RESUMO
The dorsal lateral geniculate nucleus (dLGN) contains a retinotopic map where input from the two eyes map in register to provide a substrate for binocular vision. Ten-m3, a transmembrane protein, mediates homophilic interactions and has been implicated in the patterning of ipsilateral visual projections. Ease of access to early developmental stages in a marsupial wallaby has been used to manipulate levels of Ten-m3 during the development of retinogeniculate projections. In situ hybridisation showed a high dorsomedial to low ventrolateral gradient of Ten-m3 in the developing dLGN, matching retinotopically with the previously reported high ventral to low dorsal retinal gradient. Overexpression of Ten-m3 in ventronasal but not dorsonasal retina resulted in an extension of ipsilateral projections beyond the normal binocular zone. These results demonstrate that Ten-m3 influences ipsilateral projections and support a role for it in binocular mapping.
Assuntos
Corpos Geniculados/fisiologia , Macropodidae/fisiologia , Proteínas de Membrana/metabolismo , Retina/fisiologia , Animais , Proteínas de Membrana/genéticaRESUMO
Retinal projections to the superior colliculus are organised into retinotopic maps. Binocular vision requires that inputs from the two eyes map in register with each other. Studies in mice lacking Ten-m3, a homophilic transmembrane protein, indicate that it plays a key role in this process by influencing ipsilateral projections. The postnatal, ex utero development of the wallaby allows the targeted manipulation of molecules of interest during development. The distribution of mRNA for Ten-m3 in the retina and superior colliculus of the wallaby, and the effects of its spatiotemporally restricted retinal overexpression was investigated, in particular on the mapping of ipsilateral projections. Quantitative polymerase chain reaction found that Ten-m3 mRNA is expressed at relatively higher levels in the retina and colliculus early in development. Further, it is higher in ventral than dorsal retina, and increased in the retinotopically corresponding medial compared to lateral superior colliculus. In situ hybridisation demonstrated an increasing dorsoventral gradient in retinal ganglion cells was matched to an increasing lateromedial gradient in the superior colliculus. Overexpression of Ten-m3 by in vivo retinal electroporation produced an increase in ipsilateral projections to the binocular rostromedial colliculus, fitting with the proposal that Ten-m3 mediates mapping by attractant homophilic interactions. Retrograde labelling of the projection from this region suggested that overexpression produces a shift in the axons of existing ipsilaterally projecting ganglion cells rather than a rerouting of the axons of contralaterally projecting cells. Retinal manipulation of Ten-m3 levels produces changes in ipsilateral mapping, supporting a role for it in binocular mapping.
Assuntos
Lateralidade Funcional , Proteínas do Tecido Nervoso/metabolismo , Retina/crescimento & desenvolvimento , Retina/metabolismo , Colículos Superiores/crescimento & desenvolvimento , Colículos Superiores/metabolismo , Animais , Animais Recém-Nascidos , Feminino , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Macropodidae , Masculino , Proteínas do Tecido Nervoso/genética , Vias Visuais/crescimento & desenvolvimento , Vias Visuais/metabolismoRESUMO
The visual system is characterized by precise retinotopic mapping of each eye, together with exquisitely matched binocular projections. In many species, the inputs that represent the eyes are segregated into ocular dominance columns in primary visual cortex (V1), whereas in rodents, this does not occur. Ten-m3, a member of the Ten-m/Odz/Teneurin family, regulates axonal guidance in the retinogeniculate pathway. Significantly, ipsilateral projections are expanded in the dorsal lateral geniculate nucleus and are not aligned with contralateral projections in Ten-m3 knockout (KO) mice. Here, we demonstrate the impact of altered retinogeniculate mapping on the organization and function of V1. Transneuronal tracing and c-fos immunohistochemistry demonstrate that the subcortical expansion of ipsilateral input is conveyed to V1 in Ten-m3 KOs: Ipsilateral inputs are widely distributed across V1 and are interdigitated with contralateral inputs into eye dominance domains. Segregation is confirmed by optical imaging of intrinsic signals. Single-unit recording shows ipsilateral, and contralateral inputs are mismatched at the level of single V1 neurons, and binocular stimulation leads to functional suppression of these cells. These findings indicate that the medial expansion of the binocular zone together with an interocular mismatch is sufficient to induce novel structural features, such as eye dominance domains in rodent visual cortex.
Assuntos
Mapeamento Encefálico , Dominância Ocular/genética , Lateralidade Funcional/genética , Proteínas de Membrana/deficiência , Proteínas do Tecido Nervoso/deficiência , Córtex Visual/fisiologia , Vias Visuais/fisiologia , Potenciais de Ação/fisiologia , Análise de Variância , Animais , Animais Recém-Nascidos , Autorradiografia , Regulação da Expressão Gênica/genética , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Proteínas do Tecido Nervoso/genética , Neurônios/fisiologia , Estimulação Luminosa , Proteínas Proto-Oncogênicas c-fos/metabolismo , Córtex Visual/citologia , Vias Visuais/anatomia & histologiaRESUMO
The mammalian cerebellar cortex is apparently uniform in composition, but a complex heterogeneous pattern can be revealed by using biochemical markers such as zebrin II/aldolase C, which is expressed by a subset of Purkinje cells that form a highly reproducible array of transverse zones and parasagittal stripes. The architecture revealed by zebrin II expression is conserved among many taxa of birds and mammals. In this report zebrin II immunohistochemistry has been used in both section and whole-mount preparations to analyze the cerebellar architecture of the Australian tammar wallaby (Macropus eugenii). The gross appearance of the wallaby cerebellum is remarkable, with unusually elaborate cerebellar lobules with multiple sublobules and fissures. However, despite the morphological complexity, the underlying zone and stripe architecture is conserved and the typical mammalian organization is present.
Assuntos
Antígenos/análise , Padronização Corporal , Córtex Cerebelar/anatomia & histologia , Macropodidae/anatomia & histologia , Proteínas do Tecido Nervoso/análise , Animais , Córtex Cerebelar/química , Córtex Cerebelar/imunologia , Feminino , Macropodidae/imunologia , Masculino , Mamíferos/anatomia & histologia , Mamíferos/classificação , Camundongos/anatomia & histologia , Células de Purkinje/química , Especificidade da EspécieRESUMO
In the wallaby, whisker-related patterns develop over a protracted period of postnatal maturation in the pouch. Afferents arrive simultaneously in the thalamus and cortex from postnatal day (P) 15. Whisker-related patterns are first seen in the thalamus at P50 and are well formed by P73, before cortical patterns first appear (P75) or are well developed (P85). This study used the slow developmental sequence and accessibility of the pouch young to investigate the effect of nerve lesions before afferent arrival, or at times when thalamic patterns are obvious but cortical patterns not yet formed. The left infraorbital nerve supplying the whiskers was cut at P0-93 and animals were perfused at P112-123. Sections through the thalamus (horizontal plane) and cortex (tangential) were reacted for cytochrome oxidase to visualize whisker-related patterns. Lesions of the nerve at P2-5, before innervation of the thalamus or cortex, resulted in an absence of patterns at both levels. Lesions from P66-77 also disrupted thalamic and cortical patterns, despite the fact that thalamic patterns are normally well established by P73. Lesions from P82-93 resulted in normal thalamic and cortical patterns. Thus, despite the wallaby having clearly separated times for the development of patterns at different levels of the pathway, these results suggest a single critical period for the thalamus and cortex, coincident with the maturation of the cortical pattern. Possible mechanisms underpinning this critical period could include dependence of the thalamic pattern on corticothalamic activity or peripheral signals to allow consolidation of thalamic barreloids.
Assuntos
Córtex Cerebral/anatomia & histologia , Macropodidae/anatomia & histologia , Tálamo/anatomia & histologia , Nervo Trigêmeo/crescimento & desenvolvimento , Vibrissas/crescimento & desenvolvimento , Vias Aferentes , Animais , Mapeamento Encefálico , Córtex Cerebral/enzimologia , Córtex Cerebral/ultraestrutura , Período Crítico Psicológico , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Doenças do Sistema Nervoso/patologia , Tálamo/enzimologia , Tálamo/ultraestrutura , Nervo Trigêmeo/enzimologia , Vibrissas/enzimologia , Vibrissas/ultraestruturaRESUMO
The expression of EphB1 and B2 receptors and ephrins-B1, -B2 and -B3 in the retina and superior colliculus of the wallaby (Macropus eugenii) was examined during the development of the retinocollicular projection, using reverse transcription-polymerase chain reaction and immunohistochemistry. There was an early transient differential expression of EphB2 that was higher in ventral retina and restricted to the outer neuroblast layer, whereas a high ventral to low dorsal gradient of ephrin-B2 expression occurred there throughout the study period. However, there was no dorsoventral gradient of receptors or ligands in retinal ganglion cells or a mediolateral gradient of ephrins in the colliculus. These findings suggest a limited role for these molecules in topographic mapping across the mediolateral colliculus in the wallaby. Early in retinal development there is a complementary pattern of expression of ephrin-B1 and -B2 in the outer neuroblast layer that overlaps with expression of EphB2. Ganglion and amacrine cells also express EphB2. As development proceeds subpopulations of putative horizontal and bipolar cells, also expressing EphB2, come to reside in the inner nuclear layer and ephrin-B1 is expressed throughout the outer nuclear layer. At the same time cells expressing ephrin-B2, and subpopulations of horizontal and bipolar cells come to reside in the inner nuclear layer and there is a corresponding decrease in ephrin-B2 expression in the outer nuclear layer. This pattern of coexpression of receptors and ligands suggests a role for them in cell migration and maintenance of laminar boundaries.
Assuntos
Efrinas/metabolismo , Receptores da Família Eph/metabolismo , Retina/metabolismo , Colículos Superiores/metabolismo , Envelhecimento , Animais , Animais Recém-Nascidos , Northern Blotting , Efrinas/classificação , Efrinas/genética , Regulação da Expressão Gênica no Desenvolvimento , Imuno-Histoquímica , Macropodidae/crescimento & desenvolvimento , RNA/isolamento & purificação , RNA Mensageiro/biossíntese , Receptores da Família Eph/classificação , Receptores da Família Eph/genética , Retina/anatomia & histologia , Retina/embriologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Colículos Superiores/embriologiaRESUMO
The time course of development and laminar distribution of thalamocortical synapses in the visual cortex of the marsupial mammal the wallaby (Macropus eugenii) has been studied by electron microscopy from the time of afferent ingrowth to the appearance of layer 4, the main target for thalamic axons. Axons were labeled from the thalamus by a fluorescent carbocyanine dye in fixed tissue or by transneuronal transport of horseradish peroxidase conjugated to wheat germ agglutinin from the eye. Thalamic axons first reached the cortex 2 weeks after birth and grew into the developing cortical plate without a waiting period in the subplate. The first thalamocortical synapses were detected 2 weeks later solely throughout the loosely packed zone of the cortical plate, where layer 6 cells previously have been shown to reside. As the thickness of the cortex increased with age, thalamocortical synapses were increasingly prevalent in the loosely packed zone of the cortical plate. With the appearance of layer 4, thalamocortical synapses were found there as well as in the marginal zone and layer 6. There was no evidence for an early population of thalamocortical synapses in the subplate. The first synapses made by thalamic axons were in a region containing layer 6 cells, one of their normal targets in the mature cortex.
Assuntos
Diferenciação Celular/fisiologia , Corpos Geniculados/crescimento & desenvolvimento , Cones de Crescimento/ultraestrutura , Macropodidae/crescimento & desenvolvimento , Terminações Pré-Sinápticas/ultraestrutura , Córtex Visual/crescimento & desenvolvimento , Vias Visuais/crescimento & desenvolvimento , Animais , Transporte Axonal/fisiologia , Carbocianinas , Comunicação Celular/fisiologia , Sinais (Psicologia) , Corpos Geniculados/fisiologia , Corpos Geniculados/ultraestrutura , Cones de Crescimento/fisiologia , Macropodidae/anatomia & histologia , Macropodidae/fisiologia , Microscopia Eletrônica , Terminações Pré-Sinápticas/fisiologia , Tempo de Reação , Retina/crescimento & desenvolvimento , Retina/fisiologia , Retina/ultraestrutura , Células-Tronco/fisiologia , Células-Tronco/ultraestrutura , Córtex Visual/fisiologia , Córtex Visual/ultraestrutura , Vias Visuais/fisiologia , Vias Visuais/ultraestrutura , Conjugado Aglutinina do Germe de Trigo-Peroxidase do Rábano SilvestreRESUMO
We have examined the distribution of immunoreactivity for GAP-43 in the developing and adult brain of a diprotodontid metatherian, the tammar wallaby ( Macropus eugenii). The distribution of GAP-43 immunoreactivity in the neonatal wallaby brain was strikingly heterogeneous, in contrast to that reported for the newborn polyprotodontid opossum. Immunoreactivity for GAP-43 in the developing wallaby brain showed a caudal-to-rostral spatiotemporal gradient, with the brainstem well in advance of the telencephalon throughout the first 100 days of postnatal life. In many regions examined, GAP-43 immunoreactivity passed through the following phases: 1. intense immunoreactivity in developing fiber tracts and occasional somata; 2. diffuse homogeneous immunoreactivity; 3. selective loss of immunoreactivity in particular nuclei or cortical regions. In the isocortex, selective loss of GAP-43 immunoreactivity in the somatosensory and visual cortex (at postnatal day 115) coincided with the maturation of the laminar distribution of terminal thalamocortical axonal fields. Within adult cortical regions, GAP-43 immunoreactivity was highest in layer I of all regions, lower layers (V and VI) of primary somatosensory and visual cortices, layers II/III of motor and cingulate cortex, and layer IV of entorhinal cortex. Our findings suggest that, while patterning of GAP-43 immunoreactivity in the mature brain is similar across meta- and eutheria, there may be early developmental differences in the distribution of GAP-43 immunoreactivity between poly- and diprotodontid metatheria.
Assuntos
Encéfalo/metabolismo , Proteína GAP-43/metabolismo , Macropodidae/fisiologia , Animais , Animais Recém-Nascidos , Encéfalo/crescimento & desenvolvimento , Humanos , Imuno-Histoquímica , Bulbo Olfatório/crescimento & desenvolvimento , Bulbo Olfatório/metabolismo , Gambás/fisiologia , Ratos , Especificidade da EspécieRESUMO
We have studied the development of the hypothalamus of an Australian marsupial, the tammar wallaby (Macropus eugenii), to provide an initial anatomic framework for future research on the developing hypothalamus of diprotodontid metatheria. Cytoarchitectural (hematoxylin and eosin), immunohistochemical (CD 15 and growth associated protein, GAP-43), tritiated thymidine autoradiography, and carbocyanine dye tracing techniques were applied. Until 12 days after birth (P12), the developing hypothalamus consisted of mainly a ventricular germinal zone with a thin marginal layer, but by P25, most hypothalamic nuclei were well differentiated, indicating that the bulk of hypothalamic cytoarchitectural development occurs between P12 and P25. Strong CD 15 immunoreactivity was found in radial glial fibers in the rostral hypothalamus during early developmental ages, separating individual hypothalamic compartments. Immunoreactivity for GAP-43 was used to reveal developing fiber bundles. The medial forebrain bundle was apparent by P0, and the fornix appeared at P12. Tritiated thymidine autoradiography revealed lateral-to-medial and dorsal-to-ventral neurogenetic gradients similar to those seen in rodents. Dye tracing showed that projections to the posterior pituitary arose from the supraoptic nucleus at P5 and from the paraventricular nucleus at P10. Projections to the medulla were first found from the lateral hypothalamic area at P0 and paraventricular nucleus at P10. In conclusion, the pattern of development of the wallaby hypothalamus is broadly similar to that found in eutheria, with comparable neurogenetic compartments to those identified in rodents. Because most hypothalamic maturation takes place after birth, wallabies provide a useful model for experimentally manipulating the developing mammalian hypothalamus.
Assuntos
Hipotálamo/crescimento & desenvolvimento , Macropodidae/crescimento & desenvolvimento , Animais , Autorradiografia , Carbocianinas , Corantes Fluorescentes , Proteína GAP-43/análise , Proteína GAP-43/imunologia , Hipotálamo/citologia , Imuno-Histoquímica , Antígenos CD15/análise , Antígenos CD15/imunologia , Neurônios/química , Timidina , TrítioRESUMO
Recordings from direction-selective neurons in the nucleus of the optic tract (NOT) of the marsupial wallaby, Macropus eugenii, show that 53% of cells are sensitive to visual stimulation of both eyes. Anatomical tracing studies using horseradish peroxidase reveal many retinal terminals in the contralateral NOT but very few in the ipsilateral nucleus. There was no convincing evidence of cortical inputs to the ipsilateral NOT despite large injections of tracer into the visual cortex. During visual stimulation in the visual field of the contralateral eye with moving patterns, the excitatory responses in the NOT generated by ipsiversive motion (right-to-left when recording from the left NOT) were usually larger than the inhibitory responses produced by contraversive motion. Conversely, during ipsilateral eye stimulation, the negative motion components to contraversive motion were usually larger than the positive components to ipsiversive motion. This response pattern resembles that observed in the NOT of the American opossum, Didelphis aurita, where binocularity appears to arise through a commissural subcortical pathway that connects the two nuclei and inverts the directional tuning of the transmitted signals. We propose that the lack of significant input from the ipsilateral eye and cortex in the wallaby suggests that binocularity must arise from another pathway, possibly a commissural route between the nuclei. As directional information appears not to be carried by the internucleus pathway in rats and cats, our results suggest that binocularity in the NOT arises from different sources in marsupials as compared to eutherians.