The development and activity-dependent expression of aggrecan in the cat visual cortex.

The Cat-301 monoclonal antibody identifies aggrecan, a chondroitin sulfate proteoglycan in the cat visual cortex and dorsal lateral geniculate nucleus (dLGN). During development, aggrecan expression increases in the dLGN with a time course that matches the decline in plasticity. Moreover, examination of tissue from selectively visually deprived cats shows that expression is activity dependent, suggesting a role for aggrecan in the termination of the sensitive period. Here, we demonstrate for the first time that the onset of aggrecan expression in area 17 also correlates with the decline in experience-dependent plasticity in visual cortex and that this expression is experience dependent. Dark rearing until 15 weeks of age dramatically reduced the density of aggrecan-positive neurons in the extragranular layers, but not in layer IV. This effect was reversible as dark-reared animals that were subsequently exposed to light showed normal numbers of Cat-301-positive cells. The reduction in aggrecan following certain early deprivation regimens is the first biochemical correlate of the functional changes to the γ-aminobutyric acidergic system that have been reported following early deprivation in cats.

Visual resolution and experience: acuity deficits in cats following early selective visual deprivation.

Cats reared during the first 5 months of life in environments that contain contours of a single orientation show a diminished ability to resolve gratings of the orthogonal orientation in later life. It is argued that these perceptual deficits result from changes in the organization of the visual cortex induced by the selected early visual input.

A neural effect of partial visual deprivation in humans.

Certain human subjects have considerable sensitivity differences in the visual resolution of vertical gratings as compared to horizontal gratings. Although only subjects with pronounced ocular astigmatism exhibit this effect, the differences are of neural, rather than optical, origin. It is argued that the resolution anisotropies result from early abnormal visual input caused by astigmatism. This abnormal input permanently modifies the brain.

Visual recovery after monocular deprivation is driven by absolute, rather than relative, visually evoked activity levels.

It is now well established that the anatomical and functional development of the central visual pathways of a number of higher mammalian species is activity-dependent [1-3]. This dependence was revealed by the functional effects of an early period of monocular deprivation, where one eye of a young animal was deprived for a time of patterned visual input. Subsequently, most cells in the visual cortex (area 17) could be excited only by visual stimuli delivered to the non deprived eye [4-6] and the animal appeared blind through the deprived eye [7,8]. These effects have been attributed to a competitive activity-dependent mechanism in development, whereby the two eyes compete for control of cortical cells [9,10]. There are, however, suggestions that the substantial recovery that can occur after monocular deprivation may be mediated by a different mechanism. Here, insight into the nature of this mechanism has been provided by monitoring the speed of changes in the vision of the deprived eye of a kitten after 6 days of monocular deprivation. Although both eyes were open during the recovery period, the kitten was able to see with its deprived eye only 2 hours after visual input was restored to this eye. The visual acuity of this eye improved rapidly in the first 24 hours and continued in an orderly way for 6 weeks. In contrast to the effects during monocular deprivation, which depend upon a competitive activity-dependent process, we propose that the events that follow deprivation rely on a mechanism driven by the absolute level of visually evoked activity through the formerly deprived eye.

Remodeling of lesioned kitten visual cortex after xenotransplantation of fetal mouse neopallium.

Remodeling of the mechanically injured cerebral cortex of kittens was studied in the presence of a neural xenograft taken from mouse fetuses. Solid neural tissue from the neopallium of a 14-day-old fetus was transferred into a cavity prepared in visual cortical area 18 of 33-day-old kittens. Injections of bromodeoxyuridine (BrdU) were used to monitor postoperative cell proliferation. Two months after transplantation, the presence of graft tissue in the recipient brain was assessed by Thy-1 immunohistochemistry. Antibodies specific for neurons, astrocytes, and oligodendrocytes and hematoxylin staining for endothelial cells were used for the characterization of proliferating (BrdU+) cells. The following were the major observations: 1) Of ten transplanted kittens, four had the cavity completely filled with neural tissue that resembled the intact cerebral cortex in its cytoarchitecture, whereas, in four other kittens, the cavity was partially closed. In two kittens, the cavity remained or became larger, which was also the case with all four sham-operated (lesioned, without graft) animals. 2) A substantial part of the remodeled tissue was of host origin. Only a few donor cells survived and dispersed widely in the host parenchyme. 3) In the remodeled region of transplanted animals, the densities of nerve, glial, and endothelial cells were similar to those in intact animals. 4) Cell proliferation increased after transplantation but only within a limited time, because, 2 months after the operation, the number of mitotic cells in the grafted cerebral cortex did not differ from that in intact controls. Our data suggest that the xenograft evokes repair processes in the kitten visual cortex that lead to structural recovery from a mechanical insult. The regeneration seems to rely on a complex interplay of many different mechanisms, including attenuation of necrosis, cell proliferation, and immigration of host cells into the wounded area.

Immunohistochemical study of the pattern of rapid expression of C-Fos protein in the visual cortex of dark-reared kittens following initial exposure to light.

Brief alterations to the nature of the visual input during critical periods in the early life of cats and monkeys can result in rapid anatomical and physiological changes in the central visual pathways. The immediate early genes (IEGs) represent a possible way in which these changes could be mediated since the protein products of a number of these genes have been shown to be induced rapidly in neurons in response to a variety of transsynaptic stimuli. Immunohistochemical methods were employed to examine the tempo and pattern of expression of Fos, the protein product of the c-fos gene, induced in the visual cortex of kittens dark-reared from birth to 30 days of age by brief periods of binocular visual exposure. In visual cortical area 17, the number of Fos immunoreactive cells increased rapidly from virtually zero in control kittens that received no visual exposure, to reach high levels in animals that received between 1 and 2 hours of visual experience. Immunoreactive cells were absent in the dorsal lateral geniculate nucleus, but were numerous in the ventral lateral geniculate nucleus, and in area 17, were most numerous in the extragranular layers (2, 3 and 6) but sparse in lower layer 4 and layer 5, and virtually absent in layer 1. Substantial constitutive Fos immunoreactivity was observed in area 17 of normal 30-day-old kittens but very few immunopositive cells were evident in adult animals. However, Fos immunoreactivity was observed in the visual cortex of a dark-reared (for 30 days) adult animal following a brief period of visual exposure, a finding that suggests that Fos might serve other roles in the visual cortex in addition to those it might play uniquely during development. It is suggested that Fos, in combination with the protein products of a select number of other IEGs, may mediate a variety of rapid changes in the visual cortex including those that underlie visual system plasticity during early postnatal life.

Effects of early periods of monocular deprivation and reverse lid suture on the development of Cat-301 immunoreactivity in the dorsal lateral geniculate nucleus (dLGN) of the cat.

During certain sensitive periods early in postnatal life, the anatomical and physiological development of the central visual pathways of cats and monkeys can be affected by the nature of an animal's early visual experience. In the last few years, studies have been started on some of the molecular and biochemical events that underlie the many functional changes induced by early selected visual deprivation in the visual cortex of kittens. In this respect, the monoclonal antibody Cat-301 provides a potentially powerful tool, because it recognizes in the cat dorsal lateral geniculate nucleus (dLGN) a proteoglycan associated with the surface of a particular class of cells, namely Y cells. In the dLGN, the Cat-301 proteoglycan appears late in postnatal development, and it expression has been shown to be experience dependent in both the dLGN and visual cortex (M. Sur, D. Frost, and S. Hockfield, 1988, J. Neurosci. 8:874-882; A. Guimaraes, S. Zaremba, and S. Hockfield, 1990, J. Neurosci. 10:3014-3024). We have explored further the experience-dependent nature of Cat-301 expression in the dLGN with a view to establishing a biochemical correlate of the many functional changes induced by early monocular deprivation and its reversal in the kitten visual system. In addition to demonstrating differences in Cat-301 expression between deprived and nondeprived laminae of the dLGNs of kittens monocularly deprived to only 4 or 5 weeks of age, the magnitude of the laminar difference was found to increase as the period of deprivation was extended. Moreover, monocularly deprived kittens that subsequently received long periods of reverse lid suture exhibited a reversal of the pattern of immunoreactivity, so that the greatest immunoreactivity occurred in laminae innervated by the initially deprived eye. However, possibly the most surprising and important finding was the extremely low levels of immunoreactivity observed in both A laminae of monocularly deprived animals that had received relatively short periods of reverse lid suture. These data suggest that Y cell development can be drastically altered depending on the time of initiation of the period of reverse lid suture and its duration.

Gene expression patterns during enhanced periods of visual cortex plasticity.

During a critical period in its postnatal development the mammalian visual cortex displays susceptibility to experience-dependent alterations of neuronal response properties. Plasticity represents an integrated set of developmental processes controlled by a transcriptional hierarchy that coordinates the action of many genes. To illuminate the expression of these critical genes, we examined gene expression patterns of 18371 non-redundant cDNAs in the visual cortex of cats at birth, at eye opening, at the peak of the critical period of eye dominance plasticity and in the adult cat using filter-based cDNA arrays and software-based hierarchical cluster analysis. We identified a small set of genes that were selectively expressed during the peak of the critical period for plasticity. We further examined the patterns of expression of these genes by analyzing the gene expression pattern of dark-reared chronologically older animals that are known to retain this ocular dominance plasticity beyond the chronologically defined critical period. This additional cluster assessment allowed us to separate age-related changes in the patterns of gene expression from plasticity-related changes, thus identifying a subset of genes that we define as plasticity candidate genes. Those plasticity candidate genes that have previously characterized functions include participants in second messenger systems, in cell adhesion, in transmitter recycling and cytokines, among others. Comparison of cDNA array quantitation with reverse transcription-polymerase chain reaction showed almost identical expression profiles for three genes that we examined. The expression pattern of one identified gene, opioid binding cell adhesion molecule, from the cDNA array analysis, is also in agreement with immunocytochemical results. We conclude that the approach of high-density cDNA array hybridization can be used as a useful tool for examining a complex phenomenon of developmental plasticity since it is amenable to multiple developmental stage gene expression comparisons.

The effect of minimal occlusion therapy on binocular visual functions in amblyopia.

The binocular visual functions of amblyopic children were studied during treatment involving brief weekly periods of occlusion of the unaffected eye while the child performed demanding visuomotor tasks against either a background of rotating gratings or a stationary uniform gray stimulus. The gains in stereoacuity were quite significant and in most cases more obvious than the rather small gains in letter visual acuity. On initial presentation only 21 of the 60 patients showed evidence of stereopsis and of these only seven possessed a stereoacuity of 100 secs or better. Following six treatment sessions the number of patients that demonstrated stereopsis increased to 36 of whom 17 possessed reasonably good stereoacuity (100 secs or better). However, there was no difference in the degree of improvement exhibited by those patients that viewed rotating grating patterns during treatment and others from the control group that viewed the uniform gray stimulus. Thus, there was no evidence that any of the visual gains were enhanced or promoted by active visual stimulation of the amblyopic eye with rotating gratings during the brief periods of occlusion of the unaffected eye. Finally, a comparison of the scores of the children on various stereo-tests suggest that tests comprised of small figure elements that are present in high density may be best for screening purposes. On the other hand, for quantifying the stereoacuity of children known to possess abnormal binocular vision it may be more appropriate to employ tests that use large figure elements that provide strong fusion cues.

Contrast thresholds for sine gratings of children with amblyopia.

Contrast threshold functions were measured on 26 amblyopic children before and after minimal occlusion therapy. On initial presentation the relative contrast sensitivity deficit the amblyopic eye was in every case much less than that predicted from the deficit in visual acuity for letters. In fact, in seven of the children the contrast sensitivity functions for the amblyopic and fellow nonamblyopic eye were indistinguishable despite the expression of substantial amblyopia on letter charts. Only four children exhibited a substantial contrast sensitive loss in the amblyopic eye with a cut-off spatial frequency below 30 cycles/degree. The majority of children who showed deficits in contrast sensitivity did so only at medium and high spatial frequencies. On the basis of these findings it appears that measurements of contrast thresholds for single sinusoidal gratings do not probe fully the deficits of spatial resolution in amblyopia. Finally, among the few children who exhibited a sizeable initial deficit, only two showed obvious improvement in contrast sensitivity in response to minimal occlusion therapy.

The efficacy of brief periods of reverse occlusion in promoting recovery from the physiological effects of mononuclear deprivation in kittens.

The relative effect of short daily periods of reverse occlusion in promoting recovery from the physiological effect of monocular deprivation in kittens were examined with a view to identifying a neurophysiological basis for the visual improvement observed with minimum occlusion therapy in amblyopia. Kittens were monocularly deprived from near birth until 5 weeks of age, at which time they were reverse-sutured and housed in total darkness. Each kitten received a short period of visual exposure through its initially deprived eye each day for either a fixed number of days or for a constant total visual exposure spread over a different number of exposure sessions. Electrophysiological recordings from single cells in the visual cortex were made the day after the last visual exposure. Kittens that received daily periods of reverse occlusion as brief as 30 min for 20 days showed a substantial degree of reversal of cortical ocular dominance. Other experiments indicated that 20 hr of reverse occlusion distributed over a number of brief daily sessions was far more effective in promoting physiological recovery than the same total period of exposure imposed in only two sessions. In general these results suggest that a given period of reverse occlusion may be more effective in promoting recovery with distributed than with massed periods of occlusion.

The permanence of the visual recovery that follows reverse occlusion of monocularly deprived kittens.

While the behavioral and physiologic effects of an early period of monocular deprivation can be extremely severe, they are not necessarily irreversible. Considerable recovery can occur if visual input is restored to the deprived eye sufficiently early, particularly if the nondeprived eye is occluded at the same time (reverse occlusion). This study examines the permanence of the visual recovery promoted by a period of reverse occlusion in kittens that were monocularly deprived from near birth for periods ranging from 3 to 18 weeks. During the period of reverse occlusion, the vision of the initially deprived eye improved from apparent blindness to good levels of acuity. However, upon restoring visual input to the formerly nondeprived eye a surprisingly rapid and reciprocal change occurred in the visual acuity of both eyes. Much of the substantial gain in the vision of the initially deprived eye that occurred during reverse occlusion was lost within 3 weeks, while at the same time the vision of the initially nondeprived eye improved substantially. Nevertheless, in many animals the acuity of the initially nondeprived eye did not recover to levels it had reached prior to reverse occlusion. These results hold important implications for the nature of the mechanisms responsible for the dramatic physiologic effects of monocular occlusion and reverse occlusion on the visual cortex. The results also may help elucidate recent observations on patching therapy in human amblyopia.

An evaluation of the Gore-Tex surgical membrane for the prevention of postoperative peritoneal adhesions.

The Gore-Tex surgical membrane, expanded polytetrafluoroethylene, has been used as a pericardial graft with minimal adhesion formation reported. The purpose of this study was to assess its efficacy as a barrier method for diminishing postoperative peritoneal adhesions in an animal model. Fifteen New Zealand white rabbits underwent laparotomy, with scrape and cut lesions created bilaterally on the uterine body and horns, respectively. On one side, the lesions were covered with the graft using 7-0 Gore-Tex suture; the contralateral side served as an internal control. After four weeks, the adhesions were graded and mean adhesion scores were calculated. The Gore-Tex score was 4.1 times higher than the control for scrape lesions and 1.9 times higher for cut lesions, but the difference was not statistically significant in either case. Overall, the Gore-Tex mean adhesion score was 2.3 times higher than the control, a statistically significant difference. The Gore-Tex surgical membrane did not appear to be an effective adjuvant for postoperative adhesion prophylaxis in this animal model.

The enzymes in cyclooxygenase and lipoxygenase pathways of arachidonic acid metabolism in human corpora lutea: dependence on luteal phase, cellular and subcellular distribution.

Eicosanoids synthesized within corpus luteum are presumed to regulate luteal function in women. However, the potential cellular source(s) of the eicosanoids, whether small and large luteal cells differ in eicosanoid synthesis and whether eicosanoids other than prostaglandin (PG)E2, PGF2 alpha and 6-keto-PGI1 alpha can be synthesized, have not been investigated. The present immunocytochemical studies were undertaken to answer these questions using mono and polyclonal antibodies to several enzymes in arachidonic acid metabolism by cyclooxygenase and lipoxygenase pathways. Human corpora lutea from early (n = 5), mid (n = 6) and late (n = 3) luteal phases were specifically immunostained for all the enzymes. All the enzymes were present in small and large luteal cells as well as in non luteal cells. However, small luteal cells contained more immunoreactive 5-lipoxygenase, PGD2 and PGF2 alpha synthases; large luteal cells contained more TXA2 synthase and 12-lipoxygenase; small and large luteal cells contained similar amounts of cyclooxygenase and PGI2 synthase. In all the cells, immunoreactive PGD2, PGI2 and TXA2 synthases increased from early to mid luteal phase and then declined in late luteal phase. Cyclooxygenase, 5- and 12-lipoxygenases and PGF2 alpha synthase, on the other hand, increased from early to mid and mid to late luteal phases. Immunoreactive cyclooxygenase and 5- and 12-lipoxygenases were present primarily in rough endoplasmic reticulum (ER) and/or smooth ER and cytoplasm. Quite unexpectedly, all three enzymes were also found in nuclear membranes, condensed chromatin and especially at the perimeter of condensed chromatin. Dispersed chromatin contained very little or no immunoreactive enzyme. These results indicate that regulation of human luteal function by eicosanoids synthesized within the corpus luteum is complex involving perhaps a) small and large luteal as well as non luteal cells, b) eicosanoids which have not been previously considered to play a role in luteal function and c) coordinate regulation of more than one enzyme in the pathways of arachidonic acid metabolism.

Grating acuity of cats measured with detection and discrimination tasks.

In order to help resolve the widely discrepant claims for the grating acuity of the cat, behavioral measurements were made of the acuities of two normal adult cats using two different tasks. In one, a conventional detection task, the cats were required to discriminate between high contrast vertical gratings and a uniform field of the same space-averaged luminance. The second, less commonly-used task, required the cats to discriminate between vertical and horizontal gratings of the same spatial frequency. Both cats obtained thresholds of between 8.5 and 9 cycles/degree, with no difference between tasks. These results suggest that detection of aliased patterns is not a likely factor contributing to the wide range of published acuities, and they provide support for one of two competing models of beta-ganglion cell sampling in the retina.

Behavioral deficits in cats following early selected visual exposure to contours of a single orientation.

The ability of adult cats, whose early visual experience was confined to contours of a single orientation (either vertical or horizontal), to resolve gratings of different orientations was studied by operant methods. Following selective visual exposure during part or all of the first 4 months of life, the cats were trained on a simultaneous discrimination between gratings of various orientations and blank fields of the same mean luminance. The spatial frequency of the gratings was systematically altered in order to obtain an estimate of acuity based upon extrapolation to chance levels of performance. Selectively deprived cats performed as well as normally reared cats on gratings having the same orientation as that of the stripes they saw as kittens, but their performance on gratings orthogonal to these was poorer. The deficits in acuity for gratings perpendicular to the experienced orientation varied between 0.26 and 0.87 of an octave. On the other hand, control cats whose early visual experience alternated between vertical and horizontal stripes, or who were reared in an environment containing randomly oriented contours, failed to show any difference in their acuity for vertical and horizontal gratings. The acuity deficits shown by the selectively deprived animals are long-standing since they remain unchanged even after 30 months of normal visual exposure. It is argued that these perceptual deficits are a consequence of the changes in cortical physiology that other investigators have described in cats who had undergone similar early visual deprivation. Taken together, these findings provide a basis for explaining a number of human perceptual disorders.

Period of susceptibility of kitten visual cortex to the effects of monocular deprivation extends beyond six months of age.

The duration of the sensitive period of the kitten visual cortex to the effects of monocular deprivation was explored by studies of the behavioral and physiological recovery from extended periods of monocular occlusion imposed from birth, and by examination of the physiological effects of a 3 month period of monocular occlusion imposed on animals at either 4, 5, 6, 7 or 8 months of age. Animals monocularly deprived until 4 months of age eventually show considerable behavioral and physiological recovery from the severe deficits observed immediately following termination of the period of deprivation. The conclusion that binocular connectivity may still be altered by the nature of the animal's visual input beyond 4 months of age was supported by the results obtained from animals that were monocularly deprived at 4 months of age or older. Animals deprived at either 4, 5 or 6 months showed a clear shift of cortical ocular dominance in favour of the non-deprived eye, but those deprived at 7 or 8 months showed approximately normal ocular dominance distributions. It is concluded that the sensitive period lasts at least twice as long as previously thought, to between 6 and 8 months of age.

Depth perception, eye alignment and cortical ocular dominance of dark-related cats.

On first exposure to light, animals that have been reared from birth until about 4 months of age in total darkness exhibit substantial visual and visuomotor deficits, which decline in severity during the first few months following exposure to light. In order to determine whether dark-reared animals eventually acquire stereoscopic vision following exposure to light we examined the binocular status of 5 dark-reared animals two of which developed convergent eye alignment. The binocular status was assessed behaviorally by measurements of the ability of each animal to perceive depth using either one or both eyes, and physiologically by documentation of the distribution of cortical ocular dominance of a sample of visual acuity, their binocular depth perception remained very poor, comparable to the monocular performance of normal cats. In marked contrast to normal animals none of the dark-reared animals, even those with normal eye alignment, performed substantially better binocularly than monocularly, a result indicating the absence of a uniquely binocular mechanism for depth perception in these animals. Although the dark-reared animals were found to retain a substantial (but reduced) complement of binocularly influenced cortical neurons, the tuning of these cells for retinal disparity must be insufficiently precise to mediate depth perception under binocular viewing conditions that is superior to that which can be achieved monocularly.

Visual resolution of retinal ganglion cells in monocularly-deprived cats.

The spatial resolution of 87 on-centre retinal ganglion cells was measured at different retinal eccentricities in the retina of the deprived eye of two monocularly-deprived cats. Intra-ocular recording techniques were employed in order that particular attention could be placed on ganglion cells in the region of the area centralis. The spatial cut-off frequencies of both major classes on-centre ganglion cells were comparable to those observed in normal retinas at similar retinal eccentricities. In particular there was no evidence of any loss of spatial resolution of either class of cell in the vicinity of the area centralis as has been reported in the retina of the deviating eye of strabismic cats.

The role of beta-endorphins and catechol estrogens on the hypothalamic-pituitary axis in female athletes.

Competitive swimmers were followed over a 2-year period when they trained at different levels of exercise which coincided with distinct changes in their menstrual history. Oligomenorrhea was identified in 5 of 13 of these athletes when they swam approximately 100,000 yards per week. Weight and percentage of body fat were not significantly different between the period of oligomenorrhea and regular menstrual function (P = 0.24). Mean and median levels of luteinizing hormone, follicle-stimulating hormone, prolactin, and 17beta-estradiol were decreased and catechol estrogens and beta-endorphins were increased in serum during the strenuous, when compared with the moderate, training period. The serum levels of the steroid and protein hormones were similar to those of normal cycling, nonexercising control subjects during moderate exercise (60,000 yards per week). The significant differences between beta-endorphins and catechol estrogens during periods of strenuous exercise suggest an explanation for oligomenorrhea in female athletes. These hormonal changes result in hypothalamic anovulation, which appears to be reversible, because the hormone levels and menstrual cycles return to normal when the exercise is reduced.