Gabor Halasi

Publications of Gabor Halasi

• Proliferation and recapitulation of developmental patterning associated with regulative regeneration of the spinal cord neural tube.

  Authors: Gabor Halasi, Anne Mette Søviknes, Olafur Sigurjonsson, Joel C Glover

  Developmental biology. 02/2012; 365(1):118-32.

  Developmental patterning during regulative regeneration of the chicken embryo spinal neural tube was characterized by assessing proliferation and the expression of transcription factors specific toDevelopmental patterning during regulative regeneration of the chicken embryo spinal neural tube was characterized by assessing proliferation and the expression of transcription factors specific to neural progenitor and postmitotic neuron populations. One to several segments of the thoracolumbar neural tube were selectively excised unilaterally to initiate regeneration. The capacity for regeneration depended on the stage when ablation was performed and the extent of tissue removed. 20% of surviving embryos exhibited complete regulative regeneration, wherein the missing hemi-neural tube was reconstituted to normal size and morphology. Fate-mapping of proliferative adjacent tissue indicated contributions from the opposite side of the neural tube and potentially from the ipsilateral neural tube rostral and caudal to the lesion. Application of the thymidine analog EdU (5-ethynyl-2'-deoxyuridine) demonstrated a moderate increase in cell proliferation in lesioned relative to control embryos, and quantitative PCR demonstrated a parallel moderate increase in transcription of proliferation-related genes. Mathematical calculation showed that such modest increases are sufficient to account for the amount of regenerated tissue. Within the regenerated neural tube the expression pattern of progenitor-specific transcription factors was recapitulated in the separate advancing ventral and dorsal fronts of regeneration, with no evidence of abnormal mixing of progenitor subpopulations, indicating that graded patterning mechanisms do not require continuity of neural tube tissue along the dorsoventral axis and do not involve a sorting out of committed progenitors. Upon completion of the regeneration process, the pattern of neuron-specific transcription factor expression was essentially normal. Modest deficits in the numbers of transcription factor-defined neuron types were evident in the regenerated tissue, increasing particularly in dorsal neuron types with later lesions. These results confirm the regulative potential of the spinal neural tube and demonstrate a capacity for re-establishing appropriate cellular patterning despite a grossly abnormal morphogenetic situation.

• Xenotransplantation of human stem cells into the chicken embryo.

  Authors: Jean-Luc Boulland, Gabor Halasi, Nedim Kasumacic, Joel C Glover

  Journal of visualized experiments : JoVE. 01/2010;

  The chicken embryo is a classical animal model for studying normal embryonic and fetal development and for xenotransplantation experiments to study the behavior of cells in a standardized in vivoThe chicken embryo is a classical animal model for studying normal embryonic and fetal development and for xenotransplantation experiments to study the behavior of cells in a standardized in vivo environment. The main advantages of the chicken embryo include low cost, high accessibility, ease of surgical manipulation and lack of a fully developed immune system. Xenotransplantation into chicken embryos can provide valuable information about cell proliferation, differentiation and behavior, the responses of cells to signals in defined embryonic tissue niches, and tumorigenic potential. Transplanting cells into chicken embryos can also be a step towards transplantation experiments in other animal models. Recently the chicken embryo has been used to evaluate the neurogenic potential of human stem and progenitor cells following implantation into neural anlage. In this video we document the entire procedure for transplanting human stem cells into the developing central nervous system of the chicken embryo. The procedure starts with incubation of fertilized eggs until embryos of the desired age have developed. The eggshell is then opened, and the embryo contrasted by injecting dye between the embryo and the yolk. Small lesions are made in the neural tube using microsurgery, creating a regenerative site for cell deposition that promotes subsequent integration into the host tissue. We demonstrate injections of human stem cells into such lesions made in the part of the neural tube that forms the hindbrain and the spinal cord, and into the lumen of the part of the neural tube that forms the brain. Systemic injections into extraembryonic veins and arteries are also demonstrated as an alternative way to deliver cells to vascularized tissues including the central nervous system. Finally we show how to remove the embryo from the egg after several days of further development and how to dissect the spinal cord free for subsequent physiological, histological or biochemical analyses.

• Chimeric animal models in human stem cell biology.

  Authors: Joel C Glover, Jean-Luc Boulland, Gabor Halasi, Nedim Kasumacic

  ILAR journal / National Research Council, Institute of Laboratory Animal Resources. 01/2009; 51(1):62-73.

  The clinical use of stem cells for regenerative medicine is critically dependent on preclinical studies in animal models. In this review we examine some of the key issues and challenges in the use ofThe clinical use of stem cells for regenerative medicine is critically dependent on preclinical studies in animal models. In this review we examine some of the key issues and challenges in the use of animal models to study human stem cell biology-experimental standardization, body size, immunological barriers, cell survival factors, fusion of host and donor cells, and in vivo imaging and tracking. We focus particular attention on the various imaging modalities that can be used to track cells in living animals, comparing their strengths and weaknesses and describing technical developments that are likely to lead to new opportunities for the dynamic assessment of stem cell behavior in vivo. We then provide an overview of some of the most commonly used animal models, their advantages and disadvantages, and examples of their use for xenotypic transplantation of human stem cells, with separate reviews of models involving rodents, ungulates, nonhuman primates, and the chicken embryo. As the use of human somatic, embryonic, and induced pluripotent stem cells increases, so too will the range of applications for these animal models. It is likely that increasingly sophisticated uses of human/animal chimeric models will be developed through advances in genetic manipulation, cell delivery, and in vivo imaging.

• Dendrodendritic and dendrosomatic contacts between oculomotor and trochlear motoneurons of the frog, Rana esculenta.

  Authors: Tímea Bacskai, Gabor Veress, Gabor Halasi, Adam Deak, Eva Racz, György Szekely, Clara Matesz

  Brain research bulletin. 04/2008; 75(2-4):419-23.

  Gaze fixation requires very fast movements of the eye during body displacement. The morphological and physiological background of the very fine and continuous tuning of gaze fixation is not yet fullyGaze fixation requires very fast movements of the eye during body displacement. The morphological and physiological background of the very fine and continuous tuning of gaze fixation is not yet fully understood. In a previous study we have shown that the dendrites of oculomotor neurons form bundles which invade the trochlear nucleus, and vice versa, trochlear dendritic bundles invade the oculomotor nucleus. Earlier physiological observations demonstrating electrotonic coupling between dendrites of spinal motoneurons in the frog suggest a similar mechanism between the oculomotor and trochlear motoneurons. We studied a possible morphological basis of gaze fixation. The experiments were carried out on common water frogs, Rana esculenta. The trochlear and oculomotor nerves were cut, and their proximal stumps were labeled simultaneously with different retrograde fluorescent tracers. Using confocal laser scanning microscope we detected a large number of close contacts in both nuclei, the majority of them were dendrodendritic apposition. The distance between the adjacent profiles suggested close membrane appositions without intercalating glial or neuronal elements. At the ultrastructural level, the dendrodendritic and dendrosomatic contacts did not show any morphological specialization; the long membrane appositions may provide ephaptic interactions between the neighboring profiles. This electrotonic coupling between the oculomotor and trochlear nerve motoneurons may promote the co-activation of the muscles responsible for vertical eye movements.

• Organization of dye-coupled cerebellar granule cells labeled from afferent vestibular and dorsal root fibers in the frog Rana esculenta.

  Authors: Eva Rácz, Timea Bácskai, Gabor Halasi, Endre Kovács, Clara Matesz

  The Journal of comparative neurology. 06/2006; 496(3):382-94.

  Application of neurobiotin to the nerves of individual labyrinthine organs and dorsal root fibers of limb-innervating segments of the frog resulted in labeling of granule cells in the cerebellumApplication of neurobiotin to the nerves of individual labyrinthine organs and dorsal root fibers of limb-innervating segments of the frog resulted in labeling of granule cells in the cerebellum showing a significant overlap with a partial segregation in the related areas of termination. In different parts of the cerebellum, various combinations of different canal and otolith organ-related granule cells have been discerned. The difference in the extension of territories of vertical canals vs. horizontal canals may reflect their different involvement in the vestibuloocular and vestibulospinal reflex. Dye-coupled cells related to the lagenar and saccular neurons were localized in more rostral parts of the cerebellum, whereas cells of the utricle were represented only in its caudal half. This separation is supportive of the dual function of the lagena and the saccule. The territories of granule cells related to the cervical and lumbar segments of the spinal cord were almost completely separated along the rostrocaudal axis of cerebellum, whereas their territories were almost entirely overlapping in the mediolateral and ventrodorsal directions. The partial overlap of labyrinthine organ-related and dorsal root fiber-related granule cells are suggestive of a convergence of sensory modalities involved in the sense of balance. We propose that the afferent input of vestibular and proprioceptive fibers mediated by gap junctions to the cerebellar granule cells subserve one of the possible morphological correlates of a very rapid modification of the motor activity in the vestibulocerebellospinal neuronal circuit.

• Connections of the superior vestibular nucleus with the oculomotor and red nuclei in the rat: an electron microscopic study.

  Authors: Gabor Halasi, Timea Bácskai, Clara Matesz

  Brain research bulletin. 10/2005; 66(4-6):532-5.

  Phaseolus vulgaris leucoagglutinin (PHA-L) was injected into the individual vestibular nuclei of the rat to study their efferent connections. One of the major differences between the connections ofPhaseolus vulgaris leucoagglutinin (PHA-L) was injected into the individual vestibular nuclei of the rat to study their efferent connections. One of the major differences between the connections of these nuclei was found at the level of the mesencephalon: the eye-moving cranial nerve nuclei received the densest projection from the superior vestibular nucleus (SVN). In the present electron microscopic study, we have found that terminals of SVN origin established symmetric synaptic contacts in the oculomotor nucleus. More than two-thirds of PHA-L-labeled boutons terminated on dendrites, the rest of them established axosomatic contacts. Most of the labeled terminals were GABA-positive, supporting the results of previous physiological experiments, which showed inhibitory effects. In the mesencephalon, the other termination area was found in the red nucleus. The PHA-L-labeled boutons of SVN origin were in close contact with the perikarya and proximal dendrites of the magnocellular part of the red nucleus. The types of synaptic contacts and distribution of terminals of SVN origin were similar to those found in the oculomotor nucleus. Our results indicate that the SVN can modify the activity of the cerebellorubral and corticorubral pathways, exerting inhibitory action on the neurons of the red nucleus.

• Extracellular matrix molecules and their possible roles in the regeneration of frog nervous system.

  Authors: Clara Matesz, Laszlo Modis, Gabor Halasi, Zsuzsa M Szigeti, Szabolcs Felszeghy, Timea Bacskai, György Szekely

  Brain research bulletin. 10/2005; 66(4-6):526-31.

  Recent biochemical and histochemical analyses explored different components of the extracellular matrix (ECM) in the nervous system, and either permissive or non-permissive roles in neuronalRecent biochemical and histochemical analyses explored different components of the extracellular matrix (ECM) in the nervous system, and either permissive or non-permissive roles in neuronal development and regeneration were suggested. The aim of this study was to detect the distribution pattern of a few of these molecules in the nervous system of intact frogs and during nerve regeneration. The hyaluronan (HA) and tenascin C reactions were negative in the peripheral nerves, but appeared in their entry zones. In the CNS, different populations of neurons were surrounded with HA and tenascin C-positive material, forming a perineuronal net (PN). The phosphacan reaction was weakly positive in the PNS, and a moderate intensity was detected in the entry zone and in the PN. Laminin and fibronectin immunoreactivity was strong in the PNS, but laminin could not be detected in the CNS. In animals with cut and regenerating vestibulocochlear nerve, the distribution of the ECM molecules in the CNS and PNS characteristically changed from that of the normal pattern. Our results showed a non-homogenous distribution of ECM components in the frog nervous system that could be associated with their different roles in physiological and pathological processes.