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Expression of TrkC receptors in the developing brain of the Monodelphis opossum and its effect on the development of cortical cells.

Bartkowska K, Gajerska M, Turlejski K, Djavadian RL - PLoS ONE (2013)

Bottom Line: We found that in different neocortical areas located both at the rostral and caudal regions of the cortex, up to 98% of BrdU-labeled cells forming cortical layers (II-VI) had prominently expressed TrkC.The shRNAs were individually tested in transfected cortical progenitor cells grown on culture plates for 2 days.The effects of the shRNA-TrkC constructs were similar: blockade of TrkC receptors decreased the number of Ki67-positive and apoptotic cells, and it did not change the number of TUJ-positive neurons in vitro.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cellular Neurobiology, Nencki Institute of Experimental Biology, Warsaw, Poland.

ABSTRACT
In this study, we investigated the distribution, localization and several various functions of TrkC receptors during development of the Monodelphisopossum brain. Western blotting analysis showed that two different forms of the TrkC receptor, the full-length receptor and one of its truncated forms, are abundantly expressed in the opossum brain. The expression of TrkC receptors was barely detected in the brain of newborn opossums. At postnatal day (P) 3, the expression of full-length TrkC remained at low levels, while moderate expression of the TrkC truncated form was detected. The expression levels of both forms of this protein gradually increased throughout development, peaking at P35. We found that in different neocortical areas located both at the rostral and caudal regions of the cortex, up to 98% of BrdU-labeled cells forming cortical layers (II-VI) had prominently expressed TrkC. To assess which developmental processes of cortical cells are regulated by TrkC receptors, three different shRNAs were constructed. The shRNAs were individually tested in transfected cortical progenitor cells grown on culture plates for 2 days. The effects of the shRNA-TrkC constructs were similar: blockade of TrkC receptors decreased the number of Ki67-positive and apoptotic cells, and it did not change the number of TUJ-positive neurons in vitro. Thus, the lack of TrkC receptors in cultured progenitor cells provided insight on the potential role of these receptors in the regulation of proliferation and cell survival but not in the differentiation of cortical cells.

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Coexpression of TrkC with the markers Tle4 and NeuN in neocortical cells at P7.A-D – triple labeling for DAPI (A), Tle4 (B) and TrkC (C) or DAPI (E), NeuN (F) and TrkC (G), and all three labels were merged in D and H, respectively. Analysis of the images using a confocal microscope showed that in both cases, nearly all the Tle4 or NeuN cells expressed TrkC. In A and E, the cortical layers are marked. Scales in D and H refer to all images.
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pone-0074346-g006: Coexpression of TrkC with the markers Tle4 and NeuN in neocortical cells at P7.A-D – triple labeling for DAPI (A), Tle4 (B) and TrkC (C) or DAPI (E), NeuN (F) and TrkC (G), and all three labels were merged in D and H, respectively. Analysis of the images using a confocal microscope showed that in both cases, nearly all the Tle4 or NeuN cells expressed TrkC. In A and E, the cortical layers are marked. Scales in D and H refer to all images.

Mentions: To characterize whether the cells forming cortical layers have a neuronal phenotype, we stained developing opossum brains for Tle4 and TrkC or NeuN. Nearly all Tle4-labeled cells coexpressed TrkC and NeuN (Figure 6).


Expression of TrkC receptors in the developing brain of the Monodelphis opossum and its effect on the development of cortical cells.

Bartkowska K, Gajerska M, Turlejski K, Djavadian RL - PLoS ONE (2013)

Coexpression of TrkC with the markers Tle4 and NeuN in neocortical cells at P7.A-D – triple labeling for DAPI (A), Tle4 (B) and TrkC (C) or DAPI (E), NeuN (F) and TrkC (G), and all three labels were merged in D and H, respectively. Analysis of the images using a confocal microscope showed that in both cases, nearly all the Tle4 or NeuN cells expressed TrkC. In A and E, the cortical layers are marked. Scales in D and H refer to all images.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3760877&req=5

pone-0074346-g006: Coexpression of TrkC with the markers Tle4 and NeuN in neocortical cells at P7.A-D – triple labeling for DAPI (A), Tle4 (B) and TrkC (C) or DAPI (E), NeuN (F) and TrkC (G), and all three labels were merged in D and H, respectively. Analysis of the images using a confocal microscope showed that in both cases, nearly all the Tle4 or NeuN cells expressed TrkC. In A and E, the cortical layers are marked. Scales in D and H refer to all images.
Mentions: To characterize whether the cells forming cortical layers have a neuronal phenotype, we stained developing opossum brains for Tle4 and TrkC or NeuN. Nearly all Tle4-labeled cells coexpressed TrkC and NeuN (Figure 6).

Bottom Line: We found that in different neocortical areas located both at the rostral and caudal regions of the cortex, up to 98% of BrdU-labeled cells forming cortical layers (II-VI) had prominently expressed TrkC.The shRNAs were individually tested in transfected cortical progenitor cells grown on culture plates for 2 days.The effects of the shRNA-TrkC constructs were similar: blockade of TrkC receptors decreased the number of Ki67-positive and apoptotic cells, and it did not change the number of TUJ-positive neurons in vitro.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cellular Neurobiology, Nencki Institute of Experimental Biology, Warsaw, Poland.

ABSTRACT
In this study, we investigated the distribution, localization and several various functions of TrkC receptors during development of the Monodelphisopossum brain. Western blotting analysis showed that two different forms of the TrkC receptor, the full-length receptor and one of its truncated forms, are abundantly expressed in the opossum brain. The expression of TrkC receptors was barely detected in the brain of newborn opossums. At postnatal day (P) 3, the expression of full-length TrkC remained at low levels, while moderate expression of the TrkC truncated form was detected. The expression levels of both forms of this protein gradually increased throughout development, peaking at P35. We found that in different neocortical areas located both at the rostral and caudal regions of the cortex, up to 98% of BrdU-labeled cells forming cortical layers (II-VI) had prominently expressed TrkC. To assess which developmental processes of cortical cells are regulated by TrkC receptors, three different shRNAs were constructed. The shRNAs were individually tested in transfected cortical progenitor cells grown on culture plates for 2 days. The effects of the shRNA-TrkC constructs were similar: blockade of TrkC receptors decreased the number of Ki67-positive and apoptotic cells, and it did not change the number of TUJ-positive neurons in vitro. Thus, the lack of TrkC receptors in cultured progenitor cells provided insight on the potential role of these receptors in the regulation of proliferation and cell survival but not in the differentiation of cortical cells.

Show MeSH
Related in: MedlinePlus