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Lipid metabolism is a key central, systemic and gut microbial feature of the decline in rat hippocampal function during middle age.

Middle age is emerging as a turning point in brain ageing, prognostic of future cognitive health and amenable to intervention. Metabolic and proteomic differences during this period are not yet fully understood and may potentially influence functions of the hippocampus, a brain area that regulates memory and anxiety. While the gut microbiota is implicated in brain ageing, the relationship between the gut microbiota, the metabolic state, and hippocampal proteome in middle age has not been investigated. We hypothesise that peripheral metabolic or protein features are associated with hippocampal vulnerability in middle age. Therefore, young adult and middle-aged rats were assessed for behavioural, proteomic, metabolic, and gut microbiota differences. Proteomic profiling of the hippocampus revealed differential expression of proteins indicative of altered synaptic signalling. Concurrently, adult hippocampal neurogenesis was decreased in middle age. Hippocampal microglia exhibited a lipid rich, inflammatory phenotype in middle age which correlated with poorer memory performance. CSF and serum proteomic and metabolomic analyses identified dysregulated lipid-related pathways potentially contributing to hippocampal vulnerability in middle age. Furthermore, 16S rRNA sequencing revealed reduced abundance of bacteria involved in lipid metabolism regulation. However, faecal microbiota transfer from young to middle aged rats was not sufficient to robustly improve hippocampus-dependent spatial memory. Together, these findings highlight dysfunctional lipid metabolism as a key feature of middle age that may contribute to decline in hippocampal function. Given that the scope for intervention is limited during older age, targeting biomarkers involved in metabolic and lipid homeostasis may be pivotal for the development of pharmacological or lifestyle-based interventions during middle age which could ultimately delay future cognitive ageing.

Animals↗

Estimating the electrostatic potential at the acetylcholine receptor agonist site using power saturation EPR.

Continuous wave EPR power saturation was used to measure electrostatic potentials at spin-labeled sites. Membrane surface potentials were estimated by power saturating the EPR spectrum of a membrane bound 14N spin-labeled amphiphile in the presence of a neutral or positively charged 15N labeled aqueous spin probe. The potentials that are measured are in good agreement with other probe measurements and with the predictions of the Gouy-Chapman-Stern theory, indicating that this is a valid approach to determine electrostatic potentials. A spin-labeled affinity probe based on maleimidobenzyltrimethylammonium was synthesized and could be derivatized to a sulfhydryl near either agonist site on the nicotinic acetylcholine receptor. The amplitudes of motion of the spin-probe on the ns time scale are significantly different when the two labeled sites are compared, and the probe is more restricted in its motion when attached to the more easily labeled site. When attached to this agonist site, power saturation EPR yields an electrostatic potential of -15 mV. Two other sulfhydryl-specific probes were used to label this site in reconstituted receptor containing membranes. These probes show less contact with the receptor and reduced electrostatic potentials, indicating that there is a strong spatial dependence to the potential at the agonist site. This work demonstrates that power saturation EPR provides a general method that can be used to estimate electrostatic potentials at any specifically spin-labeled macromolecular site.

Binding Sites↗

Quasi-elastic light scattering from migrating chemotactic bands of Escherichia coli.

We report the observation of migrating chemotactic bands of Escherichia coli in a buffer solution. The temporal development of the bacterial density profile is observed by the scattered light intensity as the band migrates through a stationary laser beam. We have made a preliminary analysis of the observed band profile with help of the Keller-Segel theory. The model accounts for only some aspects of the observed time evolution of the density profile. The microscopic motility characteristics of the E. coli in the band are simultaneously studied by photon correlation. The measured correlation functions are analyzed to obtain the spatial dependence of the half-width within the band. A simple analytical model is proposed to account for the contribution of the twiddle motion to the correlation function. By analyzing the correlation function as a superposition of straight-line and twiddle motions, we obtain a satisfactory agreement between the theory and the measured angular dependence of the line shape. As a consequence we are able to extract a parameter beta, which measures the average fraction of twiddling bacteria in the center of the band at a given time.

Cell Movement↗

Surface-induced lamellar orientation of multilayer membrane arrays. Theoretical analysis and a new method with application to purple membrane fragments.

The orientation of membrane fragments into a lamellar array by a flat surface is analyzed. This analysis includes processes such as centrifugation and drying and physical effects due to membrane fragment steric interactions, finite size, elasticity, and thermal fluctuations. Several model calculations of optimal orientational order in multilayer membrane arrays are presented. The predictions of a smectic A model agree quantitatively with the measured spatial dependence of the fluctuations in layer orientation in a multilamellar arrays. A new technique, based in part on this analysis, for the preparation of well-oriented multilamellar arrays of natural and artificial membranes, isopotential spin-dry centrifugation, is described. The method involves the use of specially designed inserts for the buckets of a standard vacuum ultracentrifuge. The membrane fragments to be oriented are sedimented from solution or suspension onto a substrate of a convenient material which forms a gravitational isopotential surface at high g. Sedimentation is accompanied by removal of the suspending medium at high g to produce oriented films with a selected degree of solvation. In addition, a method is described whereby small solute molecules can be maintained in constant concentration with the membrane fragments during this process. Initial application of the method to the orientation of purple membrane fragments is described. The degree of orientation obtained in this system is evaluated using freeze-fracture and scanning electron microscopy, optical birefringence, linear dichroism, and microscopy.

Bacteriorhodopsins↗

Current injection into a two-dimensional anisotropic bidomain.

A two-dimensional sheet of anisotropic cardiac tissue is represented with the bidomain model, and the finite element method is used to solve the bidomain equations. When the anisotropy ratios of the intracellular and extracellular spaces are not equal, the injection of current into the tissue induces a transmembrane potential that has a complicated spatial dependence, including adjacent regions of depolarized and hyperpolarized tissue. This behavior may have important implications for the electrical stimulation of cardiac tissue and for defibrillation.

Animals↗

Stochastic versions of the Hodgkin-Huxley equations.

A Hodgkin-Huxley model algorithm for the numerical simulation of noise in neurons is contracted from a master equation description (cellular automoton) into a Langevin description. This reduction reduces the time required for a simulation by about two orders of magnitude. Earlier work is summarized, condensed, and made explicit to make the algorithm transparent and facilitate applications. Two approximate treatments are reported. An extension of this approach is presented that includes spatial dependence and the propagation of a noisy action potential along an axon.

Axons↗

Simulation of the fertilization Ca2+ wave in Xenopus laevis eggs.

In the preceding paper Fontanilla and Nuccitelli (Biophysical Journal 75:2079-2087 (1998)) present detailed measurements of the shape and speed of the fertilization Ca2+ wave in Xenopus laevis eggs. In order to help interpret their results, we develop here a computational technique based on the finite element method that allows us to carry out realistic simulations of the fertilization wave. Our simulations support the hypothesis that the physiological state of the mature egg is bistable, i.e., that its cytoplasm can accommodate two alternative physiological Ca2+ concentrations: a low concentration characteristic of the prefertilization state and a greatly elevated concentration characteristic of the state following the passage of the wave. We explore this hypothesis by assuming that the bistability is due to the release and re-uptake properties of the endoplasmic reticulum (ER) as determined by inositol trisphosphate (IP3) receptor/Ca2+ channels and sarcoendoplasmic reticulum calcium ATPase (SERCA) pumps. When combined with buffered diffusion of Ca2+ in the cytoplasm, our simulations show that inhomogeneities in the Ca2+ release properties near the plasma membrane are required to explain the temporal and spatial dependences of the shape and speed of these waves. Our results are consistent with an elevated IP3 concentration near the plasma membrane in the unfertilized egg that is augmented significantly near the site of fertilization. These gradients are essential in determining the concave shape of the Ca2+ fertilization wave front.

Animals↗

Allopregnanolone inhibits learning in the Morris water maze.

The progesterone metabolite allopregnanolone (3alpha-OH-5alpha-pregnane-20-one) inhibits neural functions, enhancing the GABA induced GABA(A) receptor activation. This effect is benzodiazepine like and benzodiazepines are known to impair memory. Acute effects of allopregnanolone on the hippocampus dependent spatial learning in the Morris water maze have not been studied. Adult male Wistar rats where injected (i.v.) with allopregnanolone (2 mg/kg), or vehicle, daily for 11 days. At 8 or 20 min after each injection, studies of place navigation were performed in the Morris water maze. Allopregnanolone concentrations in plasma and in nine different brain areas where analyzed by radioimmunoassay. The latency to find the platform was increased 8 min after the allopregnanolone injection, while normal learning was seen after 20 min. Swim speed did not differ between groups. A higher number of rats were swimming close to the pool wall (thigmotaxis) in the 8 min allopregnanolone group compared to the other groups. Allopregnanolone concentrations in the brain tissue at 8 min were 1.5 to 2.5 times higher then at 20 min after the allopregnanolone injections. After vehicle injections the brain concentrations of allopregnanolone were at control levels. Plasma concentrations of allopregnanolone followed the same pattern as in the brain, with the exception of an increase 8 min after vehicle injections. The natural progesterone metabolite allopregnanolone can inhibit learning in the Morris water maze, an effect not caused by motor impairment. The learning impairment might be due to a combination of changed swimming behavior and difficulties in navigation.

Anesthetics↗

The effects of blur and size on monocular and stereoscopic localization.

Monocular localization of non-abutting stimuli and stereoscopic localization of the same second-order targets are performed with the same precision (Wilcox, L.M. & Hess, R.F. (1996) Is the site of non-linear filtering in stereopsis before or after binocular combination? Vision Research, 36, 391-399). Further, both tasks show a similar dependence on the scale of the stimulus. Since prior studies used Gaussian-enveloped stimuli, modifications of stimulus scale produced concurrent changes in edge blur. The experiments reported here assess the relative contributions of size and blur to the observed dependence on envelope scale for both monocular localization and stereoacuity. Stereoacuity for first-order targets was found to be an order of magnitude better than stereoacuity for second-order targets and monocular acuity for both first- and second-order targets. Further, while first-order stereopsis was found to depend solely on blur, second-order stereoacuity and monocular acuity were affected by both size and blur. These results suggest that while stereoacuity for first-order stimuli may be determined by a correlative process limited by early additive noise, stereoacuity for second-order stimuli and monocular acuity for non-abutting targets are more likely limited by stimulus-dependent spatial subsampling.

Depth Perception↗

A comparison of saccadic and blink suppression in normal observers.

Recent research suggests that blink and saccadic suppression are produced by the same mechanism (Volkmann, 1986; Uchikawa & Sato, 1995; Ridder & Tomlinson, 1993, 1995). These studies demonstrated that blink and saccadic suppression have the same effect on various visual functions. However, none of these studies made a comparison of blink and saccadic suppression in the same individual. The purpose of this study was to compare the effects of blink and saccadic suppression on contrast sensitivity functions in the same subject. The effect of saccadic suppression on the contrast sensitivity function in three normal observers was determined. Employing a two-alternative, forced-choice technique, thresholds were measured for seven spatial frequencies. At each spatial frequency, the threshold was determined immediately following detection of a voluntary saccade. The magnitude of suppression was taken as the log ratio of the contrast sensitivities obtained while foveating the stimulus and those obtained during saccades. The magnitude of saccadic suppression was found to increase as the saccade amplitude increased and to be spatial-frequency dependent. Low spatial frequencies were suppressed more than high spatial frequencies. The blink suppression data have been measured previously (Ridder & Tomlinson, 1993). Saccadic and blink suppression were qualitatively similar. A vertical shift of the data brought the saccadic and blink suppression data into register. These results suggest that blink and saccadic suppression are produced by the same or similar mechanisms.

Blinking↗

Inducible and reversible enhancement of learning, memory, and long-term potentiation by genetic inhibition of calcineurin.

The threshold for hippocampal-dependent synaptic plasticity and memory storage is thought to be determined by the balance between protein phosphorylation and dephosphorylation mediated by the kinase PKA and the phosphatase calcineurin. To establish whether endogenous calcineurin acts as an inhibitory constraint in this balance, we examined the effect of genetically inhibiting calcineurin on plasticity and memory. Using the doxycycline-dependent rtTA system to express a calcineurin inhibitor reversibly in the mouse brain, we find that the transient reduction of calcineurin activity facilitates LTP in vitro and in vivo. This facilitation is PKA dependent and persists over several days in vivo. It is accompanied by enhanced learning and strengthened short- and long-term memory in several hippocampal-dependent spatial and nonspatial tasks. The LTP and memory improvements are reversed fully by suppression of transgene expression. These results demonstrate that endogenous calcineurin constrains LTP and memory.

Animals↗

Uncertainty analysis of the external gamma-dose rate due to the variability of the vertical distribution of 137Cs in the soil.

The external gamma-dose rate at 1 m height above a flat area due to the presence of fallout radiocesium in the soil is frequently calculated from the observed depth profile of the 137Cs activity as well as the soil mass per unit area. At a given site, these depth profiles may, however, vary considerably, thus introducing an uncertainty to the external gamma-dose calculated in this way. To assess this source of uncertainty for a typical grassland site, the activity of Chernobyl-derived 137Cs and the wet bulk density in the three upper soil layers at 100 plots in a 100 m x 100 m pasture were determined. Analysis of these data shows that the frequency distribution of the dose rates calculated from the corresponding depth profiles of all plots is similar to a log-normal distribution (mean 25 nGy h-1, median 22 nGy h-1, standard deviation 11 nGy h-1; range 1.6-56 nGy h-1). The various sources which contribute to the uncertainty of the dose rate are quantified. The semi-variogram indicates that any spatial dependence of the dose rates occurs on this pasture only over distances that are smaller than the shortest sampling interval (here about 10 m). It is estimated which errors have to be expected for the median dose rate when the depth profiles of 137Cs and of the wet bulk density are determined only for a small number of plots. It is preferable to calculate the mean dose rate as a mean from the n individual dose rates rather than from an averaged 137Cs depth profile of the n plots.

Agriculture↗

Loss of medial septal modulation of dentate gyrus physiology in young mice overexpressing human beta-amyloid precursor protein.

Mice overexpressing the human mutant beta-amyloid precursor protein (hbetaAPP; PDAPP mice) show deficits in hippocampal-dependent spatial learning and hippocampal short- and long-term plasticity at ages preceding Abeta plaque deposition. We determined whether young PDAPP mice also exhibit alterations in septohippocampal function in vivo, which plays an important role in cognitive function. Electrical stimulation of the medial septum significantly increased neuronal excitability and reduced paired-pulse facilitation in the dentate gyrus. Medial septal-induced facilitation of dentate neuronal excitability was reduced in PDAPP mice. The inhibitory effects of medial septum stimulation on dentate paired-pulse facilitation were also attenuated in PDAPP mice. Young mice overexpressing hbetaAPP exhibit early abnormalities in neural circuits implicated in cognitive function that may play an important role in the more profound deficits observed in aged PDAPP mice.

Age Factors↗

Partial reversal of the effect of maternal care on cognitive function through environmental enrichment.

Maternal care influences hippocampal development in the rat. The offspring of mothers that exhibit increased levels of pup licking/grooming and arched-back nursing (High LG-ABN mothers) show increased hippocampal N-methyl-D-aspartate (NMDA) receptor binding and enhanced hippocampal-dependent spatial learning. In these studies we examined whether environmental enrichment from days 22-70 of life might reverse the effects of low maternal care. Environmental enrichment eliminated the differences between the offspring of High and Low LG-ABN mothers in both Morris water maze learning and object recognition. However, enrichment did not reverse the effect of maternal care on long-term potentiation in the dentate gyrus or on hippocampal NMDA receptor binding. In contrast, peripubertal enrichment did reverse the effects of maternal care on hippocampal alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor binding. These findings provide evidence for the reversal of the effects of reduced maternal investment in early life on cognitive function in adulthood. Such effects might involve compensatory changes associated with peripubertal enrichment.

Animals↗

Effects of aging on the hippocampal formation in a naturally occurring animal model of mild cognitive impairment.

Vertical integration is being used to great advantage in neurobiological research on the basis of age-related cognitive decline. Such research bridges analysis between the molecular and cellular levels and the outcome of impaired psychological functions. Current use of animals models within this paradigm has defined mild cognitive impairment in a subpopulation of outbred aged Long-Evans rats by assessment of hippocampal-dependent spatial cognition. Aged rats with cognitive impairment exhibited no loss of neurons in the hippocampus. Current research is focused on the functional alterations in neurons by methods which assess transcriptional mechanisms and signaling pathways.

Aging↗

Finite size disc gradient coil set for open vertical field magnets.

A new analytical approach is used in the design of disc-like gradient coils suitable for magnet geometries with main field direction perpendicular to the surface of the disc. An inverse procedure is used to optimize the coil's characteristics, subject to the restrictions imposed by the desired field behavior over a certain set of constraint points inside a predetermined imaging volume. Excellent agreement between the expected values of the gradient magnetic field and the numerical values generated by applying the Biot-Savart law to a discrete current pattern of the perspective disc coil was found. A Finite Element Analysis package was used to predict the fringe gradient field levels for a non-shielded axial disc coil and for a self-shielded transverse disc coil in the vicinity of the magnet poles. The numerical results indicate that for the self-shielded design the gradient fringe field is 1000 times smaller than the corresponding fringe field for the non-shielded disc case. Also no significant spatial dependence was noticed for the shielded coil's fringe field.

Algorithms↗

A technique for single-channel MR brain tissue segmentation: application to a pediatric sample.

A segmentation method is presented for gray matter, white matter, and cerebrospinal fluid (CSF) in thin-sliced single-channel brain magnetic resonance (MR) scans. The method is based on probabilistic modeling of intensity distributions and on a region growing technique. Interrater and intrarater reliabilities for the method were high, and comparison with phantom studies and hand-traced results from an experienced rater indicated good validity. The method was designed to account for spatially dependent image intensity inhomogeneities. Segmentation of MR brain scans of 105 (56 male and 49 female) healthy children and adolescents showed that although the total brain volume was stable over age 4-18, white matter increased and gray matter decreased significantly. There were no sex differences in total gray and white matter growth after correction for total brain volume. White matter volume increased the most in superior and posterior regions and laterality effects were seen in hemisphere tissue volumes. These findings are consistent with other reports, and further validate the segmentation technique.

Adolescent↗

Mapping eddy current induced fields for the correction of diffusion-weighted echo planar images.

Diffusion-weighted echo-planar magnetic resonance imaging is potentially of great importance as a diagnostic imaging tool; however, the technique currently suffers a number of limitations, including the image distortion caused by the eddy current induced fields when the diffusion-weighting magnetic field gradient pulses are applied. The distortions cause mis-registration between images with different diffusion-weighting, that then results in artifacts in quantitative diffusion images. A method is presented to measure the magnetic fields generated from the eddy currents for each of three orthogonal gradient pulse vectors, and then to use these to ascertain the image distortion that occurs in subsequent diffusion-weighted images with arbitrary gradient pulse vector amplitude and direction, and image plane orientation. The image distortion can then be reversed. Both temporal and spatial dependence of the residual eddy current induced fields are included in the analysis. Image distortion was substantially reduced by the correction scheme, for arbitrary slice position and angulation. This method of correction is unaffected by the changes in image contrast that occur due to diffusion weighting, and does not need any additional scanning time during the patient scan. It is particularly suitable for use with single-shot echo planar imaging.

Anisotropy↗