Inhibition of astrocytes promotes long-distance growing nerve fibers in ventral mesencephalic cultures.
Tyrosine hydroxylase-positive nerve fiber formation occurs in two diverse morphological patterns in rat fetal ventral mesencephalic slice cultures; one is non-glial-associated and the other is glial-associated. The aim of this study was to characterize the non-glial-associated nerve fibers and its relation to migration of astrocytes. Organotypic slice cultures were prepared from embryonic days 12, 14, and 18 rat fetuses and maintained for 5, 7 or 14 days in vitro. Inhibition of cell proliferation using cytosine beta-D-arabinofuranoside was conducted in embryonic day 14 ventral mesencephalic cultures. The treatment impaired astrocytic migration at 7 and 14 days in vitro. The reduced migration of astrocytes exerted a negative effect on the glial-associated tyrosine hydroxylase-positive nerve fibers, reducing the outgrowth from the tissue slice. The non-glial-associated outgrowth was, however, positively affected by reduced astrocytic migration, reaching distances around 3mm in 2 weeks, and remained for longer time in culture. Co-cultures of fetal ventral mesencephalon and frontal cortex revealed the cortex as a target for the non-glial-associated tyrosine hydroxylase-positive outgrowth. The age of the fetal tissue at plating affected the astrocytes such that older tissue increased the length of astrocyte migration. Younger tissue at plating promoted the presence of non-glial-associated outgrowth and long radial-glia-like processes, while older tissue promoted migration of neurons instead of formation of nerve fiber network. In conclusion, inhibition of astrocytic proliferation promotes the persistence of long-distance growing tyrosine hydroxylase-positive nerve fibers in ventral mesencephalic slices cultures. Furthermore, the long-distance growing nerve fibers target the frontal cortex and are absent in cultures derived from older tissue.
af Bjerkén
Sara
S
Department of Integrative Medical Biology, Umeå University, S-901 87 Umeå, Sweden.
Marschinke
Franziska
F
Strömberg
Ingrid
I
D000367:Age Factors; D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000502:physiology*; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D016380:Brain Tissue Transplantation / Q000379:methods; D002450:Cell Communication / Q000502:physiology*; D002465:Cell Movement / Q000502:physiology; D002478:Cells, Cultured; D004298:Dopamine / Q000378:metabolism; D005911:Gliosis / Q000503:physiopathology / Q000517:prevention & control; D006085:Graft Survival / Q000502:physiology*; D020439:Growth Cones / Q000502:physiology* / Q000648:ultrastructure; D006131:Growth Inhibitors / Q000494:pharmacology; D008636:Mesencephalon / Q000166:cytology / Q000196:embryology* / Q000502:physiology*; D009434:Neural Pathways / Q000166:cytology / Q000196:embryology / Q000502:physiology; D055495:Neurogenesis / Q000502:physiology*; D009924:Organ Culture Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley; D013378:Substantia Nigra / Q000166:cytology / Q000196:embryology / Q000502:physiology; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis / Q000378:metabolism; D017557:Ventral Tegmental Area / Q000166:cytology / Q000196:embryology / Q000502:physiology
D015415:Biomarkers; D006131:Growth Inhibitors; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine