Cell necrosis, intrinsic apoptosis and senescence contribute to the progression of exencephaly to anencephaly in a mice model of congenital chranioschisis

Cell Death Dis. 2019 Sep 26;10(10):721. doi: 10.1038/s41419-019-1913-6.

Abstract

Exencephaly/anencephaly is one of the leading causes of neonatal mortality and the most extreme open neural tube defect with no current treatments and limited mechanistic understanding. We hypothesized that exencephaly leads to a local neurodegenerative process in the brain exposed to the amniotic fluid as well as diffuse degeneration in other encephalic areas and the spinal cord. To evaluate the consequences of in utero neural tissue exposure, brain and spinal cord samples from E17 exencephalic murine fetuses (maternal intraperitoneal administration of valproic acid at E8) were analyzed and compared to controls and saline-injected shams (n = 11/group). Expression of apoptosis and senescence genes (p53, p21, p16, Rbl2, Casp3, Casp9) was determined by qRT-PCR and protein expression analyzed by western blot. Apoptosis was measured by TUNEL assay and PI/AV flow cytometry. Valproic acid at E8 induced exencephaly in 22% of fetuses. At E17 the fetuses exhibited the characteristic absence of cranial bones. The brain structures from exencephalic fetuses demonstrated a loss of layers in cortical regions and a complete loss of structural organization in the olfactory bulb, hippocampus, dental gyrus and septal cortex. E17 fetuses had reduced expression of NeuN, GFAP and Oligodendrocytes in the brain with primed microglia. Intrinsic apoptotic activation (p53, Caspase9 and 3) was upregulated and active Caspase3 localized to the layer of brain exposed to the amniotic fluid. Senescence via p21-Rbl2 was increased in the brain and in the spinal cord at the lamina I-II of the somatosensory dorsal horn. The current study characterizes CNS alterations in murine exencephaly and demonstrates that degeneration due to intrinsic apoptosis and senescence occurs in the directly exposed brain but also remotely in the spinal cord.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amniotic Fluid / metabolism
  • Anencephaly / chemically induced
  • Anencephaly / embryology
  • Anencephaly / pathology*
  • Animals
  • Apoptosis* / drug effects
  • Apoptosis* / genetics
  • Brain / cytology
  • Brain / embryology
  • Brain / pathology*
  • Caspase 3 / metabolism
  • Caspase 9 / metabolism
  • Cellular Senescence / drug effects
  • Cellular Senescence / genetics
  • Cyclin-Dependent Kinase Inhibitor p16 / metabolism
  • Cyclin-Dependent Kinase Inhibitor p21 / genetics
  • Cyclin-Dependent Kinase Inhibitor p21 / metabolism
  • Disease Models, Animal
  • Disease Progression
  • Female
  • Mice
  • Microglia / cytology
  • Microglia / drug effects
  • Microglia / metabolism
  • Microglia / pathology
  • Necrosis / embryology
  • Necrosis / metabolism
  • Necrosis / pathology*
  • Neural Tube Defects / pathology*
  • Neurons / cytology
  • Neurons / pathology
  • Retinoblastoma-Like Protein p130 / genetics
  • Retinoblastoma-Like Protein p130 / metabolism
  • Spinal Cord / cytology
  • Spinal Cord / embryology
  • Spinal Cord / metabolism
  • Spinal Cord / pathology*
  • Tumor Suppressor Protein p53 / metabolism
  • Up-Regulation
  • Valproic Acid

Substances

  • Cyclin-Dependent Kinase Inhibitor p16
  • Cyclin-Dependent Kinase Inhibitor p21
  • Rbl2 protein, mouse
  • Retinoblastoma-Like Protein p130
  • Trp53 protein, mouse
  • Tumor Suppressor Protein p53
  • Valproic Acid
  • Casp3 protein, mouse
  • Casp9 protein, mouse
  • Caspase 3
  • Caspase 9