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Genes induced in programmed cell death of neuronal PC12 cells and developing sympathetic neurons in vivo
S Wang1, A J Dibenedetto, R N Pittman
1Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, 19104, USA.
Abstract:
To identify primary response genes induced during early stages of neuronal programmed cell death (PCD), we screened by differential hybridization a subtracted cDNA library prepared from neuronal PC12 cells deprived of NGF for 6 hr in the presence of cycloheximide. Eight induced cDNA sequences were identified and designated message up-regulated during death (mud)-1-8. To determine which cloned sequences might be involved in neuronal PCD in vivo, expression of mud genes was analyzed in developing rat superior cervical ganglia (SCG) undergoing programmed cell death, using a combination of reverse Southern, reverse transcription polymerase chain reaction (RT-PCR), and in situ hybridization. Five sequences (mud-1, -3, -5/8, -6, and -7) are induced in SCG undergoing cell death in vivo, and induction of at least three of these (mud-3, -6, and -7) occurs in neurons. Partial sequence analysis reveals that mud-1 corresponds to annexin VI; mud-3 corresponds to rat PC3, mouse TIS21; mud-4 appears to be the rat homolog of human TAFII70; mud-5 and -8 are >85% identical members of the rodent gene family of B2-transcribed repeats; and mud-6 appears to be the rat homolog of human Ring 3 and Drosophila female sterile homeotic (fsh). Mud-2 and mud-7 encode novel sequences. These new candidate genes provide markers for early stages of neuronal PCD, are potentially involved in the cell death process, and serve to expand our view of cell death control in the developing nervous system.
Insights
Researchers identified eight novel genes, termed "message up-regulated during death" (mud) genes, that are activated during early neuronal programmed cell death (PCD). Five of these mud genes are induced in developing neurons undergoing PCD in vivo, offering new insights into cell death regulation.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- Neuronal programmed cell death (PCD) is crucial for nervous system development.
- Identifying genes involved in early PCD is essential for understanding neuronal development and disease.
Purpose of the Study:
- To identify novel genes that are rapidly induced during the initial phases of neuronal PCD.
- To validate the expression of these candidate genes in vivo during neuronal cell death.
Main Methods:
- Differential hybridization screening of a subtracted cDNA library from NGF-deprived PC12 cells.
- Analysis of gene expression in developing rat superior cervical ganglia (SCG) using reverse Southern, RT-PCR, and in situ hybridization.
Main Results:
- Eight "message up-regulated during death" (mud) sequences were identified.
- Five mud genes (mud-1, -3, -5/8, -6, -7) showed induction during in vivo neuronal PCD in SCG.
- mud-3, -6, and -7 were specifically induced in neurons undergoing cell death.
- Sequence analysis identified known genes (annexin VI, PC3/TIS21, TAFII70, B2 repeats, Ring 3/fsh) and novel sequences (mud-2, -7).
Conclusions:
- Several novel genes are identified as early markers of neuronal PCD.
- These genes are potentially involved in regulating neuronal cell death during development.
- The findings expand the understanding of molecular mechanisms controlling cell death in the nervous system.