Immunocytochemical localization of a chondroitin sulfate proteoglycan in nervous tissue. I. Adult brain, retina, and peripheral nerve.

Abstract

Monospecific antibodies were prepared to a previously characterized chondroitin sulfate proteoglycan of brain and used in conjunction with the peroxidase-antiperoxidase technique to localize the proteoglycan by immunoelectron microscopy. The proteoglycan was found to be exclusively intracellular in adult cerebellum, cerebrum, brain stem, and spinal cord. Some neurons and astrocytes (including Golgi epithelial cells and Bergmann fibers) showed strong cytoplasmic staining. Although in the central nervous system there was heavy axoplasmic staining of many myelinated and unmyelinated fibers, not all axons stained. Staining was also seen in retinal neurons and glia (ganglion cells, horizontal cells, and M~iller cells), but several central nervous tissue elements were consistently unstained, including Purkinje cells, oligodendrocytes, myelin, optic nerve axons, nerve endings, and synaptic vesicles. In sympathetic ganglion and peripheral nerve there was no staining of neuronal cell bodies, axons, myelin, or Schwann cells, but in sciatic nerve the Schwann cell basal lamina was stained, as was the extracellular matrix surrounding collagen fibrils. Staining was also observed in connective tissue surrounding the trachea and in the lacunae of tracheal hyaline cartilage. These findings are consistent with immunochemical studies demonstrating that antibodies to the chondroitin sulfate proteoglycan of brain also cross-react to various degrees with certain connective tissue proteoglycans.We have previously described the isolation and properties of a 6.5S chondroitin sulfate proteoglycan from a PBS extract of rat brain (19). This proteoglycan has a relatively high protein content (56% by weight), in addition to 24% glycosaminoglycans (predominantly chondroitin 4-sulfate) and 20% glycoprotein oligosaccharides, including a series of novel O-glycosidically linked oligosaccharides that contain mannose at their proximal ends (16, 28). The brain proteoglycan differs from the prototypical cartilage proteoglycans in a number of respects including its higher protein content, smaller monomer size, and its relatively limited ability to interact with hyaluronic acid to produce larger size aggregates.Biochemical analyses (for reviews, see references 27 and 28) demonstrated the presence of significant amounts of chondroitin sulfate in neuronal cell bodies, axons, and astrocytes (isolated in bulk from brain) as well as in purified nuclei, whereas there is little or no chondroitin sulfate in oligodendroglia, myelin, mitochondria, or nerve endings (synaptosomes). Approximately half of the chondroitin sulfate is found in the soluble fraction after high speed centrifugation of a brain homogenate, and much of the remainder is loosely associated with microsomal membranes from which it is easily extractable by mild washing procedures (19,29).When neurons are isolated in bulk from rat brain and lysed by a change in tonicity or pH, 82% of the chondroitin sulfate is released together with >90% of the lactate dehydrogenase (but only 20-25% of th...

Collapsin Response Mediator Proteins of Neonatal Rat Brain Interact with Chondroitin Sulfate

“…8, B and D) are in agreement with published data (9,48,11) and presumably represent extracellular localizations of the glycosaminoglycan. No cytoplasmic CS was detected in the present study, and in the literature, evidence for cytoplasmic proteoglycans in the CNS is limited to the adult stage (49). On the other hand, CRMP-4 was found (i) in cells of the cortical plate mainly in nuclei, (ii) in the marginal zone with a reticular staining pattern very similar to the CS-staining, and (iii) in the subplate and prospective white matter (Fig.…”

Do neurons in the vertebrate CNS migrate on laminin?

“…Fibronectin has some neurite-promoting activity in vitro, but this effect appears to be limited to neurones of the peripheral nervous system (Baron-Van Evercooren et al, 1982;Rogers et al, 1983). The remaining known connective tissue and basement membrane components investigated, such as type IV collagen, which is produced by Schwann-cells (Bunge et al, 1980) and proteoglycans, which are present in the brain in vivo (Aquino et al, 1984), do not influence neurite outgrowth (Carbonetto et al, 1983;Manthorpe et al, 1983 vations suggest, together with the reported transient induction (Terranova et al, 1980;Foidart et al, 1980;Carlsson et al., of laminin in astrocytes of adult rat brain after injury (Liesi et 1981), and has an adhesive function . Since al., 1984a), that laminin may also play a role in neuronal embryonic brain and the external granule cell layer of post-natal regeneration.…”

“…There is at present no information about the interaction of laminin with neuronal cell surface molecules, such as the neural cell adhesion molecule (Edelman, 1984;Rutishauser, 1984), the neuronal surface antigen (Ng-CAM), possibly mediating neuronal attachment to glial cells in vitro (Grummet et al, 1984), the neuronal LI-antigen (Rathjen and Schachner, 1984), or proteoglycans (Aquino et al, 1984). If some of these molecules turn out to interact with laminin, they differ considerably from the 68 000 mol.…”

Transient Expression of PG‐M/Versican, a Large Chondroitin Sulfate Proteoglycan in Developing Chicken Retina

“…Several chondroitin sulfate proteoglycans have been identified and characterized in the nervous system (for review, see Small et al, 1996) and in the retina (Morris and Ting, 1981;Aquino et al, 1984;Morris, 1984;Morris et al, 1984Morris et al, , 1987Needham et a!., 1988;Brittis et al, 1992;Ring et a!., 1995 to extracellular matrix molecules, such as fibronectin, and that the activity can be attributed to chondroitin sulfate chains (Yamagata et al, 1989). Indeed, PG-MI versican-like chondroitin sulfate proteoglycan purified from sciatic nerve inhibited adhesion of several nerve cells and affected neurite outgrowth (Braunewell et al, 1995).…”