Immunoreactivity of subunits of the (Na+ + K+)-ATPase. Cross-reactivity of the alpha, alpha + and beta forms in different organs and species.

The immunologic cross-reactivity of the alpha and alpha + forms of the large subunit and the beta subunit of the (Na+ + K+)-ATPase from brain and kidney preparations was examined using rabbit antiserum prepared against the purified holo lamb kidney enzyme. As previously reported by Sweadner ((1979) J. Biol. Chem. 254, 6060-6067) phosphorylation of the large subunit of the (Na+ + K+)-ATPase in the presence of Na+, Mg2+, and [gamma-32P]ATP revealed that dog and, very likely, rat brain contain two forms of the large subunit (designated alpha and alpha +) while dog, rat, and lamb kidney contain only one form (alpha). The cross-reactivity of the alpha and alpha + forms in these preparations was investigated by resolving the subunits by SDS-polyacrylamide gel electrophoresis. The separated polypeptides were transferred to unmodified nitrocellulose paper and reacted with rabbit anti-lamb kidney serum, followed by detection of the antigen-antibody complex with 125I-labeled protein A and autoradiography. By this method, the alpha and alpha + forms of rat and dog brain, as well as the alpha form found in kidney, were shown to cross-react. In addition, membranes from human cerebral cortex were shown to contain two immunoreactive bands corresponding to the alpha and alpha + forms of dog brain. In contrast, the brain of the insect Manduca sexta contains only one immunoreactive polypeptide with a molecular weight intermediate to the alpha and alpha + forms of dog brain. The beta subunit from lamb, dog and rat kidney and from dog and rat brain cross-reacts with anti-lamb kidney (Na+ + K+)-ATPase serum. The mobility of the beta subunit from dog and rat brain on SDS-polyacrylamide electrophoresis gels is greater than the mobility of the beta subunit from lamb, rat or dog kidney.

Chemotherapy for brain tumors.

Traditionally, cytotoxic drugs have played a limited role in the treatment of brain tumors, but important advances in chemotherapy have occurred during the past decade. Certain central nervous system (CNS) malignancies are remarkably chemosensitive. These include primary CNS lymphoma, medulloblastoma, oligodendroglioma, and intracranial germ-cell tumors. This review focuses on advances in the chemotherapy of these chemosensitive tumors and also discusses the potential use of chemotherapeutic agents, both cytotoxic and cytostatic, in other brain tumors, such as glioblastomas and anaplastic astrocytomas. In addition, a brief description of future directions that may hold promise, including high-dose chemotherapy with stem-cell rescue, blood-brain barrier disruption, and regional treatment using controlled-release biodegradable polymers, is included.

Immunocytochemical localization of NA+, K+-ATPase in primary cultures of rat retina.

Conditions have been established which allow growth of embryonic rat retinal cells in dissociated cell culture for up to one month. Na+, K+-ATPase localization was studied in both neuronal and mixed neuronal-glial (flat cell) cultures. High Na+, K+-ATPase-like-immunoreactivity was associated with plasma membranes of neuronal cell bodies and their processes. Markedly lower immunoreactivity was found in the underlying flat cells in mixed cultures. Staining was generally uniform over perikaryal plasma membranes and showed a bead-like appearance in neuronal processes, supporting previous studies in brain tissue which used histocytochemical procedures specific for the Na+, K+-ATPase. This system should be useful for examining distribution of the enzyme in developing nerve and glial cells and may help to resolve questions regarding Na+-K+ homeostasis by neurons and glia.

Growth cone migration across extracellular matrix components depends on integrin, but migration across glioma cells does not.

To promote neurite elongation, nerve growth cones must adhere to other surfaces. A complex of integral membrane glycoproteins mediates cell binding to the extracellular glycoproteins fibronectin and laminin (Horwitz et al., J Cell Biol 101:2134-2144, 1985). The receptor complex, named integrin, binds to fibronectin by recognition of a specific peptide sequence, Arg-Gly-Asp-Ser (RGDS), in the fibronectin molecule (Pierschbacher and Ruoslahti, Proc Natl Acad Sci USA 81:5985-5988, 1984). We have used antibodies to integrin and an RGDS synthetic peptide to probe the functions of integrin in the migration of growth cones extended from sensory and spinal cord neurons of chick embryos. Analyses of time lapse videotapes of growth cone migration before and after adding RGDS indicated that 2 mM RGDS rapidly inhibits growth cone movement on substrata coated with fibronectin or a fragment of fibronectin containing the RGDS sequence. RGDS has no effect on growth cone movement on laminin or on a surface coated with material deposited from heart conditioned medium. However, a monclonal antibody to the integrin complex (10 micrograms/ml CSAT) completely blocks growth cone movement on substrata treated with fibronectin, laminin, or heart conditioned medium. Thus integrin may be involved in growth cone adhesion to several extracellular molecules, although the selective effects of RGDS indicate that the integrin complex may have heterogeneous sites for interaction with different components of the extracellular matrix. CSAT antibody has no discernible effect, however, on growth cone migration across the upper surfaces of C6 glioma cells. These data indicate that the surfaces of nerve growth cones contain multiple binding molecules that mediate different adhesive interactions during migration.