Detection of apoptosis during planarian regeneration by the expression of apoptosis-related genes and TUNEL assay

Jung Shan Hwang, Chiyoko Kobayashi, Kiyokazu Agata, Kazuho Ikeo, Takashi Gojobori*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

78 Scopus citations

Abstract

Apoptosis is a tightly organized cell death process that plays a crucial role in metazoan development, but it has not yet been revealed whether apoptotic events are involved in the process of regeneration. Here, we tried to detect apoptotic cells during planarian regeneration using the TdT-mediated dUTP nick-end labeling (TUNEL) assay as well as the expression of apoptosis-related genes. Three novel cDNAs were isolated from a planarian cDNA library and shown to be closely related to other metazoan caspases at the amino acid sequence level. One of these cDNAs, Caspase-like gene 3 (DjClg3), was expressed primarily in apoptotic cells by double detections with the TUNEL assay. Whole mount in situ studies indicated that DjClg3 was expressed in the cells of the mesenchymal space and also around the pharynx of the intact body. Its expression in the regenerating head piece was seen in the blastema and less significantly in the brain, while in the regenerating tail piece, DjClg3 expression was detected uniformly throughout the entire region. In parallel experiments, we performed in situ TUNEL assays to localize the regions where cell death occurred during regeneration and comparable results to the DjClg3 expression patterns were obtained. This is the first report to show that planarians have apoptosis-related genes and the results suggest that the apoptotic mechanism probably takes place to a large extent in normal intact worms as well as during their regeneration. We hypothesize that the presence of apoptosis in planarians may have a role in controlling cell numbers, eliminating unnecessary tissues or cells and remodeling the old tissues of regenerating body parts.

Original languageEnglish (US)
Pages (from-to)15-25
Number of pages11
JournalGene
Volume333
Issue numberSUPPL.
DOIs
StatePublished - May 26 2004

Keywords

  • Blastema
  • Caspase
  • EST
  • NBT
  • Platyhelminthes
  • Programmed cell death
  • RACE
  • Remodeling
  • TUNEL
  • TdT
  • TdT-mediated dUTP nick-end labeling
  • expressed sequence tag
  • rapid amplification of cDNA ends
  • terminal deoxynucleotidyl transferase

ASJC Scopus subject areas

  • Genetics

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