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Bio lab 1 > Efferoctyosis > Flashcards

Efferoctyosis Flashcards

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Biology 101 flashcards
1
Q

Efferocytosis

A

Removal of STRICTLY apostolic cells by phagocytes

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2
Q

What are we doing in this experiment

A

Using cultured mouse macrophages to investigate the efficiency of macrophage engulfment of plastic beads. Two different effectors will be tested to see if the rate of phagocytosis can be increased

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3
Q

Phagocytosis

A

Macrophage engulfment

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4
Q

Effector

A

A molecule that binds to a protein (often an enzyme) and affects function of protein. These molecules can enhance phagocytosis

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5
Q

What is the purpose of lab this week

A

To demonstrate that macrophages can detect and engulf foreign substances. In this case, the macrophages will engulf plastic beads so we can know that we have a good model system for next week. Also, we will alter the concentration of different effectors to alter efficiency of process. Lastly, we willtitrate the ideal concentration of effector.

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6
Q

Apoptosis

A

Programmed cell death

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7
Q

Necrosis

A

Cell rupture

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8
Q

Steps when the apoptotic cell has clearance

A

Before it becomes necrotic, live macrophage efferocytoses dead cell. No release of cell contents

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9
Q

Resolution

A

When dead cell is consumed by macrophage and there is no release of cell contents

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10
Q

What causes inflammation

A

Cell being necrotic and release contents

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11
Q

Phagocytes

A

Broader term. Cells that engulf and digest foreign invaders (bacteria) and other cells (cancer cells and dead cells)

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12
Q

M1 macrophage is pro inflammatory or pro resolving

A

pro inflammatory

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13
Q

M2 macrophage is pro inflammatory or pro resolving

A

pro resolving

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14
Q

Cytokines

A

Signaling molecules that can be pro or anti inflammatory. They are small protein(s) significant in immune response signaling

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15
Q

M1 has incresased/decreased bactericidal activity, pro/anti inflammatory cytokines and high/low efferocytosis

A

Increased, pro, low

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16
Q

M2 has incresased/decreased bactericidal activity, pro/anti inflammatory cytokines and high/low efferocytosis

A

Decreased, anti, high

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17
Q

How does live macrophage deal with toxins after engulfing dead cell

A

Efflux: move toxins out
Metabolism: break them down

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18
Q

Steps of efficient dead cell clearance

A

Find me signal release from dead cell and phagocyte recruitment
Exposed eat me signal on dead cell and phagocytic receptors recognize signals
Actin filament of phagocyte push out plasma membrane to engulf dead cell
Dead cell digests and phagocyte releases cytokines that are pro inflammatory or pro resolving

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19
Q

How does efferocytosis help in resolution and repair

A

Prevents necrosis, which terminates inflammatory response, encourages body to not attack own cells, activation of pro resolving pathways

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20
Q

Efferoctyosis defective diseases it can cause

A

Autoimmune, atherosclerosis, myocardial infarction, obesity and diabetes

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21
Q

IL

A

Largest kind of cytokine

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22
Q

SPMs

A

Specialized proresolving mediators (effectors). These enhance efferoctyosis

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23
Q

SPMs do what

A

Resolve tissue inflammation

24
Q

Kind of effectors we’re using

A

Resolvins, IL-10, dexamethasone

25
Q

Cell plasticity

A

Macro expresses genes for both m1 and m2 macrophages

26
Q

LPS

A

Pushes macrophages toward m1 by changing gene expression. So then it can engulf bacteria of beads better

27
Q

IL-4

A

Makes macrophage more like m2 so it can engulf dead cells better

28
Q

Steps to pipette

A

To set at right volume:
Spin dial at top of pipette
Steps:
Put tip on pipette
Open tube
Push down on plunger to first notch
Put tip in liquid, pull plunger up
Take new tube
Put pipette near bottom of tube
Push plunger to first stop to get rid of liquid then 2nd stop
Eject tip in waste

29
Q

How to draw up liquid from well

A

Tilt dish at 30 degrees, put tips at bottom, draw up liquid

30
Q

EvOS microscope

A

Identifties problems after plate reader data. Looks at confirmation that only engulfed beads are left

31
Q

Plate reader

A

Scans for fluorescence at each well by exciting and emitting at wavelengths

32
Q

Phosphatidylserine

A

Eat me signal

33
Q

Leukotrienes

A

stimulate the production of proinflammatory cytokines

34
Q

When resolvin:Leuko ratio is high, are there more m1 or m2 macrophages. And how is plaque affected

A

More resolvin=higher resolution, less inflammation=more m2 macrophages and decreased plaque size

35
Q

Inflammasome

A

cytosolic proteins (composed of many subunits) responsible for the activation of the inflammatory response

36
Q

Gas6

A

Bridging molecule between eat me signal and mertk (a scavenger receptor found on surface of macrophage)

37
Q

Mertk

A

Scavenger receptor on macrophage

38
Q

How does efferocytosis become more defective. By what factor

A

Age.

39
Q

How does cell engulf dead cell while in plate

A

The cell pushes out protrusions at its leading edge
• Protrusions adhere (anchor) to the “crawling” surface
• Rest of cell drags itself forward by traction on the anchorage points
Cell stays on plate

40
Q

Do spms and cytokines bind to different cell surface receptors

A

Yes

41
Q

Purpose of lab

A

To see how different concentrations of inhibitors affect the inhibition of efferocytosis. Also seeing how combinations of different resolvins enhance efferocytosis

42
Q

CD36

A

Scavenger receptor

43
Q

Thbs-1

A

bridging protein for CD36

44
Q

CD47

A

“Don’t eat me” signal

45
Q

Annexin A1

A

inhibits prostaglandin synthesis – anti-inflammatory effect

46
Q

FPR2

A

resolvin receptor

47
Q

IL-10

A

anti-inflammatory cytokine

48
Q

TNF-alpha

A

pro-inflammatory cytokine

49
Q

TGF-beta

A

anti-inflammatory cytokine

50
Q

Rac1

A

activates actin polymerization/reorganization

51
Q

LC-3

A

autophagy protein that is recruited to the phagosome (main marker for phagocytosis)

52
Q

Rubicon

A

negative regulator of autophagy/phagocytosis

53
Q

ATG16L

A

downstream LAP machinery

54
Q

Control/housekeeping gene for experiment 4

A

Glyceraldehyde 3-phosphate dehydrogenase (GAPDH)

55
Q

Formula for relative quantification (RQ)

A

2^(-🔼🔼CT)
Triangle means delta
CT is threshold cycle .
Aka, subtract first 🔼CT from 2nd 🔼CT, switch sign, and raise 2 to this

Bio lab 1 (1 decks)

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