Genes Induced in Programmed Cell Death of Neuronal PC12 Cells and Developing Sympathetic Neuronsin Vivo

AbstractTo 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 designatedmessage up-regulated during death (mud)-1–8.To determine which cloned sequences might be involved in neuronal PCDin vivo,expression ofmudgenes 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), andin situhybridization. Five sequences (mud-1, -3, -5/8, -6,and-7) are induced in SCG undergoing cell deathin vivo,and induction of at least three of these (mud-3, -6, and -7) occurs in neurons. Partial sequence analysis reveals thatmud-1corresponds toannexin VI; mud-3corresponds to ratPC3,mouseTIS21; mud-4appears to be the rat homolog of humanTAFII70; mud-5and-8are >85% identical members of the rodent gene family of B2-transcribed repeats; andmud-6appears to be the rat homolog of humanRing 3and Drosophilafemale sterile homeotic(fsh).Mud-2andmud-7encode 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