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- Englund Johansson, Ulrica, et al.
(författare)
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Migration patterns and phenotypic differentiation of long-term expanded human neural progenitor cells after transplantation into the adult rat brain
- 2002
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Ingår i: Developmental Brain Research. - : Elsevier. - 0165-3806 .- 1872-6755. ; 134:1-2, s. 123-141
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Tidskriftsartikel (refereegranskat)abstract
- We have examined long-term growth-factor expanded human neural progenitors following transplantation into the adult rat brain. Cells, obtained from the forebrain of a 9-week old fetus, propagated in the presence of epidermal growth factor, basic fibroblast growth factor, and leukemia inhibitory factor were transplanted into the striatum, subventricular zone (SVZ), and hippocampus. At 14 weeks, implanted cells were identified using antisera recognizing human nuclei and the reporter gene green fluorescent protein. Different migration patterns of the grafted cells were observed: (i) target-directed migration of doublecortin (DCX, a marker for migrating neuroblasts)-positive cells along the rostral migratory stream to the olfactory bulb and into the granular cell layer following transplantation into the SVZ and hippocampus, respectively; (ii) non-directed migration of DCX-positive cells in the grey matter in striatum and hippocampus, and (iii) extensive migration of above all nestin-positive/DCX-negative cells within white matter tracts. At the striatal implantation site, neuronal differentiation was most pronounced at the graft core with axonal projections extending along the internal capsule bundles. In the hippocampus, cells differentiated primarily into interneurons both in the dentate gyrus and in the CA1-3 regions as well as into granule-like neurons. In the striatum and hippocampus, a significant proportion of the grafted cells differentiated into glial cells, some with long processes extending along white matter tracts. Although the survival time was over 3 months in the present study a large fraction of the grafted cells remained undifferentiated in a stem or progenitor cell stage as revealed by the expression of nestin and/or GFAP.
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- Nomiya, Shigenobu, et al.
(författare)
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Appearance and distribution of two Ca2+-binding proteins during development of the cochlea in the musk shrew
- 1998
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Ingår i: Developmental Brain Research. - 0165-3806 .- 1872-6755. ; 110:1, s. 7-19
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Tidskriftsartikel (refereegranskat)abstract
- In the developing cochlea of the musk shrew, Suncus murinus, the localization of two Ca2+-binding protein, calbindin and calmodulin, which are thought to play different roles in the nervous system, was examined during gestational and postpartum periods. Calbindin is thought to play a Ca2+ buffering role, while calmodulin activates other proteins. Cochleae from the musk shrews sacrificed from gestational day (GD) 15 to postnatal day (PP) 9 and as adults, were immunohistochemically analyzed. The localization and order of appearance of calmodulin in sensorineural elements were similar to those of calbindin, except for timing of appearance. Calmodulin-staining was recognized first in the spiral ganglion neurons on GD21, followed by the inner hair cells (IHCs) on GD23 and outer hair cells (OHCs) on GD26, while calbindin immunoreactivity in the spiral ganglion neurons on GD19, the IHCs on GD21 and the OHCs on GD23. In hair cells, during development, immunostaining of calbindin and calmodulin was initially seen in the cytoplasm, followed by the cuticular plate. Cytoplasmic staining then decreased in mature hair cells. Non-sensorineural components also showed positivity for both calbindin and calmodulin. The lateral wall of the cochlear duct was positive for calbindin, while the stria vascularis was positive for calmodulin. Immunoreactivity for calbindin was present earlier than that of calmodulin in sensorineural elements, suggesting that in the developing cochlea, calbindin and calmodulin have different functions and that Ca2+ buffering capacity, which is regulated by Ca2+ buffer proteins, such as calbindin, may be required before trigger proteins, such as calmodulin, function.
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6. |
- Ekström, Peter, et al.
(författare)
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Ontogenetic development of serotoninergic neurons in the brain of a teleost, the three-spined stickleback. An immunohistochemical analysis
- 1985
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Ingår i: Developmental Brain Research. - : Elsevier BV. - 0165-3806. ; 17:1-2, s. 209-224
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Tidskriftsartikel (refereegranskat)abstract
- The ontogenetic development of serotoninergic neurons in the brain of the stickleback was investigated with the indirect immunocytochemical peroxidase-antiperoxidase technique, using a specific antibody to serotonin (5-hydroxytryptamine, 5-HT). Formation of neuronal populations takes place during embryonic development. By 80 h after fertilization, the first 5-HT perikarya have appeared in the ventricular zone of the hypothalamus (nucleus recessus lateralis) and the raphe region. At 108 h the first 5-HT perikarya can be observed in area praetectalis. At 118 h a transient group of 5-HT neurons appears rostral to the nucleus recessus lateralis, and at this same age the first 5-HT perikarya may be visualized in nucleus recessus posterioris. A group of 5-HT neurons appears in the dorsolateral tegmentum at 166 h (one day after hatching, which occurs at 120-144 h after fertilization). Differentiation of the neuronal populations, in terms of migration and formation of subdivisions, starts between 80 h and 94 h, and seems to be completed between 1 and 5 days after hatching. Raphe nuclei form an anterior group comprising nuclei raphe dorsalis, raphe medialis and a ventrolateral group, and a posterior group comprising a nucleus raphe pallidus/obscurus complex, a lateral nucleus reticularis paragigantocellularis and a ventromedial nucleus raphe magnus. The posterior and ventral raphe nuclei, which are well developed at the time of hatching, have not been visualized in the adult stickleback. While formation of 5-HT neuronal systems, as well as their primary efferent pathways, takes place during early ontogenetic development, the establishment of terminal areas and their subsequent differentiation apparently takes place during later ontogenetic stages. Most presumptive target areas are penetrated by 5-HT axons at hatching, although terminal formation does not seem to start until later. A considerable number of 5-HT neuronal groups present in the embryonic and newly hatched stickleback have not been visualized in the adult stickleback. This may be due to selective cell death, changes in transmitter phenotype or maturation of axonal transport processes during development.
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7. |
- Englund Johansson, Ulrica, et al.
(författare)
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Migration patterns and phenotypic differentiation of long-term expanded human neural progenitor cells after transplantation into the adult rat brain.
- 2002
-
Ingår i: Developmental Brain Research. - 0165-3806. ; 134:1-2, s. 123-141
-
Tidskriftsartikel (refereegranskat)abstract
- We have examined long-term growth-factor expanded human neural progenitors following transplantation into the adult rat brain. Cells, obtained from the forebrain of a 9-week old fetus, propagated in the presence of epidermal growth factor, basic fibroblast growth factor, and leukemia inhibitory factor were transplanted into the striatum, subventricular zone (SVZ), and hippocampus. At 14 weeks, implanted cells were identified using antisera recognizing human nuclei and the reporter gene green fluorescent protein. Different migration patterns of the grafted cells were observed: (i) target-directed migration of doublecortin (DCX, a marker for migrating neuroblasts)-positive cells along the rostral migratory stream to the olfactory bulb and into the granular cell layer following transplantation into the SVZ and hippocampus, respectively; (ii) non-directed migration of DCX-positive cells in the grey matter in striatum and hippocampus, and (iii) extensive migration of above all nestin-positive/DCX-negative cells within white matter tracts. At the striatal implantation site, neuronal differentiation was most pronounced at the graft core with axonal projections extending along the internal capsule bundles. In the hippocampus, cells differentiated primarily into interneurons both in the dentate gyrus and in the CA1-3 regions as well as into granule-like neurons. In the striatum and hippocampus, a significant proportion of the grafted cells differentiated into glial cells, some with long processes extending along white matter tracts. Although the survival time was over 3 months in the present study a large fraction of the grafted cells remained undifferentiated in a stem or progenitor cell stage as revealed by the expression of nestin and/or GFAP.
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8. |
- Forsell, Johan, et al.
(författare)
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Molecular identification and developmental expression of UV and green opsin mRNAs in the pineal organ of the Atlantic halibut.
- 2002
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Ingår i: Developmental Brain Research. - 0165-3806. ; 136:1, s. 51-62
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Tidskriftsartikel (refereegranskat)abstract
- The pineal organ is the only differentiated photoreceptor organ present in embryos and early larvae of the Atlantic halibut (Hippoglossus hippoglossus). We investigated the molecular identity of opsins in the pineal organ, and their expression during different life stages. Using RT-PCR we identified two 681-bp gene sequences, named HPO1 and HPO4, in cDNA from adult pineal and whole embryos. The predicted amino acid sequences showed highest identity to the transmembrane regions of teleostean RH2 green cone opsins (HPO1, 72-91%) and SWS-1 UV cone opsins (HPO4, 71-83%). In situ hybridization revealed expression of HPO1 and HPO4 mRNA transcripts in photoreceptors in the pineal organ of embryos, larvae and adults. HPO1 and HPO4 mRNA transcripts were also expressed in the larval retina. Our study provides molecular evidence for short and middle wavelength light sensitive photoreceptors in the pineal organ of Atlantic halibut throughout life, and suggests that pineal photoreception may play an important role during embryonic and larval life stages, especially at the time when the retina does not possesses corresponding photoreceptor capacity.
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- Johansson, Kjell, et al.
(författare)
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Developmental expression of DCC in the rat retina
- 2001
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Ingår i: Developmental Brain Research. - 0165-3806. ; 130:1, s. 8-133
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Tidskriftsartikel (refereegranskat)abstract
- The protein product of the deleted in colorectal cancer (DCC) gene possesses netrin-binding activity and may be involved in axonal guidance during retinal development. The temporal and spatial expression of DCC was analyzed in developing rat retina by means of immunoblotting and immunohistochemistry as well as by reverse transcription-polymerase chain reaction. Transient DCC protein expression is evident on ganglion cell axons in embryonic and neonatal retina. Double labeling experiments demonstrate DCC immunolabeling on processes that stratify in the inner plexiform layer and are derived from cholinergic amacrine cells. This pattern is maintained during the early postnatal period. DCC immunolabeling in the inner plexiform layer declines with age and is not observed in adult retina. The down-regulation of the DCC protein is confirmed by Western blot analysis. mRNA for DCC is expressed in embryonic, postnatal and adult retina and shows no correlation with the protein down-regulation. We suggest that DCC expression may be correlated with the functional segregation of the inner plexiform layer.
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- Paquet-Durand, Francois, et al.
(författare)
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Turning teratocarcinoma cells into neurons: rapid differentiation of NT-2 cells in floating spheres
- 2003
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Ingår i: Developmental Brain Research. - 0165-3806. ; 142:2, s. 161-167
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Tidskriftsartikel (refereegranskat)abstract
- Cells from the human teratocarcinoma line NTera-2 can be induced to terminally differentiate into postmitotic neurons when treated with retinoic acid. However, this differentiation process is rather time consuming as it takes between 42 and 54 days. Here, we propose a modified differentiation protocol which reduces the time needed for differentiation considerably without compromising the quantity of the neurons obtained. The introduction of a proliferation step as free floating cell spheres cuts the total time needed to obtain high yields of purified NT-2 neurons to about 24-28 days. The cells obtained show neuronal morphology and migrate to form ganglion-like cell conglomerates. Differentiated cells express neuronal polarity markers such as the cytoskeleton associated proteins MAP2 and Tau. Moreover, the generation of neurons in sphere cultures induced immunoreactivity to the ELAV-like neuronal RNA-binding proteins HuC/D, which have been implicated in mechanisms of nerve cell differentiation.
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