Cell death in the lateral geniculate nucleus, and its possible relationship with nicotinic receptors and sudden infant death syndrome (SIDS).

The role of the lateral geniculate nucleus (LGN) in vision has been extensively studied, yet its extraretinal capacities are still being investigated, including its role in arousal from sleep. The ß2 nicotinic acetylcholine receptor (nAChR) subunit is involved in the laminal organisation of the LGN with magnocellular (MC) and parvocellular (PC) neurons. Sudden infant death syndrome (SIDS) occurs during a sleep period and, neuropathologically, is associated with increased neuronal cell death and altered nAChRs. A recent qualitative pilot study from our group implicates the possibility of increased neuronal death/apoptosis in the SIDS LGN. The present study used quantitative analysis to report the baseline expression of apoptotic and nAChR subunits α7 and ß2 in the PC and MC layers of the LGN, to determine correlations amongst these markers within layers and across layers, and to evaluate changes in the expression of these markers in the LGN of SIDS infants, along with associations with SIDS risk factors, such as age, sex, cigarette smoke exposure, bed-sharing, and presence of an upper respiratory tract infection (URTI). Tissue was immunohistochemically stained for cell death markers of active caspase-3 (Casp-3) and TUNEL, and for the α7 and ß2 nAChR subunits. Amongst 43 cases of sudden and unexpected deaths in infancy (SUDI), classifications included explained deaths (eSUDI, n = 9), SIDS I (n = 5) and SIDS II (n = 29). Results indicated a strong correlation of the apoptotic markers and ß2 nAChR subunit between the LGN layers, but not across the markers within the layers. Amongst the diagnostic groups, compared to eSUDI, the SIDS II cases had decreased Casp-3 expression while ß2 nAChR expression was increased in both PC and MC layers. Amongst the SIDS risk factors, URTI and bed-sharing were associated with changes in neuronal death but not in the α7 and ß2 markers. In conclusion, our findings do not support a role for the α7 and ß2 nAChRs in apoptotic regulation of the LGN layers during infancy. However, for SIDS victims, an inverse correlation between the changes for markers of apoptosis and the ß2 nAChR subunit expression suggests altered LGN function.

Medial geniculate body and primary auditory cortex differentially contribute to striatal sound representations.

The dorsal striatum has emerged as a key region in sensory-guided, reward-driven decision making. A posterior sub-region of the dorsal striatum, the auditory striatum, receives convergent projections from both auditory thalamus and auditory cortex. How these pathways contribute to auditory striatal activity and function remains largely unknown. Here we show that chemogenetic inhibition of the projections from either the medial geniculate body (MGB) or primary auditory cortex (ACx) to auditory striatum in mice impairs performance in an auditory frequency discrimination task. While recording striatal sound responses, we find that transiently silencing the MGB projection reduced sound responses across a wide-range of frequencies in striatal medium spiny neurons. In contrast, transiently silencing the primary ACx projection diminish sound responses preferentially at the best frequencies in striatal medium spiny neurons. Together, our findings reveal that the MGB projection mainly functions as a gain controller, whereas the primary ACx projection provides tuning information for striatal sound representations.

Relationship between monocularly deprivation and amblyopia rats and visual system development.

OBJECTIVE: To explore the changes of lateral geniculate body and visual cortex in monocular strabismus and form deprived amblyopic rat, and visual development plastic stage and visual plasticity in adult rats. METHODS: A total of 60 SD rats ages 13 d were randomly divided into A, B, C three groups with 20 in each group, group A was set as the normal control group without any processing, group B was strabismus amblyopic group, using the unilateral extraocular rectus resection to establish the strabismus amblyopia model, group C was monocular form deprivation amblyopia group using unilateral eyelid edge resection + lid suture. At visual developmental early phase (P25), meta phase (P35), late phase (P45) and adult phase (P120), the lateral geniculate body and visual cortex area 17 of five rats in each group were exacted for C-fos Immunocytochemistry. Neuron morphological changes in lateral geniculate body and visual cortex was observed, the positive neurons differences of C-fos expression induced by light stimulation was measured in each group, and the condition of radiation development of P120 amblyopic adult rats was observed. RESULTS: In groups B and C, C-fos positive cells were significantly lower than the control group at P25 (P<0.05), there was no statistical difference of C-fos protein positive cells between group B and group A (P>0.05), C-fos protein positive cells level of group B was significantly lower than that of group A (P<0.05). The binoculus C-fos protein positive cells level of groups B and C were significantly higher than that of control group at P35, P45 and P120 with statistically significant differences (P<0.05). CONCLUSIONS: The increasing of C-fos expression in geniculate body and visual cortex neurons of adult amblyopia suggests the visual cortex neurons exist a certain degree of visual plasticity.

Glaucoma alters the expression of NGF and NGF receptors in visual cortex and geniculate nucleus of rats: effect of eye NGF application.

We investigated the effect of glaucoma (GL) on nerve growth factor (NGF) presence in two brain visual areas. Rats with elevated intraocular pressure (EIOP), induced by hypertonic saline injection in the episcleral vein, were treated with eye topical application of saline or NGF. Rats were subsequently sacrificed, and brain tissues were used for immunohistochemical, biochemical, and molecular analyses. We found that GL alters the basal level of NGF and NGF receptors in brain visual centers and that NGF eye application normalized these deficits. These findings demonstrate that the reduced presence of NGF can arise due to degenerative events in retinal and brain visual areas.

Increased c-fos-like immunoreactivity in the superior colliculus and lateral geniculate nucleus of the rd mouse.

In most subcortical visual centers in normal mice maintained for a period in the dark, very few neurons express fos-like immunoreactivity (FLI), most likely reflecting c-fos expression, but if an animal is exposed to a flashing light, there is transient increase in the number of FLI-expressing cells. In dark-maintained retinal degeneration (rd) mice, with an inherited photoreceptor defect, numbers of FLI-positive cells, identified immunohistochemically, are anomalously elevated in the superior colliculus (SC) and lateral geniculate nucleus (LGN). Eye removal largely prevents the elevated counts. The difference in number of FLI-positive cells in the SC of rd mice and nondystrophic controls is highly significant (p<0.001). Because we have previously found a similar phenomenon in Royal College of Surgeons (RCS) rats, in which photoreceptor loss is caused by a retinal pigment cell defect, it argues for an effect related to photoreceptor loss rather than its cause.

Early events of target deprivation/axotomy-induced neuronal apoptosis in vivo: oxidative stress, DNA damage, p53 phosphorylation and subcellular redistribution of death proteins.

The mechanisms of injury- and disease-associated apoptosis of neurons within the CNS are not understood. We used a model of cortical injury in rat and mouse to induce retrograde neuronal apoptosis in thalamus. In this animal model, unilateral ablation of the occipital cortex induces apoptosis of corticopetal projection neurons in the dorsal lateral geniculate nucleus (LGN), by 7 days post-lesion, that is p53 modulated and Bax dependent. We tested the hypothesis that this degenerative process is initiated by oxidative stress and early formation of DNA damage and is accompanied by changes in the levels of pro-apoptotic mediators of cell death. Immunoblotting revealed that the protein profiles of Bax, Bak and Bad were different during the progression of neuronal apoptosis in the LGN. Bax underwent a subcellular redistribution by 1 day post-lesion, while Bak increased later. Bad showed an early sustained increase. Cleaved caspase-3 was elevated maximally at 5 and 6 days. Active caspase-3 underwent a subcellular translocation to the nucleus. A dramatic phosphorylation of p53 was detected at 4 days post-lesion. DNA damage was assessed immunocytochemically as hydroxyl radical adducts (8-hydroxy-2-deoxyguanosine) and single-stranded DNA. Both forms of DNA damage accumulated early in target-deprived LGN neurons. Transgenic overexpression of superoxide dismutase-1 provided significant protection against the apoptosis but antioxidant pharmacotreatments with trolox and ascorbate were ineffective. We conclude that overlapping and sequential signaling pathways are involved in the apoptosis of adult brain neurons and that DNA damage generated by superoxide derivatives is an upstream mechanism for p53-regulated, Bax-dependent apoptosis of target-deprived neurons.

Fos immunoreactivity in rat subcortical visual shell in response to illuminance changes.

Immediate early gene expression has been used frequently as a marker of activity in the circadian visual system. Recent evidence suggests that the pretectum participates in orchestrating sleep and circadian responses to light. Lesions of the pretectum eliminate dark shift-induced rapid eye movement sleep triggering in albino rats, and compromise circadian phase shifts in hamsters. We hypothesized that regions of the pretectum respond to light with robust and region-specific Fos activation, similar to the suprachiasmatic nucleus and intergeniculate leaflet. We used Fos expression, the protein product of the immediate early gene c-fos, as a functional marker to measure the responses of neurons following acute lighting changes. Rats maintained on a 12:12 light-dark cycle were subjected to a shift from light-to-dark or from dark-to-light at midday (Zeitgeber time 6) or midnight (Zeitgeber time 18). Fos expression was visualized with immunocytochemistry and quantified with an automated scoring system. We found three regions in the pretectum (the olivary pretectal nucleus, posterior limitans, and a region homologous to the hamster commissural pretectal nucleus), and two regions in the lateral geniculate complex (the intergeniculate leaflet and ventral lateral geniculate nucleus) that demonstrated significant Fos activation in response to light. Furthermore, the olivary pretectal nucleus, the posterior limitans, and the ventral lateral geniculate nucleus showed preferential Fos activation after acute light onset rather than following chronic exposure to light at midday, whereas at midnight these nuclei showed Fos activation following both chronic light exposure and acute light onset. Given the extensive anatomical connections between pretectal nuclei and other nuclei in the subcortical visual shell, as well as with centers for sleep and arousal, it is highly plausible that these pretectal nuclei integrate information about changes in illuminance, and aid in the coordination of acute behavioral responses to light.

Induction of Fos-like immunoreactivity in the ventral thalamus after electrical or chemical stimulation of various subcortical centres of rats.

We have examined the patterns of Fos-like immunoreactivity in the ventral thalamus (thalamic reticular nucleus (Rt), zona incerta (ZI) and ventral lateral geniculate nucleus (LGv)) after electrical or chemical stimulation of nuclei in either the brainstem (midbrain reticular nucleus), basal forebrain (substantia innominata) or dorsal thalamus (parafascicular nucleus). Sprague-Dawley rats were anaesthetised with Halothane or Ketamil/Rompun and the above mentioned centres were stimulated either electrically or chemically (using kainic acid). Brains were then processed for Fos-like immunocytochemistry using standard methods. We detected no major differences in the labelling patterns after either electrical or chemical stimulation or after using Halothane or Ketamil/Rompun anaesthesia. After brainstem or dorsal thalamic stimulations, many Fos-like immunoreactive cells were seen within the rostral pole of the Rt, the dorsal sector of the ZI and the parvocellular lamina of the LGv. After basal forebrain stimulations, many Fos-like immunoreactive cells were seen in the rostral pole of the Rt and rostral sector of the ZI, but very few were apparent in the LGv. Overall, our results show that distinct groups of cells in the ventral thalamus show increased levels of Fos-like immunoreactivity after stimulation of different subcortical centres. These activated cells of the ventral thalamus, are in turn, in a position to influence particular thalamocortical pathways through their dorsal thalamic projections.

Immunocytochemical mapping of NPY and VIP neuronal elements in the cat subcortical visual nuclei, with special reference to the pretectum and accessory optic system.

The aim of this study was to describe the distribution patterns of neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP)-immunoreactive (ir) neuronal elements in subcortical visual centers of the cat. Numerous NPY-ir neurons were present in the feline nucleus of the optic tract and in the anterior pretectal nucleus. Only a few NPY-ir neurons were found in the posterior, medial and olivary pretectal nuclei and in the accessory optic nuclei. Diffuse and heavily beaded NPY-ir fiber plexuses were observed throughout the superior colliculus, pretectum, and accessory optic system. Extensively arborising NPY-ir fibers were present in the mesencephalon and ventral lateral geniculate nucleus, while the dorsal visual thalamic nuclei contained only a few NPY-ir fibers. VIP-ir cells were present mainly in the accessory optic nuclei, and they were absent in the dorsal visual thalamus. Both NPY- and VIP-ir neurons were multipolar and fusiform in shape in the regions studied. Enucleation did not alter the appearance of NPY- and VIP-containing neuronal elements in the superior colliculus and pretectum while in the thalamus a subset of NPY-ir fiber population disappeared, indicating their retinal origin. Although there is a partial overlap in the topographical localization of the NPY- and VIP-ergic neurons in the pretectum, the colocalization of the two peptides could not be demonstrated. The present observations demonstrate the existence of two different and separate peptidergic (NPY and VIP) neuronal populations in the pretectum.

Estrogen receptor beta and progesterone receptor mRNA in the intergeniculate leaflet of the female rat.

The present study was undertaken to explore the possibility that the integration of hormonal cues in the regulation of neuroendocrine mechanisms may occur outside of the hypothalamus at the level of the lateral geniculate body. In situ hybridization for mRNA encoding estrogen receptor beta and progesterone receptor was carried out on sections containing the lateral geniculate body using [35S]-labeled antisense riboprobes. Labeled cells were present in different limbic and hypothalamic sites as described previously. Populations of cells distributed homogeneously in the ventral lateral geniculate nucleus and intergeniculate leaflet were also found to express mRNA for estrogen receptor beta and progesterone receptor. The dorsal lateral geniculate nucleus lacked specific labeling for either type of gonadal steroid hormone receptor mRNA. The present observation together with the recent demonstration of a direct pathway between the intergeniculate leaflet and hypothalamic neuroendocrine cells indicate that integration of hormonal and photic stimuli in the central regulation of endocrine mechanisms occurs outside of the hypothalamus in the lateral geniculate body.

The effectiveness of light on the circadian clock is linked to its emotional value.

Studies carried out within the primary visual system have shown that neural responses to light stimuli transmitted via the retinogeniculate pathway are significantly altered when these stimuli are made aversive through conditioning. The effect of such aversive conditioning on neural responses to light transmitted within the circadian visual system has not been investigated. In mammals, the principal projection of the circadian visual system, the retinohypothalamic tract, is functionally and anatomically distinct from the primary visual pathway allowing for direct transmission of light from the retina to the suprachiasmatic nucleus of the hypothalamus, the circadian clock. Light transmitted within this pathway is essential for entrainment of circadian rhythms providing the critical stimulus for resetting the circadian clock. We asked whether the response of neural elements within the suprachiasmatic nucleus to a resetting light stimulus would be altered if that stimulus had acquired aversive properties through conditioning. To study this we assessed the effect of a light stimulus made aversive through pairings with footshock on a cellular correlate of clock resetting, the expression of the transcription factor Fos in neurons of the suprachiasmatic nucleus. We show that Fos expression in the suprachiasmatic nucleus in response to light previously paired with footshock is significantly suppressed. This finding provides the first evidence that the effectiveness of a light as a resetting stimulus can be modulated by its conditioned aversive properties.

Delayed loss of p75 neurotrophin receptor-immunoreactivity in the rat suprachiasmatic nucleus and intergeniculate leaflet after binocular enucleation.

The rat suprachiasmatic nucleus (SCN) contains a dense plexus of low-affinity p75 neurotrophin receptor (p75NTR)-immunoreactivity. Scattered patches of p75NTR immunoreactivity are present in the intergeniculate leaflet (IGL). Both SCN and IGL receive a direct retinal input. After binocular enucleation, there is a delayed loss of p75NTR-immunoreactivity in the SCN and IGL beginning at, respectively, 4 and 8 weeks post-enucleation, with complete loss occurring in both nuclei by week 12. This delayed loss may be due to an up-regulation of growth factor secretion by local cells in response to retinal axon degeneration.

Constant light induces persistent Fos expression in rat intergeniculate leaflet.

Fos protein expression in retinorecipient suprachiasmatic nucleus (SCN) neurons is a marker of photic entrainment of circadian rhythms. Light-induced Fos in neurons of the intergeniculate leaflet (IGL) is not well-characterized. We compared Fos immunoreactivity (Fos-IR) in SCN and IGL neurons of rats housed under various lighting conditions and sacrificed at different phases of the circadian period. IGL neurons of rats that received 1 h-3 weeks of light exposure prior to sacrifice displayed Fos-IR, whereas the IGL of animals exposed only to darkness displayed little if any staining. In contrast with light-induced Fos in SCN neurons, Fos-IR was observed in the IGL regardless of circadian time. This work supports the idea that the IGL is involved in transmission of photic information to the SCN in rats.

Unilateral enucleation induces an increase of 160 kd neurofilament in lateral geniculate nuclei synapses.

Ultrastructural synaptic changes of retinal origin in the pars dorsalis lateral geniculate nuclei (dLGN) after enucleation have been studied in this laboratory, showing a filamentous hypertrophy with maximal expression at 4-6 days post-lesion in monkeys (Pecci Saavedra et al., 1970, 1971). The aim of this work was to elucidate the nature of the newly formed filament in dLGN in post-enucleated rats. Male Wistar rats were fixed with 4% paraformaldehyde plus 0.25% glutaraldehyde in 0.1M phosphate buffer, through the abdominal aorta after 3, 5, and 7 days postenucleation. Sections obtained were incubated with antibodies to the phosphorylated portion of the 160 Kd neurofilaments (1:3000) and anti-GFAP (1:25000). There was an increase in 160 Kd neurofilament staining in axons and degenerating nerve endings in dLGN, as well as a typical astroglial immunostained reaction. Our results show that the newly formed neurofilaments after deafferentation are of the 160 Kd type, commonly present in normal axons.

Cellular colocalization of Fos and neuropeptide Y in the intergeniculate leaflet after nonphotic phase-shifting events.

Nonphotic and photic stimuli that phase shift circadian rhythms were presented to hamsters, Mesocricetus auratus. The nonphotic stimulus was a 3-h pulse of novelty-induced wheel running starting at circadian time 4-5. The photic stimulus used was a 0.5 h light pulse starting at circadian time 18. Double immunocytochemistry was used to determine the neurochemical phenotype of cells in the intergeniculate leaflet that were activated by these stimuli. Both the nonphotic and the photic phase-shifting stimuli induced the expression of c-fos in the intergeniculate leaflet compared to unstimulated controls. However, after nonphotic stimulation, Fos-like immunoreactivity was common in neurons that also were NPY positive. Such colocalization of Fos and NPY after photic stimuli was rare. These findings suggest that the NPY pathway from the intergeniculate leaflet to the suprachiasmatic nucleus carries information about nonphotic events.

Reduced retinal activity increases GFAP immunoreactivity in rat lateral geniculate nucleus.

Dynamic regulation of astrocytic processes by the electrical activity of local neurons has been previously described in chick cochlear nucleus. The present study extends this observation by showing that astrocytes in the rat lateral geniculate nucleus (LGN) also increase their immunoreactivity for glial fibrillary acidic protein (GFAP) soon after deprivation of afferent visual neuronal activity. Within 6 h of enucleation, which eliminates a major source of afferent input to the contralateral LGN, GFAP immunoreactivity increases relative to the ipsilateral LGN. A similar increase in GFAP immunoreactivity can be induced by intraocular injections of tetrodotoxin, demonstrating that a reversible manipulation of optic nerve electrical activity is sufficient to regulate LGN astrocytes. This rapid response to activity deprivation is less dramatic than the gliotic reaction observed 3 weeks following deafferentation, by which time afferent terminals have degenerated. These results support the notion that regulation of astrocytic processes by neural activity may play an important role in activity-dependent synaptic regulations in the various sensory systems of vertebrates.

Effects of eye-enucleation on substance P-immunoreactive fibers of some retinorecipient nuclei of the rat in relation to their origin from the superior colliculus.

We have previously shown that retinal deafferentation causes a decrease in immunoreactive dendrites of substance P-positive neurons of the superficial superior colliculus of the rat. Since some retinorecipient thalamic and pretectal nuclei are putative targets for substance P-containing cells of the superior colliculus, the present study attempted to ascertain whether substance P-immunoreactive fibers in these nuclei are also affected by retinal denervation. We found that unilateral eye removal produced a progressive increase in fibrous substance P immunoreactivity in the nucleus of the optic tract, lateral posterior nucleus, and lateral geniculate nucleus of the side contralateral to the enucleation. On the other hand, unilateral lesions to the superficial layers of the superior colliculus produced a dramatic reduction in substance P immunoreactivity in the ipsilateral nucleus of the optic tract, lateral posterior nucleus, and dorsal and ventral lateral geniculate nuclei. In bilaterally enucleated animals, unilateral lesion to the superior colliculus produced, as expected, loss of immunoreactive fibers only in the lateral posterior nucleus and the retinorecipient nuclei ipsilateral to the lesion. These results suggest that transneuronal changes in the distribution of substance P in collicular neurons observed after enucleation could be reflected in their projections to the other primary visual centers and to the lateral posterior nucleus.

Proteins of axonal transport: investigation of solubility characteristics and behaviour in gel filtration.

Rapid axonal transport of proteins in retinal ganglion cells of the rabbit was studied following intraocular injections of labelled amino acids. Approximately 10% of the transported radioactivity was found in the supernatant following homogenization and high-speed centrifugation of the nerve terminal region. Relatively simple manipulations with ionic strength, pH and the presence of a chelating agent could solubilize an equivalent amount of radioactivity from the pellet. Lithium diiodosalicylate solubilized most rapidly transported membrane proteins. Gel filtration of readily soluble rapidly transported radioactivity gave a main macromolecular radioactive peak with an approximate mol. wt. of 500,000 dalton as determined on Sephadex G-200. However, gel filtration on Sepharose CL-6B gave a mol. wt. of about 160,000 for the same radioactive peak. SDS polyacrylamide gel electrophoresis of rapidly transported soluble proteins and fractions derived from these proteins via gel filtration and ion exchange chromatography revealed in all cases a very complex picture of labelled polypeptides. Thus rapid axonal transport of soluble proteins in this system seems to involve many different macromolecules.