RESUMEN
Recently, a Y727C variant in the dual-specific 3',5'-cyclic nucleotide phosphodiesterase 11A (PDE11A-Y727C) was linked to increased sleep quality and reduced myopia risk in humans. Given the well-established role that the PDE11 substrates cAMP and cGMP play in eye physiology and sleep, we determined if (1) PDE11A protein is expressed in the retina or other eye segments in mice, (2) PDE11A-Y7272C affects catalytic activity and/or subcellular compartmentalization more so than the nearby suicide-associated PDE11A-M878V variant, and (3) Pde11a deletion alters eye growth or sleep quality in male and female mice. Western blots show distinct protein expression of PDE11A4, but not PDE11A1-3, in eyes of Pde11a WT, but not KO mice, that vary by eye segment and age. In HT22 and COS-1 cells, PDE11A4-Y727C reduces PDE11A4 catalytic activity far more than PDE11A4-M878V, with both variants reducing PDE11A4-cAMP more so than PDE11A4-cGMP activity. Despite this, Pde11a deletion does not alter age-related changes in retinal or lens thickness or axial length, nor vitreous or anterior chamber depth. Further, Pde11a deletion only minimally changes refractive error and sleep quality. That said, both variants also dramatically alter the subcellular compartmentalization of human and mouse PDE11A4, an effect occurring independently of dephosphorylating PDE11A4-S117/S124 or phosphorylating PDE11A4-S162. Rather, re-compartmentalization of PDE11A4-Y727C is due to the loss of the tyrosine changing how PDE11A4 is packaged/repackaged via the trans-Golgi network. Therefore, the protective impact of the Y727C variant may reflect a gain-of-function (e.g., PDE11A4 displacing another PDE) that warrants further investigation in the context of reversing/preventing sleep disturbances or myopia.
Asunto(s)
3',5'-GMP Cíclico Fosfodiesterasas , Miopía , Humanos , Masculino , Femenino , Animales , Ratones , 3',5'-GMP Cíclico Fosfodiesterasas/metabolismo , Hidrolasas Diéster Fosfóricas/genética , Hidrolasas Diéster Fosfóricas/metabolismo , Calidad del Sueño , Western BlottingRESUMEN
Recently, a Y727C variant in the dual-specific 3',5'-cyclic nucleotide phosphodiesterase 11A (PDE11A-Y727C) was linked to increased sleep quality and reduced myopia risk in humans. Given the well-established role that the PDE11 substrates cAMP and cGMP play in eye physiology and sleep, we determined if 1) PDE11A protein is expressed in the retina or other eye segments in mouse, 2) PDE11A-Y7272C affects catalytic activity and/or subcellular compartmentalization more so than the nearby suicide-associated PDE11A-M878V variant, and 3) Pde11a deletion alters eye growth or sleep quality in male and female mice. Western blots show distinct protein expression of PDE11A4, but not PDE11A1-3, in eyes of Pde11a WT-but not KO mice-that vary by eye segment and age. In HT22 and COS-1 cells, PDE11A4-Y727C reduces PDE11A4 catalytic activity far more than PDE11A4-M878V, with both variants reducing PDE11A4-cAMP more so than PDE11A4-cGMP activity. Despite this, Pde11a deletion does not alter age-related changes in retinal or lens thickness, axial length, nor vitreous or anterior chamber depth. Further, Pde11a deletion only minimally changes refractive error and sleep quality. That said, both variants also dramatically alter the subcellular compartmentalization of human and mouse PDE11A4, an effect occurring independently of dephosphorylating PDE11A4-S117/S124 or phosphorylating PDE11A4-S162. Rather, re-compartmentalization of PDE11A4-Y727C is due to the loss of the tyrosine changing how PDE11A4 is packaged/repackaged via the trans-Golgi network. Therefore, the protective impact of the Y727C variant may reflect a gain-of-function (e.g., PDE11A4 displacing another PDE) that warrants further investigation in the context of reversing/preventing sleep disturbances or myopia.
RESUMEN
In Nyxnob mice, a model for congenital nystagmus associated with congenital stationary night blindness (CSNB), synchronous oscillating retinal ganglion cells (RGCs) lead to oscillatory eye movements, i.e. nystagmus. Given the specific expression of mGluR6 and Cav 1.4 in the photoreceptor to bipolar cell synapses, as well as their clinical association with CSNB, we hypothesize that Grm6nob3 and Cav 1.4-KO mutants show, like the Nyxnob mouse, oscillations in both their RGC activity and eye movements. Using multi-electrode array recordings of RGCs and measurements of the eye movements, we demonstrate that Grm6nob3 and Cav 1.4-KO mice also show oscillations of their RGCs as well as a nystagmus. Interestingly, the preferred frequencies of RGC activity as well as the eye movement oscillations of the Grm6nob3 , Cav 1.4-KO and Nyxnob mice differ among mutants, but the neuronal activity and eye movement behaviour within a strain remain aligned in the same frequency domain. Model simulations indicate that mutations affecting the photoreceptor-bipolar cell synapse can form a common cause of the nystagmus of CSNB by driving oscillations in RGCs via AII amacrine cells. KEY POINTS: In Nyxnob mice, a model for congenital nystagmus associated with congenital stationary night blindness (CSNB), their oscillatory eye movements (i.e. nystagmus) are caused by synchronous oscillating retinal ganglion cells. Here we show that the same mechanism applies for two other CSNB mouse models - Grm6nob3 and Cav 1.4-KO mice. We propose that the retinal ganglion cell oscillations originate in the AII amacrine cells. Model simulations show that by only changing the input to ON-bipolar cells, all phenotypical differences between the various genetic mouse models can be reproduced.
Asunto(s)
Miopía , Ceguera Nocturna , Nistagmo Congénito , Ratones , Animales , Ceguera Nocturna/genética , Ceguera Nocturna/metabolismo , Miopía/genética , Miopía/metabolismo , Células Ganglionares de la Retina/fisiología , Mutación , ElectrorretinografíaRESUMEN
PURPOSE: To study the effectiveness of high-dose atropine for reducing eye growth in Mendelian myopia in children and mice. METHODS: We studied the effect of high-dose atropine in children with progressive myopia with and without a monogenetic cause. Children were matched for age and axial length (AL) in their first year of treatment. We considered annual AL progression rate as the outcome and compared rates with percentile charts of an untreated general population. We treated C57BL/6J mice featuring the myopic phenotype of Donnai-Barrow syndrome by selective inactivation of Lrp2 knock out (KO) and control mice (CTRL) daily with 1% atropine in the left eye and saline in the right eye, from postnatal days 30-56. Ocular biometry was measured using spectral-domain optical coherence tomography. Retinal dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DOPAC) were measured using high-performance liquid chromatography. RESULTS: Children with a Mendelian form of myopia had average baseline spherical equivalent (SE) -7.6 ± 2.5D and AL 25.8 ± 0.3 mm; children with non-Mendelian myopia had average SE -7.3 ± 2.9 D and AL 25.6 ± 0.9 mm. During atropine treatment, the annual AL progression rate was 0.37 ± 0.08 and 0.39 ± 0.05 mm in the Mendelian myopes and non-Mendelian myopes, respectively. Compared with progression rates of untreated general population (0.47 mm/year), atropine reduced AL progression with 27% in Mendelian myopes and 23% in non-Mendelian myopes. Atropine significantly reduced AL growth in both KO and CTRL mice (male, KO: -40 ± 15; CTRL: -42 ± 10; female, KO: -53 ± 15; CTRL: -62 ± 3 µm). The DA and DOPAC levels 2 and 24 h after atropine treatment were slightly, albeit non-significantly, elevated. CONCLUSIONS: High-dose atropine had the same effect on AL in high myopic children with and without a known monogenetic cause. In mice featuring a severe form of Mendelian myopia, atropine reduced AL progression. This suggests that atropine can reduce myopia progression even in the presence of a strong monogenic driver.
Asunto(s)
Atropina , Miopía Degenerativa , Humanos , Masculino , Femenino , Animales , Ratones , Ácido 3,4-Dihidroxifenilacético , Ratones Endogámicos C57BL , Atropina/farmacología , Refracción Ocular , Retina , Progresión de la Enfermedad , Soluciones OftálmicasRESUMEN
Congenital nystagmus is a condition where the eyes of patients oscillate, mostly horizontally, with a frequency of between 2 and 10 Hz. Historically, nystagmus is believed to be caused by a maladaptation of the oculomotor system and is thus considered a disease of the brain stem. However, we have recently shown that congenital nystagmus associated with congenital stationary night blindness is caused by synchronously oscillating retinal ganglion cells. In this perspective article, we discuss how some details of nystagmus can be accounted for by the retinal mechanism we propose.
RESUMEN
Refractive errors are common eye disorders characterized by a mismatch between the focal power of the eye and its axial length. An increased axial length is a common cause of the refractive error myopia (nearsightedness). The substantial increase in myopia prevalence over the last decades has raised public health concerns because myopia can lead to severe ocular complications later in life. Genomewide association studies (GWAS) have made considerable contributions to the understanding of the genetic architecture of refractive errors. Among the hundreds of genetic variants identified, common variants near the gap junction delta-2 (GJD2) gene have consistently been reported as one of the top hits. GJD2 encodes the connexin 36 (Cx36) protein, which forms gap junction channels and is highly expressed in the neural retina. In this review, we provide current evidence that links GJD2(Cx36) to the development of myopia. We summarize the gap junctional communication in the eye and the specific role of GJD2(Cx36) in retinal processing of visual signals. Finally, we discuss the pathways involving dopamine and gap junction phosphorylation and coupling as potential mechanisms that may explain the role of GJD2(Cx36) in refractive error development.
Asunto(s)
Conexinas , Miopía , Errores de Refracción , Conexinas/genética , Conexinas/metabolismo , Uniones Comunicantes/metabolismo , Humanos , Miopía/genética , Miopía/metabolismo , Errores de Refracción/genética , Errores de Refracción/metabolismo , Retina/metabolismo , Proteína delta-6 de Union ComunicanteRESUMEN
Myopia is the most common developmental disorder of juvenile eyes, and it has become an increasing cause of severe visual impairment. The GJD2 locus has been consistently associated with myopia in multiple independent genome-wide association studies. However, despite the strong genetic evidence, little is known about the functional role of GJD2 in refractive error development. Here, we find that depletion of gjd2a (Cx35.5) or gjd2b (Cx35.1) orthologs in zebrafish, cause changes in the biometry and refractive status of the eye. Our immunohistological and scRNA sequencing studies show that Cx35.5 (gjd2a) is a retinal connexin and its depletion leads to hyperopia and electrophysiological changes in the retina. These findings support a role for Cx35.5 (gjd2a) in the regulation of ocular biometry. Cx35.1 (gjd2b) has previously been identified in the retina, however, we found an additional lenticular role. Lack of Cx35.1 (gjd2b) led to a nuclear cataract that triggered axial elongation. Our results provide functional evidence of a link between gjd2 and refractive error.
Asunto(s)
Conexinas/genética , Modelos Animales de Enfermedad , Proteínas del Ojo/genética , Mutación , Errores de Refracción/genética , Proteínas de Pez Cebra/genética , Pez Cebra/genética , Animales , Catarata/genética , Conexinas/metabolismo , Proteínas del Ojo/metabolismo , Perfilación de la Expresión Génica/métodos , Humanos , Miopía/genética , RNA-Seq/métodos , Retina/metabolismo , Retina/patología , Epitelio Pigmentado de la Retina/metabolismo , Epitelio Pigmentado de la Retina/patología , Análisis de la Célula Individual/métodos , Pez Cebra/metabolismo , Proteínas de Pez Cebra/metabolismoRESUMEN
Congenital nystagmus, involuntary oscillating small eye movements, is commonly thought to originate from aberrant interactions between brainstem nuclei and foveal cortical pathways. Here, we investigated whether nystagmus associated with congenital stationary night blindness (CSNB) results from primary deficits in the retina. We found that CSNB patients as well as an animal model (nob mice), both of which lacked functional nyctalopin protein (NYX, nyx) in ON bipolar cells (BCs) at their synapse with photoreceptors, showed oscillating eye movements at a frequency of 4-7 Hz. nob ON direction-selective ganglion cells (DSGCs), which detect global motion and project to the accessory optic system (AOS), oscillated with the same frequency as their eyes. In the dark, individual ganglion cells (GCs) oscillated asynchronously, but their oscillations became synchronized by light stimulation. Likewise, both patient and nob mice oscillating eye movements were only present in the light when contrast was present. Retinal pharmacological and genetic manipulations that blocked nob GC oscillations also eliminated their oscillating eye movements, and retinal pharmacological manipulations that reduced the oscillation frequency of nob GCs also reduced the oscillation frequency of their eye movements. We conclude that, in nob mice, synchronized oscillations of retinal GCs, most likely the ON-DCGCs, cause nystagmus with properties similar to those associated with CSNB in humans. These results show that the nob mouse is the first animal model for a form of congenital nystagmus, paving the way for development of therapeutic strategies.
Asunto(s)
Enfermedades Hereditarias del Ojo/fisiopatología , Enfermedades Genéticas Ligadas al Cromosoma X/fisiopatología , Miopía/fisiopatología , Ceguera Nocturna/fisiopatología , Nistagmo Congénito/etiología , Células Ganglionares de la Retina/fisiología , Animales , Preescolar , Modelos Animales de Enfermedad , Femenino , Humanos , Lactante , Masculino , Ratones NoqueadosRESUMEN
OBJECTIVE: Disrupting thalamocortical activity patterns has proven to be a promising approach to stop generalized spike-and-wave discharges (GSWDs) characteristic of absence seizures. Here, we investigated to what extent modulation of neuronal firing in cerebellar nuclei (CN), which are anatomically in an advantageous position to disrupt cortical oscillations through their innervation of a wide variety of thalamic nuclei, is effective in controlling absence seizures. METHODS: Two unrelated mouse models of generalized absence seizures were used: the natural mutant tottering, which is characterized by a missense mutation in Cacna1a, and inbred C3H/HeOuJ. While simultaneously recording single CN neuron activity and electrocorticogram in awake animals, we investigated to what extent pharmacologically increased or decreased CN neuron activity could modulate GSWD occurrence as well as short-lasting, on-demand CN stimulation could disrupt epileptic seizures. RESULTS: We found that a subset of CN neurons show phase-locked oscillatory firing during GSWDs and that manipulating this activity modulates GSWD occurrence. Inhibiting CN neuron action potential firing by local application of the γ-aminobutyric acid type A (GABA-A) agonist muscimol increased GSWD occurrence up to 37-fold, whereas increasing the frequency and regularity of CN neuron firing with the use of GABA-A antagonist gabazine decimated its occurrence. A single short-lasting (30-300 milliseconds) optogenetic stimulation of CN neuron activity abruptly stopped GSWDs, even when applied unilaterally. Using a closed-loop system, GSWDs were detected and stopped within 500 milliseconds. INTERPRETATION: CN neurons are potent modulators of pathological oscillations in thalamocortical network activity during absence seizures, and their potential therapeutic benefit for controlling other types of generalized epilepsies should be evaluated.
Asunto(s)
Potenciales de Acción/fisiología , Núcleos Cerebelosos/fisiopatología , Epilepsia Tipo Ausencia/fisiopatología , Neuronas/fisiología , Potenciales de Acción/efectos de los fármacos , Animales , Canales de Calcio Tipo N/genética , Núcleos Cerebelosos/efectos de los fármacos , Modelos Animales de Enfermedad , Femenino , Antagonistas del GABA/farmacología , Agonistas de Receptores de GABA-A/farmacología , Masculino , Ratones , Ratones Endogámicos C3H , Ratones Transgénicos , Neuronas/efectos de los fármacos , Optogenética , Tálamo/efectos de los fármacos , Tálamo/fisiopatologíaRESUMEN
In addition to the well-known signals of retinal image slip, floccular complex spikes (CSs) also convey nonvisual signals. We recorded eye movement and CS activity from Purkinje cells in awake rabbits sinusoidally oscillated in the dark on a vestibular turntable. The stimulus frequency ranged from 0.2 to 1.2 Hz, and the velocity amplitude ranged from 6.3 to 50°/s. The average CS modulation was evaluated at each combination of stimulus frequency and amplitude. More than 75% of the Purkinje cells carried nonvisual CS signals. The amplitude of this modulation remained relatively constant over the entire stimulus range. The phase response of the CS modulation in the dark was opposite to that during the vestibulo-ocular reflex (VOR) in the light. With increased frequency, the phase response systematically shifted from being aligned with contraversive head velocity toward peak contralateral head position. At fixed frequency, the phase response was dependent on peak head velocity, indicating a system nonlinearity. The nonvisual CS modulation apparently reflects a competition between eye movement and vestibular signals, resulting in an eye movement error signal inferred from nonvisual sources. The combination of this error signal with the retinal slip signal in the inferior olive results in a net error signal reporting the discrepancy between the actual visually measured eye movement error and the inferred eye movement error derived from measures of the internal state. The presence of two error signals requires that the role of CSs in models of the floccular control of VOR adaption be expanded beyond retinal slip.
Asunto(s)
Potenciales de Acción/fisiología , Cerebelo/citología , Células de Purkinje/fisiología , Vestíbulo del Laberinto/fisiología , Animales , Biofisica , Femenino , Funciones de Verosimilitud , Masculino , Modelos Estadísticos , Vías Nerviosas/fisiología , Nistagmo Optoquinético/fisiología , Conejos , Reflejo Vestibuloocular/fisiología , Vestíbulo del Laberinto/inervaciónRESUMEN
Humans with Down syndrome (DS) and Ts65Dn mice both show a reduced volume of the cerebellum due to a significant reduction in the density of granule neurons. Recently, cerebellar hypoplasia in Ts65Dn mice was rescued by a single treatment with SAG, an agonist of the Sonic hedgehog pathway, administered on the day of birth. In addition to normalizing cerebellar morphology, this treatment restored the ability to learn a spatial navigation task, which is associated with hippocampal function. It is not clear to what extent this improved performance results from restoration of the cerebellar architecture or a yet undefined role of Sonic hedgehog (Shh) in perinatal hippocampal development. The absence of a clearly demonstrated deficit in cerebellar function in trisomic mice exacerbates the problem of discerning how SAG acts to improve learning and memory. Here we show that phase reversal adaptation and consolidation of the vestibulo-ocular reflex is significantly impaired in Ts65Dn mice, providing for the first time a precise characterization of cerebellar functional deficits in this murine model of DS. However, these deficits do not benefit from the normalization of cerebellar morphology following treatment with SAG. Together with the previous observation that the synaptic properties of Purkinje cells are also unchanged by SAG treatment, this lack of improvement in a region-specific behavioral assay supports the possibility that a direct effect of Shh pathway stimulation on the hippocampus might explain the benefits of this potential approach to the improvement of cognition in DS.
Asunto(s)
Cerebelo/fisiopatología , Ciclohexilaminas/uso terapéutico , Síndrome de Down/tratamiento farmacológico , Aprendizaje/efectos de los fármacos , Memoria/efectos de los fármacos , Tiofenos/uso terapéutico , Animales , Cerebelo/patología , Ciclohexilaminas/farmacología , Modelos Animales de Enfermedad , Proteínas Hedgehog/agonistas , Ratones , Células de Purkinje/patología , Reflejo Vestibuloocular/efectos de los fármacos , Sinapsis/patología , Tiofenos/farmacologíaRESUMEN
Compensatory eye movements (CEMs) stabilize the field of view enabling visual sharpness despite self-induced motion or environmental perturbations. The vestibulocerebellum makes it possible to adapt these reflex behaviors to perform optimally under novel circumstances that are sustained over time. Because of this and the fact that the eye is relatively insensitive to fatigue and musculoskeletal aging effects, CEMs form an ideal motor system to assess aging effects on cerebellar motor learning. In the present study, we performed an extensive behavioral examination of the impact of aging on both basic CEMs and oculomotor-based learning paradigms spanning multiple days. Our data show that healthy aging has little to no effect on basic CEM performance despite sensory deterioration, suggesting a central compensatory mechanism. Young mice are capable of adapting their oculomotor output to novel conditions rapidly and accurately, even to the point of reversing the direction of the reflex entirely. However, oculomotor learning and consolidation capabilities show a progressive decay as age increases.
Asunto(s)
Adaptación Ocular/fisiología , Adaptación Fisiológica , Envejecimiento/fisiología , Reflejo Vestibuloocular/fisiología , Animales , Movimientos Oculares/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Actividad Motora/fisiología , Factores de TiempoRESUMEN
Thoracic aortic aneurysm and dissection (TAAD) are associated with connective tissue disorders like Marfan syndrome and Loeys-Dietz syndrome, caused by mutations in the fibrillin-1, the TGFß-receptor 1- and -2 genes, the SMAD3 and TGFß2 genes, but have also been ascribed to ACTA2 gene mutations in adults, spread throughout the gene. We report on a novel de novo c.535C>T in exon 6 leading to p.R179C aminoacid substitution in ACTA2 in a toddler girl with primary pulmonary hypertension, persistent ductus arteriosus, extensive cerebral white matter lesions, fixed dilated pupils, intestinal malrotation, and hypotonic bladder. Recently, de novo ACTA2 R179H substitutions have been associated with a similar phenotype and additional cerebral developmental defects including underdeveloped corpus callosum and vermis hypoplasia in a single patient. The patient here shows previously undescribed abnormal lobulation of the frontal lobes and position of the gyrus cinguli and rostral dysplasis of the corpus callosum; she died at the age of 3 years during surgery due to vascular fragility and rupture of the ductus arteriosus. Altogether these observations support a role of ACTA2 in brain development, especially related to the arginine at position 179. Although all previously reported patients with R179H substitution successfully underwent the same surgery at younger ages, the severe outcome of our patient warns against the devastating effects of the R179C substitution on vasculature.
Asunto(s)
Actinas/genética , Aneurisma de la Aorta Torácica/genética , Conducto Arterioso Permeable/genética , Sustitución de Aminoácidos , Aneurisma de la Aorta Torácica/diagnóstico por imagen , Aneurisma de la Aorta Torácica/cirugía , Trastornos Cerebrovasculares/diagnóstico por imagen , Trastornos Cerebrovasculares/genética , Preescolar , Cuerpo Calloso/diagnóstico por imagen , Cuerpo Calloso/cirugía , Anomalías del Sistema Digestivo/genética , Conducto Arterioso Permeable/diagnóstico por imagen , Conducto Arterioso Permeable/cirugía , Femenino , Estudios de Asociación Genética , Genotipo , Heterocigoto , Humanos , Hipertensión Pulmonar , Vólvulo Intestinal/genética , Mutación Missense , Midriasis/genética , Fenotipo , Radiografía , Vasos Retinianos/patologíaRESUMEN
The function of sensory hair cells of the cochlea and vestibular organs depends on an influx of K(+) through apical mechanosensitive ion channels and its subsequent removal over their basolateral membrane. The KCNQ4 (Kv7.4) K(+) channel, which is mutated in DFNA2 human hearing loss, is expressed in the basal membrane of cochlear outer hair cells where it may mediate K(+) efflux. Like the related K(+) channel KCNQ5 (Kv7.5), KCNQ4 is also found at calyx terminals ensheathing type I vestibular hair cells where it may be localized pre- or postsynaptically. Making use of Kcnq4(-/-) mice lacking KCNQ4, as well as Kcnq4(dn/dn) and Kcnq5(dn/dn) mice expressing dominant negative channel mutants, we now show unambiguously that in adult mice both channels reside in postsynaptic calyx-forming neurons, but cannot be detected in the innervated hair cells. Accordingly, whole cell currents of vestibular hair cells did not differ between genotypes. Neither Kcnq4(-/-), Kcnq5(dn/dn) nor Kcnq4(-/-)/Kcnq5(dn/dn) double mutant mice displayed circling behavior found with severe vestibular impairment. However, a milder form of vestibular dysfunction was apparent from altered vestibulo-ocular reflexes in Kcnq4(-/-)/Kcnq5(dn/dn) and Kcnq4(-/-) mice. The larger impact of KCNQ4 may result from its preferential expression in central zones of maculae and cristae, which are innervated by phasic neurons that are more sensitive than the tonic neurons present predominantly in the surrounding peripheral zones where KCNQ5 is found. The impact of postsynaptic KCNQ4 on vestibular function may be related to K(+) removal and modulation of synaptic transmission.
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Sordera/metabolismo , Canales de Potasio KCNQ/metabolismo , Animales , Oído Interno/metabolismo , Electrofisiología/métodos , Femenino , Genotipo , Células Ciliadas Auditivas Internas/metabolismo , Masculino , Ratones , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , Ratones Noqueados , Microscopía Fluorescente/métodos , Modelos Biológicos , Neuronas/metabolismo , Canales de Potasio/metabolismo , Transmisión Sináptica , Vestíbulo del Laberinto/metabolismoRESUMEN
PURPOSE: To study three-dimensional optokinetic eye movements of wild-type C57BL/6J mice, the most commonly used mouse in oculomotor physiology. Optokinetic eye movements are reflexive eye movements that use visual feedback to minimize image motion across the retina. These gaze-stabilizing reflexes are a prominent model system for studying motor control and learning. They are three dimensional and consist of a horizontal, vertical, and torsional component. METHODS: Eye movements were evoked by sinusoidally rotating a virtual sphere of equally spaced dots at six frequencies (0.1-1 Hz), with a fixed amplitude of 5 degrees . Markers were applied to the mouse eye and video oculography was used to record its movements in three dimensions. In addition, marker tracking was compared with conventional pupil tracking of horizontal optokinetic eye movements. RESULTS: Gains recorded with marker and pupil tracking are not significantly different. Optokinetic eye movements in mice are equally well developed in all directions and have a uniform input-output relation for all stimuli, including stimuli that evoke purely torsional eye movements, with gains close to unity and minimal phase differences. CONCLUSIONS: Optokinetic eye movements of C57Bl6 mice largely compensate for image motion over the retina, regardless of stimulus orientation. All responses are frequency-velocity dependent: gains decrease and phase lags increase with increasing stimulus frequency. Mice show strong torsional responses, with high gains at low stimulus frequency.
Asunto(s)
Percepción de Movimiento/fisiología , Nistagmo Optoquinético/fisiología , Animales , Fijación Ocular/fisiología , Movimientos de la Cabeza/fisiología , Imagenología Tridimensional , Masculino , Ratones , Ratones Endogámicos C57BL , Orientación/fisiologíaRESUMEN
PURPOSE: To measure contrast sensitivity in C57BL/6, the most commonly used mouse in behavioral neuroscience, and to study the effect of sex, age, and miotic drugs on the contrast sensitivity function. In addition, the authors tested a mutant in which plasticity in the cerebellum is impaired by expressing a protein kinase C inhibitor. This inhibitor is also expressed in the retina, possibly affecting vision. METHODS: The gain of the optokinetic reflex (OKR) decreases as stimuli become more difficult to see. Recording OKR gains evoked by moving sine gratings shows whether the stimulus was distinguished from a homogeneous background and how well the stimulus was distinguished. RESULTS: Female mice have lower OKR gains than male mice (both groups: n = 10, P = 0.001). A similar difference was observed between 4-month-old (n = 10) and 9-month-old (n = 5) C57Bl/6 mice (P = 0.001). These differences could not be detected with earlier dichotomic tests. C57BL/6 mice are able to see contrasts as low as 1%, well below the previously reported 5% threshold. Pilocarpine had no significant effect on contrast sensitivity (both groups: n = 10, P = 0.89). Vision in L7-PKCi mutants was unaffected (both groups: n = 10, P = 0.82). CONCLUSIONS: OKR gains decrease as stimuli become more difficult to see, making the OKR a powerful tool to quantify contrast sensitivity. In C57BL/6 these response magnitudes vary greatly between sexes and between mice that differ only a few months in age. Therefore, it is important to match groups according to age and sex in experiments that require unimpaired vision. Otherwise, impaired vision can be misinterpreted as a learning or motor problem.
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Factores de Edad , Sensibilidad de Contraste/fisiología , Nistagmo Optoquinético/fisiología , Factores Sexuales , Animales , Femenino , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Mutantes , Ratones Transgénicos , Mióticos/administración & dosificación , Plasticidad Neuronal/fisiología , Pilocarpina/administración & dosificación , Proteína Quinasa C/antagonistas & inhibidores , Pupila/efectos de los fármacosRESUMEN
The cerebellum is divided into multiple parasagittally organized modules, which are thought to represent functional entities. How individual modules participate in cerebellar control of complex movements such as locomotion remains largely unknown. To a large extent, this is caused by the inability to study the contribution of individual modules during locomotion. Because of the architecture of modules, based on narrow, elongated cortical strips that may be discontinuous in the rostrocaudal direction, lesion of a complete module, without affecting neighboring modules, has not been possible. Here, we report on a new method for inducing a selective dysfunction of spatially separated parts of a single module using a small cortical injection of a retrogradely transported neurotoxin, cholera toxin b-subunit-saporin. We show that such a local injection into the C1 module results in climbing fiber and partial mossy fiber deafferentation of functionally related areas of this module, thereby resulting in a severe impairment of the whole module without affecting neighboring modules. A subsequent functional analysis indicates that such an impairment of the hindlimb part of the C1 module did not have a significant impact on skilled walking or overall stepping pattern. However, the modulation of cutaneously induced reflexes during stepping was severely diminished. We propose that the C1 module is specifically involved in the adaptive control of reflexes.