Phenotype comparisons of ALDH1L1 BAC-TRAP mice under control and neurotoxic (MPTP) conditions

Details

• Personal Author:
  Kelly KA ; Locker AR ; Michalovicz LT ; Miller DB ; O'Callaghan JP
• Description:
  A central problem in neurotoxicology is detecting selective and unpredictable damage to specific cells produced by toxic agents/mixtures. With few biomarkers of damage known, evaluating astrogliosis by measuring GFAP overcomes this problem as reactive astrocytes show the location of toxicant-induced damage occurring anywhere in the CNS. Thus, determining specific in vivo transcriptomic profiles of astrocytes under control and reactive conditions will identify additional astrogliosis biomarkers. Heintz and Greengard (2008) introduced BAC-TRAP (translating ribosome affinity purification) technology that allows the characterization of the actively translating transcriptome of a particular cell type. Using multiple models of neurotoxicity we confirmed that ALDH1L1 is an astrocyte specific "housekeeping" gene/protein. Thus, ALDH1L1 BAC-TRAP (ALB-T) mice can be used to characterize the transcriptome of astrocytes under various conditions. Here, we characterize the phenotype of the 0, 1, and 2 copy ALB-T mouse (18 - 26 weeks old) and C57BL6/J (WT) mice, in both sexes, under control conditions and in response to neurotoxic damage using the well characterized dopaminergic neurotoxicant MPTP (12.5 mg/kg, s.c.) at 72 hrs post dosing. Brain, thymus, spleen, liver, kidney, adrenal gland, testes, uterus and ovaries were removed and weights recorded. Additionally, the olfactory bulbs, cerebellum, hypothalamus, hippocampus, striatum, cortex, midbrain, brain stem and pituitary brain regions were dissected and weights recorded. Striatum was analyzed for astrogliosis (GFAP), nerve terminal damage (tyrosine hydroxylase (TH) and dopamine (DA) and results indicate no basal differences in GFAP, TH, or DA in control 0, 1, or 2 copy ALB-T when compared to WT mice. Although ALB-T mice showed greater increases in GFAP protein after MPTP exposure when compared to WT the TH and DA decreases were less than those of WT mice. Striatal tissue obtained at 48 hrs post MPTP was subjected to TRAP to isolate actively translating RNA in astrocytes and changes in the transcriptome determined by microarray (Illumina) and the dataset interrogated with Ingenuity Pathway Analysis. MPTP induced robust transcriptome changes revealing genes both expected to change with astrogliosis and not previously attributed to astrocytes. Our data indicate that BAC-TRAP technology can be used to identify additional biomarkers of astrogliosis and will aid in characterizing various astrocyte phenotypes. [Description provided by NIOSH]
• Subjects:
  Animals Biomarkers Laboratories Nervous System Nervous System Disorders Occupational Health Safety
• Keywords:
  Analytical-processes Animal-studies Central-nervous-system Central-nervous-system-disorders Laboratory-animals Neurological-reactions Neurological-system Neurophysiology Neurotoxic-effects Neurotoxicity Neurotoxicology Neurotoxins

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• Publisher:
  Society for Neuroscience
• Document Type:
  Text
• Genre:
  Abstract or Summary
• Place as Subject:
  OSHA Region 3 ; West Virginia
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Division:
  HELD - Health Effects Laboratory Division
• Topic:
  Workplace Safety & Health
• Location:
  North America
• NIOSHTIC Number:
  nn:20047012
• Citation:
  Neuroscience 2015, October 17-21, 2015, Chicago, Illinois. Washington, DC: Society for Neuroscience, 2015 Oct; :692.07/L30
• Federal Fiscal Year:
  2016
• Peer Reviewed:
  False
• Source Full Name:
  Neuroscience 2015, October 17-21, 2015, Chicago, Illinois
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:ccf87ef34232e2658357e7854f86586be7703f0309e4b6b46fdc3eb4f254feb9ae76909a13cafa0b89b447167a78588d9a374a1a126dcda2d8ee90f122010794
• Download URL:
  https://stacks.cdc.gov/view/cdc/202249/cdc_202249_DS1.pdf
• File Type:
  [PDF - 77.16 KB ]