy are tightly coupled to changes in muscle size during development. It has been proposed that a signal from muscle to motor neuron maintains this coupling. Here we show, by genetically manipulating muscle innervation, that there are two independent mechanisms by which muscle regulates synaptic efficacy at the terminals of single motor neurons. Increased muscle innervation results in a compensatory, target-specific decrease in presynaptic transmitter release, implying a retrograde regulation of presynaptic release. Decreased muscle innervation results in a compensatory increase in quantal size.     

Sequence of single neuron changes in CA1 hippocampus of rabbits during acquisition of trace eyeblink conditioned responses

The sequence of changes in single neuron activity in the CA1 area of the rabbit hippocampus was examined during daily sessions (80 trials/session) of hippocampally dependent nonspatial trace eyeblink (i.e., nictitating membrane response) conditioning. Each trial for trace conditioned animals (n = 7) consisted of a tone conditioned stimulus (CS; 6 kHz; 90 dB, 100 ms) followed by a 500-ms silent trace period, then a corneal airpuff unconditioned stimulus (US; 3.0 psi; 150 ms). Control animals (n = 5) received unpaired CSs and USs. Most pyramidal (n = 309) and theta (n = 21) cells were recorded for a single day of training. The activity of cells for each animal were grouped according to: the day of training that CRs began to increase and the day of training that CR performance became asymptotic. Pyramidal cells from trace conditioned animals demonstrated several stages of learning-related activity: large increases in activity after both the CS and US early in conditioning on the day of training when CRs began to increase, smaller moderate increases in activity on the following days of training, and decreases in activity after the US during asymptotic CRs. Pyramidal cell-increases declined significantly across the trials of each daily session. Theta cells showed an activity pattern opposite to the pyramidal cells, consistent with the notion that theta cells have an inhibitory influence on pyramidal cells. Single pyramidal cells also were categorized into response profiles. Most pyramidal response profiles showed increases in activity specific to the day of initial CRs. Two of the pyramidal response profiles may be involved in assessing the temporal properties of the CS-US trace conditioning trial.     

The chronometry of single neuron activity: Testing discrete and continuous models of information processing.

The authors propose to study information transmission by comparing the effects of experimental factors on reaction time (RT) with the latency of the changes in activity of single-neurons. An experiment was conducted in which a monkey (Macaca mulatta) performed a tactilo-manual 2-choice RT task and the compatibility of the stimulus–response mapping was manipulated. Task-related neurons were recorded in the monkey's primary somesthetic and motor cortices. The changes in activity of 105 of these neurons were classified either as sensory-like or as motor-like. The sensory-like changes occurred before the motor-like ones. The stimulus–response mapping exerted its entire effect on the RT after the sensory-like changes and before the motor-like ones. These findings suggest that the information was transmitted discretely from the processes affected by the mapping to the processes implemented by the motor-like changes. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

A single amino acid substitution in yeast eIF-5A results in mRNA stabilization

Most factors known to function in mRNA turnover are not essential for cell viability. To identify essential factors, approximately 4000 temperature-sensitive yeast strains were screened for an increase in the level of the unstable CYH2 pre-mRNA. At the non-permissive temperature, five mutants exhibited decreased decay rates of the CYH2 pre-mRNA and mRNA, and the STE2, URA5 and PAB1 mRNAs. Of these, the mutant ts1159 had the most extensive phenotype. Expression of the TIF51A gene (encoding eIF-5A) complemented the temperature-sensitive growth and mRNA decay phenotypes of ts1159. The tif51A allele was rescued from these cells and shown to encode a serine to proline change within a predicted alpha-helical segment of the protein. ts1159 also exhibited an approximately 30% decrease in protein synthesis at the restrictive temperature. Measurement of amino acid incorporation in wild-type cells incubated with increasing amounts of cycloheximide demonstrated that a decrease in protein synthesis of this magnitude could not account for the full extent of the mRNA decay defects observed in ts1159. Interestingly, the ts1159 cells accumulated uncapped mRNAs at the non-permissive temperature. These results suggest that eIF-5A plays a role in mRNA turnover, perhaps acting downstream of decapping.     

Amplification of specific mRNA from a single human renal glomerulus, with an approach to the separation of epithelial cell mRNA

PURPOSE: To evaluate endothelial cell morphology and density after excimer laser photorefractive keratectomy (PRK). METHODS: We used a noncontact specular microscope to examine the central corneal endothelium of 50 eyes of 50 patients who had PRK for an attempted correction between -2.5 and -17.0 diopters (D) (mean -7.8 D) beginning 18 to 24 hours postoperatively. RESULTS: After a follow-up of 11.4 +/- 6.1 months (mean +/- standard deviation), mean endothelial cell density was 2577.6 +/- 402.0 cells/mm2 with rare signs of polymegathism and pleomorphism. Preoperative and untreated fellow eye endothelial cell density values were used as a control. Paired Student's t-test and analysis of variance results were not significant (P > .05). CONCLUSION: Excimer laser PRK did not significantly change cell density and morphology.     

Analysis of the response of a pacemaker neuron model to transient inputs

The response of a pacemaker neuron model to a train of transient inhibitory impulsive perturbations is examined. The model reproduces the heterogeneous discharge forms and abrupt switchings displayed by the crayfish slowly adapting stretch receptor organ (Segundo et al., 1994, Neuroscience 62(2), pp. 459-480). The non-monotonous aspect of the instantaneous firing rate of the model reflects the fact that in some regimes input and output rates are both increasing, despite the inhibitory effect of the former. We determine how such paradoxical acceleration takes place by analyzing the response of the model using its phase transition curve. We show that paradoxical acceleration results from the fact that the phase transition curve exhibits a large slowly increasing, almost linear section similar to that of living preparations.     

Superposition of binaural influences on single neuron activity in the medial superior olive elicited by electrical stimulation of the osseous spiral laminae

The single unit response of medial superior olive (accessory nucleus) neurons was investigated, when elicited by electrical stimulation of nervous processes in the osseous spiral laminae in both cochleae and also by antidromic stimulation from the inferior colliculi. The osseous spiral laminae were stimulated at the first, second or third turns with grounding of one of the unstimulated turns. Time of signal arrival differences (delta tau) at the two cochleae and also intensity differences (deltai) were varied. (2) It is concluded that (A) binaural interaction in the MSO is not based on totally excitatory or inhibitory influences from either ear, but on cycles of excitation and inhibition; (B) these influences approximately obey the laws of superposition, i.e., the influences add linearly to determine discharge. (3) It is demonstrated that: (a) electrical stimulation, unlike acoustical stimulation, elicits a regular neural response; (b) the neural response functions elicited by changes in delta tau are deltai exhibit maxima and minima and are not smooth functions; (c) fine latency changes in the neural response are correlated with changes in delta tau and deltai; (d) the stimulation of different turns of the osseous spiral laminae (producing different current flows) elicits a different neural response depending on the turn stimulated, an effect reflecting the anatomical tonotopic arrangement of the medial superior olive; (e) changes in the stimulus differences delta tau and deltai are correlated with the ratio of the spike count in a short period following the stimulus, to the spike count in a longer total period; (f) changes in the stimulus differences delta tau and deltai elicit a neural response which varies according to the turn stimulated.     

Analysis of single trials in pigeons' timing performance.

Response distributions of pigeons on the peak procedure, averaged over many trials, were well fitted by a Gaussian function plus either a straight line or a ramp. But, each of 11 distributions analyzed showed a systematic positive skew. Response distributions on individual trials showed break–run–break patterns with abrupt transitions in the rate of responding. Within the run phase was an inner run phase at yet a higher rate. Intercorrelations among the starting time, ending time, middle, duration, and rate of the run showed a similar pattern in the outer and inner runs. A positive correlation between starting and ending times suggests variance across trials in clock speed, delay to start the clock, or expected time of reward. A negative correlation between starting time and duration suggests variance across trials in the threshold or thresholds to start and end the run. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Motor neuron syndromes in cancer patients

Previous reports indicate that motor neuron disease (MND) may rarely be associated with systemic cancer. We have encountered 14 patients with MND and cancer who formed three distinct groups. Group 1: Three patients developed a rapidly progressive MND, less prominent symptoms of involvement of other areas of the nervous system, and anti-Hu antibodies. Group 2: Five women developed signs of upper motor neuron (UMN) disease, initially resembling primary lateral sclerosis (PLS), and breast cancer. In 4, symptoms of UMN occurred within 3 months of cancer diagnosis or tumor recurrence. They had no metastases or spinal cord compression. Serum anti-neuronal antibodies were negative. Three patients are alive (follow-up of 156, 15, and 12 months), and 2 remain without lower motor neuron signs. Group 3: Six patients developed MND resembling amyotrophic lateral sclerosis between 47 months before and 48 months after their cancer diagnosis. In group 1, the MND associated with the anti-Hu antibody is unequivocally paraneoplastic. In group 2, the proximate onset of MND with the diagnosis of cancer or its recurrence, its pure or long-lasting UMN signs, and its association with breast cancer, suggest that the disorder may be paraneoplastic. Although for most cancer patients who develop MND the occurrence of both disorders is probably coincidental, in some patients with MND a careful search for an underlying cancer is warranted (ie, patients in groups 1 and 2).     

Oculomotor abnormalities in motor neuron disease

OBJECTIVE: To assess whether giving a leaflet containing norms of self-control to diabetics receiving insulin treatment results in a metabolic improvement. The HbA1c was established prior and subsequent to giving out the leaflet. The written norms contained in the leaflet were highlighted from the wider range of instructions obtained from diabetes education. DESIGN: A longitudinal intervention study with no random allocation. SETTING: Hospital care in a specialised Endocrinology clinic. PATIENTS AND OTHER PARTICIPANTS: 122 types 1 and 2 diabetics, treated with various insulin diets, who had received prior diabetes education and carried out blood self-controls. Cases of meta-diabetic syndrome, serious illnesses and those without apparent hypoglycaemia symptoms were excluded. INTERVENTIONS: Giving out of a leaflet with written norms of self-control. MEASUREMENTS AND MAIN RESULTS: The averages, standard deviations and 95% confidence intervals of the basal HbA1c and of those at four-monthly check-ups were calculated. Basal 8.07 (CI 7.75 - 8.39); 4th month 6.88 (CI 6.74 - 7.17); 8th month 6.59 CI 6.29 - 6.90); 12th month 6.60 (CI 6.23 - 6.94); 16th month 6.06 (CI 5.63 - 6.49); 20th month 5.40 (CI 3.04 - 7.75). Averages were compared by the Student T test and all values had p < 0.005 against the basal. CONCLUSIONS: Following the written norms produced a clear metabolic improvement, represented by the significant fall of HbA1c. Therefore, though without ignoring the overall context of treatment and diabetes education, it is proposed that the relationship of the most practical features of insulin dosing to written norms of self-control should be emphasised.     

Analysis of cytokine mRNA profiles in the lungs of Pneumocystis carinii-infected mice

Severe combined immunodeficient (scid) mice lack functional CD4+ lymphocytes, and therefore develop life-threatening Pneumocystis carinii infection. However, when scid mice are immunologically reconstituted with spleen cells, including CD4+ cells, a protective inflammatory response is mounted against the organism. To determine whether these lymphocytes induce elevated cytokine mRNA levels in response to P. carinii infection, steady-state levels of cytokine mRNAs were measured in the lungs of both reconstituted and unaltered scid mice. Despite significant numbers of organisms and the presence of functional alveolar macrophages in the lungs of 8- and 10-wk-old scid mice, there was neither evidence of pulmonary inflammation, nor increased proinflammatory cytokine expression. However, when 8-wk-old scid mice were immunologically reconstituted, signs of intense, focal pulmonary inflammation were observed, and levels of interleukin (IL)-1alpha, IL-1beta, IL-3, IL-6, interferon-gamma (IFN-gamma), tumor necrosis factor (TNF)-alpha, and TNF-beta mRNAs were all significantly elevated. Cytokine expression was increased at day 10 post-reconstitution (PR), maximal at day 12 PR, and returned to baseline by day 22 PR. In situ hybridization demonstrated that at day 12 PR, increased IL-1beta and TNF-alpha expression was localized to sites of intense inflammation and focal P. carinii colonization. Many of the cells expressing high levels of IL-1beta and TNF-alpha in these regions were in direct contact with organisms, or contained degraded organisms within their cytoplasm. Thus, even though functional macrophages are present in scid mice, CD4+ T cells are required for proinflammatory cytokine expression, which is associated with the generation of a protective inflammatory response at sites of P. carinii infection.     

Postradiation motor neuron syndrome

We report an unusual case of selective lower motor neuron syndrome (MNS) complicating whole neuraxis radiation therapy. Only three well-documented similar cases have been found in a thorough review of the literature. The syndrome has a stereotyped time course and is self-limited. We discuss here possible pathogenetic mechanisms and their relationship to motor neuron disease.     

Central motor conduction in motor neuron disease

Central motor conduction time (CMCT) to abductor digiti minimi (ADM) and tibialis anterior (TA) was measured in 21 patients of motor neuron disease (MND). In the upper limb, the motor pathways were inexcitable in 13 and central motor conduction time (CMCT-ADM) was prolonged in 7 sides. In the lower limbs the motor pathways were inexcitable in 10 and CMCT-TA was prolonged in 14 sides. The CMCT abnormalities did not follow a constant pattern but were randomly distributed and were asymmetric in the upper limbs in 7 and lower limbs in 3 patients. Asymmetric and randomly focal abnormalities in central motor conduction in our patients are consistent with asymmetric and focal neuronopathy in MND.