J. A. De Carlos

THE TELENCEPHALIC VESICLES ARE INNERVATED BY OLFACTORY PLACODE-DERIVED CELLS: A POSSIBLE MECHANISM TO INDUCE NEOCORTICAL DEVELOPMENT

During early embryonic development, the olfactory placode is the source of different cell types migrating toward the telencephalic vesicle. Among these cell types are the ensheathing cells, the luteinizing hormone-releasing hormone-producing cells and the olfactory marker protein-immunoreactive cells. We have identified a novel group of olfactory placode-derived migratory cells using an antibody against beta-tubulin to label neurons and acetylcholinesterase histochemistry to label posmitotic cells. In this paper we describe the morphology, migration and fate of this novel group of cells. The first neurons detected in the rostral prosencephalon with acetylcholinesterase and anti-beta-tubulin antibody are localized in the olfactory placodes at embryonic day 11 in the rate. At embryonic day 12, anti-beta-tubulin antibody-positive cells were observed in the mesenchymal tissue between the olfactory pit and the rostral pole of the telencephalic vesicle. Anti-beta-tubulin antibody-positive cells were seen running superficially over the pial (dorsal) side of the telencephalic vesicle at embryonic day 13. The majority of these cells have a bipolar profile with short leading and trailing processes, suggesting that they are migratory elements. However, some of these cells showed elaborate processes extending for quite long distances, overlying the pial surface of the telencephalic vesicle. A mass of cells extending over the telencephalic vesicle from the developing olfactory epithelium were observed at embryonic day 13 using acetylcholinesterase histochemistry. Some of these acetylcholinesterase-positive cells were identified as neurons with the specific neuronal marker anti-beta-tubulin antibody. On embryonic day 12, neurons from the olfactory epithelium send axonal fibers toward the telencephalic vesicles. Most of these fibers spread over the anteroventral pole of the vesicles but others entered deep into the telencephalon, reaching the germinal ventricular zone. We also show that fibers run rostrocaudally over the surface of the telencephalic vesicles. We suggest that these cells and fibers, apparently originating in the olfactory placode and migrating through non-conventional routes, might play a significant role in the earliest stages of telencephalic vesicle development.

Early onset of the rat olfactory bulb projections

Using the fluorescent carbocyanine tracer DiI, we examined in detail the early development of the projections emanating from the rat olfactory bulb. The study commenced at embryonic day 13 when the first fibres can be detected and ended at embryonic day 20, when all major fibre systems have been established. The first axons arising from the prospective olfactory bulb area are seen at embryonic day 13. Labelled fibres are provided with elaborate axonal growth cones advancing through the ventrolateral part of the telencephalic vesicle. At embryonic day 14, while the main fibre tract has not developed much further, some isolated fibres are located quite distally from the prospective olfactory bulb. These early fibres apparently course within a narrow cell-free space that extends caudally along the ventrolateral part of the telencephalic vesicle. At embryonic day 15, a number of labelled fibres form a compact bundle, corresponding to the lateral olfactory tract, that ultimately reaches the prospective primary olfactory cortex. The fibres do not stop growing, but continue to extend caudally at embryonic day 17. The results of this study provide new information on the development of axonal tracts in the olfactory system. We show that the olfactory tract projection develops earlier than the morphological appearance of the olfactory bulbs. This suggests that the early development of olfactory projections might not depend on the arrival of the olfactory epithelium axons and thus, could be governed by factors intrinsic to the neurons and/or cues present in the target environment.

Further studies on cortical tangential migration in wild type and Pax-6 mutant mice

In this study we present new data concerning the tangential migration from the medial and lateral ganglionic eminences (MGE and LGE) to the cerebral cortex during development. We have used Calbindin as a useful marker to follow the itinerary of tangential migratory cells during early developmental stages in wild-type and Pax-6 homozygous mutant mice. In the wild-type mice, at early developmental stages, migrating cells advance through the intermediate zone (IZ) and preplate (PP). At more advanced stages, migrating cells were present in the subplate (SP) and cortical plate (CP) to reach the entire developing cerebral cortex. We found that, in the homozygous mutant mice (Pax-6Sey-Neu/Pax-6Sey-Neu), this tangential migration is severely affected at early developmental stages: migrating cells were absent in the IZ, which were only found some days later, suggesting that in the mutant mice, there is a temporal delay in tangential migration. We have also defined some possible mechanisms to explain certain migratory routes from the basal telencephalon to the cerebral cortex. We describe the existence of two factors, which we consider to be essential for the normal migration; the first one is the cell adhesion molecule PSA-NCAM, whose role in other migratory systems is well known. The second factor is Robo-2, whose expression delimits a channel for the passage of migratory cells from the basal telencephalon to the cerebral cortex.

Vasoactive intestinal polypeptide-immunoreactive neurons in the main olfactory bulb of the hedgehog (Erinaceus europaeus)

Vasoactive intestinal polypeptide (VIP) immunoreactivity was localized by the indirect antibody enzyme method (PAP technique) in the main olfactory bulb of the hedgehog. Most VIP-immunoreactive cells were located in the glomerular layer and throughout the external plexiform layer. Fewer cells were observed in the granule cell layer. At the morphological level they exhibit the characteristics of periglomerular, external tufted, superficial short axon, horizontal and Van Gehuchten cells. It should be mentioned that another specific neuronal type was found in the inner third of the external plexiform layer, which is not described in other animals. These results revealed that a high number of intrinsic neuronal types of the olfactory bulb of the hedgehog display a strong VIP immunoreactivity.

Extracortical origin of some murine subplate cell populations.

Significance Early-generated subplate cells play important roles in development of the cerebral cortex. Although it has been assumed that most subplate cells are generated in the cortical germinative zone, here we describe an alternative site of neurogenesis, the rostromedial telencephalic wall. The rostromedial telencephalic wall gives rise to both subplate projection neurons and GABAergic interneurons. Both populations migrate tangentially to the cortex and strongly express the Lpar1-eGFP transgene. This unusual extracortical source of subplate projection neurons and interneurons is challenging the currently accepted scheme of cortical development. The subplate layer, the deepest cortical layer in mammals, has important roles in cerebral cortical development. The subplate contains heterogeneous cell populations that are morphologically diverse, with several projection targets. It is currently assumed that these cells are generated in the germinative zone of the earliest cortical neuroepithelium. Here we identify a pallial but extracortical area located in the rostromedial telencephalic wall (RMTW) that gives rise to several cell populations. Postmitotic neurons migrate tangentially from the RMTW toward the cerebral cortex. Most RMTW-derived cells are incorporated into the subplate layer throughout its rostrocaudal extension, with others contributing to the GABAergic interneuron pool of cortical layers V and VI.

Peripheral contributions to olfactory bulb cell populations (migrations towards the olfactory bulb).

The olfactory system represents one of the most suitable models to study interactions between the peripheral and central nervous systems. The developing olfactory epithelium (olfactory placode and pit) gives rise to several cell populations that migrate towards the telencephalic vesicle. One of these cell populations, called the Migratory Mass (MM), accompanies the first emerging olfactory axons from the olfactory placode, but the fate of these cells and their contribution to the Olfactory Bulb (OB) populations has not been properly addressed. To asses this issue we performed ultrasound‐guided in utero retroviral injections at embryonic day (E) 11 revealing the MM as an early source of Olfactory Ensheathing Cells in later postnatal stages. Employing a wide number of antibodies to identify the nature of the infected cells we described that those cells generated within the MM at E11 belong to different cell populations both in the mesenchyma, where they envelop olfactory axons and express the most common glial markers, and in the olfactory bulb, where they are restricted to the Olfactory Nerve and Glomerular layers. Thus, the data reveal the existence of a novel progenitor class within the MM, potentially derived from the olfactory placode which gives rise to different neural cell population including some CNS neurons, glia and olfactory ensheathing cells.

Distribution and morphology of Alz-50-immunoreactive cells in the developing visual cortex of kittens.

SummaryThe population of interstitial cells of the white matter in the postlateral gyrus of the cat was studied at different postnatal ages using the antibody Alz-50. These neurons are among the first cells to develop in the cortex, and many of them are transitory, disappearing by cell death during the first postnatal days. In the present study, we found that immunoreactivity to Alz-50 is expressed during the first three postnatal weeks, and that positive neurons were not detected after postnatal day (P) 23. In addition to marking cells in the white matter, Alz-50 also recognizes many pyramidal cells in the cortical layers II-III and V of the visual cortex at postnatal day 4. The staining of cortical cells was not observed at other ages. We found that the number of positive cells in the white matter decreases by postnatal days 12 and 16, showing an apparent increase in number at postnatal day 23.In this study we also attempted to correlate the morphology of Alz-50-immunoreactive cells with the interstitial cells of the white matter, as seen in Golgi preparations. We conclude that Alz-50 immunoreactivity may be related to specific developmental changes and not particularly associated to the occurrence of cell death.

Morphological Characterization of ALZ-50 Immunoreactive Cells in the Developing Neocortex of Kittens

Alz-50 is a monoclonal antibody isolated from brain tissue of patients with Alzheimer’s disease (Wolozin et al., 1986). This antibody recognizes an antigenic protein of 68 kilodaltons in neurons involved in the formation of neuritic plaques and neurofibrillary tangles. It has been reported recently that Alz-50 immunoreactive (ir) neurons are found in normal fetal and neonatal human brains and in brain tissue from neonatal individuals with Down’s syndrome (Wolozin et al., 1988).