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Parasitology Quiz 2 Study Guide.docx

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Boston College
BIOL 2000

Nicola Kahale Parasitology Quiz 2 Guide Adult Somatic Cells in the human parasite Schistosoma mansoni • Problem: Schistosomes can live and reproduce in human host for decades • Solution/Goal: Finding the mechanism that promote their longetivity • Hypothesis: Stem Cells are able to renew tissue and sustain longetivity • Pluripotent cells called neoblasts in flatworms (planaria) • Planaria can pretty much regenerate entire body because stem cells throughout entire body Experiment 1: Special Neoblasts or Omnipotent cNeoblasts • EdU labeling to examine distribution of S phase cells • Next to Neoblasts, germ line cells in reproductive organs are also expected to be labeled o EdU modified base used instead of thymidine, contains alkyl group modified by click chemistry (add azide group = green fluorescence) • As expected cells lit up green in reproductive organs and other tissues (PSCs) • What and Where are these cells? o Co stain with Phalloidin which detects F-actin and outlines worm to see where cells are o Remove male testes for less “noise”, use enzymes to loosen cells, and find only globular cells labeled by EdU+  Supporting cNeoblasts (omnipotent)  Stained with h2b and see tails on associated with stem cells in planaria Experiment 2: What genes are expressed in PSCs • Insight from planaria – gamma radiation kills neoblasts because are fast dividing cells (makes mutatins in DNA) • Compare mRNAtranscriptome of untreated and irradiated worms (male non productive part only) - 128 genes were down regulated o 1) Isolate mRNA o 2) High throughput sequencing of mRNA o 3) Computational comparison  Transcripts expressed in differentiating cell types remain same (CathepsinB)  Transcripts expressed in dividing cells types – decrease substantially (histone, protein regulator of cytokinesis, etc.)  Other genes include stem cell associated genes (FGF, NANOS,AGO) • FGF promote prolif
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