Testican-1 inhibits attachment of Neuro-2a cells

Testican-1 is a highly conserved, multidomain, chondroitin sulfate proteoglycan that is most abundantly transcribed in the brain by neurons. This testican messenger RNA is not detected in normal quiescent astrocytes, but is up regulated when these cells are activated in response to injury such as cerebral stroke. Other chondroitin sulfate proteoglycans found in glial scars, including neurocan, have been shown to inhibit neural cell attachment and neurite extensions and may thus impede axonal regeneration. Here we report the expression and purification of a proteoglycan form of recombinant testican and its effects on neuron-derived cells in culture. We demonstrate that testican inhibits attachment of Neuro-2a cells and their ability to form neurite extensions. Both testican proteoglycan and the core glycoprotein that has been depleted of chondroitin sulfate inhibit cell attachment. Pre-treatment of the culture substratum with testican inhibits Neuro-2a attachment, but pre-treatment of the cells with testican does not inhibit their attachment. Testican, therefore, blocks attachment sites on cultureware and may also block attachment sites in the extracellular matrix of the brain.