Immunohistochemical detection of L-DOPA-derived dopamine within serotonergic fibers in the striatum and the substantia nigra pars reticulata in Parkinsonian model rats.

On the basis of our previous studies in the normal rat [Arai, R., Karasawa, N., Geffard, M., Nagatsu, I., 1995. L-DOPA is converted to dopamine in serotonergic fibers of the striatum of the rat: a double-labeling immunofluorescence study. Neurosci. Lett. 195, 195-198; Arai, R., Karasawa, N., Nagatsu, I., 1996a. Aromatic L-amino acid decarboxylase is present in serotonergic fibers of the striatum of the rat. A double-labeling immunofluorescence study. Brain Res. 706, 177-179; Arai, R., Karasawa, N., Nagatsu, I., 1996b. Dopamine produced from L-DOPA is degraded by endogenous monoamine oxidase in neurons of the dorsal raphe nucleus of the rat: an immunohistochemical study. Brain Res. 722, 181-184] we have assumed that exogenously administered L-dihydroxyphenylalanine (L-DOPA) is converted into dopamine (DA) in serotonergic (5-HT) fibers within the striatum (ST) and the substantia nigra pars reticulata (SNR). In the present study, an attempt was made to confirm the assumptions in Parkinsonian rats, which were produced by unilateral injections of 6-hydroxydopamine (6-OHDA) into the substantia nigra pars compacta (SNC). The rats exhibiting more than 150 total controversial circles were regarded as satisfactory models of Parkinson disease (PD). Using a dual immunofluorescence histochemistry, we examined DA-immunoreactivity in the 5-HT fibers within the ST and the SNR of the PD model rats after L-DOPA was injected intraperitoneally. In experimental cases with the L-DOPA administration, DA-immunoreactivity was detected in 5-HT fibers in both the ST and the SNR on the 6-OHDA injection side; no DA-immunoreactivity was found in 5-HT fibers in the ST or the SNR in control cases without the L-DOPA administration. The results support the assumption that exogenously administered L-DOPA may be converted into DA within the 5-HT fibers in the ST and SNR of the PD model rats.

Changes of immunocytochemical localization of vesicular glutamate transporters in the rat visual system after the retinofugal denervation.

To clarify which vesicular glutamate transporter (VGluT) is used by excitatory axon terminals of the retinofugal system, we examined immunoreactivities and mRNA signals for VGluT1 and VGluT2 in the rat retina and compared immunoreactivities for VGluT1 and VGluT2 in the retinorecipient regions using double immunofluorescence method, anterograde tracing, and immunoelectron microscopy. Furthermore, the changes of VGluT1 and VGluT2 immunoreactivities were studied after eyeball enucleation. Intense immunoreactivity and mRNA signal for VGluT2, but not for VGluT1 immunoreactivity, were observed in most perikarya of ganglion cells in the retina. Immunoelectron microscopy revealed that VGluT1- and VGluT2-immunolabeled terminals made asymmetrical synapses, suggesting that they were excitatory synapses, and that VGluT1-immunolabeled terminals were smaller than VGluT2-labeled ones in many retinorecipient regions, such as the dorsal lateral geniculate nucleus (LGd) and superior colliculus (SC). Double immunofluorescence study further revealed that almost no VGluT2 immunoreactivity was colocalized with VGluT1 in the retinorecipient regions. After wheat germ agglutinin (WGA) injection into the eyeballs, WGA immunoreactivity was colocalized in the single axon terminals of LGd and SC with VGluT2 but not VGluT1 immunoreactivity. After unilateral enucleation, VGluT2 immunoreactivity in the LGd, SC, nucleus of the optic tract, and nuclei of the accessory optic tract in the contralateral side of the enucleated eye was clearly decreased. Although only a small change of VGluT2 immunoreactivity was observed in the contra- and ipsilateral suprachiasmatic nuclei, olivary pretectal nucleus, anterior pretectal nucleus, and posterior pretectal nucleus, moderate reduction of VGluT2 was found in these regions after bilateral enucleation. On the other hand, almost no change in VGluT1 immunoreactivity was found in the structures examined in the present enucleation study. Thus, the present results support the notion that the retinofugal pathways are glutamatergic, and indicate that VGluT2, but not VGluT1, is employed for accumulating glutamate into synaptic vesicles of retinofugal axons.

Stabilization technique for real-time high-resolution vascular ultrasound using frequency domain interferometry.

We have proposed an ultrasound imaging method based on frequency domain interferometry (FDI) with an adaptive beamforming technique to depict real-time high-resolution images of human carotid artery. Our previous study has investigated the performance of the proposed imaging method under an ideal condition with a high signal-to-noise ratio (SNR). In the present study, we propose a technique that has the potential to improve accuracy in estimating echo intensity using the FDI imaging method. We investigated the performance of the proposed technique in a simulation study that two flat interfaces were located at depths of 15.0 and 15.2 mm and white noise was added. Because the -6 dB bandwidth of the signal used in this simulation study is 2.6 MHz, the conventional B-mode imaging method failed to depict the two interfaces. Both the conventional and proposed FDI imaging methods succeeded to depict the two interfaces when the SNR ranged from 15 to 30 dB. However, the average error of the estimated echo intensity at the interfaces using the conventional FDI imaging method ranged from 7.2 to 10.5 dB. In contrast, that using the FDI imaging method with the proposed technique ranged from 2.0 to 2.2 dB. The present study demonstrates the potential of the FDI imaging method in depicting robust and high-range-resolution ultrasound images of arterial wall, indicating the possibility to improve the diagnosis of atherosclerosis in early stages.

Real-time high-resolution vascular ultrasound using frequency domain interferometry with the ROI-division process.

We have proposed a high-range-resolution ultrasound imaging method for human carotid artery using an adaptive beamforming technique based on frequency domain interferometry (FDI). The method assumes that the received signal consists of multiple echoes of targets and noise, where the waveform of each echo is similar to that of the reference signal. In this study, we examine the dependence of the echo waveform on the target depth, and investigate the proper measurement-range for the FDI imaging method using a reference signal. Furthermore, we propose a ROI-division process, where each sub-ROI has a proper measurement-range for the application of the FDI imaging method. Simulation and experimental results show the efficiency of the ROI-division process in improving the image quality of human carotid artery acquired using the FDI imaging method. We believe that the modified FDI imaging method with the ROI-division process has the potential to facilitate significant progress in the detection of vessel stenosis and in the assessment of cardiovascular disease risk.

High range resolution ultrasonographic vascular imaging using frequency domain interferometry with the Capon method.

For high range resolution ultrasonographic vascular imaging, we apply frequency domain interferometry with the Capon method to a single frame of in-phase and quadrature (IQ) data acquired using a commercial ultrasonographic device with a 7.5 MHz linear array probe. In order to tailor the adaptive beam forming algorithm for ultrasonography we employ four techniques: frequency averaging, whitening, radio-frequency data oversampling, and the moving average. The proposed method had a range resolution of 0.05 mm in an ideal condition, and experimentally detected the boundary couple 0.17 mm apart, where the boundary couple was indistinguishable from a single boundary utilizing a B-mode image. Further, this algorithm could depict a swine femoral artery with a range beam width of 0.054 mm and an estimation error for the vessel wall thickness of 0.009 mm, whereas using a conventional method the range beam width and estimation error were 0.182 and 0.021 mm, respectively. The proposed method requires 7.7 s on a mobile PC with a single CPU for a 1×3 cm region of interest. These findings indicate the potential of the proposed method for the improvement of range resolution in ultrasonography without deterioration in temporal resolution, resulting in enhanced detection of vessel stenosis.

High range resolution medical acoustic vascular imaging with frequency domain interferometry.

For high range resolution acoustic vascular imaging we apply frequency domain interferometry and Capon method to a few frames of in-phase and quadrature (IQ) data acquired by a commercial ultrasonographic device. To suit the adaptive beamforming algorithm to medical acoustic imaging we employ three techniques; frequency averaging, whitening, and pseudo-double RF data conversion. The proposed method detected two couples of boundaries 0.26 and 0.19 mm apart using a single frame and two frames of IQ data, respectively, where each couple of boundaries is indistinguishable from a single boundary utilizing B-mode images. Further this algorithm could depict a swine femoral artery with higher range resolution than conventional B-mode imaging. These results indicate the potential of the proposed method for the range resolution improvement in ultrasonography, originating the progress in detection of vessel stenosis.