L7 Neural development and regeneration Flashcards

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1
Q

CNS

A

brain, brainstem, spinal cord, eye

CNS controls higher order thoughts, integrates and mounts co-ordinated responses to environmental signals

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2
Q

Neuron

A

Nerve cell

define information flow: dendrite (input), soma (integration), axon (action potential - output), synapse (connection)

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3
Q

Neural circuit

A

function of each neuron depends entirely on its connections i.e. neural circuitry, which can be depicted in different ways

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4
Q

neurodevelopmental disorders

A

see onenote

genetic disorder that affect the normal central nervous system

some genes are important in the proper generation of neurons, specific regions/neural types or neural circuits

depends on stage of neurogenesis and whether the gene function is confined to a particular part of the CNS

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5
Q

neurodevelopmental vs neurodegenerative

A

genetic disorder that affect the normal central nervous system

term indicates when the phenotype occurs during development or is only observed later in life
- gives us a hint to the function of the affected gene(s)

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6
Q

Proliferation, fate, migration, terminal differentiation

A

see onenote

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7
Q

Retina as neural development model

A

see onenote

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8
Q

Multipotency

A

see onenote

Cells can generate multiple types

fate specification: which fate genes tell these progenitors what neuron type to make?

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9
Q

Gene networks

A

see onenote slides

What combination in what sequence makes what neuron?

combine transgenes to watch gene expression with live imaging

cannot be examined in species where we have to rely on post-mortem analysis of fixed non-living tissue/cells

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10
Q

Lineage experiments

A

see onenote slides

Lineage describes gene networks in each progenitor over time - allows us to determine combination and sequence of genes

Lineage experiment: examine temporal sequence of gene expression in progenitors, does each progenitor only turn on 1 fate gene?

Anthonal 5 to make photoreceptor and ganglion cells

Visual homeobox genes 1 and 2 to make bipolar cells

Pancreas TF 1 a to make horizontal amacrine cells?

live imagine revealed more complex and very stereotypical gene interactions within progenitors refuting the one gene one fate theory

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11
Q

Gene loss of function

A

see onenote slides

Necessity of a gene (reverse genetics): mutant or morphant

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12
Q

Gene loss of function - Assessing cell death

A

Activated caspase 3 staining labels apoptopic cells, key converging factor that triggers DNA fragmentation

Terminal dUTP nick-end labelling (TUNEL) marks DNA breaks (fragmentation)

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13
Q

Combine transgenesis to mark all cells that activate the promoter with MO knockdown

A

see onenote

can visualise what happens to cells that turned on the promoter but unable to generate the resulting protein

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14
Q

Assess fate of wrongly located cells using neuron type markers

A

see onenote slides

transgenes or immunostaining with antibodies against proteins

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15
Q

Gene gain of function

A

see onenote slides

Sufficiency of a gene (reverse genetics): GAL4/UAS

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16
Q

Cell autonomy

A

see onenote

genes can function within a given cell (autonomous) or influence surrounding cells (non-autonomous e.g. secreted factor)

embryo transplantation studies to generate chimeras

17
Q

Studying neural development

A

see onenote

Wt cells transplanted into WT or morpholino

Do they grow like normal WT or are they influenced by their surroundings?
E.g. if any of a cell type is missing, the body will make more of those - responding to the surroundings, how the brain makes the right sub-types in the right proportions

18
Q

Cell cycle and proliferation

A

see onenote slides

DNA replication inhibitor geminin shows phase specific expression - fusion to transgenes allows us to visualise this

labelling DNA histones shows DNA localisation and shape and identifies mitosis

DNA replication factor cdt1 shows differential phase dependent expression to geminin

specific proteins expressed at distinct phases and can be stained for immunihistochemically

19
Q

Spinal cord regeneration

A

see onenote slides

Mode: Spinal cord - role of fibroblast growth factor (Fgf)
Relevance: Human astrocyte culture

Fgf has multiple roles:
1. proliferation (first 6 days)
2. glia bridge formation (3 weeks)
Distinguished by timing using inducible heat shock

Fgf increases proliferation, supports “bridge” morphology