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ANAT 365 - Cellular Trafficking > Cell Polarity and Regulated Secretion > Flashcards

Cell Polarity and Regulated Secretion Flashcards

1
Q

How many plasma membranes do epithelial cells have?

A

Two

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2
Q

What are the sides of the cell?

A

Apical and basolateral

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3
Q

Apical side faces the _____________ world and basolateral side faces the _____________ world

A

Outside; inside

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4
Q

Why are plasma membrane and secreted proteins distinct?

A

To face different environments

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5
Q

What separates apical and basolateral sides?

A

Tight junction

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6
Q

Tight junctions made of _______________ proteins tightly linked to the ______________ prevent leakage through epithelial layer and prevent _____________ of membrane proteins from one membrane to another

A

Transmembrane; cytoskeleton; diffusion

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7
Q

Which concept does each prompt describe?
i. Components of apical and basolateral membranes are sorted into distinct vesicles in the TGN or in endosomes that then fuse specifically to their target membranes
ii. After fusion, proteins are endocytosed and reinserted, but are not endocytosed from the correct compartment

A

i. Vectorial sorting
ii. Selective retention

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8
Q

What are the three directions of vectorial sorting?

A

Transport from TGN to the apical or basolateral compartment
Transport from TGN to recycling endosomes
Proteins targeted to the wrong compartment transported to early endosomes and RE to the correct area

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9
Q

What is basolateral sorting?

A

Sorting in TGN into vesicle that is directed to basolateral plasma membrane

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10
Q

Basolateral signals are found in the ___________

A

Cytoplasm

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11
Q

What are the motifs of the basolateral signals?

A

Tyrosine-based motif, di-leucine motif

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12
Q

What are the adaptor proteins of basolateral sorting?

A

AP1 and AP2

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13
Q

Special subunit of AP1 is only found in ___________ cells that contains different m subunit

A

Polarized

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14
Q

The m subunit of AP1B binds to __________

A

Cargo

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15
Q

True or false: AP1B and AP1 are colocalized

A

False, AP1B is at recycling endosomes and AP1 is at TGN

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16
Q

AP1B is only important for basolateral proteins with ______________-based motif

A

Tyrosine

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17
Q

Apical sorting occurs directly through ___________ vesicles

A

Transport

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18
Q

Where are apical signals found?

A

In the membrane or the lumen

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19
Q

What are the motifs of the apical signals?

A

Lipid linkage (GPI), glycosylation signals, transmembrane domain, segregation into rafts, some in cytoplasmic domain

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20
Q

What is the use of raft clustering in apical sorting?

A

Budding of the membrane

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21
Q

What do the two types of apical transport vesicles aggregate with?

A

One with annexin in lipid rafts and the other with a leptin (Gal3)

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22
Q

Identify the vectorial route with the SNARE protein:
i. Cellubrevin
ii. TI-VAMP
iii. VAMP8
iv. Syntaxin 3
v. Syntaxin 4

A

i. Basolateral
ii. Apical
iii. Apical
iv, Apical
v. Basolateral

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23
Q

Which cells have more polarity than other cells?

A

Neurons

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24
Q

Neurons are very complicated cells with ______________ unique compartments

A

Multiple

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25
Q

Axons in neurons represent the _____________ side and dendrites represent the ______________ side

A

Apical; basal

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26
Q

What is a good model for understanding how polarity is generated?

A

Axon determination

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27
Q

What are the three rules of axon determination?

A

Longest neurite becomes axon
Neurite with greatest amounts of dynamic actin becomes axon
Cutting an axon shorter than another neurite allows other neurite to become axon

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28
Q

Kinesins are useful for sorting ____________ and moving the _____________ on microtubules

A

Vesicles; vesicles

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29
Q

The special isoform of kinesin only transports vesicles down ______________

A

Axons

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30
Q

What protein is used to signal to kinesin to the axon?

A

PI-3 kinase

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31
Q

Actin and microtubule cytoskeletal dynamics are ________________ of PI-3 kinase pathway

A

Downstream

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32
Q

True or false: Only negative feedback mechanisms are important for only one neurite to become an axon

A

False, positive and negative feedback mechanisms are important for only one neurite to become an axon

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33
Q

How do dynamic microtubules tell kinesin where the axon is found?

A

The kinesin isoform going down axons recognizes post-translational modifications on microtubules (detyrosinated microtubules)

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34
Q

What happens if the enzyme that forms the tyrosine on microtubules is removed?

A

Multiple axons form

35
Q

Kinesins that select axons prefer tubulin without _______________; the tyrosine will be replaced by an enzyme _______________

A

Tyrosine; TTI

36
Q

After axon determination, __________________ and actin filaments form at axon ________________, which generates a diffusion barrier between axon and ____________

A

Ankyrin G; hillock; soma

37
Q

What proteins can make it through the diffusion barrier?

A

Motor proteins

38
Q

What is the advantage of the motor proteins passing through the diffusion barrier?

A

It limits transport down the axon

39
Q

Cytoplasmic region in cytoplasmic loop between segments ___________ and _______________ is sufficient for axonal hillock localization

A

II and III

40
Q

What are the two motifs for full sorting in selective retention?

A

One required for association with ankyrin G
One leads to endocytosis

41
Q

Proteins sorted to dendrites have specific sorting signals for vesicles that are targeted to ___________________

A

Dendrites

42
Q

What kind of motor proteins are good for dendrite sorting?

A

Retrograde motor proteins

43
Q

Associate the following descriptions to either classical or peptide neurotransmitters:
i. Synthesized in the cytoplasm
ii. Synthesized by translation and processed through ER and Golgi
iii. Packaged into regulated secretory vesicles at the TGN
iv. Packaged into synaptic vesicles by transporters

A

i. Classical
ii. Peptide
iii. Peptide
iv. Classical

44
Q

Classical transmitters are released at the _______________ _______________ and may mediate ____________ or _____________ neurotransmission

A

Active zone; slow; fast

45
Q

Neuropeptides are released from ___________ the active zone and mediate ______________ neurotransmission

A

Outside; slow

46
Q

Fast neurotransmission is linked to _____________ ion channels and slow neurotransmission is linked to ____________ receptors

A

Ligand-gated; G-protein

47
Q

How are synaptic vesicles made?

A

After endocytosis from plasma membrane or endosomes through budding of clathrin-coated vesicles

48
Q

Where are dense core vesicles sorted?

A

TGN into special type of vesicle

49
Q

When are dense core vesicles released?

A

After a stimulus, either calcium entry in neurons or cyclic nucleotides in some endocrine cells

50
Q

True or false: Dense core vesicles can partially or completely fuse

A

True

51
Q

True or false: Dense core vesicles are released by partial fusion, but biogenic amines cannot be released

A

False, DCVs are not released by partial fusion, but biogenic amines can be released

52
Q

Dense core vesicles can also store ______________ transmitters

A

Classical

53
Q

What are two ways that proteins are sorted into regulated secretory vesicles?

A

Sorting by entry and by retention

54
Q

Aggregation of dense core vesicles is formed by binding to _____________ or lipid ______________

A

Receptor; rafts

55
Q

Aggregation of dense core is facilitated by _________________

A

Chromogranin

56
Q

The sorting receptor for the formation of the dense core is _____________________ binding or di-basic residue binding _______________ convertase

A

Carboxypeptidase E; prohormone

57
Q

Chromogranins are ___________ proteins that aggregate in the ____________ environment of the TGN

A

Acidic; acidic

58
Q

Removing chromogranin A using siRNA _____________ number of DCVs

A

Decreases

59
Q

Which of the following is found in the neuropeptide precursor structure?
i. Signal sequence
ii. Cleavage sites
iii. Glycine for amidation

A

All of the above

60
Q

What are some cleavage sites in neuropeptide precursor?

A

Furin site B and PC1/PC2 (basic residues)

61
Q

Furin cleaves at pH of _____________ and PC1 and PC2 cleave in immature ___________ granules

A

TGN; secretory

62
Q

The egg laying hormone precursor has how many furin sites?

A

One

63
Q

What is sufficient for DCV entry?

A

Aggregation

64
Q

Does aggregation explain the sorting of transmembrane proteins to DCVs?

A

No, instead, cytoplasmic signals are required for sorting into DCVs

65
Q

Where does sorting of transmembrane proteins into DCVs occur?

A

Vesicles budding from endosome and fusing with immature dense core vesicles

66
Q

What proteins are involved in endosomal trafficking for neuropeptide secretion?

A

RAb2, Vtia, EARP complex

67
Q

How are neurotransmitters packaged?

A

Through transporters

68
Q

Where are classical neurotransmitters synthesized?

A

In the cytoplasm

69
Q

Associate the transporter to the classical neurotransmitter
i. VMAT
ii. VGAT
iii. Vacht
iv. Vglut

A

i. Dopamine, noradrenaline, serotonin, histamine, GABA
ii. Glycine, GABA
iii. Acetylcholine
iv. Glutamate

70
Q

True or false: The regulated secretion of ATP and glycine started from DCVs before going to synaptic vesicles

A

False, ATP and glutamate

71
Q

Vesicular transporters can be found in __________

A

DCVs

72
Q

Which transporter regulates the entry and exit of potassium into the cell?

A

Plasma membrane transporter

73
Q

Which transporter uses ATP to put H+ ions into the vesicle?

A

Vesicular protein ATPase

74
Q

What determines which neuropeptide a neuron uses?

A

Expression of the peptide itself

75
Q

What determines which classical transmitter a neuron will use?

A

Specific synthetic enzymes that make the transmitter

76
Q

Which transmitters are present in all cells?

A

Glycine and glutamate

77
Q

Dale’s hypothesis states that neurons use how many transmitters?

A

One

78
Q

Is Dale’s hypothesis true for both classical transmitters and neuropeptides?

A

No, it is true for classical transmitters, but not for neuropeptides that are co-transmitters

79
Q

Which transmitters are released from some neurons in the same vesicle?

A

GABA and glycine

80
Q

Biogenic amines use the same ___________

A

Transporter

81
Q

Which transmitter’s transporters are found in more than just glutaminergic neurons?

A

Glutamate

82
Q

What does the presence of glutamate transporters in other neuronal cells suggest?

A

Glutamate is co-released at many synapses

83
Q

Which vesicles can be re-used?

A

Synaptic vesicles

ANAT 365 - Cellular Trafficking (5 decks)

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