Gigliola Fontanesi

Changing distribution of GABA-like immunoreactivity in pigeon visual areas during the early posthatching period and effects of retinal removal on tectal GABAergic systems

The distribution of GABA-like immunoreactivity in the pigeon visual system was studied during the first 9 days after hatching using a mouse monoclonal antibody, mAb 3A12, to glutaraldehyde linked GABA (Matute & Streit, 1986). GABA-like immunoreactivity was seen in cell bodies as well as in neuropil at the level of both the retina and central visual regions at any posthatching age. However, the distribution of putative GABAergic cells and processes varied with age reaching the adult pattern at around 9 days. As a general observation, almost no cell bodies in the retina (except for some perikarya in the ganglion cell layer) were labeled at hatching but densely packed immunostained processes were present in the inner plexiform layer. During the next few days, GABA-immunoreactive amacrine and horizontal cells appeared and the adult distribution of GABA-like immunoreactivity was reached at around 9 days. In the other visual regions examined, the general trend in the variation of GABA-like immunoreactivity included: (1) a progressive decrease in the density of immunostained cell bodies and (2) an almost parallel increase in the concentration of stained neuropil. Since in pigeons the adult organization of visual pathways and the final distribution of putative GABAergic systems are reached at around the same age, we suggest the possibility that incoming ganglion cell axons play a role in regulating the distribution of GABA-like immunoreactivity in visual areas. This hypothesis is supported by the fact that the distribution of GABA-like immunoreactivity in the superficial layers of the optic tectum was altered following ablation of the contralateral retina immediately after hatching.

Nerve growth factor (NGF) uptake and transport following injection in the developing rat visual cortex

Recent investigations have shown that cortical nerve growth factor (NGF) infusions during the critical period inhibit ocular-dominance plasticity in the binocular portion of the rat visual cortex. The mechanisms underlying the effects of NGF on visual cortical plasticity are still unclear. To investigate whether during normal development intracortical and/or extracortical cells possess uptake/transport mechanisms for the neurotrophin, we injected 125I-NGF into the occipital cortex of rats at different postnatal ages. Within the cortex, only a few labelled cells were observed. These cells were confined to the vicinity of the injection site and their number depended on the animals age at the time of injection. Labelled cells were absent at postnatal day (PD) 10 but could be detected between PD 14 and PD 18. They then decreased in number over the following period and were not detected in adult animals. Outside the cortex, neurons of the lateral geniculate nucleus (LGN) were not observed to take up and retrogradely transport NGF at any age after birth. In contrast, retrogradely labelled neurons were found in the basal forebrain. Labelled cells were first observed here at PD 14 and then increased in number until reaching the adult pattern. Our results show that intrinsic and extrinsic neurons are labelled following intracortical injections of iodinated NGF. In both neuronal populations, the uptake and transport of NGF is present over a period corresponding to the critical period for visual cortical plasticity. These findings suggest that NGF may play a role, both intra and extracortically, in plasticity phenomena.

Binocularity in the little owl, Athene noctua. II. Properties of visually evoked potentials from the Wulst in response to monocular and binocular stimulation with sine wave gratings.

Visually evoked potentials (VEPs) have been recorded from the Wulst surface of the little owl, Athene noctua, in response to counterphase-reversal of sinusoidal gratings with different contrast, spatial frequency and mean luminance, presented either monocularly or binocularly. Monocular full-field stimuli presented to either eye evoked VEPs of similar amplitude, waveform and latency. Under binocular viewing, VEPs approximately doubled in amplitude without waveform changes. VEPs with similar characteristics could be obtained in response to stimulation of the contralateral, but not ipsilateral, hemifield. These results suggest that a 50% recrossing occurs in thalamic efferents and that different ipsilateral and contralateral regions converge onto the same Wulst sites. The VEP amplitude progressively decreased with increase of the spatial frequency beyond 2 cycles/degree, and the high spatial frequency cut-off (VEP acuity) was under binocular viewing (8 cycles/degree) higher than under monocular (5 cycles/degree) viewing (200 cd/m2, 45% contrast). The VEP acuity increased with increase in the contrast and decreased with reduction of the mean luminance. The binocular gain in both VEP amplitude and VEP acuity was largest at the lowest luminance levels. Binocular VEP summation occurred in the medium-high contrast range. With decreased contrast, both monocular and binocular VEPs progressively decreased in amplitude and tended to the same contrast threshold. The VEP contrast threshold depended on the spatial frequency (0.6-1.8% in the range 0.12-2 cycles/degree). Binocular VEPs often showed facilitatory interaction (binocular/monocular amplitude ratio greater than 2), but the binocular VEP amplitude did not change either by changing the stimulus orientation (horizontal vs. vertical gratings) or by inducing different retinal disparities.(ABSTRACT TRUNCATED AT 250 WORDS)

The electroretinogram of the little owl (Athene noctua)

Electroretinographic responses (ERGs) have been recorded from the cornea of the little owl (Athene noctua) in response to single light flashes and to alternating sinusoidal gratings (pattern) at different levels of light adaptation. Both flash- and pattern-evoked ERGs show scotopic as well as photopic components. The pattern evoked ERG is spatially tuned with tuning functions which shift towards lower frequencies by reducing the mean luminance. The retinal acuity is about 6 c/deg at 2.3 log cd/m2 and decreases progressively by reducing the mean luminance. No pattern ERG can be recorded beyond -6.7 log cd/m2 at any spatial frequency. The pattern ERG amplitude decreases progressively by reducing the contrast. The extrapolated contrast threshold is about 1%. Acuity and contrast sensitivity ERG values are in the range of those obtained by operant techniques in other species with duplex retinae such as owls and cats.

Postnatal development of somatostatin 2A (sst2A) receptors expression in the rabbit retina

In the retina, somatostatin (SRIF) acts as a neuromodulator by interacting with specific SRIF subtype (sst) receptors. Aim of this investigation was to determine the cellular localization of the sst2A receptor isoform in the postnatal rabbit retina. Receptor immunoreactivity was localized using the antiserum K-230, directed to the C-terminus of the human sst2A receptor. In the postnatal rabbit retina, sst2A receptors were abundantly expressed without significant regional differences. They were localized predominantly to rod bipolar cells, identified with a protein kinase C (PKC) antibody, to amacrine cells, some of which also containing tyrosine hydroxylase (TH), and to presumed rare horizontal cells. Quantitative analysis showed that sst2A-immunoreactive (-IR) bipolar and amacrine cells reached their maximum density and absolute number at the time of eye opening, when the expression pattern of sst2A receptors was similar to that in adult retinas. In the adult retina, 68% of the PKC-IR rod bipolars and 34% of the TH-IR amacrine cells were observed to also express sst2A receptors. The appearance of sst2A receptor immunolabeling prior to eye opening and the developmental profile of sst2A receptor expression are compatible with a role of SRIF in the maturation of retinal circuitries. The partial expression of sst2A receptors in PKC-IR rod bipolar cells and in TH-IR amacrine cells may suggest some type of heterogeneity within these cell populations.

Postnatal development of functional properties of visual cortical cells in rats with excitotoxic lesions of basal forebrain cholinergic neurons

In the rat, visual cortical cells develop their functional properties during a period termed as critical period, which is included between eye opening, i.e. postnatal day (PD) 15, and PD40. The present investigation was aimed at studying the influence of cortical cholinergic afferents from the basal forebrain (BF) on the development of functional properties of visual cortical neurons. At PD15, rats were unilaterally deprived of the cholinergic input to the visual cortex by stereotaxic injections of quisqualic acid in BF cholinergic nuclei projecting to the visual cortex. Cortical cell functional properties, such as ocular dominance, orientation selectivity, receptive-field size, and cell responsiveness were then assessed by extracellular recordings in the visual cortex ipsilateral to the lesioned BF both during the critical period (PD30) and after its end (PD45). After the recording session, the rats were sacrificed and the extent of both cholinergic lesion in BF and cholinergic depletion in the visual cortex was determined. Our results show that lesion of BF cholinergic nuclei transiently alters the ocular dominance of visual cortical cells while it does not affect the other functional properties tested. In particular, in lesioned animals recorded during the critical period, a higher percentage of visual cortical cells was driven by the contralateral eye with respect to normal animals. After the end of the critical period, the ocular dominance distribution of animals with cholinergic deafferentation was not significantly different from that of controls. Our results suggest the possibility that lesions of BF cholinergic neurons performed during postnatal development only transiently interfere with cortical competitive processes.

Somatostatin-like immunoreactivity in the pigeon visual system: developmental expression and effects of retina removal.

The distribution of somatostatin (SS)-containing neurons was investigated by immunocytochemical methods in the central visual system of adult, developing, and retina-ablated pigeons. In normal adult brains, SS-positive cells and processes were present in the optic tectum, the nucleus of the basal optic root, the visual Wulst, and the ectostriatum. During development, progressive increase or decrease in the numerical density and the total number of SS-containing neurons occurred as determined by quantitative analysis. Changes in SS immunoreactivity also occurred as a consequence of unilateral and bilateral retina removal immediately after hatching, i.e. before retinofugal connections have been established. In spite of the segregation of visual inputs due to the almost completely crossed retinal projections, unilateral and bilateral deafferentation differentially affected SS-containing visual regions. In addition, different effects were observed on the relative packing density of labeled cells as compared to their total number. A possible role of retinal axons in regulating the distribution of SS immunoreactivity was suggested by its altered expression induced by retinal deafferentation. In addition, parallels with the distribution of SS immunoreactivity in the pigeons visual system were used to suggest possible equivalence between cell populations in the avian and the mammalian brains.

Transient somatostatin-immunoreactive ganglion cells in the developing rat retina

The peptide somatostatin (SRIF) is likely to play important roles in neuronal differentiation and maturation. In the mammalian retina, it is reported to be expressed by populations of amacrine and/or displaced amacrine cells and, in some species, by some ganglion cells. Previous studies have shown that in the rat retina the maturation of somatostatinergic systems encompasses late prenatal and early postnatal periods, suggesting a role of SRIF in maturative events of the retina. SRIF-expressing ganglion cells have not been reported in the rat retina at any developmental age. In the present study, we re-evaluated the postnatal development of SRIF-containing neurons of the rat retina focusing on the analysis of SRIF-containing cells in the ganglion cell layer (GCL), to test the possibility that SRIF is expressed by some ganglion cells during development. To this aim we combined immunocytochemical staining of SRIF-positive neurons with retrograde tracing of ganglion cell bodies through Fluoro-Gold injections into the superior colliculus. Double-labelling experiments revealed the presence of SRIF-containing ganglion cells at postnatal day (PND) 10. They accounted for 14% of the total SRIF-containing cells in the GCL. Such double-labelled cells were not observed either before (PND 7) or after (PND 15 and PND 45) this period. This transient expression of SRIF in retinal ganglion cells suggests that SRIF may be a factor regulating the maturation of retinocollicular projections in a restricted period of postnatal development.

Chronic exposure to either somatostatin (SS) or octreotide, a long-lasting SS analogue, affects SS expression in the postnatal visual cortex of the rat.

The peptide somatostatin (SS) is widely distributed in the mammalian brain where it modulates neuronal activity through interactions with specific membrane-bound receptor subtypes (ssts). Five different ssts were characterized so far (sst1-5) and their selective agonists were developed on the basis of their binding specificity. SS and ssts are transiently expressed in the developing brain, suggesting a functional role of somatostatinergic systems in neuronal maturation. In the present study, we investigated the effects of chronic exposure to either the SS synthetic analogue, SS-14 or octreotide (a long-acting sst2-preferring analogue) on the maturation of SS-immunoreactivity (-ir) in the primary visual cortex of the rat. SS-ir maturation was investigated both by an evaluation of the number of SS-immunoreactive cells and by radioimmunoassay (RIA) to measure the levels of SS in the postnatal visual cortex. In the visual cortex of normal rats, the number of SS-positive cells markedly increased during the second postnatal week and then significantly decreased until the adult value was reached at the third week. Early and repeated intracerebroventricular (i.c.v.) injections of either SS-14 or octreotide prevented the increase in the number of SS-positive cells, with adult values reached at the end of the first postnatal week. Similarly, administration of either SS-14 or octreotide significantly decreased the SS content of the visual cortex, measured at the end of the second postnatal week. These results show that high local concentrations of either SS-14 or octreotide interfere with SS expression in developing cortical neurons in a restricted postnatal period.

Maturation and Plasticity of Neuropeptides in the Visual System

A large number of peptides have been recently described within the vertebrate central nervous system. Some of them were previously localized in non-neural vertebrate tissue as well as lower species in which they may serve as primitive elements of intercellular communication (Krieger, 1983).