Normal Kittens Research Articles

Expression of early growth responsive gene-1 in the lateral geniculate body of kittens with amblyopia caused by monocular form deprivation.

The expression of early growth responsive gene-1 (Egr-1) in the lateral geniculate body in the normal kittens and those affected with amblyopia caused by monocular visual deprivation was compared to explore the potential significance of Egr-1 in the pathogenesis of amblyopia. A total of 30 healthy kittens were equally and randomly divided into the control (n = 15) and the deprivation group (n = 15). The kittens were raised in natural light and the right eyes of the deprived kittens were covered with a black opaque covering. Pattern visual evoked potential (PVEP) was measured before and 1, 3, and 5 weeks after covering. Five kittens from each group were randomly selected and euthanized with 2% sodium pentobarbital (100 mg/kg) during the 1st, 3rd and 5th week after covering. The expression of Egr-1 in the lateral geniculate body in the two groups was compared by performing immunohistochemistry and in situ hybridization. After three weeks of covering, PVEP detection indicated that the P100 wave latency in the deprivation group was significantly higher than that in the control group (P < 0.05), whereas the amplitude decreased markedly (P < 0.05). The number of the positive cells (P < 0.05) and mean optical density (P < 0.05) of Egr-1 protein expression in the lateral geniculate body of the deprivation group were found to be substantially lower in comparison to the normal group, as well as the number (P < 0.05) and mean optical density of Egr-1 mRNA-positive cells (P < 0.05). However, with increase of age, positive expression of Egr-1 in the control group showed an upward trend (P < 0.05), but this trend was not noted in the deprivation group (P > 0.05). Monocular form deprivation can lead to substantially decreased expressions of Egr-1 protein and mRNA in the lateral geniculate body, which in turn can affect the normal expression of neuronal functions in the lateral geniculate body, thereby promoting the occurrence and development of amblyopia.

ARC/Arg3.1 expression in the lateral geniculate body of monocular form deprivation amblyopic kittens

PurposeThe present study compared the expression of activity-regulated cytoskeleton-associated protein (ARC/Arg3.1) in the lateral geniculate body between form deprivation amblyopia kittens and normal kittens to examine the significance of ARC/Arg3.1 in the lateral geniculate body in the pathogenesis of amblyopia.MethodsTwenty kittens were randomly divided into an experimental group (n = 10) and a control group (n = 10). Black opaque covering cloth was used to cover the right eye of kittens in the experimental group. Pattern visual evoked potentials (PVEP) were detected weekly in all kittens. The expression of the ARC/Arg3.1 gene was detected by immunohistochemistry and in situ hybridization, and apoptosis of lateral geniculate body cells was detected by TUNEL.ResultsPVEP detection showed that at the age of 5 and 7 weeks, the latency of P100 in the right eye of the experimental group was higher than that of the other three groups (P < 0.05), and the amplitude of P100 was lower than that of the other three groups (P < 0.05). The expression of ARC/Arg3.1 protein (P < 0.05) and mRNA (P < 0.05) in the lateral geniculate body of the experimental group was significantly lower than that of the control group. The level of neuronal apoptosis in the experimental group was higher than that in the control group (P < 0.05). The expression of the ARC/Arg3.1 gene was negatively correlated with the apoptosis level of lateral geniculate body neurons.ConclusionsThe expression of ARC/Arg3.1 is associated with monocular form deprivation amblyopia and apoptosis of lateral geniculate body cells.

Documentation of the Development of Various Visuomotor Responses in Typically Reared Kittens and Those Reared With Early Selected Visual Exposure by Use of a New Procedure.

A new procedure was used to study the development of gaze (responses to moving targets or laser spots in normal kittens, those that had been reared in total darkness to 6 weeks of age, and others that received a period of monocular deprivation (MD). Gaze responses were observed to all stimuli in normal kittens at between 25–30 days of age and striking responses occurred on the same day or the next. Despite slow acquisition of spatial vision in the dark reared kittens over 3 months, they were able to follow and even strike at moving visual stimuli within a day of their initial exposure to light. By contrast, for a week following a period of MD, kittens showed no gaze or striking responses to moving stimuli when using their previously deprived eye. The very different profiles of acquisition of visuomotor skills and spatial vision in visually deprived kittens point to a dissociation between the neuronal populations that support these functions.

Expression of early growth responsive gene-1 in the visual cortex of monocular form deprivation amblyopic kittens

PurposeThe present study compared the expression of early growth responsive gene-1 (Egr-1) in visual cortex between amblyopia kittens and normal kittens, and to explore the role of Egr-1 in the pathogenesis of amblyopia.MethodsA total of 20 healthy kittens were randomly divided into deprivation group and control group with 10 kittens in each group. Raised in natural light, and covered the right eye of the deprived kittens with a black opaque covering cloth. Pattern visual evoked potentials (PVEP) were measured before and at the 1st, 3rd and 5th week after covering in all kittens. After the last PVEP test, all kittens were killed. The expression of Egr-1 in the visual cortex of the two groups was compared by immunohistochemistry and in situ hybridization.ResultsPVEP detection showed that at the age of 6 and 8 weeks, the P100 wave latency in the right eye of deprivation group was higher than that in the left eye of deprivation group (P < 0.05) and the right eye of control group (P < 0.05), while the amplitude decreased (P < 0.05). The number of positive cells (P < 0.05) and mean optical density (P < 0.05) of Egr-1 protein expression in visual cortex of 8-week-old deprivation group were lower than those of normal group, as well as the number (P < 0.05) and mean optical density of Egr-1 mRNA-positive cells (P < 0.05).ConclusionsMonocular form deprivation amblyopia can lead to the decrease of Egr-1 protein and mRNA expression in visual cortex, and then promote the occurrence and development of amblyopia.

Expression of brain-derived neurotrophic factor in the lateral geniculate body of monocular form deprivation amblyopic kittens.

The present study compared the expression of brain-derived neurotrophic factor (BDNF) in the lateral geniculate body between form deprivation amblyopia kittens and normal kittens to examine the significance of BDNF in the lateral geniculate body in the pathogenesis of amblyopia. Twenty kittens were divided into control group (n = 10) and deprivation group (n = 10). A black opaque eye mask was placed to cover the right eye of the deprivation group. Pattern visual-evoked potentials (PVEPs) were detected weekly in all kittens .After the kittens in the deprivation group developed monocular amblyopia, the lateral geniculate bodies of all kittens were removed. The expression of BDNF in the lateral geniculate body of the two groups was compared by immunohistochemistry and Western blotting. The latency of the P100 wave in the right eye of the deprivation group was longer than that of the left eye and that of the right eye of the control group (p < 0.05), and the amplitude decreased (p < 0.05). The number and average optical density of BDNF-positive cells in the deprivation group were lower than those in the control group (p < 0.05), and the expression of BDNF in the deprivation group was lower than that in the control group (p < 0.05). The expression of BDNF in the lateral geniculate body of the amblyopic kittens decreased, and the decrease in BDNF promoted the development of amblyopia. These results demonstrate that BDNF in the lateral geniculate body plays an important role in visual development.

The post-natal development of intraocular pressure in normal domestic cats (Felis catus) and in feline congenital glaucoma

Intraocular pressure (IOP) is the most consistent risk factor for progressive vision loss in glaucoma. Cats with recessively inherited feline congenital glaucoma (FCG) exhibit elevated IOP with gradual, painless progression of glaucoma similar to humans and are studied as a model of glaucoma in humans and animals. Here, post-natal development of IOP was characterized in normal domestic cats and in cats with FCG caused by a homozygous LTBP2 mutation. Rebound tonometry (TonoVet®, ICare Oy, Finland) was used to measure IOP non-invasively, 2–3 times weekly in 63 FCG and 33 normal kittens, of both sexes, from eyelid opening until 3–6 months of age. IOPs in the left and right eyes of both FCG and normal kittens were compared by paired t-test and linear regression. One-way ANOVA and Tukey-Kramer post-tests were used to compare IOP of cats grouped by age and disease status. A p-value <0.05 was considered significant. In the second week of life, mean IOP was 7.16 mmHg (SD = 1.3) in normal kittens and 8.72 mmHg (SD = 1.4) in kittens with FCG. Mean IOP at age 10 weeks was significantly higher in FCG (19.8 mmHg; 95% CI = 17.7, 21.9 mmHg) than in normal kittens (13.2 mmHg; 95% CI = 11.9, 14.5 mmHg). At 3 months of age, IOP in normal cats reached adult values while IOP in FCG cats continued to increase through at least six months of age. These results provide ranges for normal IOP values in young kittens and confirm that IOP is significantly higher than normal by 10wks of age in this spontaneous feline glaucoma model.

Binocular pattern deprivation interferes with the expression of proteins involved in primary visual cortex maturation in the cat.

BackgroundBinocular pattern deprivation from eye opening (early BD) delays the maturation of the primary visual cortex. This delay is more pronounced for the peripheral than the central visual field representation within area 17, particularly between the age of 2 and 4 months [Laskowska-Macios, Cereb Cortex, 2014].ResultsIn this study, we probed for related dynamic changes in the cortical proteome. We introduced age, cortical region and BD as principal variables in a 2-D DIGE screen of area 17. In this way we explored the potential of BD-related protein expression changes between central and peripheral area 17 of 2- and 4-month-old BD (2BD, 4BD) kittens as a valid parameter towards the identification of brain maturation-related molecular processes. Consistent with the maturation delay, distinct developmental protein expression changes observed for normal kittens were postponed by BD, especially in the peripheral region. These BD-induced proteomic changes suggest a negative regulation of neurite outgrowth, synaptic transmission and clathrin-mediated endocytosis, thereby implicating these processes in normal experience-induced visual cortex maturation. Verification of the expression of proteins from each of the biological processes via Western analysis disclosed that some of the transient proteomic changes correlate to the distinct behavioral outcome in adult life, depending on timing and duration of the BD period [Neuroscience 2013;255:99-109].ConclusionsTaken together, the plasticity potential to recover from BD, in relation to ensuing restoration of normal visual input, appears to rely on specific protein expression changes and cellular processes induced by the loss of pattern vision in early life.Electronic supplementary materialThe online version of this article (doi:10.1186/s13041-015-0137-7) contains supplementary material, which is available to authorized users.

Ten days of darkness causes temporary blindness during an early critical period in felines.

Extended periods of darkness have long been used to study how the mammalian visual system develops in the absence of any instruction from vision. Because of the relative ease of implementation of darkness as a means to eliminate visually driven neural activity, it has usually been imposed earlier in life and for much longer periods than was the case for other manipulations of the early visual input used for study of their influences on visual system development. Recently, it was shown that following a very brief (10 days) period of darkness imposed at five weeks of age, kittens emerged blind. Although vision as assessed by measurements of visual acuity eventually recovered, the time course was very slow as it took seven weeks for visual acuity to attain normal levels. Here, we document the critical period of this remarkable vulnerability to the effects of short periods of darkness by imposing 10 days of darkness on nine normal kittens at progressively later ages. Results indicate that the period of susceptibility to darkness extends only to about 10 weeks of age, which is substantially shorter than the critical period for the effects of monocular deprivation in the primary visual cortex, which extends beyond six months of age.

The Expression of Vasoactive Intestinal Polypeptide in Visual Cortex-17 in Normal Visual Development and Formation of Anisometropic Amblyopia

Aims: To document the expression of vasoactive intestinal polypeptide (VIP) in the visual cortex-17 of kittens with anisometropic amblyopia, and to investigate the relationship between VIP and the development of the visual system. Methods: Sixteen normal kittens (4-wk of age) were randomly divided into two groups: control and amblyopic. Amblyopia was produced by atropinization of one eye in eight kittens. Four (2 normal and 2 amblyopia) kittens were sacrificed at weeks 3, 6, 9, or 12 post-treatment respectively. Expression of VIP-mRNA in the visual cortex-17 was detected through in-situ hybridization. Neurons in the visual cortex were visualized by transmission electron microscopy (TEM). The number of neurons was analyzed via light microscopy (LM). Results: VIP-mRNA expression was increased with age in control kittens but remained nearly static in age-matched anisometropic amblyopic kittens (p < 0.05). The number of VIP-positive cells of amblyopic kittens decreased dramatically when compared to normal age-matched kittens (p < 0.05). The total comparison between different positive ranks suggested a significant difference. The degree of expression between these two groups was significantly different. Ultrastructurally, in the control group, the nuclear membrane of most neurons was discernable and chromatin was evenly distributed within the nucleus. Abundant cytoplasm and tubular-shaped mitochondria were observed. These cells were also rich in Golgi bodies, ribosomes, and endoplasmic reticulum. In amblyopic kittens, nuclei of most neurons were aggregated, the number of ribosomes and Golgi bodies was reduced, mitochondria were swollen, and mitochondrial cristae were shortened or even absent. The endoplasmic reticulum was distended and reduced in magnitude. Conclusions: VIP appears to play an important role in visual development, and its mRNA expression is affected by visual experiences. Visual dysfunction may down-regulate the expression of VIP-mRNA by impairing the structure and function of the neurons in the visual cortex, finally leading to amblyopia.

Synaptic plasticity after chemical deafening and electrical stimulation of the auditory nerve in cats.

The effects of deafness on brain structure and function have been studied using animal models of congenital deafness that include surgical ablation of the organ of Corti, acoustic trauma, ototoxic drugs, and hereditary deafness. This report describes the morphologic plasticity of auditory nerve synapses in response to ototoxic deafening and chronic electrical stimulation of the auditory nerve. Normal kittens were deafened by neonatal administration of neomycin that eliminated auditory receptor cells. Some of these cats were raised deaf, whereas others were chronically implanted with cochlear electrodes at 2 months of age and electrically stimulated for up to 12 months. The large endings of the auditory nerve, endbulbs of Held, were studied because they hold a key position in the timing pathway for sound localization, are readily identifiable, and exhibit deafness-associated abnormalities. Compared with those of normal hearing cats, synapses of ototoxically deafened cats displayed expanded postsynaptic densities, a 35.4% decrease in synaptic vesicle (SV) density, and a reduction in the somatic size of spherical bushy cells (SBCs). In comparison with normal hearing cats, ototoxically deafened cats that received cochlear stimulation had endbulbs that expressed postsynaptic densities (PSDs) that were statistically identical in size, showed a 48.1% reduction in SV density, and whose target SBCs had a 25.5% reduction in soma area. These results demonstrate that electrical stimulation via a cochlear implant in chemically deafened cats preserves PSD size but not other aspects of synapse morphology. This determination further suggests that the effects of ototoxic deafness are not identical to those of hereditary deafness.

Congenital Thyroid Hypoplasia and Seizures in 2 Littermate Kittens

Congenital Thyroid Hypoplasia and Seizures in 2 Littermate Kittens

Some aspects of the modular organization of the primary visual cortex of the cat: Patterns of cytochrome oxidase activity

The distribution of the enzyme cytochrome oxidase (CO) in continuous series of parasagittal sections from field 17 and frontal sections of the dorsal nucleus of the lateral geniculate body (LGB) from normal kittens and adult cats was studied. In all cats apart from neonates, layer IV showed regularly alternating areas with above-background levels of CO activity ("spots"). There was a significant increase in the contrast of the "spots" from days 13 to 21, which was followed by a significant decrease from days 48 to 93. These changes coincided with ontogenetic changes in the level of visual system plasticity. There were no differences in CO activity between layers A and A1 of the dorsal nucleus of the LGB. It is suggested that the non-uniform distribution of the level of functional activity of neurons in field 17 reflects the formation of columnar cortical structures during the critical period of postnatal ontogenesis.

Neonatal deafness results in degraded topographic specificity of auditory nerve projections to the cochlear nucleus in cats

We previously examined the early postnatal maturation of the primary afferent auditory nerve projections from the cat cochlear spiral ganglion (SG) to the cochlear nucleus (CN). In normal kittens these projections exhibit clear cochleotopic organization before birth, but quantitative data showed that their topographic specificity is less precise in perinatal kittens than in adults. Normalized for CN size, projections to the anteroventral (AVCN), posteroventral (PVCN), and dorsal (DCN) subdivisions are all significantly broader in neonates than in adults. By 6-7 postnatal days, projections are proportionate to those of adults, suggesting that significant refinement occurs during the early postnatal period. The present study examined SG projections to the CN in adult cats deafened as neonates by ototoxic drug administration. The fundamental organization of the SG-to-CN projections into frequency band laminae is clearly evident despite severe auditory deprivation from birth. However, when normalized for the smaller CN size in deafened animals, projections are disproportionately broader than in controls; AVCN, PVCN, and DCN projections are 39, 26, and 48% broader, respectively, than predicted if they were precisely proportionate to projections in normal hearing animals. These findings suggest that normal auditory experience and neural activity are essential for the early postnatal development (or subsequent maintenance) of the topographic precision of SG-to-CN projections. After early deafness, the basic cochleotopic organization of the CN is established and maintained into adulthood, but the CN is severely reduced in size and the topographic specificity of primary afferent projections that underlies frequency resolution in the normal central auditory system is significantly degraded.

Cerebral ventricular size in developing normal kittens measured by ultrasonography.

To evaluate the age of fontanelle closure of normal kittens and the size of their lateral ventricles, 50 ultrasonographic examinations using the bregmatic fontanelle as an acoustic window were made. Seven kittens, laboratory animals, were included in the study. To verify the location of the lateral ventricle, two of the kittens were sacrificed as neonates. In one of them ink was injected prior to autopsy into one lateral ventricle under sonographic guidance. In a longitudinal study of five of the kittens, the skull depth and the depth of the central part of the lateral ventricle reproduced in a longitudinal view could be measured up to the age of about 5 months. During that period, the skull depth increased from a median value of 1.95 cm (1.92-1.98) in a seven-day-old cat to 2.58 cm (2.52-2.59) in a 154-day-old cat, while the afore-mentioned ventricle values increased from 0.3 mm to 1.1 mm.

Improved growth of lipoprotein lipase deficient kittens by feeding a low-fat, highly digestible diet

Adult domestic cats homozygous with a naturally occurring Gly412Arg LPL gene mutation are good models for the study of LPL deficiency. Previous studies report that homozygous LPL deficient kittens have reduced growth rates and develop subnormal body fat mass. It was hypothesized in the present study that homozygote kittens would have normal growth if provided a standard low fat, highly digestible diet at weaning and that their body fat would be increased by provision of a diet high in protein. When fed a nutritionally complete, 10% fat, purified or commercial extruded diet, the body weights of homozygous (n = 24), heterozygous (n = 25) and normal (n = 16) kittens were determined at birth, 2, 3, 4, 6, 12 and 18 weeks of age. Male homozygote kittens from homozygote dams had reduced weight gains (p < 0.05) compared to normal males at 2, 3 and 4 weeks. Female heterozygotes and homozygotes from homozygote and heterozygote dams had reduced weight gains (p < 0.05) compared to normal females at 2, 3, 4 and 6 weeks. By 6 weeks for males and 18 weeks for females, genotype related differences in weight gain were not observed. At 30 weeks, homozygotes and heterozygotes were given either a 60 or 30% (dry matter) protein diet for two months. As indicated by deuterium dilution estimation of body composition, cats eating the 30% protein diet (n = 12) tended to have a lower increase in lean body mass (p = 0.057) and a greater increase in fat mass (p = 0.092) compared to cats eating the 60% protein diet (n = 12). Increase in lean body mass among homozygotes tended to be not as great as that observed in heterozygotes (p = 0.057). Poor postweaning gains previously reported in homozygotes probably reflected inappropriate selection of diet for this genotype. The high protein diet increased the rate of lean body mass development but not body fat mass.

Brain-Derived Neurotrophic Factor Expands Ocular Dominance Columns in Visual Cortex in Monocularly Deprived and Nondeprived Kittens But Does Not in Adult Cats

Segregation and stabilization of thalamocortical afferents to eye-specific patches, so-called "ocular dominance (OD) columns," in visual cortex are hypothesized to be based on activity-dependent competition for trophic factors such as brain-derived neurotrophic factor (BDNF) between afferents representing the two eyes during the critical period of postnatal development. To test this hypothesis we observed effects of an intracortical infusion of BDNF on OD columns in monocularly deprived kittens and also compared effects between normal kittens and adult cats. BDNF had a hypertrophic action on afferents irrespective of visual inputs so that it desegregated OD columns in the visual cortex of deprived and normal kittens, but this action was not seen in the adults, substantiating its hypothesized trophic role in plasticity of OD columns in the developing visual cortex.

Feline anaemia 1. Clinical signs and investigation

ANAEMIA is a decrease in the number of erythrocytes, concentration of haemoglobin or haematocrit volume and, as such, should be regarded as a clinical sign rather than a disease. The...

Photoreceptor allografts in a feline model of retinal degeneration.

Photoreceptor transplants provide a potential means to restore function in a degenerate retina and/or rescue degenerating host photoreceptors by trophic influences. We have examined photoreceptor allografts in the Abyssinian cat model of hereditary photoreceptor degeneration to determine the viability and influence of such transplants on the host retina. Small pieces of 3- to 5-day-old normal kitten retina containing undifferentiated photoreceptors were injected into the subretinal space of adult Abyssinian cats at an early stage of retinal degeneration using standard vitreo-retinal surgical techniques. The retinas were examined by ophthalmoscopy and fundus photography, then by light and electron microscopy at different times after surgery. Such allografts survive for at least 6 months after surgery. The photoreceptors develop outer segments, invariably in rosettes. The transplants gradually integrate with the host retina but detach the host photoreceptor layer from the retinal pigment epithelium (RPE), which tends to reduce the number of host photoreceptors over the transplant. There is no slowing of the photoreceptor degeneration in neighboring non-detached retina. Inflammation or rejection was not detected. Undifferentiated, neonatal photoreceptor allografts survive and develop outer segments in the subretinal space of the Abyssinian mutant feline retina. The allografts gradually integrate with the host neural retina without inducing rejection. In the vicinity of the transplant there is increased loss of host photoreceptors, considered to be due to their detachment from the RPE layer. There is no evidence of any rescue of host photoreceptors elsewhere in this mutant retina.

Effect of sensory disuse on geniculate afferents to cat visual cortex.

In the kitten, as little as a week of monocular lid suture during early life causes a remarkable remodeling of the geniculocortical projections serving the deprived eye (Antonini & Stryker, 1993a, 1996). While the physiological effects of monocular deprivation have been shown to be due to competitive interactions between the projections serving the two eyes, it is not known whether these morphological changes are due to competitive interactions or to sensory disuse. We addressed this question by analyzing the morphology of geniculocortical arbors in kittens deprived of patterned vision by binocular lid suture for 1 week or 2 weeks ending at 6 weeks of age. Such deprivation would be expected to affect the afferents serving the two eyes equally, giving neither eye a competitive advantage. The arbors were anterogradely filled with Phaseolus lectin iontophoretically injected into lamina A of the lateral geniculate nucleus. The lectin was visualized immunohistochemically, and single geniculocortical arbors were serially reconstructed in three dimensions. Arbors reconstructed in binocularly deprived animals were compared with arbors serving the deprived and nondeprived eye in animals monocularly deprived by lid suture of one eye for a week and with arbors obtained in age-matched normal controls. Geniculocortical arbors in binocularly deprived animals did not suffer the drastic remodeling of the deprived arbors in monocularly deprived animals. Indeed, arbors in binocularly deprived animals were indistinguishable from arbors in normal kittens or nondeprived arbors in short-term monocularly deprived animals. These results support the notion that competitive mechanisms rather than sensory disuse are responsible for gross morphological remodeling of geniculocortical arbors.

Characterization of anti-feline CD8 monoclonal antibodies

We generated three monoclonal antibodies (mAbs) (2D7, 10C7 and 12A3) reactive to the α-chain of feline CD8 (fCD8) molecule. Further we showed that reference anti-fCD8 mAbs, FT2, 3.357 and vpg9 recognize the β-chain, α-chain and αβ-complex epitope, respectively. Flow cytometric analysis using these mAbs suggested that fCD8 α + β - cells were present in lymphocytes of spleen, but not significantly in those of thymus, lymph nodes and peripheral blood of normal kittens.