Brain angiogenesis: Variations in vascular basement membrane glycoprotein immunoreactivity

Abstract

Changes in the distribution and quantity of laminin and fibronectin within the basement membranes of developing or regenerating CNS blood vessels were investigated using two immunocytochemical techniques. Three models of angiogenesis were studied: normal pre- and postnatal development, wound healing, and vascularization of fetal neocortical transplants placed in the adult rat brain. Although all brain vessels were stained in enzymatically pretreated immunoreacted paraffin sections, those associated with wound and transplant sites were the most intensely reactive with both antisera during the first postoperative week. When 40-μm vibratome sections of normal adult brains were immunoprocessed, only the meninges and vessels of the circumventricular organs were stained. The remainder of the brain vasculature was immunoreactive only if sections were enzymatically treated prior to immunoprocessing. In contrast, the nascent vasculature in developing brain and the regenerating vessels at wound and transplant sites were reactive to both antisera without enzymatic pretreatment of the sections. This immunoreactivity decreased by 11 days postnatal in normal animals and 4 weeks postoperative in experimental animals, coinciding with the period of astrocytic contact and complete vascular basement membrane formation in both cases. The variations in staining pattern and intensity may be reflections of differences in the quantity of laminin and fibronectin within the basement membranes of proliferating and/or non-blood-brain barrier vasculature. However, the results of the different experimental protocols suggest that immature vascular basement membranes may have a molecular configuration that does not require an enzymatic unmasking step to react with the antisera. Alternatively, the looseness of the surrounding neuropil inherent in developing and injured CNS could allow the antisera greater access to basement membrane antigens.