Cell lineage tracing approaches

jambomber's version from 2015-05-18 10:56


Question Answer
What does lineage tracing allow us to do?Follow destinies of a cells progeny and assesses a cells potency - limits of its developmental potential
Why can tissue specific makers alone not be used to follow cell fate?Increasingly recognised as plastic- cells moving in and out of the expression domain alter their cell surface to local environment
Where was lineage tracing investigatedEarly pigemented blastomeres of Tunicates by Whitman + Wilson
Why is it diffuclt to lineage trace in c elegansNo pigmentation or morpho basis for discriminatin b/w
What hinders direct observations usefulness?becomes difficult to unambiguously det a cells descendants especiall w/in oparque regions
What serves as an alternative to vital dyesTrithiated thymidine 3H radio labels
What is a weakness about vital dyes?Since they are water soluble can exchange b/w cells
How used pigmentation to address fate?Spemann
What solved problem of water solubilityDi) and I are lipophlllcs
What is a problem with dye labelling? How solvedDifficult to apply to single cell. Can only track fate of groups. Use flourodextrans +HRP instead
What is the problem w/ retroviral tranfectionRetrospective mapping - cant select a cell to be labelled or be certain that independant events
What is the main conclusion: No univerasally perfect method. Choice method is dictated by question being addressed.
Give a use for DiI and OQuickly found applications first for the tracing of axon pathways
What could be said about the trend?Evolivng in sophistication
Why was quail chick so important quail cells can be followed long-term in the chick environment, this technique allowed for not only the mapping of neural crest migratory pathways but also the delineation of the full range of derivatives that formed from the neural crest in amniote embryos
Disadvantage to quail chickequires surgical grafting between non-identical species, which may introduce artifacts
What is a big plus of DiO/I they can be applied to many different vertebrates, allowing for comparative analysis across species.
How NC pops labelled in Z fish The availability of transgenic zebrafish lines that label neural crest populations with Sox10 (Rodrigues et al., 2012) or FoxD3 (Hochgreb-Hagele and Bronner, 2013) circumvents this difficulty and enable long-term studies of neural crest migration and contributions to subsets of derivatives
Example of a Cre lox pop trace Pax3 lineage labels a population of melanocyte precursors, that then require repression of Pax3 to execute the full melanocytic program
What is the most recent disc from lineage tracingR mice hdemonstrated that much of the parasympathetic system arises from neural crest-derived glial cells, which appear to be Schwann cell precursors that are resident in nerves. These bi-potent progenitors appear to be able to generate both glia and neurons. In addition, the precursors appear to contribute to melanocytes (Adameyko et al., 2009) and thus may represent a type of neural crest stem cell that remains resident along peripheral nerves.
Where next for studiesvisualize the process by which peripheral nervous system precursors move from their site of origin to their final sites in the developing PNS.
what advances tech imaging technologies like two photon light sheet microscopy have advanced enormously, increasing resolution as well as depth of focus into intact tissue.
What is wrong with cre loxmany of the promoters used to study early neural crest formation control the expression of genes that are expressed only after neural crest induction, or are not specific to the neural crest cells. Importantly, the most widely used Cre line (Wnt1-Cre) recently has been shown to cause activation of Wnt signaling in the midbrain

Recent badges