Biochemical Studies of Neuronal and Other Cell Types

Brenneman, D E.

Abstract

Cell cultures from the fetal mammalian central nervous system were used to study the regulation of neurodevelopment by neuropeptides, trophic factors and electrical activity. Vasoactive intestinal peptide was shown to increase the number of astorcytes during development in culture. Radioligand binding studies on whole cell and membrane preparations of astrocytes indicated the presence of high affinity receptors for VIP. Inositol phospholipid turnover was shown to be increased in astrocytes after stimulation with norepinephrine and bradykinin, but not with VIP treatment. Viral pentapeptides (TTSYT and TINYT) with sequence homology to VIP(7-11) were shown to exhibit VIP-like increases in neuronal survival during electrical blockade. Feasibility studies using DEAE sephacel, affinity column (wheat germ agglutinin, Concanavalin A and heparin), and Amicon filtration indicated that an activity-related neuron survival factor and a choline acetyltransferase (CAT)-stimulating factor did retain biological activity through these separation procedures. A number of possible sources for the factors have been found and potential second messenger systems have been identified. The survival of spinal cord neurons was shown to be influenced by NMDA antagonists. Activity blockade with TTX and the GABAergic agonist muscimol were shown to produce a reversible and developmentally sensitive decrease in the cells immunoreactive to methionine-enkephalin in spinal cord cultures.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
• Type: Intramural Research (Z01)
• Project #: 1Z01HD000047-19
• Application #: 3942006
• Support Year: 19
• Fiscal Year: 1987

Institution

• Name: U.S. National Inst/Child Hlth/Human Dev
• Country: United States

Publications

Romano, Jacob; Beni-Adani, Liana; Nissenbaum, Orlev Levy et al. (2002) A single administration of the peptide NAP induces long-term protective changes against the consequences of head injury: gene Atlas array analysis. J Mol Neurosci 18:37-45

Steingart, Ruth A; Heldenberg, Eitan; Pinhasov, Albert et al. (2002) A vasoactive intestinal peptide receptor analog alters the expression of homeobox genes. Life Sci 71:2543-52

Leker, Ronen R; Teichner, Angella; Grigoriadis, Nikolas et al. (2002) NAP, a femtomolar-acting peptide, protects the brain against ischemic injury by reducing apoptotic death. Stroke 33:1085-92

Ashur-Fabian, O; Giladi, E; Furman, S et al. (2001) Vasoactive intestinal peptide and related molecules induce nitrite accumulation in the extracellular milieu of rat cerebral cortical cultures. Neurosci Lett 307:167-70

Zamostiano, R; Pinhasov, A; Gelber, E et al. (2001) Cloning and characterization of the human activity-dependent neuroprotective protein. J Biol Chem 276:708-14

Spong, C Y; Abebe, D T; Gozes, I et al. (2001) Prevention of fetal demise and growth restriction in a mouse model of fetal alcohol syndrome. J Pharmacol Exp Ther 297:774-9

Servoss, S J; Lee, S J; Gibney, G et al. (2001) IGF-I as a mediator of VIP/activity-dependent neurotrophic factor-stimulated embryonic growth. Endocrinology 142:3348-53

Porter, A E; Auth, J; Prince, M et al. (2001) Optimization of cytokine stability in stored amniotic fluid. Am J Obstet Gynecol 185:459-62

Beni-Adani, L; Gozes, I; Cohen, Y et al. (2001) A peptide derived from activity-dependent neuroprotective protein (ADNP) ameliorates injury response in closed head injury in mice. J Pharmacol Exp Ther 296:57-63

Zia, H; Leyton, J; Casibang, M et al. (2000) (N-stearyl, norleucine17) VIP hybrid inhibits the growth of pancreatic cancer cell lines. Life Sci 66:379-87