title: Pax6 influences expression patterns of genes involved in neuro- degeneration.
abstract: BACKGROUND
  Pax6, a highly conserved multifunctional transcription factor, has been critical for neurogenesis and neuronal plasticity. It is presumed that if level of Pax6 approaches either low or null, critical genes responsible for maintaining functional status of neurons or glia would be modulated.
  
  
  OBJECTIVE
  Therefore, it has been intended to explore possibility of either direct or indirect influence of Pax6 in neurodegeneration.
  
  
  METHODS
  The cell lines having origin of murine embryonic fibroblast (Pax6-non expressing, NIH3T3-cell line), murine neuroblastoma (Pax6-expressing brain-derived, Neuro-2a-cell line), and human glioblastoma-astrocytoma (U87MG) were cultured and maintained in a CO2 incubator at 37°C and 5% CO2 in DMEM containing 10% fetal bovine serum. The knockdown of endogenous Pax6 in Neuro-2a cells was achieved through siRNA based gene knock-down approach. The efficiency and validation of knock-down was done by real time PCR. The knock-down of Pax6 was successfully achieved.
  
  
  RESULTS
  The levels of expression of transcripts of some of the proposed putative markers of neurodegeneration like Pax6, S100β, GFAP, BDNF, NGN2, p73α, p73δ, LDH, SOD, and Catalase were analyzed in Pax6 knockdown condition for analysis of role of Pax6 in neurodegeneration. Since the Pax6 has been proposed to bind to promoter sequences of catalase, and catalase suppresses TGFβ, relative lower levels of catalase in Neuro-2a and U-87MG as compared to NIH-3T3 indicates a possible progressive dominant negative impact of Pax6. However, presence of SOD and LDH indicates alternative protective mechanism.
  
  
  CONCLUSIONS
  Presence of BDNF and TGFβ indicates association between them in glioblastoma-astrocytoma. Therefore, Pax6 seems to be involved directly with p53 and TGFβ mediated pathways and indirectly with redox-sensitive pathway regulation. The neurodegenerative markers S100β, GFAP, BDNF, NGN2, p73α, p73δ, observed downregulated in Pax6 knockdown condition suggest Pax6-mediated regulation of these markers. Observations enlighten Pax6-mediated influences on cascades of genes involved in growth, differentiation and maturation of neurons and glia.
authors.affiliation: Biochemistry and Molecular Biology Lab, Department of Zoology, Banaras Hindu University, Varanasi- 221005, INDIA.
authors.forename: Suman
authors.initials: S
authors.lastname: Mishra
authors.affiliation: Biochemistry and Molecular Biology Lab, Department of Zoology, Banaras Hindu University, Varanasi- 221005, INDIA.
authors.forename: Shashank Kumar
authors.initials: SK
authors.lastname: Maurya
authors.affiliation: Biochemistry and Molecular Biology Lab, Department of Zoology, Banaras Hindu University, Varanasi- 221005, INDIA.
authors.forename: Khushboo
authors.initials: K
authors.lastname: Srivastava
authors.affiliation: Biochemistry and Molecular Biology Lab, Department of Zoology, Banaras Hindu University, Varanasi- 221005, INDIA.
authors.forename: Sachin
authors.initials: S
authors.lastname: Shukla
authors.affiliation: Biochemistry and Molecular Biology Lab, Department of Zoology, Banaras Hindu University, Varanasi- 221005, INDIA.
authors.forename: Rajnikant
authors.initials: R
authors.lastname: Mishra
keywords: BDNF; Human glioblastoma; LDH; Pax6; SOD