PubMed Health⌕ Search

Biomedical subjects

L R Britto

Publications and source records attributed to L R Britto.

At least 37 records · Page 2Linked to original sources

Distribution of the alpha 2, alpha 3, and alpha 5 nicotinic acetylcholine receptor subunits in the chick brain.

Nicotinic acetylcholine receptors (nAChRs) are ionotropic receptors comprised of alpha and beta subunits. These receptors are widely distributed in the central nervous system, and previous studies have revealed specific patterns of localization for some nAChR subunits in the vertebrate brain. In the present study we used immunohistochemical methods and monoclonal antibodies to localize the alpha 2, alpha 3, alpha 5 nAChR subunits in the chick mesencephalon and diencephalon. We observed a differential distribution of these three subunits in the chick brain, and showed that the somata and neuropil of many central structures contain the alpha 5 nAChR subunit. The alpha 2 and alpha 3 subunits, on the other hand, exhibited a more restricted distribution than alpha 5 and other subunits previously studies, namely alpha 7, alpha 8 and beta 2. The patterns of distribution of the different nAChR subunits suggest that neurons in many brain structures may contain several subtypes of nAChRs and that in a few regions one particular subtype may determine the cholinergic nicotinic responses.

Animals↗

Distribution of calcium-binding proteins in the chick visual system.

The calcium-binding proteins calbindin (CB), calretinin (CR), and parvalbumin (PV) have been extensively studied over the last decade since they appear to be important as buffers of intracellular calcium. In the present study we investigated the distribution of these proteins in the chick visual system by means of conventional immunocytochemistry. The results indicated that CB, CR, and PV are widely distributed in retinorecipient areas of the chick brain. In some regions, all three calcium-binding proteins were present at different intensities and often in different neurons such as in the dorsolateral thalamic complex. In other areas, such as the nucleus geniculatus lateralis ventralis, only CB and CR were detected, whereas PV was absent. These results show that these three calcium-binding proteins are differentially distributed in the visual system of the chick, with varying degrees of co-localization.

Animals↗

Expression of glutamate receptor subunit genes during development of the mouse retina.

The distribution of expression during retinal differentiation of nine AMPA and kainate glutamate receptor subunit genes was studied by in situ hybridization. Transcripts encoding all subunits were detected in the inner nuclear and ganglion cell layers of the developing mouse retina, but their time courses of expression differed. The earliest expression was detected at embryonic day 14 (E14) for GluR1, GluR4 and KA-2; all the other subunits were first detected at E16. Most subunits were expressed at higher levels during development than in the adult. This suggests they play a role in the processes of neuronal differentiation and synaptogenesis that occur during the early postnatal days in the rodent retina.

Animals↗

Presumptive presynaptic nicotinic acetylcholine receptors in the chick tectum: effects of lesions of the lateral spiriform nucleus.

There are indications that nicotinic acetylcholine receptor subunits in the superficial layers of the chick tectum (Cajal's layers 1-7) may be transported from the retina. However, nicotinic receptor subunits are detectable by immunohistochemistry in all layers of the optic tectum. In this study, we performed unilateral electrolytic lesions of the lateral spiriform nucleus, which projects to the deep layers of the tectum and contains high amounts of nicotinic receptors in its perikarya. Following lesions of the lateral spiriform nucleus, both the alpha 5 and the beta 2 subunits were markedly depleted in the neuropil of the deep layers of the ipsilateral optic tectum (layers 8-13). No changes were observed in somata that contain either subunit. The present results suggest that most of the nicotinic acetylcholine receptor subunits in the chick optic tectum occur in axonal systems and could then constitute presynaptic receptors.

Animals↗

Expression of the Fos protein reveals functional subdivisions of the avian ventral lateral geniculate nucleus.

The Fos protein was immunocytochemically detected in the chick ventral lateral geniculate nucleus after novel stationary and optokinetic stimulation. Fos-positive nuclei were mainly detected in the internal part of the ventral geniculate when the animals were submitted to stationary visual stimulation. On the other hand, Fos-positive nuclei were mainly seen in the external part of the nucleus when optokinetic stimuli were used. These data reveal functional subdivisions of the avian ventral geniculate, and support the hypothesis that this nucleus is involved in several aspects of the visual function.

Animals↗

Neurogenesis of cholinoceptive neurons in the chick retina.

Immunocytochemistry and [3H]thymidine autoradiography were combined in this study to determine the neurogenesis of cholinoceptive cells in the chick retina. After injections of [3H]thymidine between embryonic days 1 and 11, the time of birth of retinal neurons containing either the alpha 3 or the alpha 8 subunit of the nicotinic acetylcholine receptors was determined. The results indicate that the alpha 3-positive neurons in the ganglion cell layer leave the cell cycle from E2 through E7, and those in the inner nuclear layer (amacrine and displaced ganglion cells) from E2 through E9. The alpha 8-positive cells in the ganglion cell layer were born from E1 through E7, and those in the inner nuclear layer (amacrine and bipolar cells) from E2 through E11. These data suggest that the time of birth of cholinoceptive neurons in the chick retina follows the general pattern of cell generation in the chick retina, and that alpha 8-positive cells in the ganglion cell layer start to leave the cell cycle almost one day earlier than the alpha 3-positive cells in the same layer.

Acetylcholine↗

Backup compass mechanisms in pigeon orientation with the sun in the zenith.

The sun is known to guide homing pigeons as a priority cue. The literature indicates that under total overcast conditions pigeons rely on a backup mechanism akin to the magnetic inclination compass for which there is much laboratory evidence in migratory birds. Total overcast conditions are not favorable for orientation research in the State of São Paulo, Brazil. The orientation of homing pigeons raised near the tropic of Capricorn was therefore observed around the time of the December solstice, when the sun culminated directly overhead, with a consequent interruption of the sun compass for a short time every day. In these experiments, carried out between 1981 and 1993, local geomagnetic field inclination was -25 degrees to -29 degrees 30', so that a functioning magnetic inclination compass should have been available to the birds. Whereas the birds released with sun to zenith angles between 10 degrees and 30 degrees were well oriented, both in the morning (99 vanishing bearings) and in the afternoon (143 vanishing bearings), those released with the sun less than 5 degrees away from the zenith showed random orientation (105 vanishing bearings), with no evidence of an alternative magnetic compass mechanism.

Animals↗

Calretinin in the mouse superior colliculus originates from retinal ganglion cells.

We investigated the origin of the calretinin-immunoreactive fibers in the mouse superior colliculus. The dense plexus of calretinin-positive fibers in the superficial layers of the colliculus was completely eliminated after eye enucleation. Retrograde tracing combined with immunohistochemistry revealed many calretinin-positive small-to-medium retinal ganglion cells projecting to the colliculus. These results indicate that calretinin-containing ganglion cells are the source of this calcium-binding protein in the superficial layers of the superior colliculus.

Animals↗

Serotonergic afferents of the pigeon accessory optic nucleus.

Injections of a retrograde tracer into the accessory optic nucleus of the basal optic root (nBOR) of the pigeon, combined with 5-HT immunochemistry, revealed that serotonergic projections to the nBOR appeared to originate mainly from the median (MR) and paramedian (PMR) raphe nuclei. These projections were confirmed by the significant decrease in 5-HT immunoreactivity observed in nBOR after lesions in MR and PMR. These data characterize distinct sources of 5-HT innervation to the pigeon nBOR and suggest that those afferents could represent part of a modulatory system that contributes to the role of the nBOR in optokinetic mechanisms.

Animals↗

Changes in expression of glutamate receptor subunits following photoreceptor degeneration in the rd mouse retina.

The effects of photoreceptor cell degeneration on the expression of nine alpha-amino-3-hydroxy-5-methyl-isoxasole-4-propionate (AMPA)/kainate glutamate receptor subunit genes were investigated in the rd mouse retina by in situ hybridization. All known AMPA/kainate receptor subunits were found to be expressed in normal mouse retina. Following retinal degeneration, the expression of KA-1 was reduced, that of the GluR7 subunit was greatly increased, and that of the other seven subunits were not significantly affected.

Animals↗

Effects of retinal lesions upon the distribution of nicotinic acetylcholine receptor subunits in the chick visual system.

Immunohistochemistry was used in this study to evaluate the effects of retinal lesions upon the distribution of neuronal nicotinic acetylcholine receptor subunits in the chick visual system. Following unilateral retinal lesions, the neuropil staining with an antibody against the beta 2 receptor subunit, a major component of alpha-bungarotoxin-insensitive nicotinic receptors, was dramatically reduced or completely eliminated in all of the contralateral retinorecipient structures. On the other hand, neuropil staining with antibodies against two alpha-bungarotoxin-sensitive receptor subunits, alpha 7 and alpha 8, was only slightly affected after retinal lesions. Decreased neuropil staining for alpha 7-like immunoreactivity was only observed in the nucleus of the basal optic root and layers 2-4 and 7 of the optic tectum. For alpha 8-like immunoreactivity, slight reduction of neuropil staining could be observed in the ventral geniculate complex, griseum tecti, nucleus lateralis anterior, nucleus lentiformis mesencephali, layers 4 and 7 of the tectum, and nucleus suprachiasmaticus. Taken together with previous data on the localization of nicotinic receptors in the retina, the present results indicate that the beta 2 subunit is transported from retinal ganglion cells to their central targets, whereas the alpha 7 and alpha 8 subunit immunoreactivity appears to have a central origin. The source of these immunoreactivities could be, at least in part, the stained perikarya that were observed to contain alpha 7 and alpha 8 subunits in all retinorecipient areas. In agreement with this hypothesis, the beta 2 subunit of the nicotinic acetylcholine receptors was not frequently found in perikarya of the same areas.

Afferent Pathways↗

Depletion and recovery of the calcium-binding proteins calbindin and parvalbumin in the pigeon optic tectum following retinal lesions.

Retinal lesions in pigeons produced a marked depletion of somata and neuropil staining for both calbindin-D28k and parvalbumin immunoreactivities in the contralateral optic tectum. Calbindin-like immunoreactivity reappeared in some tectal layers by 6 weeks postlesion, whereas paravalbumin-like immunoreactivity recovered almost completely after 5 weeks. These data indicate that the retinal input may control the expression of calbindin and parvalbumin in the pigeon optic tectum.

Animals↗

Differential development of alpha-bungarotoxin-sensitive and alpha-bungarotoxin-insensitive nicotinic acetylcholine receptors in the chick retina.

The development of cells containing neuronal nicotinic receptors (nAChRs) in the chick retina was investigated by means of immunohistochemical techniques with antibodies directed against the alpha 3 and alpha 8 nAChR subunits. The alpha 3 subunit is one of the major alpha-bungarotoxin-insensitive nicotinic receptor subunits in the chick retina, whereas alpha 8 appears to be the most common alpha-bungarotoxin-sensitive subunit in the same structure, alpha 3-like immunoreactivity (alpha 3-LI) was first detected in cells of the vitreal margin, on the embryonic day 4.5 (E4.5). alpha 8-LI was first detected in the same type of cell almost a day later. However, the processes of alpha 8-LI cells developed much faster than those of alpha 3-LI cells, generating visible stained laminae in the prospective inner plexiform layer as early as E7. alpha 3-LI was only clearly seen in laminae of the inner plexiform layer by E12. By this date, both alpha 3 and alpha 8-LI were seen in the same types of cells as in the adult retina, i.e., amacrines, displaced ganglion cells, and cells of the ganglion cell layer for alpha 3-LI; and amacrines, bipolar cells, and cells of the ganglion cell layer for alpha 8-LI. These results reveal different patterns of development of cells containing the alpha 3 and alpha 8 nAChR subunits in the chick retina and indicate that those nAChR subunits are expressed in the chick retina before choline acetyltransferase-positive cells can be detected and well before synaptogenesis. These data also suggest that nAChRs may have a developmental function in the retina.

Animals↗

Different subsets of displaced ganglion cells in the pigeon retina exhibit cholecystokinin-like and enkephalin-like immunoreactivities.

Immunohistochemical and retrograde tracing techniques were combined in order to identify chemically specific displaced ganglion cells in the pigeon retina. About 15% of the displaced ganglion cells that were retrogradely labeled following injections of different tracers into the accessory optic nucleus were shown to contain cholecystokinin8-like immunoreactivity. These cells were medium to large (15-30 microns) and located mostly in the peripheral retina. Another population of about 9% of the retrogradely labeled displaced ganglion cells was shown to contain leucine-enkephalin-like immunoreactivity. These cells were medium-sized (11-18 microns) and distributed almost evenly throughout the retina. These two types of displaced ganglion cells represent together only about 0.1% of the total number of ganglion cells in the pigeon retina. Taken together with previous results, these data indicate that the displaced ganglion cells of the avian retina may comprise several chemically specific cell types. The present results also contribute information on the chemical heterogeneity of retinal ganglion cells.

Animals↗

Three subtypes of alpha-bungarotoxin-sensitive nicotinic acetylcholine receptors are expressed in chick retina.

A recent report described the isolation of cDNA clones encoding alpha 7 and alpha 8 subunits of alpha-bungarotoxin-sensitive nicotinic ACh receptors (alpha BgtAChRs) from chick brain and demonstrated that they were related to, but distinct from, the alpha subunits of nicotinic ACh receptors (nAChRs) from muscles and neurons. Monoclonal antibodies against the two alpha BgtAChR subunits were used to demonstrate that at least two subtypes are present in embryonic day 18 chicken brain. The predominant brain subtype contains alpha 7 subunits, while a minor subtype contains both alpha 7 and alpha 8 subunits. Both subtypes may also contain other subunits. Here we report the results of immune precipitation studies and immunohistochemical studies of alpha BgtAChRs in the chick retina. In addition to the two subtypes found in brain, a new alpha BgtAChR subtype that contains alpha 8 subunits, but not alpha 7 subunits, was identified and was found to be the major subtype in chick retina. This subtype has a lower affinity for alpha-bungarotoxin (alpha Bgt) than does the subtype containing only alpha 7 subunits. Small amounts of this alpha 8 subtype were also detected in brain by labeling with higher concentrations of 125I-alpha Bgt than had been used previously. The subtype containing only alpha 7 subunits comprised 14% of the alpha BgtAChRs in hatchling chick retina. The subtype containing alpha 8 subunits (but no alpha 7 subunits) accounted for 69%, and the alpha 7 alpha 8 subtype accounted for 17%. Amacrine, bipolar, and ganglion cells displayed alpha 8 subunit immunoreactivity, and a complex pattern of labeling was evident in both the inner and outer plexiform layers. In contrast, only amacrine and ganglion cells exhibited alpha 7 subunit immunoreactivity, and the pattern of alpha 7 subunit labeling in the inner plexiform layer differed from that of alpha 8 subunit labeling. These disparities suggest that the alpha BgtAChR subunits are differentially expressed by different populations of retinal neurons. In addition, the distribution of alpha BgtAChR subunit immunoreactivity was found to differ from that of alpha-Bgt-insensitive nAChR subunits.

Animals↗

Visual discrimination performance after lesions of the ventral lateral geniculate nucleus in pigeons (Columba livia).

Pigeons were trained to perform simultaneous pattern and color discrimination tasks. After their training was completed, bilateral electrolytic lesions were made in the ventral lateral geniculate nucleus (GLv). Following the surgery, they were retrained to their preoperative performance levels. Lesions of GLv caused no deficits in pattern discrimination performance. The birds which had been trained for discrimination of red vs. magenta showed a slight decline in their performance. This impaired performance on color discrimination was not, however, as severe as that of a bird with lesions in the nucleus rotundus. These results suggest that GLv plays some role in the detection of short wavelengths of light.

Animals↗

Neurons of the chick brain and retina expressing both alpha-bungarotoxin-sensitive and alpha-bungarotoxin-insensitive nicotinic acetylcholine receptors: an immunohistochemical analysis.

Immunohistochemical methods were used to study the possible co-localization of two alpha-bungarotoxin-sensitive (alpha 7 and alpha 8) and two alpha-bungarotoxin-insensitive (beta 2 and alpha 3) subunits of the nicotinic acetylcholine receptors in neurons of the chick brain and retina. Several structures contained neurons that were doubly-labeled with antibodies against the alpha 7 subunit and the beta 2 subunit. These structures included, for example, the interpeduncular nucleus, nucleus spiriformis lateralis, optic tectum, pretectal visual nuclei, and the lateral hypothalamus. Double-labeling with antibodies against the alpha 7 and alpha 8 subunits was also seen in several regions, which included the interpeduncular nucleus, visual pretectum, lateral hypothalamus, dorsal thalamus, and the habenular complex. In the retina, many cells in the inner nuclear layer were observed to contain alpha 8 and alpha 3 subunits, whereas neurons in the ganglion cell layer were seen to contain alpha 7 and alpha 8 or, less frequently, alpha 7 and alpha 3 subunits. These results indicate that alpha-bungarotoxin-sensitive and alpha-bungarotoxin-insensitive subunits of the nicotinic receptors are co-expressed by neurons of the chick brain and retina.

Animals↗

Immunohistochemical localization of nicotinic acetylcholine receptor subunits in the mesencephalon and diencephalon of the chick (Gallus gallus).

Monoclonal antibodies against two alpha-bungarotoxin-binding subunits (alpha 7 and alpha 8) of the nicotinic acetylcholine receptors (nAChRs) were used as immunohistochemical probes to map their distribution in the chick diencephalon and mesencephalon. The distribution of the alpha 7 and alpha 8 nAChR subunits was compared to the distribution of immunoreactivity produced by a monoclonal antibody against the beta 2 structural subunit of the nAChRs. Structures that contained high numbers of alpha 7-like immunoreactive (LI) somata included the intergeniculate leaflet, nucleus intercalatus thalami, nucleus ovoidalis, organum paraventricularis, nucleus rotundus, isthmic nuclei, nucleus trochlearis, oculomotor complex, nucleus interstitio-pretecto-subpretectalis, stratum griseum centrale of the optic tectum, and nucleus semilunaris. Neuropil staining for alpha 7-LI was intense in the nucleus dorsomedialis hypothalami, nucleus geniculatus lateralis ventralis, griseum tecti, isthmic nuclei, nucleus lentiformis mesencephali, nucleus of the basal optic root, and stratum griseum et fibrosum superficiale of the tectum. High numbers of alpha 8-LI somata were found in the stratum griseum et fibrosum superficiale of the tectum and the nucleus interstitio-pretecto-subpretectalis, and intense neuropil staining for alpha 8-LI was found in the dorsal thalamus, nucleus geniculatus lateralis ventralis, lateral hypothalamus, griseum et fibrosum superficiale of the tectum. High numbers of beta 2-LI somata were found only in the nucleus spiriformis lateralis, whereas neuropil staining for beta 2-LI was intense in the nucleus geniculatus lateralis ventralis, nucleus suprachiasmaticus, nucleus lateralis anterior, nucleus habenularis lateralis, area pretectalis, griseum tecti, nucleus lentiformis mesencephalis, nucleus externus, and nucleus interpeduncularis, and in the stratum griseum centrale, stratum griseum et fibrosum superficiale, and stratum opticum of the tectum. These results indicate that there are major disparities in the localization of the alpha-bungarotoxin-binding alpha 7 and alpha 8 nAChR subunits and the beta 2 structural nAChR subunit in the chick diencephalon and mesencephalon. These nAChR subunits appear, however, to coexist in several regions of the chick brain.

Animals↗