The identification and neurochemical characterization of central neurons that target parasympathetic preganglionic neurons involved in the regulation of choroidal blood flow in the rat eye using pseudorabies virus, immunolabeling and conventional pathway tracing methods

The choroidal blood vessels of the eye provide the main vascular support to the outer retina. These blood vessels are under parasympathetic vasodilatory control via input from the pterygopalatine ganglion (PPG), which in turn receives its preganglionic input from the superior salivatory nucleus (SSN) of the hindbrain. The present study characterized the central neurons projecting to the SSN neurons innervating choroidal PPG neurons, using pathway tracing and immunolabeling. In the initial set of studies, minute injections of the Bartha strain of the retrograde transneuronal tracer pseudorabies virus (PRV) were made into choroid in rats in which the superior cervical ganglia had been excised (to prevent labeling of sympathetic circuitry). Diverse neuronal populations beyond the choroidal part of ipsilateral SSN showed transneuronal labeling, which notably included the parvocellular part of the paraventricular nucleus of the hypothalamus (PVN), the periaqueductal gray, the raphe magnus (RaM), the B3 region of the pons, A5, the nucleus of the solitary tract (NTS), the rostral ventrolateral medulla (RVLM), and the intermediate reticular nucleus of the medulla. The PRV+ neurons were located in the parts of these cell groups that are responsive to systemic blood pressure signals and involved in systemic blood pressure regulation by the sympathetic nervous system. In a second set of studies using PRV labeling, conventional pathway tracing, and immunolabeling, we found that PVN neurons projecting to SSN tended to be oxytocinergic and glutamatergic, RaM neurons projecting to SSN were serotonergic, and NTS neurons projecting to SSN were glutamatergic. Our results suggest that blood pressure and volume signals that drive sympathetic constriction of the systemic vasculature may also drive parasympathetic vasodilation of the choroidal vasculature, and may thereby contribute to choroidal baroregulation during low blood pressure

Effect of the Decrease in Luminance Noise Range on Color Discrimination of Dichromats and Trichromats

Color vision assessment can be done using pseudoisochromatic stimuli, which has a luminance noise to eliminate brightness differences between the target and background of the stimulus. It is not clear the influence of the luminance noise on color discrimination. We investigated the effect of change in the luminance noise limits on color discrimination. Eighteen trichromats and ten congenital dichromats (eight protans, two deutans) had their color vision evaluated by the Cambridge Colour Test, and were genetically tested for diagnostic confirmation. The stimuli were composed of a mosaic of circles in a 5° circular field. A subset of the circles differed in chromaticity from the remaining field, forming a letter C. Color discrimination was estimated in stimulus conditions differing in luminance noise range: (i) 6–20 cd/m2; (ii) 8–18 cd/m2; (iii) 10–16 cd/m2; and (iv) 12–14 cd/m2. Six equidistant luminance values were used within the luminance noise limits with the mean stimulus luminance maintained constant under all conditions. A four-alternative, forced-choice method was applied to feed a staircase procedure to estimate color discrimination thresholds along eight chromatic axes. An ellipse model was adjusted to the eight color discrimination thresholds. The parameters of performance were threshold vector lengths and the ellipse area. Results were compared using the Kruskal-Wallis test with a significance level of 5%. The linear function model was applied to analyze the dependence of the discrimination parameters on the noise luminance limits. The first derivative of linear function was used as an indicator of the rate of change in color discrimination as a function of luminance noise changes. The rate of change of the ellipse area as a function of the luminance range in dichromats was higher than in trichromats (p < 0.05). Significant difference was also found for individual thresholds in half of the axes we tested. Luminance noise had a greater effect on color discrimination ability of dichromats than the trichromats, especially when the chromaticities were close to their protan and deutan color confusion lines

Color discrimination is affected by modulation of luminance noise in pseudoisochromatic stimuli

Pseudoisochromatic stimuli have been widely used to evaluate color discrimination and to identify color vision deficits. Luminance noise is one of the stimulus parameters used to ensure that subject´s response is due to their ability to discriminate target stimulus from the background based solely on the hue between the colors that compose such stimuli. We studied the influence of contrast modulation of the stimulus luminance noise on threshold and reaction time color discrimination. We evaluated color discrimination thresholds using the Cambridge Color Test (CCT) at six different stimulus mean luminances. Each mean luminance condition was tested using two protocols: constant absolute difference between maximum and minimum luminance of the luminance noise (constant delta protocol, CDP), and constant contrast modulation of the luminance noise (constant contrast protocol, CCP). MacAdam ellipses were fitted to the color discrimination thresholds in the CIE 1976 color space to quantify the color discrimination ellipses at threshold level. The same CDP and CCP protocols were applied in the experiment measuring RTs at three levels of stimulus mean luminance. The color threshold measurements show that for the CDP, ellipse areas decreased as a function of the mean luminance and they were significantly larger at the two lowest mean luminances, 10 cd/m2 and 13 cd/m2, compared to the highest one, 25 cd/m2. For the CCP, the ellipses areas also decreased as a function of the mean luminance, but there was no significant difference between ellipses areas estimated at six stimulus mean luminances. The exponent of the decrease of ellipse areas as a function of stimulus mean luminance was steeper in the CDP than CCP. Further, reaction time increased linearly with the reciprocal of the length of the chromatic vectors varying along the four chromatic half-axes. It decreased as a function of stimulus mean luminance in the CDP but not in the CCP. The findings indicated that visual performance using pseudoisochromatic stimuli was dependent on the Weber´s contrast of the luminance noise. Low Weber´s contrast in the luminance noise is suggested to have a reduced effect on chromatic information and, hence, facilitate desegregation of the hue-defined target from the background.Fil: Cormenzana Méndez, Iñaki. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Exactas y Tecnología. Laboratorio de Luminotecnia; ArgentinaFil: Martín, Andrés. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Investigación en Luz, Ambiente y Visión. Universidad Nacional de Tucumán. Facultad de Ciencias Exactas y Tecnología. Instituto de Investigación en Luz, Ambiente y Visión; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Exactas y Tecnología. Laboratorio de Luminotecnia; ArgentinaFil: Charmichael, Teaire L.. Christian Brothers University; Estados UnidosFil: Jacob, Mellina M.. Universidade Federal do Pará; BrasilFil: Lacerda, Eliza M. C. B.. Universidade Federal do Pará; BrasilFil: Gomes, Bruno D.. Universidade Federal do Pará; BrasilFil: Fitzgerald, Malinda E. C.. Christian Brothers University; Estados Unidos. University of Tennessee; Estados UnidosFil: Ventura, Dora F.. Universidade de Sao Paulo; BrasilFil: Silveira, Luiz C. L.. Universidade do Ceuma; Brasil. Universidade Federal do Pará; BrasilFil: O´donell, Beatriz Maria. Universidad Nacional de Tucumán. Facultad de Ciencias Exactas y Tecnología. Laboratorio de Luminotecnia; ArgentinaFil: Souza, Givago S.. Universidade Federal do Pará; Brasi

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Stimulation of Baroresponsive Parts of the Nucleus of the Solitary Tract Produces Nitric Oxide-mediated Choroidal Vasodilation in Rat Eye

Preganglionic parasympathetic neurons of the ventromedial part of the superior salivatory nucleus (SSN) mediate vasodilation of orbital and choroidal blood vessels, via their projection to the nitrergic pterygopalatine ganglion (PPG) neurons that innervate these vessels. We recently showed that the baroresponsive part of the nucleus of the solitary tract (NTS) innervates choroidal control parasympathetic preganglionic neurons of SSN in rats. As this projection provides a means by which blood pressure signals may modulate ChBF, we investigated if activation of baroresponsive NTS evokes ChBF increases in rat eye, using Laser Doppler flowmetry to measure ChBF transclerally. We found that electrical activation of ipsilateral baroresponsive NTS and its efferent fiber pathway to choroidal SSN increased mean ChBF by about 40-80% above baseline, depending on current level. The ChBF responses obtained with stimulation of baroresponsive NTS were driven by increases in both choroidal blood volume (i.e. vasodilation) and choroidal blood velocity (presumed orbital vessel dilation). Stimulation of baroresponsive NTS, by contrast, yielded no significant mean increases in systemic arterial blood pressure. We further found that the increases in ChBF with NTS stimulation were significantly reduced by administration of the neuronal nitric oxide synthase inhibitor Nω-propyl-l-arginine (NPA), thus implicating nitrergic PPG terminals in the NTS-elicited ChBF increases. Our results show that NTS neurons projecting to choroidal SSN do mediate increase in ChBF, and thus suggest a role of baroresponsive NTS in the blood pressure-dependent regulation of ChBF

The identification and neurochemical characterization of central neurons that target parasympathetic preganglionic neurons involved in the regulation of choroidal blood flow in the rat eye using pseudorabies virus, immunolabeling and conventional pathway tracing methods

The choroidal blood vessels of the eye provide the main vascular support to the outer retina. These blood vessels are under parasympathetic vasodilatory control via input from the pterygopalatine ganglion (PPG), which in turn receives its preganglionic input from the superior salivatory nucleus (SSN) of the hindbrain. The present study characterized the central neurons projecting to the SSN neurons innervating choroidal PPG neurons, using pathway tracing and immunolabeling. In the initial set of studies, minute injections of the Bartha strain of the retrograde transneuronal tracer pseudorabies virus (PRV) were made into choroid in rats in which the superior cervical ganglia had been excised (to prevent labeling of sympathetic circuitry). Diverse neuronal populations beyond the choroidal part of ipsilateral SSN showed transneuronal labeling, which notably included the parvocellular part of the paraventricular nucleus of the hypothalamus (PVN), the periaqueductal gray, the raphe magnus (RaM), the B3 region of the pons, A5, the nucleus of the solitary tract (NTS), the rostral ventrolateral medulla (RVLM), and the intermediate reticular nucleus of the medulla. The PRV+ neurons were located in the parts of these cell groups that are responsive to systemic blood pressure signals and involved in systemic blood pressure regulation by the sympathetic nervous system. In a second set of studies using PRV labeling, conventional pathway tracing, and immunolabeling, we found that PVN neurons projecting to SSN tended to be oxytocinergic and glutamatergic, RaM neurons projecting to SSN were serotonergic, and NTS neurons projecting to SSN were glutamatergic. Our results suggest that blood pressure and volume signals that drive sympathetic constriction of the systemic vasculature may also drive parasympathetic vasodilation of the choroidal vasculature, and may thereby contribute to choroidal baroregulation during low blood pressure

Control of choroidal blood flow by the nucleus of Edinger-Westphal in pigeons: A laser doppler study. Investigative Ophthalmology and Visual

Anatomical studies in birds have suggested that choroidal blood flow may be regulated by a circuit involving the following serially-connected components: the retina-the suprachiasmatic nucleus (SCN)-the medial subdivision of the nucleus of Edinger-Westphal (mEW)-the ciliary ganglion-the choroidal blood vessels. In order to better clarify the role of this circuit, we examined the effects of electrical stimulation of EW on choroidal blood flow in the ipsilateral eye, using laser Doppler velocimetry to monitor choroidal blood flow in the superior pole of the eye. Baseline choroidal blood flow values (144-311.3 mg/min per eye) were found to be comparable to those previously reported in rabbits, cats and primates. Stimulation of EW dramatically increased choroidal blood flow. The increases were current-related and the average maximal increases ranged between 300-700% above baseline values. In contrast, EW stimulation had little or no effect on overall bodily blood flow. All EW stimulation sites were later verified histologically. These results indicate that the SCN-mEW circuit in birds may be involved in mediating increases in choroidal blood flow, possibly in response to the levels of retinal illumination. Such adaptive neural regulation of choroidal blood flow may play an important role in mitigating the potentially deleterious effects of light on the retina. Invest Ophthalmol Vis Sc

Bruton tyrosine kinase is essential for botrocetin/VWF-induced signaling and GPIb-dependent thrombus formation in vivo

Botrocetin (bt)-facilitated binding of von Willebrand factor (VWF) to the platelet membrane glycoprotein (GP) Ib-IX-V complex on platelets in suspension initiates a signaling cascade that causes αIIbβ3 activation and platelet aggregation. Previous work has demonstrated that bt/VWF-mediated agglutination activates αIIbβ3 and elicits ATP secretion in a thromboxane A2 (TxA2)-dependent manner. The signaling that results in TxA2 production was shown to be initiated by Lyn, enhanced by Src, and propagated through Syk, SLP-76, PI3K, PLCγ2, and PKC. Here, we demonstrate that the signaling elicited by GPIb-mediated agglutination that results in TxA2 production is dependent on Bruton tyrosine kinase (Btk). The results demonstrate that Btk is downstream of Lyn, Syk, SLP-76, and PI3K; upstream of ERK1/2, PLCγ2, and PKC; and greatly enhances Akt phosphorylation. The relationship(s), if any, between ERK1/2, PLCγ2, and PKC were not elucidated. The requirement for Btk and TxA2 receptor function in GPIb-dependent arterial thrombosis was confirmed in vivo by characterizing blood flow in ferric chloride-treated mouse carotid arteries. These results demonstrate that the Btk family kinase, Tec, cannot provide the function(s) missing because of the absence of Btk and that Btk is essential for both bt/VWF-mediated agglutination-induced TxA2 production and GPIb-dependent stable arterial thrombus formation in vivo

Low number of luminance levels in the luminance noise increases color discrimination thresholds estimated with pseudoisochromatic stimuli

In pseudoisochromatic stimuli the presence of spatial and luminance noise forces the subject to discriminate the target from the background solely on the basis of chromaticity difference. Color-blind subjects may show difficulty to identify the target due to the elimination of borders and brightness clues caused by the luminance and spatial noise. Few studies have fully described the features of pseudoisochromatic stimuli. Fewer investigators have focused their studies in the effects of specific pseudoisochromatic parameters on color discrimination. We used the Cambridge Color Test (CCT) to investigate the influence on color discrimination thresholds due to the number of luminance levels present in the luminance noise. The CCT default has 6 luminance steps; however, in our investigation a total of eight different conditions were tested from 2 to 16 luminance steps. It was found that the CCT provided very robust values for color discrimination thresholds, which were degraded only for very small number of luminance steps. When the number of steps was increased, the color discrimination thresholds improved from 2 to 6 luminance steps and gradually reached a plateau for 10 or more luminance steps. The area of color discrimination ellipses as a function of luminance steps matches the relative proportion of ineffective contrasts between mosaic patches as a function of luminance steps, assuming that contrast becomes ineffective for values 18.6% or less. The lower number of color and luminance interactions in these conditions could explain the measured increase of color discrimination thresholds. The primary conclusion from this investigation was that results from pseudoisochromatic tests should have their parameters described in more detail. This type of description would allow a better understanding of the results provided, interpretations, and therefore cross study comparison of results obtained from different laboratories