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In JoVE (2)
- Dissecting the Non-human Primate Brain in Stereotaxic Space
- The Gateway to the Brain: Dissecting the Primate Eye
Other Publications (9)
- Experimental Brain Research. Experimentelle Hirnforschung. Expérimentation Cérébrale
- Visual Neuroscience
- Experimental Brain Research. Experimentelle Hirnforschung. Expérimentation Cérébrale
- Experimental Brain Research. Experimentelle Hirnforschung. Expérimentation Cérébrale
- Journal of Chemical Neuroanatomy
- The Journal of Comparative Neurology
- The European Journal of Neuroscience
- Neuroscience Letters
- Experimental Neurology
Articles by Denis Boire in JoVE
JoVE General
Dissecting the Non-human Primate Brain in Stereotaxic Space
1Department of Physiology, University of Montreal, 2School of Optometry, University of Montreal, 3Département de chimie-biologie , Université du Québes à Trois-Rivières
The non-human primate is an important translational species for our understanding of the normal processing of the brain. The anatomical organization of the primate brain can provide important insights into normal and pathological conditions in humans.
JoVE General
The Gateway to the Brain: Dissecting the Primate Eye
1Department of Physiology, University of Montreal, 2School of Optometry, University of Montreal, 3Departement de chimie-biologie, Universite du Quebec a Trois-Rivieres
The non-human primate is an important translational species for our understanding of development and aging. The anatomical organization of the primate retina may provide important insights into normal and pathological conditions in humans.
Other articles by Denis Boire on PubMed
Stereological Evaluation of Neurons and Glia in the Monkey Dorsal Lateral Geniculate Nucleus Following an Early Cerebral Hemispherectomy
The effects of an early, unilateral cerebral hemispherectomy on the cytoarchitecture of the dorsal lateral geniculate nucleus (dLGN) were quantitatively evaluated in the green monkey. The dLGN ipsilateral to the lesion showed a 73% reduction in size, more than 99% neuronal cell loss, 50% increase in glial cell density, but a 50% reduction in the total number of glial cells. The total number of neural and glial cells estimated for the dLGN contralateral to the ablation did not differ from control values. Despite evidence for substantial degeneration of the ipsilateral dLGN, cytochrome oxidase histochemistry revealed a small population of surviving cells that exhibited features of neuronal cells. More surviving cells were found in the parvocellular than in the magnocellular layers, and surviving parvocellular cells had the same size-frequency distribution as Nissl-stained neurons in an intact animal. These findings suggest that the intrinsic geniculate circuitry may be able to sustain the residual interneurons that can, in turn, contribute to maintaining retinal and brainstem afferents. The remaining neurons in the dLGN following hemispherectomy appear to be insufficient in number to be importantly implicated in the residual visual functions that have been reported in some hemispherectomized patients.
Retinal Projections in the Cat: a Cholera Toxin B Subunit Study
The B fragment of cholera toxin (CTb) is a highly sensitive anterograde tracer for the labelling of retinal axons. It can reveal dense retinofugal projections to well-known retinorecipient nuclei along with sparse but distinct input to target areas that are not commonly recognized. Following a unilateral injection of CTb into the vitreous chamber of seven adult cats, we localized the toxin immunohistochemically in order to identify direct retinal projections in these animals. Consistent with previous findings, the strongest projections were observed in the superficial layers of the superior colliculus, the dorsal and ventral lateral geniculate nuclei, the pretectal nuclei, the accessory optic nuclei, and the suprachiasmatic nucleus of the hypothalamus. However, we also found labelled terminals in several other brain areas, including the zona incerta, the medial geniculate nucleus, the lateral posterior-pulvinar complex, the lateral habenular nucleus, and the anterior and lateral hypothalamic regions. The morphological characteristics of the retinal axon terminals in most of the identified novel target sites are described.
Retinal Projections to the Lateral Posterior-pulvinar Complex in Intact and Early Visual Cortex Lesioned Cats
In intact cats, it is generally considered that the lateral posterior-pulvinar complex (LP-pulvinar) does not receive direct retinal terminals, with the exception of the retino-recipient zone known as the geniculate wing. There is, however, some evidence that early lesions of the visual cortex can occasionally induce the formation of novel retinal projections to the LP nucleus. Given the importance of knowing the connectivity pattern of the LP-pulvinar complex in intact and lesioned animals, we used the B fragment of cholera toxin, a sensitive anterograde tracer, to reinvestigate the retinal projections to the LP-pulvinar in normal cats and in cats with early unilateral lesions of the visual cortex (areas 17 and 18). Immunohistochemical localization of the toxin was performed to show the distribution and morphology of retinofugal terminals. A direct bilateral but predominantly contralateral retinal projection reached the caudal portion of LPl and LPm in the form of patches located mainly along its dorsomedial surface and many scattered terminals. The distribution of retinal projections to LP-pulvinar in intact and operated cats did not differ. Contrary to what had been previously reported, we found no evidence for lesion-induced sprouting of retinal axons in these higher-order thalamic nuclei. Retinal input to the LP-pulvinar might modulate visual responses driven by primary visual cortex or superior colliculus.
Distribution of Calcium Binding Proteins in Visual and Auditory Cortices of Hamsters
The morphology and distribution of neurons immunoreactive (ir) to parvalbumin (PV), calretinin (CR) and calbindin (CB) were studied in the primary visual (V1) and auditory (A1) cortices of hamsters. Cortical cell populations were labelled immunohistochemically using a glucose oxidase-diaminobenzidine-nickel combined revelation method. Quantitative analysis revealed significant differences between V1 and A1 in the density and distribution of their neuronal population. CBir cells exhibited several typologies in both cortical regions. Most cells were multipolar even though many of them had bitufted or bipolar morphologies. These cells were distributed in layers II/III and in layer V of both A1 and V1, but were more numerous in layer V of V1. CRir cells were of the fusiform type with long bipolar dendritic arbours. These were similarly distributed in both cortices with a peak in superficial layers II/III. PVir cells were also found in both cortices and had round or oval-shaped somata with multipolar processes. They were mostly located in layer V for V1 and in layers III/IV for A1. Visual and auditory primary cortices can thus be differentiated on the basis of their immunoreactivity to specific calcium binding proteins.
Regional Analysis of Neurofilament Protein Immunoreactivity in the Hamster's Cortex
The laminar distribution of several distinct populations of neurofilament protein containing neurons has been used as a criterion for the delineation of cortical areas in hamsters. SMI-32 is a monoclonal antibody that recognizes a non-phosphorylated epitope on the medium- and high-molecular weight subunits of neurofilament proteins. As in carnivores and primates, SMI-32 immunoreactivity in the hamster neocortex was present in cell bodies, proximal dendrites and axons of some medium and large pyramidal neurons located in cortical layers III, V and VI. A small population of labeled multipolar cells was also found in layer IV. Neurofilament protein immunoreactive neurons were found throughout isocortical areas. Very few labeled cells were encountered in supplemental motor area, insular cortex, medial portion of associative visual cortex and in parietal association cortex. Our data indicate that SMI-32 immunoreactive cells can be efficiently used to trace boundaries between neocortical areas in the hamster's brain. The regional distribution SMI-32 immunoreactivity in the hamster cortex corresponds quite closely with cortical areas as defined by their cytoarchitecture and myeloarchitecture. The primary sensory cortical areas contain the most intense of SMI-32 immunoreactivity and are also those with the highest density of myelinated axons. Very low SMI-32 immunoreactivity was found in orbital, insular, perirhinal, cingulate and infralimbic cortices, which are also poor in myelinated axons. This supports the association between SMI-32 immunoreactivity and myelin contents.
Development of the Commissure of the Superior Colliculus in the Hamster
The development of the corpus callosum (CC) and the anterior commissure (CA) is well known in a wide variety of species. No study, however, has described the development of the commissure of the superior colliculus (CSC) from embryonic state to adulthood in mammals. In this study, by using the lipophylic tracer DiI, we investigated the ontogeny of this mesencephalic commissure in the hamster at various ages. The development of axonal terminals, growth cone morphologies, and axons branching were described for the superior colliculus (SC) contralateral to the tracer injection. The first CSC axons cross the midline at embryonic day 11 (E-11) and grow further into the intermediate layers of the contralateral SC between E-12 and E-14. There is little axon growth therein between E-14 and the day of birth (P-0). Growth cones at the tip of these axons adopt complex morphologies at E-12 and progressively simplify until P-0. Pioneer axons are clearly visible between E-14 and P-1. These are followed by other axons progressively more numerous between P-0 and P-5. Axons do not show any branching until P-2. Between P-3 and P-9, the axons progressively arborize in the intermediate layers. Some axons reach the superficial layers at P-5, and they become more numerous around P-11, and only a few axons remain therein by P-21. Myelinated axons appear at P11 and are very dense at P-21. Our results indicate that the CSC follows developmental schemes similar to those of the CC and the AC but that initial axon midline crossing occurs earlier.
Audition Differently Activates the Visual System in Neonatally Enucleated Mice Compared with Anophthalmic Mutants
The occipital cortex, normally visual, can be activated by auditory or somatosensory tasks in the blind. This cross-modal compensation appears after early or late onset of blindness with differences in activation between early and late blind. This could support the hypothesis of a reorganization of sensory pathways in the early blind that does not occur in later onset blindness. Using immunohistochemistry of the c-Fos protein following a white noise stimulus and injections of the anterograde tracer dextran-biotin in the inferior colliculus, we studied how the occurrence of blindness influences cross-modal compensation in the mutant anophthalmic mouse strain and in C57BL/6 mice enucleated at birth. We observed, in mutant mice, immunolabeled nuclei in the visual thalamus - the dorsal lateral geniculate nucleus - in the primary visual area (V1) and a few labeled nuclei in the secondary visual area (V2). In enucleated mice, we observed auditory activity mainly in V2 but also sparsely in V1. No labeled cells could be found in the visual thalamus. Tracing studies confirmed the difference between anophthalmic and birth-enucleated mice: whereas the first group showed inferior colliculus projections entering both the dorsal lateral geniculate and the latero-posterior nuclei, in the second, auditory fibers were found only within the latero-posterior thalamic nucleus. None was found in controls with intact eyes. We suggest that the prenatal period of spontaneous retinal activity shapes the differences of the sensory reorganization in mice.
Subcortical Auditory Input to the Primary Visual Cortex in Anophthalmic Mice
Anatomical and imaging studies show ample evidence for auditory activation of the visual cortex following early onset of blindness in both humans and animal models. Anatomical studies in animal models of early blindness clearly show intermodal pathways through which auditory information can reach the primary visual cortex. There is clear evidence for intermodal corticocortical pathways linking auditory and visual cortex and also novel connections between the inferior colliculus and the visual thalamus. A recent publication [L.K. Laemle, N.L. Strominger, D.O. Carpenter, Cross-modal innervation of primary visual cortex by auditory fibers in congenitally anophthalmic mice, Neurosci. Lett. 396 (2006) 108-112] suggested the presence of a direct reciprocal connection between the inferior colliculus and the primary visual cortex (V1) in congenitally anophthalmic ZRDCT/An mice. This implies that this mutant mouse would be the only known vertebrate having a direct tectal connection with a primary sensory cortex. The presence of this peculiar pathway was reinvestigated in the ZRDCT/An mouse with highly sensitive neuronal tracers. We found the connections normally described in the ZRDCT/An mouse between: (i) the inferior colliculus and the dorsal lateral geniculate nucleus, (ii) V1 and the superior colliculus, (iii) the lateral posterior nucleus and V1 and between (iv) the inferior colliculus and the medial geniculate nucleus. We also show unambiguously that the auditory subcortical structures do not connect the primary visual cortex in the anophthalmic mouse. In particular, we find no evidence of a direct projection from the auditory mesencephalon to the cortex in this animal model of blindness.
Protein Kinase A Modulates Retinal Ganglion Cell Growth During Development
During development, retinal ganglion cells (RGCs) extend their axons toward their thalamic and mesencephalic targets. Their navigation is largely directed by guidance cues present in their environment. Since cAMP is an important second messenger that mediates the neural response to guidance molecules and its intracellular levels seem to decrease significantly following birth, we tested whether modulation of the cAMP/protein kinase A (PKA) pathway would affect the normal development of RGC axons. At postnatal day 1, hamsters received a unilateral intraocular injection of either 0.9% saline solution, 12 mM of the membrane-permeable cAMP analogue (dibutyryl cAMP; db-cAMP), or 10 microM of the PKA inhibitor KT5720. Intraocular elevation of cAMP significantly accelerated RGC axonal growth while inhibition of PKA activity decreased it. Moreover, when highly purified RGC cultures were treated with forskolin (an activator of adenylate cyclase) or cAMP analogues (db-cAMP and Sp-cAMP), neurite length, growth cone (GC) surface area and GC filopodia number were significantly increased. This indicates that intraocular elevation of cAMP acts directly on RGCs. Since these effects were prevented by PKA inhibitors, it demonstrates that cAMP also exerts its action via the PKA pathway. Taken together, these results suggest that the cAMP/PKA cascade is essential for the normal development of retinothalamic projections.
