BIOLOGY 2B03 Lecture Notes - Lecture 11: Terminal Deoxynucleotidyl Transferase, Tunel Assay, Substantia Nigra

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27 Apr 2016
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2B03- Module 11: Programmed Cell Death
Cell birth: cell division during embryogenesis
-Cells are always added to our body by mitosis and taken away by apoptosis
-Network of proteins is required to signal and regulate apoptosis
Ex. Forming hands and feet of human embryo
-Week 6: skin forms webbing between digits on hands and feed
-Week 11: Webbing disappears and digits are resolved; without cell death, webbing remains
TUNEL Assay
-Detects nicks in the DNA of apoptotic cells
Experiment: Mouse embryos were stained with a dye so cells
undergoing apoptosis are bright green
-Seen between digits
-TUNEL assay takes advantage that DNA in apoptosis contains DNA breaks or nicks
-dUTP is added into the nicks by using the enzyme terminal deoxynucleotidyl transferase
oFluorescently labelled dUTP allows for the detection seen via bright green
Ex. Tadpoles undergo cellular apoptosis to lose the tail. Apoptosis in the tail is stimulated by increase in thyroid hormone in blood, which is
also seen in tail loss during human embryonic development
Apoptosis during Development of Nervous System
-During brain development, about half the neurons
undergo apoptosis
oThose cells are ones that don’t have synaptic
connections, or are faulty
oEssential process for development of nervous
system: matches the number of nerve cells
with target cells
Inappropriate Cell Death
-Cell death is part of normal cell turnover, but
inappropriate cell death can lead to disease
-Alzheimers: neurons in hippocampus and regions of the cerebral cortex die
-Huntingtons: neurons in the striatum die
-Parkinson’s Disease: dopamine neurons in the substantia nigra die
-Seenin Duchenne Musuclar Dystrophy: muscular cell death
Ways Cells Die
1. Necrosis
oDamage causes the cell to swell and release contents into surrounding tissues
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