title: Acetylcholine release from fetal tissue homotopically grafted to the motoneuron-depleted lumbar spinal cord. An in vivo microdialysis study in the awake rat.
abstract: Grafts of spinal cord (SC) tissue can survive and develop into the severed SC, but no conclusive data are available concerning the functional activity of transplanted neurons. In the present study, suspensions of prelabeled embryonic ventral SC tissue were grafted to the lumbar SC of rats with motoneuron loss induced by perinatal injection of volkensin. Eight to ten months post-grafting, acetylcholine (ACh) release was measured by microdialysis in awake rats, under either basal or stimulated conditions. In normal animals, baseline ACh output averaged 1.6 pmol/30 microl, it exhibited a 4-fold increase after KCl-induced depolarization or handling, and it was completely inhibited by tetrodotoxin administration. Moreover, ACh levels did not change following acute SC transection performed under anesthesia during ongoing dialysis, suggesting an intrinsic source for spinal ACh. Treatment with volkensin produced a severe (>85%) motoneuronal loss accompanied by a similar reduction in baseline ACh release and almost completely abolished effects of depolarization or handling. In transplanted animals, many motoneuron-like labeled cells were found within and just outside the graft area, but apparently in no case were they able to extend fibers towards the denervated muscle. However, the grafts restored baseline ACh output up to near-normal levels and responded with significantly increased release to depolarization, but not to handling. The present findings indicate that spinal neuroblasts can survive and develop within the motoneuron-depleted SC and release ACh in a near-normal, but apparently non-regulated, manner. This may be of importance for future studies involving intraspinal stem cell grafts.
authors.affiliation: Department of Physiological Sciences, University of Catania, Viale A. Doria 6, 95125 Catania, Italy.
authors.forename: Rosario
authors.initials: R
authors.lastname: Gulino
authors.forename: Tiziana
authors.initials: T
authors.lastname: Cataudella
authors.forename: Fiorella
authors.initials: F
authors.lastname: Casamenti
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Pepeu
authors.forename: Stefania
authors.initials: S
authors.lastname: Stanzani
authors.forename: Giampiero
authors.initials: G
authors.lastname: Leanza
mesh_terms: D000109:Acetylcholine / Q000378:metabolism*; D000818:Animals; D002452:Cell Count; D005260:Female; D016332:Fetal Tissue Transplantation; D005333:Fetus / Q000378:metabolism; D008159:Lumbar Vertebrae; D008297:Male; D017551:Microdialysis; D009046:Motor Neurons / Q000473:pathology*; D010812:Physical Stimulation; D011189:Potassium Chloride / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000187:drug effects / Q000196:embryology* / Q000473:pathology / Q000601:surgery*; D013119:Spinal Cord Injuries; D013779:Tetrodotoxin / Q000494:pharmacology
chemical_list: D013779:Tetrodotoxin; D011189:Potassium Chloride; D000109:Acetylcholine