Fibroblast-like cells from rat plantar skin and neurotrophin-transfected 3T3 fibroblasts influence neurite growth from rat sensory neurons in vitro.
Our previous finding that skin-derived and muscle-derived molecules can be used to sort regenerating rat sciatic nerve axons evoked questions concerning neuron-target interactions at the level of single cells, which prompted the present study. The results show that dorsal root ganglion (DRG) neurons co-cultured with fibroblast-like skin-derived cells emit many neurites. These have a proximal linear segment and a distal network of beaded branches in direct relation to skin-derived cells. Electron microscopic examination of such co-cultures showed bundles of neurites at some distance from the target cells and single profiles closely apposed to subjacent cells. RNase protection assay revealed that cultivated skin-derived cells express nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and neurotrophin-4 (NT-4). In co-cultures of DRG neurons and 3T3 fibroblasts overexpressing either of the neurotrophins produced by skin-derived cells the picture varied. NT-3 transfected 3T3 fibroblasts gave a growth pattern similar to that seen with skin-derived cells. Neurons co-cultured with mock-transfected 3T3 fibroblasts were small and showed weak neurite growth. In co-cultures with a membrane insert between skin-derived cells or 3T3 fibroblasts and DRG neurons few neurons survived and neurite growth was very sparse. We conclude that skin-derived cells stimulate neurite growth from sensory neurons in vitro, that these cells produce NGF, BDNF, NT-3 and NT-4 and that 3T3 fibroblasts producing NT-3 mimic the effect of skin-derived cells on sensory neurons in co-culture. Finally the results suggest that cell surface molecules are important for neuritogenesis.
Jerregård
H
H
Division of Cell Biology, Department of Biomedicine and Surgery, Faculty of Health Sciences, University of Linköping, Linköping, Sweden.
Akerud
P
P
Arenas
E
E
Hildebrand
C
C
D016475:3T3 Cells / Q000166:cytology / Q000637:transplantation*; D000818:Animals; D000831:Animals, Newborn; D002450:Cell Communication / Q000235:genetics*; D002454:Cell Differentiation / Q000502:physiology; D048429:Cell Size / Q000502:physiology; D002478:Cells, Cultured / Q000166:cytology / Q000637:transplantation*; D018920:Coculture Techniques / Q000379:methods; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism / Q000637:transplantation; D005727:Ganglia, Spinal / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism; D017612:Gold Compounds / Q000493:pharmacokinetics; D007150:Immunohistochemistry; D007633:Keratins / Q000378:metabolism; D051379:Mice; D009414:Nerve Growth Factors / Q000235:genetics / Q000378:metabolism*; D009416:Nerve Regeneration / Q000235:genetics; D016501:Neurites / Q000378:metabolism* / Q000648:ultrastructure; D009475:Neurons, Afferent / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013869:Thiolester Hydrolases / Q000378:metabolism; D016378:Tissue Transplantation; D014162:Transfection; D043222:Ubiquitin Thiolesterase
D017612:Gold Compounds; D009414:Nerve Growth Factors; D007633:Keratins; D013869:Thiolester Hydrolases; D043222:Ubiquitin Thiolesterase