title: Early differential cell death and survival mechanisms initiate and contribute to the development of OPIDN: a study of molecular, cellular, and anatomical parameters.
abstract: Organophosphorus-ester induced delayed neurotoxicity (OPIDN) is a neurodegenerative disorder characterized by ataxia progressing to paralysis with a concomitant central and peripheral, distal axonapathy. Diisopropylphosphorofluoridate (DFP) produces OPIDN in the chicken that results in mild ataxia in 7-14 days and severe paralysis as the disease progresses with a single dose. White leghorn layer hens were treated with DFP (1.7 mg/kg, sc) after prophylactic treatment with atropine (1mg/kg, sc) in normal saline and eserine (1mg/kg, sc) in dimethyl sulfoxide. Control groups were treated with vehicle propylene glycol (0.1 ml/kg, sc), atropine in normal saline and eserine in dimethyl sulfoxide. The hens were euthanized at different time points such as 1, 2, 5, 10 and 20 days, and the tissues from cerebrum, midbrain, cerebellum, brainstem and spinal cord were quickly dissected and frozen for mRNA (northern) studies. Northern blots were probed with BCL2, GADD45, beta actin, and 28S RNA to investigate their expression pattern. Another set of hens was treated for a series of time points and perfused with phosphate buffered saline and fixative for histological studies. Various staining protocols such as Hematoxylin and Eosin (H&E); Sevier-Munger; Cresyl echt Violet for Nissl substance; and Gallocynin stain for Nissl granules were used to assess various patterns of cell death and degenerative changes. Complex cell death mechanisms may be involved in the neuronal and axonal degeneration. These data indicate altered and differential mRNA expressions of BCL2 (anti apoptotic gene) and GADD45 (DNA damage inducible gene) in various tissues. Increased cell death and other degenerative changes noted in the susceptible regions (spinal cord and cerebellum) than the resistant region (cerebrum), may indicate complex molecular pathways via altered BCL2 and GADD45 gene expression, causing the homeostatic imbalance between cell survival and cell death mechanisms. Semi quantitative analysis revealed that the order of severity of damage declines from the spino-cerebellar, ventral, and dorsal tract respectively, suggesting neuroanatomical specificity. Thus, early activation of cell death and cell survival processes may play significant role in the clinical progression and syndromic clinical feature presentation of OPIDN.
authors.affiliation: Dept of Medicine, Duke University Medical Center, Durham, NC, USA. tdamodar@nccu.edu
authors.forename: T V
authors.initials: TV
authors.lastname: Damodaran
authors.forename: M K
authors.initials: MK
authors.lastname: Attia
authors.forename: M B
authors.initials: MB
authors.lastname: Abou-Donia
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001285:Atropine / Q000494:pharmacology; D015152:Blotting, Northern; D001921:Brain / Q000187:drug effects / Q000473:pathology; D016923:Cell Death / Q000187:drug effects* / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects* / Q000502:physiology; D002645:Chickens; D002800:Cholinesterase Inhibitors / Q000633:toxicity; D005260:Female; D007531:Isoflurophate / Q000633:toxicity; D020258:Neurotoxicity Syndromes / Q000378:metabolism* / Q000473:pathology; D009943:Organophosphorus Compounds / Q000633:toxicity*; D010830:Physostigmine / Q000494:pharmacology; D013116:Spinal Cord / Q000187:drug effects / Q000473:pathology; D013997:Time Factors
chemical_list: D002800:Cholinesterase Inhibitors; D009943:Organophosphorus Compounds; D007531:Isoflurophate; D001285:Atropine; D010830:Physostigmine