Phosphacan and neurocan are repulsive substrata for adhesion and neurite extension of adult rat dorsal root ganglion neurons in vitro.
Phosphacan (PC) and neurocan (NC) are major chondroitin sulfate proteoglycans (CS-PGs) in nervous tissue and are involved in the modulation of cell adhesion and neurite outgrowth during neural development and regeneration. In the present study, we examined the effects of PC and NC on the attachment and neurite extension of adult rat dorsal root ganglion (DRG) neurons in vitro. Treatment with PC and NC on poly-L-lysine (PL) significantly impaired both neuronal attachment and neurite extension in a concentration-dependent manner (10 microg/ml > 1 microg/ml >> 0.1 microg/ml), and they were partially suppressed by chondroitinase ABC (ChABC) digestion. The CS-PGs applied to culture medium (1 microg/ml) also displayed inhibitory effects on neurite extension, which were not altered by ChABC treatment. These results show that PC and NC are repulsive substrata for adhesion and neurite regeneration of adult DRG neurons in vitro and suggest that both chondroitin sulfate moieties and core proteins are responsible for the inhibitory actions of the CS-PGs. We also conducted immunohistochemical analyses with the monoclonal antibodies to core proteins of PC (mAb 6B4) and NC (mAb 1G2), which revealed that only a few neurons in the DRG section were stained with these antibodies. In contrast, most DRG neurons at different stages (12 h, 1 day, 2 days, and 4 days) in culture were immunoreactive to mAb 6B4 and mAb 1G2. Taking these findings together, it is plausible that both CS-PGs expressed in the cultured neurons may play a role in the modulation of attachment, survival, and neurite regeneration.
Sango
Kazunori
K
Department of Developmental Morphology, Tokyo Metropolitan Institute for Neuroscience, 2-6 Musashidai, Fuchu-shi, Tokyo 183-8526, Japan. kazsango@tmin.ac.jp
Oohira
Atsuhiko
A
Ajiki
Kyoko
K
Tokashiki
Akiko
A
Horie
Masao
M
Kawano
Hitoshi
H
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D019765:Chondroitin ABC Lyase / Q000494:pharmacology; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism* / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005260:Female; D005727:Ganglia, Spinal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D007150:Immunohistochemistry; D037181:Lectins, C-Type; D009419:Nerve Tissue Proteins / Q000378:metabolism* / Q000494:pharmacology; D016501:Neurites / Q000187:drug effects / Q000502:physiology*; D058580:Neurocan; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D054633:Receptor-Like Protein Tyrosine Phosphatases, Class 5
D011508:Chondroitin Sulfate Proteoglycans; D037181:Lectins, C-Type; D009419:Nerve Tissue Proteins; D058580:Neurocan; C076817:NCAN protein, human; C516895:PTPRZ1 protein, human; C516897:Ptprz1 protein, rat; D054633:Receptor-Like Protein Tyrosine Phosphatases, Class 5; D019765:Chondroitin ABC Lyase