Glutathione S-transferase Pi (Gstp) proteins regulate neuritogenesis in the developing cerebral cortex

By reverse transcription-polymerase chain reaction analysis, we found that Gstp1 was expressed beginning at E15.5 in the cortex, but Gstp2 and 3 started expressing at E18.5

Xiaonan Liu; Sara M Blazejewski; Sarah A Bennison; Kazuhito Toyo-oka

2021

Scholarcy highlights

  • Neurite formation includes neurite initiation and neurite elongation, and is the process where a neuron starts to gain morphological polarity
  • Gstp proteins are expressed during cortical development, and their polarized distribution was observed during neurite formation A previous report showed that Gstp1, 2, and 3 had different expression patterns in the mouse brain
  • We found that knockdown of Gstp1 and 2 caused significant defect in neurite number from the soma compared to the scramble neurons, reflecting the decrease of the number of basal dendrites, because we observed the correct formation of the apical dendrite in the KD neurons at P15
  • Sholl analysis showed a decrease in intersections only close to soma when Gstp1 and 2 were knocked down, and there are no defects in branching pattern at distal regions from the soma. 419 Together, these results suggest that Gstp 1 and 2 are essential for proper morphogenesis of the apical dendrite at the early stage of neurite formation and the proper initial formation of basal dendrites at the later stage. 423 Time-lapse live imaging revealed the importance of Gstp1 and 2 in the orientation of neurites in neurite initiation We found that Gstp1 and 2 are essential for neurite initiation in vitro and in vivo
  • In this study, we established that Gstp proteins are essential for neurite formation, neurite initiation
  • Our in vivo experiments indicate that Gstp 621 KD neurons were not eliminated in vivo for 19 days after in utero electroporation at E15.5, suggesting that the results obtained from Gstp1 and 2 knockdown here were not the secondary effects of an unhealthy condition. 625 Our analysis of the expression of each Gstp isoform during cortical development by using RT626 PCR with the isoform-specific primers revealed that Gstp1, but not Gstp2 or 3, is the major Gstp protein expressed during embryonic cortical development, especially at the early stages
  • KD of Gstp1 599 and 2 did not affect the velocity of growth and retraction in the neurons, by quantifying the frequency of growth and retraction of neurites, we found that the KD cells retracted neurites less frequently, indicating that this is the reason why the length of the apical dendrite caught up to the one of the scramble neurons after 250 minutes of the live imaging 603

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