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Cellular neuroscience Flashcards

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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
What is indirect IHC?
Using secondary antibody to bind to primary antibody to allow visualisation
26
What are the advantages and disadvantages of direct IHC?
Simple Fewer off-target effects Expensive Lower sensitivity Antibody must be made specific to target
27
What are the advantages and disadvantages of indirect IHC?
Cheaper Known and repeatable expression patterns High sensitivity More complex Potential off target binding
28
What does MAP2 show?
Immunofluorescence Shows blue, only in soma and dendritic tree Binds to mRNA
29
What does ankG show?
Shows red Only in initial segment of axon
30
How can we determine if a population is expression an RNA for a protein of interest? (FISH)
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
31
How can you identify the newly synthesised protein?
Add puromyocin to new protein Make antibody against the new protein If antibody for protein and puromyocin bind then fluorescence occurs
32
How are proteins made at the synapse?
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
33
What is the correlation of AD and microtubules?
Tau stabilises microtubules Tau becomes phosphorylated and dissociates from microtubule Causes neurofibrillary tangles
34
What is amyloidogenic process?
Cleavage by beta-secretase and then gamma-secretase Causes amyloid beta
35
What are key differences between glial cells and neurons?
Glial:neuron = 50:1 Glia non-excitable Glia undergo mitosis, neurons dont Incl. oligodendrocytes, ependymal, schwann, microglia, astrocytes
36
What are some advantages of myelination?
Decreases energy consumption Allows for rapid propagation down the axon Which allows for predatory movement and long term integrity
37
What is contained in the juxtaparanode?
VG K+ channels Is most lateral from node of ranvierW
38
What is contained in the paranode?
Has anchoring proteins Immediately lateral to node of ranvier
39
What is contained in the node?
Node of ranvier Contains VG Na+ channels
40
How is the node anchored to the axon?
Neurofascin-155, contactin, caspr
41
What is the process of protein synthesis in oligodendrocytes? (OG)
mRNA is transported down microtubules to the paranode They are then translated locally
42
What is a key difference between OG and schwann cells?
OG myelinate multiple axons Schwann cells myelinate one segment of an axon
43
What are the proteins of the juxtaparanode and internode?
MBP and PLP Present only in CNS Bind together to maintain lamellar structure of myelin sheath
44
What are monocarboxylate transporters (MCT)?
Extracellular membrane channels that trasnport lactate and pyruvate MCT1,2,4 in CNS
45
What does the MCT-1 do?
Present in the OG plasma membrane
46
What does the MCT-2 do?
Present in axon plasma membrane, underneath the myelin Transports lactate into axon
47
How do OG convert glucose to lactate?
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
How does the OG/SC know the axon needs energy?
NMDA on OG bind glutamate and allow Ca2+ into OG Ca2+ signals increase transport of glucose
49
How is apoptosis triggered?
P38 MAP kinase, cytochrome C, ROS All caused by excitotoxicity and excessive Ca2+ signalling
50
What are the main function of microglia?
Homeostasis, killing quiet synapses, phagocytosis, immunity
51
Where are microglia found?
Near synapses and in grey matter Have large phenotypic diversity
52
What are the different morphologies of microglia?
Amoeboid = activated Bushy = neurodegeneration and toxicity Ramified = for surveillance
53
Under TM what do microglia look like?
Bean shaped w/ elongated nuclei Scattered cisternae and rough ER + golgi at each pole
54
What is the function of microglia?
When resting its surveillant Detects injury and responds to injury by synthesis of chemokines
55
What is the role of the M2 phenotype?
Associated w/ cell growth and survival Anti-inflam. IL4, 10, 13
56
What is the role of the M1 phenotype?
Associated with inflam. and destroying cells Pro-inflam. IL6
57
What is the neuronal function of microglia?
Neurogenesis, apoptosis, induction, phagocytosis, maintenance of health
58
What is the immune function of microglia?
Pro and anti-inflam. response, cytokine and chemokine production, immuno-surveillance
59
How do TLRs work?
On the extracellular side of microbial products On the cytoplasmic side: Recruits signaling molecules Alters kinase activation Modifies gene expression
60
Function of resting microglia
Not moving Ramified and branched Immuno-surveillance
61
Function of activated microglia
Amoeboid Moves freely Phagocytoses debris Prune cells and dendrites
62
What do ependymal cells do?
For CSF synthesis and circulation Found in choroid plexus Have apical microvilli and cilia Regulates osmotic pressure
63
What happens when microglia become overactivated?
Produces cytotoxic factors As AD progresses, microglia become overactivated
64
How are reactive oxygen species produced?
NADPH oxdiase activation Activated in AD, PD and other neurological disorders
65
What is the basic shape of astrocytes?
Star-shaped, long branched processes Most numerous in glia in grey matter Non-excitable but communicable
66
How can astrocytes be distinguished?
Branching processes Dominated by int. filaments, actin, mt. and microtubules Large distal feet
67
What are the two types of astrocytes?
Fibrous: in white matter Protoplasmic: in grey matter, majority of the astrocytes, envelop synapses
68
What are the general functions of astrocytes?
Structural integrity BBB maintenance Neuronal nutrition Monitor neuron health Modulate neuronal output Tripartite synapse Overall synchronisation
69
How do astrocytes maintain these functions?
Have K+ channels that move K+ from [high] --> [low]
70
What is the tripartite synapse?
When the astrocyte directly modulates ongoing communication between synapses Uses Ca2+ as a way for astrocytes to communicate
71
How do mGluRs work?
Glutamate transmission increases intra. Ca2+ in astrocyte Gliotransmitters released Can directly affect pre. and post. signalling Act on NMDA, GABA Rs etc
72
How do astrocytes influence numerous neurons?
Astrocytes make connections to numerous neurons Eg: CB1 expressed on astrocytes and causes Ca2+ release Can cause increased release of gliotransmitters
73
How does astrocytic synchronisation work?
Astrocyte contacts a number of neurons across 100uM Results in neuronal synchronisation
74
How do astrocytes take up glucose?
Take glucose from blood at distal feet and convert to lactate Neurons take lactate and feed to mt. to make ATP
75
How does neural activity lead to vasodilation?
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
How do astrocytes turn reactive?
In response to various symptoms of neuronal dysfunction Two phenotypes A1 and A2
77
What does the A1 astrocytic phenotype do?
Cause cell death via inflammation
78
What does the A2 astrocytic phenotype do?
Repair of synapses and neuronal survival
79
What are the roles of astrocytes in PD?
Alpha-synuclein overproduced in dopaminergic cells Astrocytes consider alpha-synuclein and insult and destory the dopaminergic neurons
80
How do knock ins and knock outs work?
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
How does CRISPR work?
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
How does optogenetics work?
Include channel rhodopsin for depolarisation and halorhodopsin for hyperpolarisation Can selectively turn cells on/off
83
What does tetrode based electrophysiology allow us to measure?
Records APs fired from individual neurons Can show place cell firing
84

Neurobiology (3 decks)

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