CAMS Flashcards

1
Q

What are some examples of how adhesion is important

A

Adhesion of sperm to oocyte.

Cells in the morula undergo compaction using E cadherins to stick closely together.

Cells split into the inner cell mass and the trophoblast due to their differences in adhesion properties.

Implantation of the oocyte into the endometrium requires adhesion.

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2
Q

What are L cells

How can you use them to show homopholic sorting

A

They don’t express any cadherins and so they don’t aggregate.

You can transfer them with transgenes for different cadherins and then monitor their behaviour.

One group of cells with E cadherins and the other group with N cadherins. The cells will aggregate with eachother and and those with E go into the middle and those with N surround them. This is called homophilic sorting.

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3
Q

One group of cells with high levels of E cadherin and another group of cells with low levels of E cadherin.

A

The cells aggregate with eachother and sort into two groups.

The high E cadherin cells go into the middle and the low ones surround them.

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4
Q

Cadherins structure

Hinge regions

A

Have transmembrane domains and their extra cellular domain interacts with things at the N terminal.

Ca binds to the hinge regions on the extra cellular domain to stabilise them so they are not floppy.
This means cells can modulate how much adhesion they do by changing the Ca levels to allow cadherins to work better.
More Ca means more adhesion.

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5
Q

How are cadherins localised on the cell membrane

What is the first cadherin expresses in embryos

A

They all line up on the cell membrane.

The arrays on one cell will be perpendicular to the lines of arrays on the cells it binds to.

E cadherin

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6
Q

Where are cadherins normally expressed

A

In adherins junctions

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7
Q

Classical vs non classical cadherins

Examples

Structure

A

Classical - E, N, P, VE
Non- cadherin 23

Classical have a single transmembrane domain and then repeated hinge domains on the extracellular.

Non classical have varied structures with multiple transmembrane domains or kinase domains

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8
Q

Where are protocadherins found

A

NS

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9
Q

How did the many types of protocadherins occur and what do they do

A

Alternative splicing of the gene can give rise to various isoforms.

They play a role in specifying synapses in the brain allowing for complex connectivity.

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10
Q

What do catenins do ?

What happens when a cell is not using its cadherins compared to when it is

A

Bind to the intercellular domain of cadherins.
They link the cadherins to the cells cytoskeleton actin filaments.

The alpha catenin is in a folded state.
When a cell is interacting with another cell it creates tension and causes the catenin to go into an extended state.
This reveals a binding site for vinculin which binds to actin.

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11
Q

What does rho activation do

A

Leads to stress fibre formation where the actin filaments organise into parallel fibres.

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12
Q

What cadherins do newly formed mesoderm cells lose

What replaces E cadherin in the neural tube

Where is cadherin 6B expressed

What do neural crest cells express

A

E cadherin so they can migrate inside.

N cadherin.

In the floor and roof plate.

Cadherin 7

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13
Q

What is the structure of selectins and what do they bind to

What do they do

What is on their extra cellular domain and intracellular domain

A

Cell surface protein with a single transmembrane domain which is Ca dependant and binds to carbohydrates on neighbouring cells.

They are used in the immune system for neutrophil trapping.

EGF like domain and a lectin domain on the end of their extracellular domains.
Their intracellular domains are anchored to actin.

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14
Q

When are selectins induced

A

During inflammation in endothelial cells when white blood cells become adherent to the wall of the vessel and roll along it.

Once the cells stop rolling they leave the vessels going between cells using integrines.

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15
Q

Ca independent CAMS

WTF

A

Some are secreted and others are transmembrane.

Their extra cellular domains have many ig and fibronectin repeat domains.

There is one gene they all arise from the undergoes alternative splicing and post translational glycolysation.

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16
Q

What is the ECM made up of

A

Fibronectin, laminin, collagen and elastin which are all large secreted proteins.

17
Q

What is laminins structure

A

It is a trimer.

It has integrity interacting domains and self assembly branches.

It has a coil coiled domain.

18
Q

Integrins structure

What is on each domain

How may types of each subunit are there

A

They form heterodimers and each monomer has a single transmembrane domain.

They have cystine rich extracellular domains to interact with other extra cellular components.

At the end of the extra cellular domain they have a matrix binding domain which interacts with the ECM.

The intracellular domain is anchored to actin filaments using talin and vinculin.

There are 18 types of alpha subunit and eight types of beta making 24 variations.

19
Q

How are integrins activated

What do they look like when active and inactive

A

Binding intra or extracellular. Inside out activation or outside in.

Inactive has a folded extracellular domain.

20
Q

How is focal adhesion kinase activated

A

When integrins binds to the ECM it is activated.

It will phosphorylate tyrosine and cause actin polymerisation.

It will recycle focal adhesions to enable migration and acts in attachment dependant cell death.

21
Q

How do cells migrate and what does the cell look like

How do they sense cues

A

Lamellipodia extend at the leading edge and attach to the substratum by focal adhesions.

New adhesions are made at the front and old ones are removed from the back.

Actin polymerisation pushes the membrane out. The cell is polarised.

To sense guidance cues the lamellipodia ruffle

22
Q

What is cytochlasin B

A

It is used to study actin polymerisation

It lands on the plus end of the filament and blocks further adding of the actin.

This stops migration and actin starts to disassemble.

23
Q

How does the arp 2/3 complex work

What activates it

A

Nucleates actin.

It binds to the minus end of the filament and monomers are added to the plus end.

Arp 2 and 3 have a similar structure to actin and this is why they are able to nucleate.
Another filmament extends at 70 to the original one. This gives stability.

RAC activates it

24
Q

What two things treadmill during migration

A

Actin - as you grow fibres at the leading edge you take them away from the minus end.

Focal adhesion treadmills.

25
Q

Low density vs high density focal adhesions

A

Low require rac and cdc42. They are usually found at the front of the leading edge they are immobile and fixed.

High has many clusters close together and are dependant on rho and actin myosin interactions.
They are far from the front and can slide within the membrane.

26
Q

What are some different signals to help cell migration

A

Netrins are diffusible signals that attract cells.

CAMs and the ECM can also attract cells.

Fibronectin tails are secreted by migratory cells to lay down a trail for other cells

27
Q

What is membrane cycling

A

Membrane is removed and deposited at the leading edge.

The membrane receptors for an attractive ligand are taken into the cell if they are not on the leading edge. This is so only the leading edge will be activated and attracted to the ligand so the cell migrates in the right direction.

CAMs are also removed by endocytosis when they reach the back of the cell.