- Email: [email protected]
Developmental regulation of decorin expression in postnatal rat brain
Brain Research 793 Ž1998. 328–332
Short communication
Developmental regulation of decorin expression in postnatal rat brain Joachim Kappler a
a, )
, Christine C. Stichel a , Marc Gleichmann a , Clemens Gillen a , Ulrich Junghans a , a Hans Kresse b , Hans Werner Muller ¨
Labor fur Moorenstraße 5, D-40225 Dusseldorf, Germany ¨ molekulare Neurobiologie, Neurologische Klinik der Heinrich-Heine-UniÕersitat ¨ Dusseldorf, ¨ ¨ Institut fur Wilhelms-UniÕersitat Waldeyerstraße 15, D-48149 Munster, Germany ¨ Physiologische Chemie und Pathobiochemie, Westfalische ¨ ¨ Munster, ¨ ¨
b
Accepted 3 March 1998
Abstract Here, we report on the expression of the small chondroitinrdermatan sulfate proteoglycan decorin in the developing postnatal rat brain. Northern analysis of brain RNA demonstrated decorin transcripts with peak expression on postnatal day 3 followed by a slow decline to the lower adult level. In situ hybridization and immunohistochemistry revealed postnatal decorin expression in the grey matter of neocortex, hippocampus and thalamus, in myelinated fibre tracts and in several mesenchymal tissues Žblood vessels, pia mater and the choroid plexus.. In the neocortex, decorin is expressed in a specific laminar pattern with intense staining of the cortical plate and its derivatives, which differs remarkably from the distributions observed for other proteoglycans wB. Miller, A.M. Sheppard, A.R. Bicknese, A.L. Pearlman, Chondroitin sulfate proteoglycans in the developing cerebral cortex: the distribution of neurocan distinguishes forming afferent and efferent axonal pathways, J. Comp. Neurol. 355 Ž1995. 615–28x. Thus, decorin seems to serve yet unknown functions in the developing rat brain parenchyma in addition to its well-established role as a constituent of the mesenchymal extracellular matrix. q 1998 Elsevier Science B.V. All rights reserved. Keywords: Extracellular matrix; Glycosaminoglycan; Nervous system; Neocortical development
Decorin is a small proteoglycan of the extracellular matrix carrying in mammals a single chondroitinrdermatan sulfate side chain linked to the 40 kDa core glycoprotein. As other small proteoglycans, decorin core protein is made up of characteristic leucine-rich repeats, which are surrounded by conserved disulfide loops w14,12x. The proteoglycan binds specifically to different macromolecules including fibrillar collagens w6x, fibronectin w19x, transforming growth factor b ŽTGF-b . w11x, cellular receptor proteins w10x and the b-amyloid protein w21x. Accordingly, decorin may influence such heterogenous processes as cell
)
Corresponding author. Fax: q49-211-811-8485.
0006-8993r98r$19.00 q 1998 Elsevier Science B.V. All rights reserved. PII S 0 0 0 6 - 8 9 9 3 Ž 9 8 . 0 0 2 6 0 - 1
adhesion w16,23x, collagen fibrillogenesis and growth factor signalling Že.g., by TGF-b , see Refs. w24,9x. Consistent with a wide variety of its binding partners, decorin is found in a number of different tissues including cartilage and skin. Furthermore, decorin mRNA is expressed in the peripheral and central nervous system of rat w8x. In the adult rat brain, it is found in cortical neurons, astrocytes, in the white matter and in mesenchymal structures. After injury of the postcomissural fornix, its expression is upregulated w22x. Contrasting with these data, substantial decorin mRNA expression in the mouse brain was not detected up to embryonic day 16 w20x, and in a broad immunocytochemical survey of perinatal proteoglycan expression in the rat brain w17x, no decorin expression was found. Because of these discrepancies, the purpose of the present study was to analyze the time course and the spatial distribution of decorin mRNA and protein expres-
J. Kappler et al.r Brain Research 793 (1998) 328–332
sion in the postnatal rat brain using Northern analysis and in situ hybridization in parallel with immunohistochemical methods. Upon screening of a rat sciatic nerve cDNA library w7x with a 0.3 kB rat decorin cDNA probe w8x, approximately 1 out of 1000 clones was positive. One clone containing a 1.7-kb insert was sequenced using an ABI prism 310 genetic analyzer ŽApplied Biosystems. revealing a coding region identical to the published rat decorin cDNA sequence w1x. Northern blot analysis with the decorin 1.7 kB cDNA probe was carried out on total RNA from rat brain w3x as described w8x. Transcripts of approximately 1.7 kb were detected. The amount of decorin mRNA, however, changed considerably during development. At P1, only a faint signal was seen, whereas three days later, at P4, expression was maximal and up to postnatal day 21 ŽP21. a strong signal was detected. In the adult animal decorin, mRNA was still detectable, but the level of expression was downregulated again Žsee Fig. 1.. To survey the sites of postnatal decorin mRNA expression, in situ hybridization with decorin antisense transcript probes to frontal sections of P21 rat forebrain was carried out as described w8x using digoxigenin-labelled run-off
Fig. 1. Northern blot of rat brain total RNA Ž10 m g per lane. hybridized with decorin cDNA. Top: autoradiograph after hybridization with a 32 P-labelled probe. Time points are indicated at the top of each lane; bottom: comparison of the amounts of RNA blotted in each lane using methylene blue staining of the same filter before hybridization. Left: relative position of 28S and 18S RNA bands.
329
transcripts from the 1.7 kB decorin cDNA. Decorin mRNA was detected Ža. in the grey matter including thalamus, hippocampal allocortex and the neocortex sparing the marginal zone Žlayer I., Žb. in the white matter including subcortical fibre tracts and the corpus callosum and Žc. in mesenchymal structures, namely the choroid plexus, vessels and the pia mater Žsee Fig. 2A–C.. Immunocytochemistry with a polyclonal rabbit anti-decorin core protein was carried out at three time points ŽP1, P7 and P21. with acetone-fixed brain cryostat sections as described earlier w22x. The staining pattern was similar to the data obtained with in situ hybridization at P21. Interestingly, in the cortex ŽFig. 3., there was a specific laminar expression with a pronounced cellular staining in the cortical plate at P1A contrasting with a lower density of positive cells in the marginal and the subventricular zone ŽMZ and SV, respectively.. Intermediate immunoreactivity was observed in the neuroepithel ŽNE. and the intermediate zone ŽIZ.. On P7 Žsee Fig. 3B., the distribution was similar with highest density of stained cells in the pyramidal cell layer IV. At P21 ŽFig. 3C., the total number of labeled cells had decreased, and was more evenly distributed between layers II to VI, a pattern very similar to that observed in the adult animal w22x. Additionally, a diffuse signal developed in the white matter Žwm.. Apparently, decorin is mainly found in the permanent cells of the cortex, whereas the transient preplate-derived neurons in the marginal zone and subplate do not express this proteoglycan to the same extent. On the other hand, the brain-specific large extracellular matrix proteoglycan neurocan w18x displays a complementary immunocytochemical pattern with intense staining in the subplate and the marginal zone, and a substantially lower expression the cortical plate w17x. The majority of other proteoglycans, including phosphacan, the syndecans and NG2, however, are expressed in a non-laminar pattern w17x. The onset of decorin expression in cortical cells is presumably before birth, since there is substantial immunocytochemical staining at the neonatal stage. Since no neural expression of decorin mRNA was found in embryonic mouse brain up to day 16 w20x, onset of expression may occur after this time point. However, in rat a prenatal study, extending our results should settle this issue. The chondroitinrdermatan sulfate side chain of decorin may exert specific effects on cortical neurons like enhanced survival w13x, polarization w15x or segregation of fibre tracts w2x. Based on functional studies with cultured brain slices w4x and on immunohistochemical data ŽU. Junghans, S. Franken, A. Pommer, C. Viebahn, H.W. Muller, J. Kappler, unpublished results., the existence of ¨ chondroitin sulfate-binding molecules was postulated that co-localize with decorin in the cortical plate. Such binding molecules could mediate the neurotrophic actions listed above. Furthermore, the leucine-rich decorin core protein interacts with a number of different proteins including constituents of the extracellular matrix Že.g., fibrillar colla-
330
J. Kappler et al.r Brain Research 793 (1998) 328–332
gens, fibronectin w14x and the polypeptide growth factor TGF-b w24x. Since different members of the TGF-b family are expressed in the developing rodent brain w5x, decorin may modulate the activity and endocytosis of these growth
factors. Interestingly, a study on the binding of TGF-b to recombinant decorin core protein w11x further indicated binding of NGF Žthat was included as a control. to decorin and its homologue biglycan. Therefore, decorin may mod-
Fig. 2. In situ hybridization of P21 rat brain to decorin antisense ŽA–C. or sense ŽD. cRNA transcripts. A: neocortex with adjacent subcortical white matter. B,C and D: midline structures including the corpus callosum Žcc., choroid plexus Žcp., thalamus Žth. and the neighbouring hippocampal allocortex Žhi.. D: negative control. Scale bar: 100 m m.
J. Kappler et al.r Brain Research 793 (1998) 328–332
331
Fig. 3. Decorin immunoreactivity in the developing neocortex. ŽA. postnatal day 1. CP: cortical plate, IZ: intermediate zone, SP: subplate zone, MZ: marginal zone, NE: neuroepithel, VI: cortical layer 6, WM: white matter. ŽB. postnatal day 7, I–VI: cortical layers, ŽC.: postnatal day 21. Scale bar: 100 m m.
ulate the action of other cystine-knot growth factors like the neurotrophins in the brain.
Acknowledgements We thank R. Greiner-Petter for excellent technical assistance. Supported by the Deutsche Forschungsgemeinschaft.
References w1x S.R. Abramson, J.F. Woessner Jr., cDna sequence for rat dermatan sulfate proteoglycan-Ii Ždecorin., Biochim. Biophys. Acta 1132 Ž1992. 225–227. w2x A.R. Bicknese, A.M. Sheppard, D.D. O’Leary, A.L. Pearlman, Thalamocortical axons extend along a chondroitin sulfate proteoglycan-enriched pathway coincident with the neocortical subplate and distinct from the efferent path, J. Neurosci. 14 Ž1994. 3500–3510. w3x P. Chomczynski, N. Sacchi, Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction, Anal. Biochem. 162 Ž1987. 156–159. w4x D.E. Emerling, A.D. Lander, Inhibitors and promoters of thalamic neuron adhesion and outgrowth in embryonic neocortex: functional association with chondroitin sulfate, Neuron 17 Ž1996. 1089–1100. w5x K.C. Flanders, G. Ludecke, S. Engels, D.S. Cissel, A.B. Roberts, P. Kondaiah, R. Lafyatis, M.B. Sporn, K. Unsicker, Localization and actions of transforming growth factor-beta s in the embryonic nervous system, Development 113 Ž1991. 183–191.
w6x R. Fleischmajer, L.W. Fisher, E.D. MacDonald, L. Jacobs Jr., J.S. Perlish, J.D. Termine, Decorin interacts with fibrillar collagen of embryonic and adult human skin, J. Struct. Biol. 106 Ž1991. 82–90. w7x C. Gillen, M. Gleichmann, P. Spreyer, H.W. Muller, Differentially ¨ expressed genes after peripheral nerve injury, J. Neurosci. Res. 42 Ž1995. 159–171. w8x C.O. Hanemann, G. Kuhn, A. Lie, C. Gillen, F. Bosse, P. Spreyer, H.W. Muller, Expression of decorin mRNA in the nervous system of ¨ rat, J. Histochem. Cytochem. 41 Ž1993. 1383–1391. w9x H. Hausser, A. Groning, A. Hasilik, E. Schonherr, H. Kresse, ¨ Selective inactivity of TGF-betardecorin complexes, FEBS Lett. 353 Ž1994. 243–245. w10x H. Hausser, H. Kresse, Binding of heparin and of the small proteoglycan decorin to the same endocytosis receptor proteins leads to different metabolic consequences, J. Cell. Biol. 114 Ž1991. 45–52. w11x A. Hildebrand, M. Romaris, L.M. Rasmussen, D. Heinegard, D.R. Twardzik, W.A. Border, E. Ruoslahti, Interaction of the small interstitial proteoglycans biglycan, decorin and fibromodulin with transforming growth factor beta, Biochem. J. 302 Ž1994. 527–534. w12x R.V. Iozzo, A.D. Murdoch, Proteoglycans of the extracellular environment: clues from the gene and protein side offer novel perspectives in molecular diversity and function, FASEB J. 10 Ž1996. 598–614. w13x J. Kappler, U. Junghans, A. Koops, C.C. Stichel, H.J. Hausser, H. Kresse, H.W. Muller, Chondroitinrdermatan sulphate promotes the ¨ survival of neurons from rat embryonic neocortex, Eur. J. Neurosci. 9 Ž1997. 306–318. w14x H. Kresse, H. Hausser, E. Schonherr, Small proteoglycans, Experi¨ entia 49 Ž1993. 403–416. w15x F. Lafont, M. Rouget, A. Triller, A. Prochiantz, A. Rousselet, In vitro control of neuronal polarity by glycosaminoglycans, Development 114 Ž1992. 17–29.
332
J. Kappler et al.r Brain Research 793 (1998) 328–332
w16x K. Lewandowska, H.U. Choi, L.C. Rosenberg, L. Zardi, L.A. Culp, Fibronectin-mediated adhesion of fibroblasts: inhibition by dermatan sulfate proteoglycan and evidence for a cryptic glycosaminoglycanbinding domain, J. Cell. Biol. 105 Ž1987. 1443–1454. w17x B. Miller, A.M. Sheppard, A.R. Bicknese, A.L. Pearlman, Chondroitin sulfate proteoglycans in the developing cerebral cortex: the distribution of neurocan distinguishes forming afferent and efferent axonal pathways, J. Comp. Neurol. 355 Ž1995. 615–628. w18x U. Rauch, L. Karthikeyan, P. Maurel, R.U. Margolis, R.K. Margolis, Cloning and primary structure of neurocan, a developmentally regulated, aggregating chondroitin sulfate proteoglycan of brain, J. Biol. Chem. 267 Ž1992. 19536–19547. w19x G. Schmidt, H. Hausser, H. Kresse, Interaction of the small proteoglycan decorin with fibronectin. Involvement of the sequence NKISK of the core protein, Biochem. J. 280 Ž1991. 411–414. w20x T. Scholzen, M. Solursh, S. Suzuki, R. Reiter, J.L. Morgan, A.M. Buchberg, L.D. Siracusa, R.V. Iozzo, The murine decorin. Complete cDNA cloning, genomic organization, chromosomal assignment and
w21x
w22x
w23x
w24x
expression during organogenesis and tissue differentiation, J. Biol. Chem. 269 Ž1994. 28270–28281. A.D. Snow, M.G. Kinsella, E. Parks, R.T. Sekiguchi, J.D. Miller, K. Kimata, T.N. Wight, Differential binding of vascular cell-derived proteoglycans Žperlecan, biglycan, decorin and versican. to the beta-amyloid protein of Alzheimer’s disease, Arch. Biochem. Biophys. 320 Ž1995. 84–95. C.C. Stichel, J. Kappler, U. Junghans, A. Koops, H. Kresse, H.W. Muller, Differential expression of the small chondroitinrdermatan ¨ sulfate proteoglycans decorin and biglycan after injury of the adult rat brain, Brain Res. 704 Ž1995. 263–274. M. Winnemoller, P. Schon, ¨ P. Vischer, H. Kresse, Interactions between thrombospondin and the small proteoglycan decorin: interference with cell attachment, Eur. J. Cell. Biol. 59 Ž1992. 47–55. Y. Yamaguchi, D.M. Mann, E. Ruoslahti, Negative regulation of transforming growth factor-beta by the proteoglycan decorin, Nature 346 Ž1992. 281–284.
Short communication
Developmental regulation of decorin expression in postnatal rat brain Joachim Kappler a
a, )
, Christine C. Stichel a , Marc Gleichmann a , Clemens Gillen a , Ulrich Junghans a , a Hans Kresse b , Hans Werner Muller ¨
Labor fur Moorenstraße 5, D-40225 Dusseldorf, Germany ¨ molekulare Neurobiologie, Neurologische Klinik der Heinrich-Heine-UniÕersitat ¨ Dusseldorf, ¨ ¨ Institut fur Wilhelms-UniÕersitat Waldeyerstraße 15, D-48149 Munster, Germany ¨ Physiologische Chemie und Pathobiochemie, Westfalische ¨ ¨ Munster, ¨ ¨
b
Accepted 3 March 1998
Abstract Here, we report on the expression of the small chondroitinrdermatan sulfate proteoglycan decorin in the developing postnatal rat brain. Northern analysis of brain RNA demonstrated decorin transcripts with peak expression on postnatal day 3 followed by a slow decline to the lower adult level. In situ hybridization and immunohistochemistry revealed postnatal decorin expression in the grey matter of neocortex, hippocampus and thalamus, in myelinated fibre tracts and in several mesenchymal tissues Žblood vessels, pia mater and the choroid plexus.. In the neocortex, decorin is expressed in a specific laminar pattern with intense staining of the cortical plate and its derivatives, which differs remarkably from the distributions observed for other proteoglycans wB. Miller, A.M. Sheppard, A.R. Bicknese, A.L. Pearlman, Chondroitin sulfate proteoglycans in the developing cerebral cortex: the distribution of neurocan distinguishes forming afferent and efferent axonal pathways, J. Comp. Neurol. 355 Ž1995. 615–28x. Thus, decorin seems to serve yet unknown functions in the developing rat brain parenchyma in addition to its well-established role as a constituent of the mesenchymal extracellular matrix. q 1998 Elsevier Science B.V. All rights reserved. Keywords: Extracellular matrix; Glycosaminoglycan; Nervous system; Neocortical development
Decorin is a small proteoglycan of the extracellular matrix carrying in mammals a single chondroitinrdermatan sulfate side chain linked to the 40 kDa core glycoprotein. As other small proteoglycans, decorin core protein is made up of characteristic leucine-rich repeats, which are surrounded by conserved disulfide loops w14,12x. The proteoglycan binds specifically to different macromolecules including fibrillar collagens w6x, fibronectin w19x, transforming growth factor b ŽTGF-b . w11x, cellular receptor proteins w10x and the b-amyloid protein w21x. Accordingly, decorin may influence such heterogenous processes as cell
)
Corresponding author. Fax: q49-211-811-8485.
0006-8993r98r$19.00 q 1998 Elsevier Science B.V. All rights reserved. PII S 0 0 0 6 - 8 9 9 3 Ž 9 8 . 0 0 2 6 0 - 1
adhesion w16,23x, collagen fibrillogenesis and growth factor signalling Že.g., by TGF-b , see Refs. w24,9x. Consistent with a wide variety of its binding partners, decorin is found in a number of different tissues including cartilage and skin. Furthermore, decorin mRNA is expressed in the peripheral and central nervous system of rat w8x. In the adult rat brain, it is found in cortical neurons, astrocytes, in the white matter and in mesenchymal structures. After injury of the postcomissural fornix, its expression is upregulated w22x. Contrasting with these data, substantial decorin mRNA expression in the mouse brain was not detected up to embryonic day 16 w20x, and in a broad immunocytochemical survey of perinatal proteoglycan expression in the rat brain w17x, no decorin expression was found. Because of these discrepancies, the purpose of the present study was to analyze the time course and the spatial distribution of decorin mRNA and protein expres-
J. Kappler et al.r Brain Research 793 (1998) 328–332
sion in the postnatal rat brain using Northern analysis and in situ hybridization in parallel with immunohistochemical methods. Upon screening of a rat sciatic nerve cDNA library w7x with a 0.3 kB rat decorin cDNA probe w8x, approximately 1 out of 1000 clones was positive. One clone containing a 1.7-kb insert was sequenced using an ABI prism 310 genetic analyzer ŽApplied Biosystems. revealing a coding region identical to the published rat decorin cDNA sequence w1x. Northern blot analysis with the decorin 1.7 kB cDNA probe was carried out on total RNA from rat brain w3x as described w8x. Transcripts of approximately 1.7 kb were detected. The amount of decorin mRNA, however, changed considerably during development. At P1, only a faint signal was seen, whereas three days later, at P4, expression was maximal and up to postnatal day 21 ŽP21. a strong signal was detected. In the adult animal decorin, mRNA was still detectable, but the level of expression was downregulated again Žsee Fig. 1.. To survey the sites of postnatal decorin mRNA expression, in situ hybridization with decorin antisense transcript probes to frontal sections of P21 rat forebrain was carried out as described w8x using digoxigenin-labelled run-off
Fig. 1. Northern blot of rat brain total RNA Ž10 m g per lane. hybridized with decorin cDNA. Top: autoradiograph after hybridization with a 32 P-labelled probe. Time points are indicated at the top of each lane; bottom: comparison of the amounts of RNA blotted in each lane using methylene blue staining of the same filter before hybridization. Left: relative position of 28S and 18S RNA bands.
329
transcripts from the 1.7 kB decorin cDNA. Decorin mRNA was detected Ža. in the grey matter including thalamus, hippocampal allocortex and the neocortex sparing the marginal zone Žlayer I., Žb. in the white matter including subcortical fibre tracts and the corpus callosum and Žc. in mesenchymal structures, namely the choroid plexus, vessels and the pia mater Žsee Fig. 2A–C.. Immunocytochemistry with a polyclonal rabbit anti-decorin core protein was carried out at three time points ŽP1, P7 and P21. with acetone-fixed brain cryostat sections as described earlier w22x. The staining pattern was similar to the data obtained with in situ hybridization at P21. Interestingly, in the cortex ŽFig. 3., there was a specific laminar expression with a pronounced cellular staining in the cortical plate at P1A contrasting with a lower density of positive cells in the marginal and the subventricular zone ŽMZ and SV, respectively.. Intermediate immunoreactivity was observed in the neuroepithel ŽNE. and the intermediate zone ŽIZ.. On P7 Žsee Fig. 3B., the distribution was similar with highest density of stained cells in the pyramidal cell layer IV. At P21 ŽFig. 3C., the total number of labeled cells had decreased, and was more evenly distributed between layers II to VI, a pattern very similar to that observed in the adult animal w22x. Additionally, a diffuse signal developed in the white matter Žwm.. Apparently, decorin is mainly found in the permanent cells of the cortex, whereas the transient preplate-derived neurons in the marginal zone and subplate do not express this proteoglycan to the same extent. On the other hand, the brain-specific large extracellular matrix proteoglycan neurocan w18x displays a complementary immunocytochemical pattern with intense staining in the subplate and the marginal zone, and a substantially lower expression the cortical plate w17x. The majority of other proteoglycans, including phosphacan, the syndecans and NG2, however, are expressed in a non-laminar pattern w17x. The onset of decorin expression in cortical cells is presumably before birth, since there is substantial immunocytochemical staining at the neonatal stage. Since no neural expression of decorin mRNA was found in embryonic mouse brain up to day 16 w20x, onset of expression may occur after this time point. However, in rat a prenatal study, extending our results should settle this issue. The chondroitinrdermatan sulfate side chain of decorin may exert specific effects on cortical neurons like enhanced survival w13x, polarization w15x or segregation of fibre tracts w2x. Based on functional studies with cultured brain slices w4x and on immunohistochemical data ŽU. Junghans, S. Franken, A. Pommer, C. Viebahn, H.W. Muller, J. Kappler, unpublished results., the existence of ¨ chondroitin sulfate-binding molecules was postulated that co-localize with decorin in the cortical plate. Such binding molecules could mediate the neurotrophic actions listed above. Furthermore, the leucine-rich decorin core protein interacts with a number of different proteins including constituents of the extracellular matrix Že.g., fibrillar colla-
330
J. Kappler et al.r Brain Research 793 (1998) 328–332
gens, fibronectin w14x and the polypeptide growth factor TGF-b w24x. Since different members of the TGF-b family are expressed in the developing rodent brain w5x, decorin may modulate the activity and endocytosis of these growth
factors. Interestingly, a study on the binding of TGF-b to recombinant decorin core protein w11x further indicated binding of NGF Žthat was included as a control. to decorin and its homologue biglycan. Therefore, decorin may mod-
Fig. 2. In situ hybridization of P21 rat brain to decorin antisense ŽA–C. or sense ŽD. cRNA transcripts. A: neocortex with adjacent subcortical white matter. B,C and D: midline structures including the corpus callosum Žcc., choroid plexus Žcp., thalamus Žth. and the neighbouring hippocampal allocortex Žhi.. D: negative control. Scale bar: 100 m m.
J. Kappler et al.r Brain Research 793 (1998) 328–332
331
Fig. 3. Decorin immunoreactivity in the developing neocortex. ŽA. postnatal day 1. CP: cortical plate, IZ: intermediate zone, SP: subplate zone, MZ: marginal zone, NE: neuroepithel, VI: cortical layer 6, WM: white matter. ŽB. postnatal day 7, I–VI: cortical layers, ŽC.: postnatal day 21. Scale bar: 100 m m.
ulate the action of other cystine-knot growth factors like the neurotrophins in the brain.
Acknowledgements We thank R. Greiner-Petter for excellent technical assistance. Supported by the Deutsche Forschungsgemeinschaft.
References w1x S.R. Abramson, J.F. Woessner Jr., cDna sequence for rat dermatan sulfate proteoglycan-Ii Ždecorin., Biochim. Biophys. Acta 1132 Ž1992. 225–227. w2x A.R. Bicknese, A.M. Sheppard, D.D. O’Leary, A.L. Pearlman, Thalamocortical axons extend along a chondroitin sulfate proteoglycan-enriched pathway coincident with the neocortical subplate and distinct from the efferent path, J. Neurosci. 14 Ž1994. 3500–3510. w3x P. Chomczynski, N. Sacchi, Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction, Anal. Biochem. 162 Ž1987. 156–159. w4x D.E. Emerling, A.D. Lander, Inhibitors and promoters of thalamic neuron adhesion and outgrowth in embryonic neocortex: functional association with chondroitin sulfate, Neuron 17 Ž1996. 1089–1100. w5x K.C. Flanders, G. Ludecke, S. Engels, D.S. Cissel, A.B. Roberts, P. Kondaiah, R. Lafyatis, M.B. Sporn, K. Unsicker, Localization and actions of transforming growth factor-beta s in the embryonic nervous system, Development 113 Ž1991. 183–191.
w6x R. Fleischmajer, L.W. Fisher, E.D. MacDonald, L. Jacobs Jr., J.S. Perlish, J.D. Termine, Decorin interacts with fibrillar collagen of embryonic and adult human skin, J. Struct. Biol. 106 Ž1991. 82–90. w7x C. Gillen, M. Gleichmann, P. Spreyer, H.W. Muller, Differentially ¨ expressed genes after peripheral nerve injury, J. Neurosci. Res. 42 Ž1995. 159–171. w8x C.O. Hanemann, G. Kuhn, A. Lie, C. Gillen, F. Bosse, P. Spreyer, H.W. Muller, Expression of decorin mRNA in the nervous system of ¨ rat, J. Histochem. Cytochem. 41 Ž1993. 1383–1391. w9x H. Hausser, A. Groning, A. Hasilik, E. Schonherr, H. Kresse, ¨ Selective inactivity of TGF-betardecorin complexes, FEBS Lett. 353 Ž1994. 243–245. w10x H. Hausser, H. Kresse, Binding of heparin and of the small proteoglycan decorin to the same endocytosis receptor proteins leads to different metabolic consequences, J. Cell. Biol. 114 Ž1991. 45–52. w11x A. Hildebrand, M. Romaris, L.M. Rasmussen, D. Heinegard, D.R. Twardzik, W.A. Border, E. Ruoslahti, Interaction of the small interstitial proteoglycans biglycan, decorin and fibromodulin with transforming growth factor beta, Biochem. J. 302 Ž1994. 527–534. w12x R.V. Iozzo, A.D. Murdoch, Proteoglycans of the extracellular environment: clues from the gene and protein side offer novel perspectives in molecular diversity and function, FASEB J. 10 Ž1996. 598–614. w13x J. Kappler, U. Junghans, A. Koops, C.C. Stichel, H.J. Hausser, H. Kresse, H.W. Muller, Chondroitinrdermatan sulphate promotes the ¨ survival of neurons from rat embryonic neocortex, Eur. J. Neurosci. 9 Ž1997. 306–318. w14x H. Kresse, H. Hausser, E. Schonherr, Small proteoglycans, Experi¨ entia 49 Ž1993. 403–416. w15x F. Lafont, M. Rouget, A. Triller, A. Prochiantz, A. Rousselet, In vitro control of neuronal polarity by glycosaminoglycans, Development 114 Ž1992. 17–29.
332
J. Kappler et al.r Brain Research 793 (1998) 328–332
w16x K. Lewandowska, H.U. Choi, L.C. Rosenberg, L. Zardi, L.A. Culp, Fibronectin-mediated adhesion of fibroblasts: inhibition by dermatan sulfate proteoglycan and evidence for a cryptic glycosaminoglycanbinding domain, J. Cell. Biol. 105 Ž1987. 1443–1454. w17x B. Miller, A.M. Sheppard, A.R. Bicknese, A.L. Pearlman, Chondroitin sulfate proteoglycans in the developing cerebral cortex: the distribution of neurocan distinguishes forming afferent and efferent axonal pathways, J. Comp. Neurol. 355 Ž1995. 615–628. w18x U. Rauch, L. Karthikeyan, P. Maurel, R.U. Margolis, R.K. Margolis, Cloning and primary structure of neurocan, a developmentally regulated, aggregating chondroitin sulfate proteoglycan of brain, J. Biol. Chem. 267 Ž1992. 19536–19547. w19x G. Schmidt, H. Hausser, H. Kresse, Interaction of the small proteoglycan decorin with fibronectin. Involvement of the sequence NKISK of the core protein, Biochem. J. 280 Ž1991. 411–414. w20x T. Scholzen, M. Solursh, S. Suzuki, R. Reiter, J.L. Morgan, A.M. Buchberg, L.D. Siracusa, R.V. Iozzo, The murine decorin. Complete cDNA cloning, genomic organization, chromosomal assignment and
w21x
w22x
w23x
w24x
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