Pattern-sensitive neurons reveal encoding of complex auditory regularities in the rat inferior colliculus.

A 'pattern alternation paradigm' has been previously used in human ERP recordings to investigate the brain encoding of complex auditory regularities, but prior studies on regularity encoding in animal models to examine mechanisms of adaptation of auditory neuronal responses have used primarily oddball stimulus sequences to study stimulus-specific adaptation alone. In order to examine the sensitivity of neuronal adaptation to expected and unexpected events embedded in a complex sound sequence, we used a similar patterned sequence of sounds. We recorded single unit activity and compared neuronal responses in the rat inferior colliculus (IC) to sound stimuli conforming to pattern alternation regularity with those to stimuli in which occasional sound repetitions violated that alternation. Results show that some neurons in the rat inferior colliculus are sensitive to the history of patterned stimulation and to violations of patterned regularity, demonstrating that there is a population of subcortical neurons, located as early as the level of the midbrain, that can detect more complex stimulus regularities than previously supposed and that are as sensitive to complex statistics as some neurons in primary auditory cortex. Our findings indicate that these pattern-sensitive neurons can extract temporal and spectral regularities between successive acoustic stimuli. This is important because the extraction of regularities from the sound sequences will result in the development of expectancies for future sounds and hence, the present results are compatible with predictive coding models. Our results demonstrate that some collicular neurons, located as early as in the midbrain level, are involved in the generation and shaping of prediction errors in ways not previously considered and thus, the present findings challenge the prevailing view that perceptual organization of sound only emerges at the auditory cortex level.

The effects of nicotine on cone and rod b-wave responses in larval zebrafish.

Acetylcholine is present in and released from starburst amacrine cells in the inner plexiform layer (IPL), but its role in retinal function except, perhaps, in early development, is unclear. Nicotinic acetylcholine receptors are thought to be present on ganglion, amacrine, and bipolar cell processes in the IPL, and it is known that acetylcholine increases the spontaneous and light-evoked responses of retinal ganglion cells. The effects of acetylcholine on bipolar cells are not known, and here we report the effects of nicotine on the b-wave of the electroretinogram in larval zebrafish. The b-wave originates mainly from ON-bipolar cells, and the larval zebrafish retina is cone-dominated. Only small rod responses can be elicited with dim lights in wild-type larval zebrafish retinas, but rod responses can be recorded over a range of intensities in a mutant ( n o optokinetic response f ) fi sh that has no cone function. We fi nd that nicotine strongly enhances cone-driven b-wave response amplitudes but depresses rod driven b-wave response amplitudes without, however, affecting rod- or cone-driven b-wave light sensitivity.

A genome-wide association study for myopia and refractive error identifies a susceptibility locus at 15q25.

Myopia and hyperopia are at opposite ends of the continuum of refraction, the measure of the eye's ability to focus light, which is an important cause of visual impairment (when aberrant) and is a highly heritable trait. We conducted a genome-wide association study for refractive error in 4,270 individuals from the TwinsUK cohort. We identified SNPs on 15q25 associated with refractive error (rs8027411, P = 7.91 × 10⁻8). We replicated this association in six adult cohorts of European ancestry with a combined 13,414 individuals (combined P = 2.07 × 10⁻9). This locus overlaps the transcription initiation site of RASGRF1, which is highly expressed in neurons and retina and has previously been implicated in retinal function and memory consolidation. Rasgrf1(-/-) mice show a heavier average crystalline lens (P = 0.001). The identification of a susceptibility locus for refractive error on 15q25 will be important in characterizing the molecular mechanism responsible for the most common cause of visual impairment.

Developmental expression patterns of acetylcholinesterase and cholineacetyltransferase in zebrafish retina

During recent years a key role as morphogen has been postulated for the neurotransmitter acetylcholine in the developing Central Nervous System. Acetylcholine released from growing axons regulates growth, differentiation and plasticity. The acetylcholine distribution is frequently defined by acetylcholinesterase and choline acetyltransferase expression patterns. The cholinergic/cholinoceptive system in the adult zebrafish retina has been described. Nevertheless, there are no data regarding the developing retina. The acetylcholinesterase and choline acetyltransferase distribution patterns during zebrafish retinal development are very similar. In both cases the first positive elements appear in the plexiform layers and in later stages reactive amacrine cells have been observed in the ganglion cell layer and inner nuclear layer. In the adult retina a cholinergic and cholinoceptive neuropile band is observed in the inner plexiform layer. Displaced amacrine cells and amacrine cells positive to both markers have been observed. Transient expressions of choline acetyltransferase in the optic nerve and outer plexiform layer and of acetylcholinesterase in amacrine cells and displaced amacrine cells are observed during retinal development coinciding with the arrangement of the pioneering retinal projections into the optic tectum. The mature distribution pattern of the cholinergic/ cholinoceptive system in the adult retina is conserved along the phylogenetic scale, thus it seems to be a primary feature acquired relatively early during the evolution of vertebrates (AU)

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RasGRF1 disruption causes retinal photoreception defects and associated transcriptomic alterations.

RasGRF1 null mutant mice display impaired memory/learning and their hippocampus transcriptomic pattern includes a number of differentially expressed genes playing significant roles in sensory development and function. Odour avoidance and auditory brainstem response tests yielded normal results but electroretinographic analysis showed severe light perception impairment in the RasGRF1 knockouts. Whereas no structural alterations distinguished the retinas of wild-type and knockout mice, microarray transcriptional analysis identified at least 44 differentially expressed genes in the retinas of these Knockout animals. Among these, Crb1, Pttg1, Folh1 and Myo7a have been previously related to syndromes involving retina degeneration. Interestingly, over-expression of Folh1 would be expected to result in accumulation of its enzymatic product N-acetyl-aspartate, an event known to be linked to Canavan disease, a human cerebral degenerative syndrome often involving blindness and hearing loss. Consistently, in vivo brain nuclear magnetic resonance spectroscopy identified higher levels of N-acetyl-aspartate in our RasGRF1-/- mice and immunohistochemical analysis detected reduced levels of aspartoacylase, the enzyme which degrades N-acetyl-aspartate. These studies demonstrate for the first time the functional relevance of Ras signalling in mammalian photoreception and warrant further analysis of RasGRF1 Knockout mice as potential models to analyse molecular mechanisms underlying defective photoreception human diseases.

Chemical organization of the macaque monkey olfactory bulb: III. Distribution of cholinergic markers.

The distribution patterns of choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) were studied in the olfactory bulb (OB) of three species of macaque. AChE was detected by a histochemical method and ChAT immunoreactivity by immunocytochemistry. Similar results were observed in all species analyzed. With the exception of the olfactory nerve layer, all layers of the macaque monkey OB demonstrated a dense innervation of AChE- and ChAT-positive fibers. The distribution patterns of AChE- and ChAT-labeled fibers were similar for both cholinergic markers, although the number of AChE-labeled fibers was clearly higher than the number of ChAT-immunoreactive fibers. The highest density of AChE and ChAT-stained fibers was observed in the interface between the glomerular layer and the external plexiform layer and in the internal plexiform layer. Dense bundles of labeled fibers were observed in the caudal OB, coursing from the olfactory peduncle. All ChAT-immunopositive elements were identified as centrifugal fibers, derived from neurons caudal to the OB. Neither olfactory fibers nor intrinsic neurons were observed after ChAT immunocytochemistry. However, a few AChE-positive cells were observed in the glomerular layer and in both external and internal plexiform layers. These neurons were presumably identified as periglomerular cells, superficial short-axon cells, and/or external tufted cells and deep short-axon cells. Contrary to other neurotransmitters and neuroactive substances, the distribution patterns of ChAT and AChE activities in the macaque monkey OB closely resembled the patterns described in macrosmatic mammals and showed laminar differences with the distribution pattern observed in humans.

Development of the cholinergic system in the brain and retina of the zebrafish.

We have analyzed the distribution pattern of choline acetyltransferase (ChAT) in the zebrafish brain and retina during ontogeny. ChAT-immunoreactive (ChAT-ir) neurons are observed in the prosencephalon from 60 h postfertilization (hpf) onwards, exclusively in the preoptic area (basal plate of p6) derived from the secondary prosencephalon. In the mesencephalon, ChAT-ir cells are observed in both the optic tectum and the tegmentum. Stained cells in the tegmentum are observed from 60 hpf onwards, while in the optic tectum they appear after hatching. In the rhombencephalon, ChAT-ir cells are first observed in the isthmic region (rh1) and in the medulla oblongata (rh5-rh7) at the end of embryonic life. The rhombencephalic cholinergic cell groups develop in a gradual caudorostral sequence. Motoneurons of the spinal cord are ChAT-ir from 48 hpf onwards. The retina displays ChAT-ir neuropil in both the inner and outer plexiform layers from embryonic life, whereas stained amacrine cells are only observed after hatching. The staining in the outer plexiform layer gradually decreases during juvenile development. The optic nerve axons show a transient expression of ChAT at the end of embryonic development. The early presence of ChAT immunolabeling suggests an important neuromodulator role for acetylcholine in the first developmental stages.

Comparative analysis of the distribution of choline acetyltransferase in the central nervous system of cyprinids.

The general organization of the cholinergic system in the central nervous system is similar among vertebrates, though fish show higher variability. Thus, in zebrafish, cholinergic cells are absent from the habenula and the rhombencephalic reticular formation, where such neurons are present in most vertebrate species analyzed. In this work, we compared the distribution of choline acetyltransferase in the central nervous system of both zebrafish and tench, in order to investigate whether these divergences in the distribution of cholinergic cells in zebrafish are species-specific, or a feature shared by members of the cyprinid family. Our data show that these two cyprinid possess in common some peculiarities in their cholinergic system that are not present in the rest of fish analyzed (e.g. absence of cholinergic cells in the habenula and their presence in the descendent octaval nucleus). Nonetheless, some cholinergic cells were observed in the dorsal thalamus and rhombencephalic reticular nuclei of the tench, which were absent in the same regions in zebrafish. The comparative analysis suggests a divergent evolution of the cholinergic system among close-related cyprinid species.

Cholinergic elements in the zebrafish central nervous system: Histochemical and immunohistochemical analysis.

Recently, the zebrafish has been extensively used for studying the development of the central nervous system (CNS). However, the zebrafish CNS has been poorly analyzed in the adult. The cholinergic/cholinoceptive system of the zebrafish CNS was analyzed by using choline acetyltransferase (ChAT) immunohistochemistry and acetylcholinesterase (AChE) histochemistry in the brain, retina, and spinal cord. AChE labeling was more abundant and more widely distributed than ChAT immunoreactivity. In the telencephalon, ChAT-immunoreactive (ChAT-ir) cells were absent, whereas AChE-positive neurons were observed in both the olfactory bulb and the telencephalic hemispheres. The diencephalon was the region with the lowest density of AChE-positive cells, mainly located in the pretectum, whereas ChAT-ir cells were exclusively located in the preoptic region. ChAT-ir cells were restricted to the periventricular stratum of the optic tectum, but AChE-positive neurons were observed throughout the whole extension of the lamination except in the marginal stratum. Although ChAT immunoreactivity was restricted to the rostral tegmental, oculomotor, and trochlear nuclei within the mesencephalic tegmentum, a widespread distribution of AChE reactivity was observed in this region. The isthmic region showed abundant AChE-positive and ChAT-ir cells in the isthmic, secondary gustatory and superior reticular nucleus and in the nucleus lateralis valvulae. ChAT immunoreactivity was absent in the cerebellum, although AChE staining was observed in Purkinje and granule cells. The medulla oblongata showed a widespread distribution of AChE-positive cells in all main subdivisions, including the octavolateral area, reticular formation, and motor nuclei of the cranial nerves. ChAT-ir elements in this area were restricted to the descending octaval nucleus, the octaval efferent nucleus and the motor nuclei of the cranial nerves. Additionally, spinal cord motoneurons appeared positive to both markers. Substantial differences in the ChAT and AChE distribution between zebrafish and other fish species were observed, which could be important because zebrafish is widely used as a genetic or developmental animal model.

Distribution of calbindin D-28k in the entorhinal, perirhinal, and parahippocampal cortices of the macaque monkey.

We examined the distribution of calbindin D-28k-immunoreactive (CB-IR) neurons, fibers, and neuropil in the entorhinal (area 28), perirhinal (areas 35 and 36), and parahippocampal (areas TH and TF) cortices in the macaque monkey. Two main findings are reported. First, except for CB-IR neurogliaform cells that are only observed in the parahippocampal cortex, the morphology of CB-stained pyramidal and nonpyramidal cells were similar across the three cortical areas examined. Second, we find that the topography of CB staining differed between the three areas. The entorhinal cortex exhibits the most striking gradient of CB staining such that the most anterior and medial portions are most strongly labeled, whereas posterior and lateral areas exhibit only weak labeling. The labeling throughout the perirhinal and parahippocampal cortices is more homogeneous. Area 35 contains only lightly stained neuropil and few CB-IR cells. Area 36 and areas TH and TF of the parahippocampal cortex contain a moderate to high density of CB-IR cells and fibers throughout their full rostrocaudal extents, although each area exhibits unique laminar patterns of staining. In all areas examined, the highest density of CB-positive cells and fibers is observed in superficial layers with lower densities of CB-positive cells and fibers present in deep layers. These findings, taken together with our current understanding of the connections of these areas may have implications for understanding the circuit properties of the entorhinal, perirhinal, and parahippocampal cortices areas in both normal and disease states.

Targeted disruption of Ras-Grf2 shows its dispensability for mouse growth and development.

The mammalian Grf1 and Grf2 proteins are Ras guanine nucleotide exchange factors (GEFs) sharing a high degree of structural homology, as well as an elevated expression level in central nervous system tissues. Such similarities raise questions concerning the specificity and/or redundancy at the functional level between the two Grf proteins. grf1-null mutant mice have been recently described which showed phenotypic growth reduction and long-term memory loss. To gain insight into the in vivo function of Grf2, we disrupted its catalytic CDC25-H domain by means of gene targeting. Breeding among grf2(+/-) animals gave rise to viable grf2(-/-) adult animals with a normal Mendelian pattern, suggesting that Grf2 is not essential for embryonic and adult mouse development. In contrast to Grf1-null mice, analysis of grf2(-/-) litters showed similar size and weight as their heterozygous or wild-type grf2 counterparts. Furthermore, adult grf2(-/-) animals reached sexual maturity at the same age as their wild-type littermates and showed similar fertility levels. No specific pathology was observed in adult Grf2-null animals, and histopathological studies showed no observable differences between null mutant and wild-type Grf2 mice. These results indicate that grf2 is dispensable for mouse growth, development, and fertility. Furthermore, analysis of double grf1/grf2 null animals did not show any observable phenotypic difference with single grf1(-/-) animals, further indicating a lack of functional overlapping between the two otherwise highly homologous Grf1 and Grf2 proteins.