Axonal growth and glial migration from co-cultured hippocampal and septal slices into fibrin-fibronectin-containing matrix of peripheral regeneration chambers: a light and electron microscope study.
In order to investigate whether a fibrin-fibronectin-containing matrix of a peripheral regeneration chamber could promote the growth of central nervous system neurons, hippocampal and septal slices were co-cultured in the presence of this acellular substrate. In introducing the peripheral matrix into a 2-mm-long tube between hippocampal and septal slices, a spatio-temporal sequence of cell migration and axonal growth was described by light and electron microscopy. Axons were able to elongate directly into the flocculent material constituting the matrix and a possible neurite-promoting activity was implicated in this process as axonal growth was not detected in direct contact with rat plasma coagulated with calcium, or chicken plasma coagulated with thrombin, used as control matrices. However, in the 3 different substrates tested, astrocytes were able to migrate and dilated astroglial processes containing intermediate filaments were detected. Axonal processes were observed growing on the glial cell surface. GFAP-positive phagocytic cells, that could be of the same origin as astrocytes, were involved in matrix removing. Neuronal growth and glial migration arose from hippocampal and septum slices and acetylcholinesterase-containing fibers were seen in the bridging structure suggesting that cholinergic axons were able to progress to the hippocampal slice. This technique appeared to provide a model in which axonal growth and cell migration can be studied 'in vitro' in a 3-dimensional environment.
Knoops
B
B
Laboratoire de Biologie Cellulaire, Université Catholique de Louvain, Belgium.
Hubert
I
I
Hauw
J J
JJ
van den Bosch de Aguilar
P
P
D000110:Acetylcholinesterase / Q000032:analysis; D000818:Animals; D001369:Axons / Q000502:physiology* / Q000648:ultrastructure; D002450:Cell Communication; D002465:Cell Movement; D002478:Cells, Cultured; D005337:Fibrin / Q000494:pharmacology*; D005353:Fibronectins / Q000494:pharmacology*; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D006624:Hippocampus / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D009416:Nerve Regeneration / Q000187:drug effects*; D009457:Neuroglia / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D051381:Rats; D011919:Rats, Inbred Strains; D012584:Sciatic Nerve / Q000502:physiology*; D012688:Septum Pellucidum / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure
D005353:Fibronectins; D005904:Glial Fibrillary Acidic Protein; D005337:Fibrin; D000110:Acetylcholinesterase