Extracerebral Region Research Articles

IN SILICO APPROACHES ON PHENYLALANINE HYDROXYLASE INHIBITOR-RELATED COMPOUNDS USED IN PARKINSON’S DISEASE TREATMENT

Objective: In Parkinson’s disease, Levodopa with Carbidopa addresses dopamine deficiency. Phenylalanine hydroxylase catalyzes phenylalanine to tyrosine conversion crucial for dopamine synthesis. Inhibiting phenylalanine hydroxylase may enhance Carbidopa's effects, preventing peripheral dopamine synthesis. The study used virtual scanning, molecular docking, and dynamics simulation to explore phenylalanine hydroxylase interactions with Carbidopa and similar ligands. ADME/T assessments and drug similarity tests were conducted to evaluate therapeutic potential in biological systems. Material and Method: A molecular docking study was performed on the structures obtained from the PubChem database and human PAH (PDB ID: 6PAH) using Autodock Vina within Chimera 1.16. Furthermore, the ligands underwent ADME/T assays, which are crucial aspects in drug development. Result and Discussion: The study suggests that 2-(2-Aminohydrazinyl)-3-(3,4-dihydroxyphenyl)-2-methylpropanoic acid shows promise as a phenylalanine hydroxylase inhibitor for Parkinson's disease treatment, but further research is needed to assess its safety, efficacy, and specificity, particularly in extracerebral regions, while also exploring its potential to improve the effectiveness of Levadopa/Carbidopa combination therapy.

Reliability of fNIRS for noninvasive monitoring of brain function and emotion in sheep

The aim of this work was to critically assess if functional near infrared spectroscopy (fNIRS) can be profitably used as a tool for noninvasive recording of brain functions and emotions in sheep. We considered an experimental design including advances in instrumentation (customized wireless multi-distance fNIRS system), more accurate physical modelling (two-layer model for photon diffusion and 3D Monte Carlo simulations), support from neuroanatomical tools (positioning of the fNIRS probe by MRI and DTI data of the very same animals), and rigorous protocols (motor task, startling test) for testing the behavioral response of freely moving sheep. Almost no hemodynamic response was found in the extra-cerebral region in both the motor task and the startling test. In the motor task, as expected we found a canonical hemodynamic response in the cerebral region when sheep were walking. In the startling test, the measured hemodynamic response in the cerebral region was mainly from movement. Overall, these results indicate that with the current setup and probe positioning we are primarily measuring the motor area of the sheep brain, and not probing the too deeply located cortical areas related to processing of emotions.

Measurement of [123I]FP-CIT binding to the dopamine transporter (DAT) in healthy mouse striatum using dedicated small animal SPECT imaging: feasibility, optimization and validation.

In-vivo imaging of dopamine transporter (DAT), a reliable marker of degeneration of nigrostriatal dopaminergic innervation, has gained increasing interest in preclinical neurodegenerative research for studying disease mechanisms and testing new therapeutic strategies. We assessed the feasibility and the reliability of in vivo and ex vivo quantification of Methyl (3S,4S,5R)-8-(3-fluoropropyl)-3-(4-iodophenyl)-8-azabicyclo[3.2.1]octane-4-carboxylate ([123I]FP-CIT) binding to striatal DAT sites in mouse brain. Dedicated small animal single-photon emission computed tomography (SPECT) images of [123I]FP-CIT binding were obtained in 3 groups of healthy mice: untreated (N.=6), pre-treated with lugol solution (N.=4), and pre-treated with selective dopamine transporter uptake inhibitor GBR12909 (N.=4). Ex-vivo autoradiography studies were performed at the end of SPECT studies with phosphor image system in 4 out of the 6 untreated mice and in all mice pre-treated with lugol. Regions of interest were defined over the striatum. The specific binding (SB) was calculated using the cerebral cortex as reference region. SPECT images in untreated mice showed high [123I]FP-CIT uptake in the striatum and extracerebral regions. Lugol pretreatment improved striatal images quality decreasing salivary and thyroid glands uptake. SB was higher (P<0.0001) in mice pre-treated with lugol (5.97±0.60) than in untreated mice (2.25±0.28). Autoradiography showed similar SB findings in untreated (2.27±0.33) and lugol-treated (4.27±0.57) mice (P<0.0001). In-vivo striatal 123I-FP-CIT SB and ex-vivo striatal 123I-FP-CIT SB were significantly correlated (r=0.87; P<0.0001). SPECT competition studies showed a significant (P<0.0001) reduction of [123I]FP-CIT SB in the striatum after GBR12909. We demonstrated the feasibility of [123I]FP-CIT imaging of the normal mouse brain using small-animal SPECT without pinhole collimators. The reliability of quantitative measurement of striatal [123I]FP-CIT SB is supported by competition studies showing measurable inhibition of uptake induced by GBR12909 and by the strong correlation between in vivo and ex vivo striatal [123I]FP-CIT SB. Our data also demonstrate that pre-treatment with lugol might improve striatal [123I]FP-CIT SB in mice.

Impact of partial volume effect correction on cerebral β-amyloid imaging in APP-Swe mice using [18F]-florbetaben PET

We previously investigated the progression of β-amyloid deposition in brain of mice over-expressing amyloid-precursor protein (APP-Swe), a model of Alzheimer's disease (AD), in a longitudinal PET study with the novel β-amyloid tracer [18F]-florbetaben. There were certain discrepancies between PET and autoradiographic findings, which seemed to arise from partial volume effects (PVE). Since this phenomenon can lead to bias, most especially in the quantitation of brain microPET studies of mice, we aimed in the present study to investigate the magnitude of PVE on [18F]-florbetaben quantitation in murine brain, and to establish and validate a useful correction method (PVEC).Phantom studies with solutions of known radioactivity concentration were performed to measure the full-width-at-half-maximum (FWHM) resolution of the Siemens Inveon DPET and to validate a volume-of-interest (VOI)-based PVEC algorithm. Several VOI-brain-masks were applied to perform in vivo PVEC on [18F]-florbetaben data from C57BL/6(N=6) mice, while uncorrected and PVE-corrected data were cross-validated with gamma counting and autoradiography. Next, PVEC was performed on longitudinal PET data set consisting of 43 PET scans in APP-Swe (13–20months) and age-matched wild-type (WT) mice using the previously defined masks. VOI-based cortex-to-cerebellum ratios (SUVR) were compared for uncorrected and PVE-corrected results. Brains from a subset of transgenic mice were ultimately examined by autoradiography ex vivo and histochemistry in vitro as gold standard assessments, and compared to VOI-based PET results.The phantom study indicated a FWHM of 1.72mm. Applying a VOI-brain-mask including extracerebral regions gave robust PVEC, with increased precision of the SUVR results. Cortical SUVR increased with age in APP-Swe mice compared to baseline measurements (16months: +5.5%, p<0.005; 20months: +15.5%, p<0.05) with uncorrected data, and to a substantially greater extent with PVEC (16months: +12.2% p<0.005; 20months: +36.4% p<0.05). WT animals showed no binding changes, irrespective of PVEC. Relative to autoradiographic results, the error [%] for uncorrected cortical SUVR was 18.9% for native PET data, and declined to 4.8% upon PVEC, in high correlation with histochemistry results. We calculate that PVEC increases by 10% statistical power for detecting altered [18F]-florbetaben uptake in aging APP-Swe mice in planned studies of disease modifying treatments on amyloidogenesis.

Effects of laserneedle stimulation in the external auditory meatus on brainstem and very early auditory evoked potentials in humans

Objectives: For the first time, brainstem auditory evoked potentials (BAEP) and very early auditory evoked potentials (EAEP) were investigated in 23 volunteers (mean age: 26.5 ± 3.6 years) under resting conditions and during continuous and frequency modulated (2 Hz) laser stimulation in the external auditory meatus.Methods: Using a new ear adapter, the outer region of the auditory canal was stimulated with laser light (685 nm, 4 × 30–40 mW, duration: 10 minutes).Results: The EAEPs were significantly changed during continuous (p=0.019), as well as frequency modulated (p=0.014) laser stimulation compared with control measurements.Discussion: Physiologic alterations of inner ear mechanism such as extra-cerebral changes in conductance or stimulation-dependent depolarization processes in extra-cerebral regions of the auditory system could be possible explanations for the significant difference in measurement parameters.

Ontogenesis of the GnRH neuron system in the rat. A quantitative immunohistochemical study with special reference to the extra cerebral GnRH-positive cells and the occupation of intracerebral termination fields

The distribution of GnRH immunopositive cells in the rat was studied from day 13 of fetal life to day 10 of postnatal life with the aid of a GnRH antibody (H-16) generated against the decapeptide p-Glu 1-LHRH in the rabbit and following a PAP immunohistochemical staining protocol on 70 um serial Vibratome sections. The first appearance of GnRH cells was observed simultaneously in both intra- and extra-cerebral regions on day 16 of fetal life. At all stages of development, at least 10 times more GnRH cells were found in the brain than in extracerebral areas. The population of GnRH cells was maximum in the nasal mucosa on day 20, and in intracranial, extracerebral regions (along the anterior cerebral artery and the terminal nerve) on day 22 of fetal life: it declined thereafter. The population of GnRH cells in the brain continued to increase until the day of birth and remained unchanged thereafter. Our findings are in accordance with the theory of nasal placodal origin of the GnRH neuronal system, but also suggest that in the rat, shifting of the cells from the nasal placode to the brain is somewhat preceding the onset of GnRH synthesis. Our other field of endeavor was to examine the GnRH axon terminal fields in the course of development. On day 18 of fetal life, the vascular organ of the lamina terminalis, the median eminence and the medial habenular nuclei received their GnRH axon terminals. On day 19 the subfornical organ, mammillary nuclei and the central nucleus of the amygdala, and on day 21 the periaqueductal gray and the interpeduncular nucleus became invaded by GnRH axons. These data serve the better understanding of the onset of the different functional activities of the GnRH system in the rat.

Immunocytochemical localization of the gonadotropin-releasing hormone-associated peptide portion of the LHRH precursor in the hypothalamus and extrahypothalamic regions of the rat central nervous system.

The gonadotropin-releasing hormone-associated peptide (GAP) of the LHRH precursor and the decapeptide LHRH were localized in the rat brain by immunocytochemistry in 12- to 18-day-old animals, by use of thick Vibratome sections and nickel intensification of the diaminobenzidine-reaction product. Our results indicate that the GAP portion of the LHRH precursor is present in the same population of neurons that contain LHRH in the rat brain. An important difference observed was that the GAP antiserum, in contrast to LHRH antisera, stained several perikarya in the medial basal hypothalamus. GAP-immunoreactive perikarya were observed in the following regions: the olfactory bulb and tubercle, diagonal band of Broca, medial septum, medial preoptic and suprachiasmatic areas, anterior and lateral hypothalamus, and several regions of the hippocampus. In addition to the preoptico-terminal and the septo-preoptico-infundibular pathways, we also observed GAP-immunopositive processes in several major tracts and areas of the brain, including the amygdala, stria terminals, stria medullaris thalami, fasciculus retroflexus, stria longitudinalis medialis, periventricular plexus, periaqueductal gray of the mesencephalon and extra-cerebral regions, such as the nervus terminalis and its associated ganglion. These results confirm the specificity of previous immunocytochemical results obtained with antisera to LHRH. The presence of GAP immunoreactivity in nerve terminals of the rat brain indicates that GAP or a GAP-like peptide is located in the proper site to serve as a hypophysiotropic substance and/or as a neurotransmitter or neuromodulator.

Gonadotropin-releasing hormone (GnRH) neurons and pathways in the rat brain.

Gonadotropin-releasing hormone (GnRH) neurons and their pathways in the rat brain were localized by immunocytochemistry in 6- to 18-day-old female animals, by use of thick frozen or vibratome sections, and silver-gold intensification of the diaminobenzidine reaction product. GnRH-immunoreactive perikarya were observed in the following regions: olfactory bulb and tubercle, vertical and horizontal limbs of the diagonal band of Broca, medial septum, medial preoptic and suprachiasmatic areas, anterior and lateral hypothalamus, and different regions of the hippocampus (indusium griseum, Ammon's horn). In addition to the known GnRH-pathways (preoptico-terminal, preoptico-infundibular, periventricular), we also observed GnRH-immunopositive processes in several major tracts and areas of the brain, including the medial and cortical amygdaloid complex, stria terminalis, stria medullaris thalami, fasciculus retroflexus, medial forebrain bundle, indusium griseum, stria longitudinalis medialis and lateralis, hippocampus, periaqueductal gray of the mesencephalon, and extracerebral regions, such as the lamina cribrosa, nervus terminalis and its associated ganglia. By use of the silver-gold intensification method we present Golgi-like images of GnRH perikarya and their pathways. The possible distribution of efferents from each GnRH cell group is discussed.

Study of the Cerebral Circulation by Means of Inert Diffusible Tracers

Man successfully achieved the measurement of peripheral blood flow, of cardiac output, of renal circulation and glomerular filtration rate at a time when quantitative information on the cerebral circulation had not been obtained even in lower animals. Studies began on the solubility of nitrous oxide in blood and brain. It was eventually found that the solubility of this gas was the same in brain as in blood, and moreover, the solubility was the same in the brain of dog and man, in living brain and in dead brain and was unaffected by a variety of diseases. In order to determine to what extent blood from extracerebral regions is present in the internal jugular vein at the point of sampling, studies were performed at the operating table in cases where exposure of the carotid bifurcation was necessary. As a final evaluation of the nitrous oxide method, experiments were performed in monkeys in which cerebral blood flow was measured simultaneously by this method and directly by means of the bubble flow meter. The nitrous oxide technique and its various modifications has been of value in elucidating the physiology of the circulation in the normally functioning human brain freed of such constraints as anesthesia or surgical intervention.