Visual afferentation deficit during diffuse photosensitivity period in nestlings affects morphogenesis in Wulst neurons

Golgi-stained preparations were processed with computerized morphometry to study the effect of the deprivation (eyes covered with nontransparent caps) on the development of neurons in the Wulst (the structure analogous to mammalian visual cortex) of pied flycatcher nestlings. Six-day-old nestlings that have only one form of the visual perception--diffuse photosensitivity--were studied. In the previous paper [Korneeva et al., 1994] in was shown that the Wulst of 6-day-old nestlings consisted of non-differentiated neurons and stellate-like cells at different stages of maturation; the latter group was subdivided into more- and less-mature cells. This work proved that even a 1.5-day-long deprivation (actual duration being counted off from the moment of appearance of the first retinal photoreceptors at the age of 4.5 days) resulted in significant changes in the geometry of stellate-like neurons. The changes in less-mature cells were predominantly destructive (decline of all quantitative indices of a cell, including the significant decrease in the total length of dendrites, maximal radius of the dendritic field and cell branching index), while the changes in more-mature cells were constructive (increase in all quantitative cell indices, including statistically significant increase in the soma section area, total length of dendrites, maximal radius of dendritic field and the number of foci of maximal branching). Different reactions of these cell types to the limitation of visual afferentation may be connected with differences in the afferent inflow to less- and more-mature cells or/and with different maturational stages of these neurons at the onset of deprivation.     

Age dynamics of visually guided behavior and visual evoked potentials of altricial nestlings

Comparative study has been carried out of factors of organization of visually guided feeding behaviour (observation method) and the degree of maturity of the visual mechanisms by the criterion of Wulst EPs formation and of their recovery cycles in normally developing and visually deprived nestlings. In has been established that during the period following the opening of the eyes (5-9th day) the feeding behaviour is connected with diffuse photo-sensitivity of visual mechanisms. Diffuse photo-sensitivity fully provides for natural feeding behaviour at the corresponding stage of development of visual functions. Complete visual deprivation during the whole period of diffuse photo-sensitivity does not influence subsequent development of the visual system.     

Wulst Neurons in Nestlings of the Pied Flycatcher Ficedula hypoleuca at the Pre-Visual Stage of Development

Neurons of the Wulst region, an analog of the mammalian visual cortex, were studied in Golgi-impregnated preparations of brain of non-precocial 1-day old nestlings of the pied flycatcher Ficedula hypoleuca. At this age, vision does not function in nestlings, their behavior is provided by an acoustic analyzer. Two populations of nerve cells, large and small juvenile neurons, were differentiated at visual examination. The comparative morphometry has shown these groups to differ significantly from each other by most studied parameters: the area of the profile field of their soma, the total length of dendrites, branching of the cell, the number of the maximal branching foci, the character of distribution of dendrite free endings in three concentric zones of the cell dendritic field. The distribution of dendrites in the dendritic neuron field was similar in the both groups of neurons. An increased density of dendrites was observed from the side of the afferent input. At the same time, study of orientation of the longest dendrite has shown it to be located parallel to the plane of the afferent input practically in all cells of the both groups. It is suggested that such orientation of the longest dendrite broadens the area of cell contacts, which is necessary for search for the maximal number of afferents. The predominant orientation of dendrites in the direction to the afferent input forms foundation for establishing a more extended zone of contacts with growing visual afferents.     

Formation of visual evoked potentials and development of visually guided behavior in the Ficedula hypoleuca nestling

Age dynamics of evoked potentials (EPs) of Wulst area of the dorsal hyperstriatum (functional analogue of mammalian visual cortex) was investigated in 2-7 days old nestlings. EPs in response to light stimuli, with the duration and intensity imitating natural alimentary signals for 5-7 days-old nestlings, were recorded parallel with the behaviour. It was found that the development of the visual system in precocious nestlings became completed in postembryonal ontogenesis. The most essential changes in the parameters of visual EPs occur between 2.5 and 4 days of nest life. By the end of the 4-th day the EPs parameters resemble those of the definite EPs. During the same period the visually directed form of the alimentary behaviour appears for the first time. Both the visual EPs and the visually directed alimentary behaviour first appear in the ontogenesis in response to the second component of two-fold luminosity change--the natural signal of alimentary behaviour.     

Development of Calbindin- and Calretinin-Immunopositive Neurons in the Enteric Ganglia of Rats

Calbindin D28 K (CB) and calretinin (CR) are the members of the EF-hand family of calcium-binding proteins that are expressed in neurons and nerve fibers of the enteric nervous system. CB and CR are expressed differentially in neuronal subpopulations throughout the central and peripheral nervous systems and their expression has been used to selectively target specific cell types and isolate neuronal networks. The present study presents an immunohistochemical analysis of CB and CR in the enteric ganglia of small intestine in rats of different ages (newborn, 10-day-old, 20-day-old, 30-day-old, 60-day-old, 1-year-old, and 2-year-old). The data obtained suggest a number of age-dependent changes in CB and CR expression in the myenteric and submucous plexuses. In the myenteric plexus, the lowest percentage of CB-immunoreactive (IR) and CR-IR neurons was observed at birth, after which the number of IR cells increased in the first 10 days of life. In the submucous plexus, CB-IR and CR-IR neurons were observed from 10-day-old onwards. The percentage of CR-IR and CB-IR neurons increased in the first 2 months and in the first 20 days, respectively. In all animals, the majority of the IR neurons colocalized CR and CB. From the moment of birth, the mean of the cross-sectional area of the CB-IR and CR-IR neuronal profiles was larger than that of CB- and CR-negative cells.     

Begging to differ: scrubwren nestlings beg to alarm calls and vocalize when parents are absent

Nestling birds face a dilemma: they can increase parental provisioning by begging more intensively, but by doing so may also increase their risk of predation. Nestlings could deal with this dilemma by reducing begging intensity after parents have warned them of a nearby predator. We therefore tested experimentally whether nestling scrubwrens, Sericornis frontalis, increase begging intensity with hunger but reduce it after adult alarm calls. Single 5- and 8-day-old nestlings were temporarily taken into the laboratory for playback experiments. Over a 90-min period of food deprivation we simulated parental visits every 10 min by playing back adult feeding calls. Hungrier nestlings begged louder and longer to simulated parental visits, but contrary to expectation did not beg less if they had previously heard playback of alarm calls, and even begged to the alarm calls themselves. The results were similar for both ‘mobbing’ and ‘flee’ alarm calls. Nestlings also gave distinctive calls in the 10-min interval between simulated parental visits, and the number of these calls increased with hunger and after playback of alarm calls. We suggest that nestlings acquire the ability to respond appropriately to alarm calls late in the nestling period and that therefore parents might be selected to avoid alarm calling when defending young nestlings.Copyright 2003 Published by Elsevier Science Ltd on behalf of The Association for the Study of Animal Behaviour.      

Immunocytochemical properties of stellate ganglion neurons during early postnatal development

Neurotransmitter features in sympathetic neurons are subject to change during development. To better understand the neuroplasticity of sympathetic neurons during early postnatal ontogenesis, this study was set up to immunocytochemically investigate the development of the catecholaminergic, cholinergic, and peptidergic phenotypes in the stellate ganglion of mice and rats. The present study was performed on Wistar rats and Swiss mice of different ages (newborn, 10-day-old, 20-day-old, 30-day-old, and 60-day-old). To this end, double labeling for tyrosine hydroxylase (TH), choline acetyltransferase (ChAT), vasoactive intestinal (poly)peptide (VIP), neuropeptide Y (NPY), galanin (GAL), and somatostatin (SOM) was applied. The results obtained indicate that the majority of the neurons in the stellate ganglion of both species were TH-positive from birth onward and that a large part of these neurons also contained NPY. The percentage of neurons containing TH and NPY invariably increased with age up to 60 days postnatally. A smaller portion of the stellate ganglion neurons contained other types of neuropeptides and showed a distinct chronological pattern. The proportion of VIP- and ChAT-positive neurons was maximal in 10-day-old animals and then decreased up to 60 days of age, whereas the number of SOM-positive cells in rats significantly decreased from birth onward. In newborn rats, VIP-, ChAT- and SOM-positive neurons were largely TH-positive, while their proportions decreased in 10-day-old and older rats. Accordingly, the largest part of VIP-positive neurons also expressed SOM immunoreactivity at birth, after which the number of neurons containing both peptides diminished. The VIP- and SOM-positive cells did not contain NPY in any of the age groups studied. In rats up to 10 days of life, GAL-immunoreactive (-IR) neurons were scarce, after which their number increased to reach a maximal value in 30-day-old animals and then declined again. The SOM-reactive cells had the smallest size in all rats, while the largest neurons were those containing ChAT. In the mouse stellate ganglion, VIP- and ChAT-IR neurons were larger in comparison to NPY- and TH-IR cells. Our study further revealed some species differences: compared to mice the proportion of neurons containing TH and NPY was higher in rats at all ages under study. Furthermore, no GAL-immunostained neurons were found in mice and the number of SOM-positive cells in mice was limited compared to that observed in rats. In conclusion, the development of neurotransmitter composition is complete in rats and mice by their second month of life. At this age, the percentages of immunopositive cells have become similar to those reported in adult animals.     

Cytochemical features of cortical neurons during the recovery period following early visual deprivation]

It was shown interferometrically that 10 days after the removal of rabbits, that had been exposed to early (from the virth till 2.5 month of life) visual deprivation, to the ordinary light conditions, a reliable increase in the neuron size could be noted in addition to the reliably increased protein content in the cytoplasm of the visual cortex neurons. Degrees of the above changes in neurons of laminae III, IV and V are not similar, no complete normalization occurring in any lamina. A question of the reversibility of changes caused by early visual deprivation and of the compensatory capacities of neuron in particular lamina is discussed.     

Lack of Correlation Between the Effects of Transient Exposure to Glutamate and Those of Hypoxia/Reoxygenation in Immature Neurons In Vitro

Abstract: To assess the influence of brain immaturity on the effects of oxygen deprivation and the participation of excitotoxicity, the consequences of a 6-h exposure to either hypoxia (95% N₂/5% CO₂) or 100 µ M glutamate were studied in cultured fetal rat forebrain neurons taken at two maturational stages, i.e., 6 and 13 days in vitro. Cells were examined for their morphology, viability, energy metabolism reflected by 2- d -[³H]deoxyglucose uptake, and protein synthesis assessed by [³H]leucine incorporation. Apoptosis and necrosis were scored using the fluorescent dye 4,6-diamidino-2-phenylindole. Whereas 6-day-old neurons responded to a 6-h hypoxia by transient hypermetabolism, biphasic increase in protein synthesis, and cycloheximide-sensitive apoptotic death within 72 h postexposure, glutamate did not affect cell characteristics by the same time. In 13-day-old neurons, hypoxia induced both apoptosis (8.2%) and necrosis (22.3%). At this age, glutamate definitely reduced energy metabolism (26%) and protein synthesis (17%) by the end of exposure. The percentage of necrotic neurons reached 40.7%, but the rate of apoptosis was unchanged compared with controls. Therefore, excitotoxicity cannot account for hypoxia-induced injury in immature neurons, but its participation is suggested in older cells by the suppression of the necrotic component of hypoxia by glutamate receptor antagonists at 13 days.     

Binocular visual processing in the owl's telencephalon.

Single neurons recorded from the owl's visual Wulst are surprisingly similar to those found in mammalian striate cortex. The receptive fields of Wulst neurons are elaborated, in an apparently hierarchical fashion, from those of their monocular, concentrically organized inputs to produce binocular interneurons with increasingly sophisticated requirements for stimulus orientation, movement and binocular disparity. Output neurons located in the superficial laminae of the Wulst are the most sophisticated of all, with absolute requirements for a combination of stimuli, which include binocular presentation at a particular horizontal binocular disparity, and with no response unless all of the stimulus conditions are satisfied simultaneously. Such neurons have the properties required for 'global stereopsis', including a receptive field size many times larger than their optimal stimulus, which is more closely matched to the receptive fields of the simpler, disparity-selective interneurons. These marked similarities in functional organization between the avian and mammalian systems exist in spite of a number of structural differences which reflect their separate evoluntionary origins. Discussion therefore includes the possibility that there may exist for nervous systems only a very small number of possible solutions, perhaps a unique one, to the problem of stereopsis.     

Loss of sensory input causes rapid structural changes of inhibitory neurons in adult mouse visual cortex

A fundamental property of neuronal circuits is the ability to adapt to altered sensory inputs. It is well established that the functional synaptic changes underlying this adaptation are reflected by structural modifications in excitatory neurons. In contrast, the degree to which structural plasticity in inhibitory neurons accompanies functional changes is less clear. Here, we use two-photon imaging to monitor the fine structure of inhibitory neurons in mouse visual cortex after deprivation induced by retinal lesions. We find that a subset of inhibitory neurons carry dendritic spines, which form glutamatergic synapses. Removal of visual input correlates with a rapid and lasting reduction in the number of inhibitory cell spines. Similar to the effects seen for dendritic spines, the number of inhibitory neuron boutons dropped sharply after retinal lesions. Together, these data suggest that structural changes in inhibitory neurons may precede structural changes in excitatory circuitry, which ultimately result in functional adaptation following sensory deprivation.     

Responses and properties of receptive fields of neurons in the visual projection zone of the pigeon hyperstriatum

Unit responses in the hyperstriatal region of the pigeon forebrain to the action of various visual stimuli were investigated. Particular attention was paid to the discovery of retinotopic projection in the Wulst region. It was shown that as the electrode was advanced in the caudal direction in the zone of visual projection of the hyperstriatum the receptive fields of the neurons recorded shifted in the opposite direction in the visual field. The receptive fields of neurons of the ventral and dorsal hyperstriatum lie higher in the visual field and are larger in diameter than those of neurons of the accessory hyperstriatum. Unit responses in the visual projection zone of the Wulst depend on the intensity of illumination, size, and speed and direction of movement of the test objects across the receptive field. The functional role of the retino-thalamo-telencephalic system in visual interpretation in birds is discussed and it is suggested that the Wulst region is comparable with the striatal and also with the frontal regions of the mammalian cortex.M. V. Lomonosov Moscow State University. Translated from Neirofiziologiya, Vol. 8, No. 3, pp. 230–236, May–June, 1976.     

Neurons of the Paraventricular Hypothalamic Nucleus Under Normal and Modified Illumination Conditions: Immunohistochemical and Morphometric Parallels

Using an immunohistochemical technique and densitometric analysis in experiments on rats, we examined the expression of an early-response protein, c-Fos, in the medial parvicellular subnucleus of the hypothalamic paraventricular nucleus (mpcPVN). During 7 days, the animals were kept under conditions of (i) normal photoperiod (group LD), (ii) continuous illumination (group LL), and (iii) continuous darkness (light deprivation, group DD). It was found that both normal photoperiodicity and artificial modifications of the latter result in significant changes of c-Fos immunopositivity in the nuclei of mpcPVN neurons (cells that synthesize corticotropin-releasing factor). Changes in the indices of concentration and amount of c-Fos in mpcPVN neurons were, from some aspects, similar to those observed under analogous experimental conditions in neurons of a leading pacemaker of the circadian periodicity, the suprachiasmatic nuclei (SChNs) of the hypothalamus [10], but, at the same time, demonstrated noticeable specificity from other aspects. In all experimental groups, the concentration and amount of c-Fos in mpcPVN neurons were greater during daytime than at night. Dramatic increases in the above parameters observed at daytime in rats subjected to light deprivation were the most significant effect of modifications of the photoperiod. Both continuous illumination and light deprivation resulted in some increase of the number (density) of c-Fos-immunopositive units in the mpcPVN. The nuclei of neurons of this structure demonstrated the phenomenon found earlier in SChN neuron; geometrical dimensions of the cell nuclei of these structures depended noticeably on both illumination conditions and periodicity. Probable mechanisms of the dependence of c-Fos expression in the hypothalamic nuclei on the photoperiod and its modifications, interdependence of such changes observed in the circadian pacemaker (SChN) and an important component of the system controlling the stress reactions (mpcPVN), and the relation of the above phenomena to the level of melatonin produced by the epiphysis cerebri (main neuroendocrine intermediator organizing the above-mentioned periodicity) are discussed.     

Brief daily periods of binocular vision prevent deprivation-induced acuity loss

The role of experience in the development of the central visual pathways has been explored in the past through examination of the consequences of imposed periods of continuously abnormal or biased visual input. The massive changes in the visual cortex (area 17) induced by selected early visual experience, especially monocular deprivation (MD) or experience (ME) where patterned visual input is provided to just one eye, are accompanied by profound and long-standing visual deficits. Although the use of exclusively abnormal experience permits identification of those aspects of the visual cortex and of visual function that can be influenced by visual experience during development, this approach may provide a distorted view of the nature of the role of visual experience because of the absence of any normal visual input. In this study a different approach was used whereby animals were provided daily with separate periods of normal (i.e., binocular exposure) and abnormal (monocular exposure) visual experience. We show that 2 hr of daily normal concordant binocular experience (BE) can outweigh or protect against much longer periods of monocular deprivation (MD) and permit the development of normal visual acuities in the two eyes. This result is not what would be expected if all visual input had equal influence on visual development.     

Iron deprivation induces apoptosis independently of p53 in human and murine tumour cells

Iron deprivation induces apoptosis in some sensitive cultured tumour cells, while other cells are resistant. In order to elucidate the mechanisms involved in apoptosis induction by iron deprivation, we studied the expression of p53 and the expression of selected p53-regulated genes. To discriminate between changes coupled only with iron deprivation and changes involved in apoptosis induction by iron deprivation, we compared the expression of the genes in sensitive (human Raji, mouse 38C13) versus resistant (human HeLa, mouse EL4) cells under iron deprivation. Iron deprivation was achieved by incubation in a defined iron-free medium. The level of p53 mRNA decreased significantly under iron deprivation in sensitive cells, but it did not change in resistant cells. On the contrary, the level of the p53 protein under iron deprivation was slightly increased in sensitive cells while it was not changed in resistant cells. The activity of p53 was assessed by the expression of selected p53-regulated targets, i.e. p21(WAF1/CIP1) gene, mdm2, bcl-2 and bax. We did not detect any relevant change in mRNA levels as well as in protein levels of these genes under iron deprivation with the exception of p21(WAF1/CIP1). We detected a significant increase in the level of p21 mRNA in both (sensitive and resistant) mouse cell lines tested, however, we did not find any change in both (sensitive and resistant) human cell lines. Moreover, the p21(WAF1/CIP1) protein was accumulated in mouse-sensitive 38C13 cells under iron deprivation while all other cell lines tested, including human-sensitive cell line Raji, did not show any accumulation of p21(WAF1/CIP1) protein. It seems that the p21(WAF1/CIP1) mRNA, as well as protein accumulation, is not specifically coupled with apoptosis induction by iron deprivation and that it is rather cell-line specific. Taken together, we suggest that iron deprivation induces apoptosis at least in some cell types independently of the p53 pathway.     

Age-related changes in sensory neurons containing calcitonin gene-related peptide under conditions of afferentation deficit in rats

Morphological features of calcitonin gene-related peptide (CGRP)-immunoreactive neurons were studied in the sensory ganglia of the vagus and thoracic nerves in 3-, 10-, 20-, 30-, 60-, 90-, and 180-day-old rats under conditions of chemically-induced deafferentation. We found that, in rats, CGRP-containing neurons appeared in both ganglia immediately after they were born and their number decreased with aging. Most of CGRP-immunoreactive neurons were small in size, i.e., up to 600 ??m². Administration of capsaicin modified age-related changes in the number of CGRP-immunopositive neurons. In the thoracic nerve ganglion, the mean square of these cells and their number substantially decreased, whereas, in the vagus nerve ganglion, positive cells were not observed.     

3H]-GABA selective retrograde labeling of neurons in the pigeon thalamo-Wulst pathway

Evidence is presented which is consistent with a specific retrograde labeling of GABAergic neurons following [3H]-GABA application in their zone of termination. [3H]-GABA injection in the pigeon Wulst leads to perikaryal retrograde labeling in the ipsilateral thalamic visual relay, n. dorsolateralis anterior thalami, pars lateralis (DLLv). This result gives further support to the biochemical evidence of the existence, in the pigeon, of a GABAergic projection from DLLv to the ipsilateral visual Wulst.     

Quantitative morphological characteristics of developing neurons after partial deafferentation of the sensory trigeminal nuclei

We have studied different types of neurons in the sensory trigeminal nuclei stained by the Golgi method in kittens aged 30 days with bilateral transection of the lingual nerve, made on the fifth postnatal day. We have shown that deprivation of afferent inflow from the tongue to trigeminal neurons leads to changes in the structure of all types of cells: reticular, arborescent, and bushy (68, 61, and 46% neurons respectively had changed), short-dendrite cells having changed to a lesser extent (16% changed neurons). The multipolar giant neurons hardly changed. The structural changes involved changes in the size of the bodies, number, length, and ramification of dendrites, and changes in their orientation and pattern of ramification, compared to the normal. We observed destructive changes resulting in a decrease in the quantitative parameters, and constructive changes resulting in an increase in the latter. Reticular and arborescent neurons showed both destructive and constructive changes, the short-dendritic neurons mainly constructive changes, and bushy neurons mainly destructive changes. The analysis of the differently directed rearrangements of the dendrite geometry in different types of deafferentated trigeminal neurons allowed us to put forward some proposals concerning the different functional role of these groups of cells in the system of afferent impulsation entering via the lingual nerve.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR. Institute of the Brain of the All-Union Research Center of Mental Health, Academy of Medical Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 17, No. 4, pp. 522–530, July–August, 1985.     

Multinucleated erythroid cells in blood of golden hamster embryos

Blood smears of 11 and 13-day-old embryos (20) and newborns (10) of golden hamsters were analysed. The smears were stained with brilliant cresyl blue in a moist chamber for 5--7 min. About 2--6 thousand cells were counted in 200 visual fields. By means of an ocular micrometer MOB-1 the diameters of 300 cells were measured and the curves on cell distribution were plotted according to the diameters. It was found that the blood of 11-day-old golden hamster embryos contained about 8% of binuclear and sixnuclear megaloblasts, and that of 13-day-old embryos--about 2.5% of binuclear and sixnuclear erythroblasts and megablasts of the second generation. In the bloood of newborn hamster polynuclear cells are absent.     

Relative Wulst volume is correlated with orbit orientation and binocular visual field in birds

In mammals, species with more frontally oriented orbits have broader binocular visual fields and relatively larger visual regions in the brain. Here, we test whether a similar pattern of correlated evolution is present in birds. Using both conventional statistics and modern comparative methods, we tested whether the relative size of the Wulst and optic tectum (TeO) were significantly correlated with orbit orientation, binocular visual field width and eye size in birds using a large, multi-species data set. In addition, we tested whether relative Wulst and TeO volumes were correlated with axial length of the eye. The relative size of the Wulst was significantly correlated with orbit orientation and the width of the binocular field such that species with more frontal orbits and broader binocular fields have relatively large Wulst volumes. Relative TeO volume, however, was not significant correlated with either variable. In addition, both relative Wulst and TeO volume were weakly correlated with relative axial length of the eye, but these were not corroborated by independent contrasts. Overall, our results indicate that relative Wulst volume reflects orbit orientation and possibly binocular visual field, but not eye size.