Cell attachment to frozen sections of injured adult mouse brain: effects of tenascin antibody and lectin perturbation of wound-related extracellular matrix molecules

• Published in: Journal of neuroscience methods Vol. 66; no. 2; pp. 99 - 108
• Main Authors: Laywell, Eric D., Friedman, Paul, Harrington, Kristy, Robertson, James T., Steindler, Dennis A.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.06.1996; Elsevier Science
• Subjects: Animals; Antibody perturbation; Antibody Specificity; Biological and medical sciences; Brain Injuries - metabolism; Brain wound; Carbocyanines; Cell Adhesion - physiology; Cell Culture Techniques - methods; Cells, Cultured - chemistry; Cells, Cultured - cytology; Cells, Cultured - physiology; Cryoculture; Cryopreservation; Extracellular Matrix Proteins - immunology; Extracellular Matrix Proteins - metabolism; Female; Fluorescent Dyes; Frozen Sections; Fundamental and applied biological sciences. Psychology; General aspects. Models. Methods; Immunohistochemistry; In vitro bioassay; Lectin perturbation; Lectins - pharmacology; Mice; Mice, Inbred ICR; Neutralization Tests; PC12 Cells - chemistry; PC12 Cells - cytology; PC12 Cells - physiology; Pregnancy; Rats; Tenascin - immunology; Vertebrates: nervous system and sense organs; Wound Healing - drug effects; Wound Healing - physiology; Lectin perturbation; Cryoculture; Brain wound; In vitro bioassay; Antibody perturbation; Cell culture; Neurite; Cold; Growth; Rodentia; Central nervous system; Tissue culture; Vertebrata; Mammalia; Mouse; Cryopreservation; Extracellular matrix; Lesion; Brain (vertebrata)
• ISSN: 0165-0270; 1872-678X
• DOI: 10.1016/0165-0270(96)00008-8

Previous studies describing the use of cryoculture methods have focused on the efficacy of the method for studying neuron attachment and neurite outgrowth on intact sections of nerve, and rodent and even human brain. The cryoculture method has shown promise for determining the presence of cell attachment- and neurite-growth-inhibiting molecules in such specimens, and some studies have also attempted to neutralize such molecules with antibodies to myelin inhibitory proteins, nerve growth factor, or factors present in conditioned media that may counteract the repulsiveness of some of these molecules preserved in sections of, for example, myelinated nerves or adult brain white matter. The present study describes the novel use of lesioned central nervous system cryocultures as substrates for investigating the attachment of embryonic neurons and PC12 cells. In addition to demonstrating the use of this novel scar substrate to extend previous ‘scar-in-a-dish’ models (David et al. (1990) Neuron, 5: 463–469; Rudge and Silver (1990) J. Neurosci., 10: 3594–3603; Rudge et al. (1989) Exp. Neurol., 103: 1–16), the present study also describes antibody and lectin perturbations of putative inhibitory molecules that result in an enhanced attachment of cells to cryosection cultures of brain and spinal cord wounds.