Correcting magnetic resonance k-space data for in-plane motion using an optical position tracking system.

PURPOSE: Motion is a major confound of image quality in MRI. A method of retrospectively correcting the effects of rotations and translations on the acquired k-space data is presented. METHODS: In two phantom experiments of well-controlled translation and rotation, two MRI-compatible infrared cameras recorded motion data that were used subsequently to correct the position and phase of recorded k-space samples. Motion data can be acquired with a temporal resolution of 60 Hz and spatial accuracy of 0.1 mm for translations and 0.2 degree for rotations. RESULTS: Significant improvements of image quality are demonstrated. CONCLUSIONS: The key advantages of the technique are that it is easy to implement, does not interfere with or complicate MR data acquisition, and is capable of correcting distortions within a single slice. Therefore, the technique has the potential to improve upon approaches that rely on the registration or realignment of successive imaging slices.

Volume ordering for analysis and modeling of vascular systems.

Morphological characteristics of vascular systems are commonly presented in terms of Strahler order because the logarithms of quantities such as vessel diameter and length are often linearly related to Strahler order. However, the ability to interpret Strahler order geometrically or physiologically is compromised because the precision of the order number is limited to integer values. This limitation is overcome by the volume ordering scheme, in which volume order number is defined as the logarithm of the estimated perfused tissue volume for each vascular segment. While Strahler and volume order numbers are equivalent for completely symmetrical branching trees, they deviate in the presence of asymmetries. The physiology-based definition of volume ordering offers benefits in the analysis of vascular design, fractal characterization of vascular systems, and blood flow modeling. These benefits are illustrated based on arterial kidney data that show a linear relationship of logarithmic vessel diameter and conductance as a function of both Strahler order and volume order with differing proportionality constants, which are expected to depend on the branching characteristics of the particular organ investigated.

Comparing microsphere deposition and flow modeling in 3D vascular trees.

Blood perfusion in organs has been shown to be heterogeneous in a number of cases. At the same time, a number of models of vascular structure and flow have been proposed that also generate heterogeneous perfusion. Although a relationship between local perfusion and vascular structure has to exist, no model has yet been validated as an accurate description of this relationship. A study of perfusion and three-dimensional (3D) arterial structure in individual rat kidneys is presented, which allows comparison between local measurements of perfusion and model-based predictions. High-resolution computed tomography is used to obtain images of both deposited microspheres and of an arterial cast in the same organ. Microsphere deposition is used as an estimate of local perfusion. A 3D cylindrical pipe model of the arterial tree is generated based on an image of the arterial cast. Results of a flow model are compared with local microsphere deposition. High correlation (r(2) > 0.94) was observed between measured and modeled flows through the vascular tree segments. However, the relative dispersion of the microsphere perfusion measurement was two- to threefold higher than perfusion heterogeneity calculated in the flow model. Also, there was no correlation in the residual deviations between the methods. This study illustrates the importance of comparing models of local perfusion with in vivo measurements of perfusion in the same biologically realistic vascular tree.

Estimating perfusion using microCT to locate microspheres.

The injection of microspheres into the blood stream has been a common method to measure the spatial distribution of blood flow (perfusion). A technique to conduct this kind of measurement in small animal organs is presented using silver-coated microspheres with a diameter of 16 microm and high-resolution computed tomography (microCT) to detect individual microspheres. Phantom experiments demonstrate the detectability of individual spheres. The distribution of microspheres within a rat heart is given as an example. Using non-destructive, three-dimensional imaging for microsphere detection avoids the cumbersome dissection of the organ into samples or slices and their subsequent registration. The detection of individual spheres allows high-resolution measurements of perfusion and arbitrary definition of regions of interest. These, in turn, allow for accurate statistical analysis of perfusion such as relative dispersion curves.

Branching tree model with fractal vascular resistance explains fractal perfusion heterogeneity.

Perfusion heterogeneities in organs such as the heart obey a power law as a function of scale, a behavior termed "fractal." An explanation of why vascular systems produce such a specific perfusion pattern is still lacking. An intuitive branching tree model is presented that reveals how this behavior can be generated as a consequence of scale-independent branching asymmetry and fractal vessel resistance. Comparison of computer simulations to experimental data from the sheep heart shows that the values of the two free model parameters are realistic. Branching asymmetry within the model is defined by the relative tissue volume being fed by each branch. Vessel ordering for fractal analysis of morphology based on fed or drained tissue volumes is preferable to the commonly used Strahler system, which is shown to depend on branching asymmetry. Recently, noninvasive imaging techniques such as PET and MRI have been used to measure perfusion heterogeneity. The model allows a physiological interpretation of the measured fractal parameters, which could in turn be used to characterize vascular morphology and function.

Endocytic vacuoles formed following a short pulse of K+ -stimulation contain a plethora of presynaptic membrane proteins.

It is now well established that the membrane of synaptic vesicles is recycled following exocytosis. However, little is known concerning the identity of the primary or secondary endocytic structures and their molecular composition. Using cultured rat cerebellar granule cells we combined uptake of horseradish peroxidase as a fluid phase marker and immunogold labeling for a variety of presynaptic proteins to assess the molecular identity of the stimulation-induced endocytic compartments. Short periods (5 or 30 s) of stimulation with 50 mM KCl were followed by periods of recovery for up to 30 min. Stimulation resulted in the formation of horseradish-peroxidase-filled vacuoles in the axonal varicosities as the apparent primary endocytic compartment. Horseradish peroxidase-filled synaptic vesicles were formed when stimulated cells were allowed to recover in horseradish peroxidase-free culture medium. Horseradish peroxidase-filled vacuoles as wells as vesicles contained the synaptic vesicle membrane proteins VAMP II, synaptotagmin, SV2, and synaptophysin, the vesicle-associated proteins rab 3A and synapsin I, and in addition SNAP-25. No incorporation of vesicle proteins into the plasma membrane was observed. Horseradish peroxidase-filled vesicles and vacuoles generated on incubation of unstimulated granule cells with horseradish peroxidase for prolonged periods of time were equally immunolabeled. Renewed stimulation of prestimulated granule cells with either 100 mM KCl or 30 microM Ca2+ ionophore A23187 resulted in a reduction of horseradish peroxidase-filled vacuoles suggesting that the vacuolar membrane compartment was exocytosis-competent. Our results suggest that varicosities of cultured cerebellar granule cells possess a fast stimulation-induced pathway for recycling the entire synaptic vesicle membrane compartment. The primary endocytic compartment represents not a synaptic vesicle but a somewhat larger vesicle protein-containing vacuolar entity from which smaller vesicles of identical protein composition may be regenerated. Endocytic vacuoles and synaptic vesicles share membrane and membrane-associated proteins and presumably also major functional properties.

A plethora of presynaptic proteins associated with ATP-storing organelles in cultured astrocytes.

Cultured astrocytes can release a variety of messenger substances via receptor-mediated mechanisms, implicating their potential for regulated exocytosis and the participation of proteins of the SNARE complex. Here we demonstrate the astrocytic expression and organellar association of a large variety of synaptic proteins (synaptobrevin II, synaptotagmin I, synaptophysin, rab3a, synapsin I, SNAP-25, and syntaxin I) and also of the ubiquitous cellubrevin. As revealed by immunoblotting the expression of synaptic proteins was highest within the first few days after plating. Synaptophysin and SNAP-25 showed the most significant decline with prolonged culture time. Rab3a and synaptobrevin II were retained at a high level and synaptotagmin I, synapsin I, and syntaxin I at a lower level until 20 DIV. The immunoreaction for cellubrevin was low at the beginning and increased with prolonged culture time. As revealed by light microscopical immunocytochemistry the proteins are expressed by GFAP-positive astrocytes and associated with organelles of varying size. Immunoelectron microscopical analysis allocates synaptobrevin II and synaptophysin to the membranes of vesicular organelles. Double labeling experiments for pairs of synaptic proteins reveal that individual synaptic proteins can be entirely colocalized or partly reside on different organelles. Subcellular fractionation of astrocyte cultures by sucrose density gradient centrifugation after 2, 6, 13, and 20 DIV showed that the proteins sediment with ATP containing organelles of a broad density range. Our data suggest that messenger substances may be released from cultured astrocytes via receptor-mediated, Ca2+-dependent exocytosis.

Immunocytochemical localization of synaptic proteins at vesicular organelles in PC12 cells.

The distribution of the three synaptic vesicle proteins SV2, synaptophysin and synaptotagmin, and of SNAP-25, a component of the docking and fusion complex, was investigated in PC12 cells by immunocytochemistry. Colloidal gold particle-bound secondary antibodies and a preembedding protocol were applied. Granules were labeled for SV2 and synaptotagmin but not for synaptophysin. Electron-lucent vesicles were labeled most intensively for synaptophysin but also for SV2 and to a lesser extent for synaptotagmin. The t-SNARE SNAP-25 was found at the plasma membrane but also at the surface of granules. Labeling of Golgi vesicles was observed for all antigens investigated. Also components of the endosomal pathway such as multivesicular bodies and multilamellar bodies were occasionally marked. The results suggest that the three membrane-integral synaptic vesicle proteins can have a differential distribution between electron-lucent vesicles (of which PC12 cells may possess more than one type) and granules. The membrane compartment of granules appears not to be an immediate precursor of that of electron-lucent vesicles.