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Neurobiology > Cellular neuroscience > Flashcards

Cellular neuroscience Flashcards

1
Q

What does the nissl stain show and why?

A

Stains neurons NOT glia
Nissl binds to -ve charge of mRNA found mainly in soma of neurons

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2
Q

What does the golgi stain show and why?

A

Silver-chrome rxn
Randomly stains a few neurons
Can visualise entire neuron

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3
Q

What are the pros and cons of the nissl stain?

A

PROS
Stains majority of neurons
Good for looking at gross structure

CONS
Weakly stains glia
Can only see soma clearly

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4
Q

What are the pros and cons of the golgi stain?

A

PROS
Can visualise entire neurons
Easy to determine type of neuron due to this

CONS
Only a subset of neurons take up the stain

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5
Q

What are microtubules made of?

A

Hollow tubes of tubulin
Made of alpha and beta dimers
Dimers are added on at +ve end

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6
Q

What are neurofilaments made of?

A

Made of light, medium and heavy filaments and proteins

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7
Q

What are microfilaments made of?

A

Mainly made of actin and monomeric G-actin

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8
Q

What are the functions of microtubules?

A

They are used for motor proteins to carry their vesicles and proteins down the axon
Overall for movement of cellular components

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9
Q

What is rescue factor?

A

Stabilise and destabilises microtubules

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10
Q

What occurs at the +ve end of the microtubule?

A

Dimers are added and microtubule lengthens

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11
Q

What occurs at the -ve end of the microtubule?

A

Gamma-tubulin binds to GT complex and forms the gamma-tubulin ring complex which binds -ve end and is anchored to centrosome

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12
Q

What is kinesin?

A

Kinesin is a motor protein that walks vesicles toward the synapse

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13
Q

What is anterograde transport?

A

Done by kinesin - goes towards synapse

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14
Q

What is retrograde transport?

A

Done by dynein - goes back to soma with leftover stuff

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15
Q

What is dynein?

A

Dynein is a motor protein that walks vesicles toward the soma and away from synapse

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16
Q

How do motor proteins actually walk?

A

Uses ATP for every step
Phosphate broken everytime foot swings

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17
Q

What is the function of neurofilaments?

A

Predominant component of cytoskeleton
Huge mechanical strength
Most stable part of cytoskeleton
Very important in axonal integrity and diameter

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18
Q

What is the function of microfilaments?

A

Keep organelles in place and help cell movement

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19
Q

How does actin endocytosis work?

A

F-actin forms a scaffold and transports lyosomes filled w/ endocytosed proteins and retrograde transports them to microtubule highway

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20
Q

How does actin exocytosis work?

A

Myosin transfers vesicles along actin filaments until they reach synapse and are exocytosed

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21
Q

What is the central dogma of biology?

A

DNA –> RNA –> Protein
transcript. translate

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22
Q

Why do neurons need to make so many proteins?

A

Are highly dynamic
Receptors, vesicles etc

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23
Q

How does CaMKII phosphorylate AMPA receptors?

A

Ca2+ influx activates CaMKII
Phosphorylated tails of AMPA-bound TARP bind to PSD-95, anchors receptor to membrane
Phosphorylated AMPA tails cause a conf. change in the receptor allowing more Na+ into cell

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24
Q

What is direct IHC?

A

Using primary antibody to bind
Often immunofluorescence

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25
Q

What is indirect IHC?

A

Using secondary antibody to bind to primary antibody to allow visualisation

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26
Q

What are the advantages and disadvantages of direct IHC?

A

Simple
Fewer off-target effects

Expensive
Lower sensitivity
Antibody must be made specific to target

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27
Q

What are the advantages and disadvantages of indirect IHC?

A

Cheaper
Known and repeatable expression patterns
High sensitivity

More complex
Potential off target binding

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28
Q

What does MAP2 show?

A

Immunofluorescence
Shows blue, only in soma and dendritic tree
Binds to mRNA

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29
Q

What does ankG show?

A

Shows red
Only in initial segment of axon

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30
Q

How can we determine if a population is expression an RNA for a protein of interest? (FISH)

A

Fluorescence in situ hybridisation
Must know sequence for protein
Make complementary RNA sequence
Attach a fluorescent protein to manufactured sequence
If cell is expressing target RNA, fluorescence occurs

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31
Q

How can you identify the newly synthesised protein?

A

Add puromyocin to new protein
Make antibody against the new protein
If antibody for protein and puromyocin bind then fluorescence occurs

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32
Q

How are proteins made at the synapse?

A

DNA transcribed to mRNA and is either translated in soma or mRNA binds to a protein and translation is repressed
mRNA docks with kinesin
Synaptic activation causes mRNA to be unrepressed
Translated at synapse

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33
Q

What is the correlation of AD and microtubules?

A

Tau stabilises microtubules
Tau becomes phosphorylated and dissociates from microtubule
Causes neurofibrillary tangles

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34
Q

What is amyloidogenic process?

A

Cleavage by beta-secretase and then gamma-secretase
Causes amyloid beta

35
Q

What are key differences between glial cells and neurons?

A

Glial:neuron = 50:1
Glia non-excitable
Glia undergo mitosis, neurons dont
Incl. oligodendrocytes, ependymal, schwann, microglia, astrocytes

36
Q

What are some advantages of myelination?

A

Decreases energy consumption
Allows for rapid propagation down the axon
Which allows for predatory movement and long term integrity

37
Q

What is contained in the juxtaparanode?

A

VG K+ channels
Is most lateral from node of ranvierW

38
Q

What is contained in the paranode?

A

Has anchoring proteins
Immediately lateral to node of ranvier

39
Q

What is contained in the node?

A

Node of ranvier
Contains VG Na+ channels

40
Q

How is the node anchored to the axon?

A

Neurofascin-155, contactin, caspr

41
Q

What is the process of protein synthesis in oligodendrocytes? (OG)

A

mRNA is transported down microtubules to the paranode
They are then translated locally

42
Q

What is a key difference between OG and schwann cells?

A

OG myelinate multiple axons
Schwann cells myelinate one segment of an axon

43
Q

What are the proteins of the juxtaparanode and internode?

A

MBP and PLP
Present only in CNS
Bind together to maintain lamellar structure of myelin sheath

44
Q

What are monocarboxylate transporters (MCT)?

A

Extracellular membrane channels that trasnport lactate and pyruvate
MCT1,2,4 in CNS

45
Q

What does the MCT-1 do?

A

Present in the OG plasma membrane

46
Q

What does the MCT-2 do?

A

Present in axon plasma membrane, underneath the myelin
Transports lactate into axon

47
Q

How do OG convert glucose to lactate?

A

Glucose enters via GLUT-1
Glycolysis turns it to pyruvate
Pyruvate –> Lactate via LDH
MCT-2 takes up lactate into axon
Lactate turned back to pyruvate and metabolised into ATP

48
Q

How does the OG/SC know the axon needs energy?

A

NMDA on OG bind glutamate and allow Ca2+ into OG
Ca2+ signals increase transport of glucose

49
Q

How is apoptosis triggered?

A

P38 MAP kinase, cytochrome C, ROS
All caused by excitotoxicity and excessive Ca2+ signalling

50
Q

What are the main function of microglia?

A

Homeostasis, killing quiet synapses, phagocytosis, immunity

51
Q

Where are microglia found?

A

Near synapses and in grey matter
Have large phenotypic diversity

52
Q

What are the different morphologies of microglia?

A

Amoeboid = activated
Bushy = neurodegeneration and toxicity
Ramified = for surveillance

53
Q

Under TM what do microglia look like?

A

Bean shaped w/ elongated nuclei
Scattered cisternae and rough ER + golgi at each pole

54
Q

What is the function of microglia?

A

When resting its surveillant
Detects injury and responds to injury
by synthesis of chemokines

55
Q

What is the role of the M2 phenotype?

A

Associated w/ cell growth and survival
Anti-inflam. IL4, 10, 13

56
Q

What is the role of the M1 phenotype?

A

Associated with inflam. and destroying cells
Pro-inflam. IL6

57
Q

What is the neuronal function of microglia?

A

Neurogenesis, apoptosis, induction, phagocytosis, maintenance of health

58
Q

What is the immune function of microglia?

A

Pro and anti-inflam. response, cytokine and chemokine production, immuno-surveillance

59
Q

How do TLRs work?

A

On the extracellular side of microbial products

On the cytoplasmic side:
Recruits signaling molecules
Alters kinase activation
Modifies gene expression

60
Q

Function of resting microglia

A

Not moving
Ramified and branched
Immuno-surveillance

61
Q

Function of activated microglia

A

Amoeboid
Moves freely
Phagocytoses debris
Prune cells and dendrites

62
Q

What do ependymal cells do?

A

For CSF synthesis and circulation
Found in choroid plexus
Have apical microvilli and cilia
Regulates osmotic pressure

63
Q

What happens when microglia become overactivated?

A

Produces cytotoxic factors
As AD progresses, microglia become overactivated

64
Q

How are reactive oxygen species produced?

A

NADPH oxdiase activation
Activated in AD, PD and other neurological disorders

65
Q

What is the basic shape of astrocytes?

A

Star-shaped, long branched processes
Most numerous in glia in grey matter
Non-excitable but communicable

66
Q

How can astrocytes be distinguished?

A

Branching processes
Dominated by int. filaments, actin, mt. and microtubules
Large distal feet

67
Q

What are the two types of astrocytes?

A

Fibrous: in white matter
Protoplasmic: in grey matter, majority of the astrocytes, envelop synapses

68
Q

What are the general functions of astrocytes?

A

Structural integrity
BBB maintenance
Neuronal nutrition
Monitor neuron health
Modulate neuronal output
Tripartite synapse
Overall synchronisation

69
Q

How do astrocytes maintain these functions?

A

Have K+ channels that move K+ from [high] –> [low]

70
Q

What is the tripartite synapse?

A

When the astrocyte directly modulates ongoing communication between synapses
Uses Ca2+ as a way for astrocytes to communicate

71
Q

How do mGluRs work?

A

Glutamate transmission increases intra. Ca2+ in astrocyte
Gliotransmitters released
Can directly affect pre. and post. signalling
Act on NMDA, GABA Rs etc

72
Q

How do astrocytes influence numerous neurons?

A

Astrocytes make connections to numerous neurons
Eg: CB1 expressed on astrocytes and causes Ca2+ release
Can cause increased release of gliotransmitters

73
Q

How does astrocytic synchronisation work?

A

Astrocyte contacts a number of neurons across 100uM
Results in neuronal synchronisation

74
Q

How do astrocytes take up glucose?

A

Take glucose from blood at distal feet and convert to lactate
Neurons take lactate and feed to mt. to make ATP

75
Q

How does neural activity lead to vasodilation?

A

Astrocytes detect energy demand thru GluRs
Ca2+ transient develops and travels to end feet
Release of vasodilators
Increase blood flow = more glucose for demands

76
Q

How do astrocytes turn reactive?

A

In response to various symptoms of neuronal dysfunction
Two phenotypes A1 and A2

77
Q

What does the A1 astrocytic phenotype do?

A

Cause cell death via inflammation

78
Q

What does the A2 astrocytic phenotype do?

A

Repair of synapses and neuronal survival

79
Q

What are the roles of astrocytes in PD?

A

Alpha-synuclein overproduced in dopaminergic cells
Astrocytes consider alpha-synuclein and insult and destory the dopaminergic neurons

80
Q

How do knock ins and knock outs work?

A

Use enzymatic cutting to splice out a piece of specific DNA
OR
Encoding for a protein not usually expressed, DNA recombines w/ donor DNA and cell expresses new proteins

81
Q

How does CRISPR work?

A

Viral DNA captured and integrated into bacteriums DNA
Viral DNA transcribed to RNA
RNA integrated into Cas as a guide
Guide RNA recognises viral sequence
Cas cuts viral DNA, disrupting it

82
Q

How does optogenetics work?

A

Include channel rhodopsin for depolarisation and halorhodopsin for hyperpolarisation
Can selectively turn cells on/off

83
Q

What does tetrode based electrophysiology allow us to measure?

A

Records APs fired from individual neurons
Can show place cell firing

84
Q
A

Neurobiology (3 decks)

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