title: Dicyclomine, an M1 muscarinic antagonist, reduces biomarker levels, but not neuronal degeneration, in fluid percussion brain injury.
abstract: Recent studies indicate that alphaII-spectrin breakdown products (SBDPs) have utility as biological markers of traumatic brain injury (TBI). However, the utility of SBDP biomarkers for detecting effects of therapeutic interventions has not been explored. Acetylcholine plays a role in pathological neuronal excitation and TBI-induced muscarinic cholinergic receptor activation may contribute to excitotoxic processes. In experiment I, regional and temporal changes in calpain-mediated alpha-spectrin degradation were evaluated at 3, 12, 24, and 48 h using immunostaining for 145-kDa SBDP. Immunostaining of SBDP-145 was only evident in the hemisphere ipsilateral to TBI and was generally limited to the cortex except at 24 h when immunostaining was also prominent in the dentate gyrus and striatum. In Experiment II, cerebral spinal fluid (CSF) samples were analyzed for various SBDPs 24 h after moderate lateral fluid percussion TBI. Rats were administered either dicyclomine (5 mg/kg i.p.) or saline vehicle (n = 8 per group) 5 min prior to injury. Injury produced significant increases (p < 0.001) of 300%, 230%, and >1000% in SBDP-150, -145, and -120, respectively in vehicle-treated rats compared to sham. Dicyclomine treatment produced decreases of 38% (p = 0.077), 37% (p = 0.028), and 63% (p = 0.051) in SBDP-150, -145, and -120, respectively, compared to vehicle-treated injury. Following CSF extraction, coronal brain sections were processed for detecting degenerating neurons using Fluoro-Jade histofluorescence. Stereological techniques were used to quantify neuronal degeneration in the dorsal hippocampus CA2/3 region and in the parietal cortex. No significant differences were detected in numbers of degenerating neurons in the dorsal CA2/3 hippocampus or the parietal cortex between saline and dicyclomine treatment groups. The percent weight loss following TBI was significantly reduced by dicyclomine treatment. These data provide additional evidence that, as TBI biomarkers, SBDPs are able to detect a therapeutic intervention even in the absence of changes in neuronal cell degeneration measured by Fluoro-jade.
authors.affiliation: Department of Neurological Surgery, University of California at Davis, Davis, California 95616-8797, USA.
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Cox
authors.forename: Eric J
authors.initials: EJ
authors.lastname: West
authors.forename: Ming Cheng
authors.initials: MC
authors.lastname: Liu
authors.forename: Kevin K W
authors.initials: KK
authors.lastname: Wang
authors.forename: Ronald L
authors.initials: RL
authors.lastname: Hayes
authors.forename: Bruce G
authors.initials: BG
authors.lastname: Lyeth
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000134:cerebrospinal fluid; D015153:Blotting, Western; D001930:Brain Injuries / Q000134:cerebrospinal fluid* / Q000188:drug therapy* / Q000473:pathology; D002154:Calpain / Q000378:metabolism; D020169:Caspases / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D004025:Dicyclomine / Q000627:therapeutic use*; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007150:Immunohistochemistry; D008297:Male; D018727:Muscarinic Antagonists / Q000627:therapeutic use*; D009410:Nerve Degeneration / Q000188:drug therapy* / Q000473:pathology; D009930:Organic Chemicals; D051381:Rats; D017207:Rats, Sprague-Dawley; D043643:Receptor, Muscarinic M1 / Q000037:antagonists & inhibitors*; D013049:Spectrin / Q000378:metabolism
chemical_list: D015415:Biomarkers; D005452:Fluoresceins; D005456:Fluorescent Dyes; D018727:Muscarinic Antagonists; D009930:Organic Chemicals; D043643:Receptor, Muscarinic M1; C435731:fluoro jade; D013049:Spectrin; D004025:Dicyclomine; D002154:Calpain; D020169:Caspases