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ANAT242 > Lecture 12- Cell Biology of Neurons 2 > Flashcards

Lecture 12- Cell Biology of Neurons 2 Flashcards

1
Q

What is the central dogma of biology?

A
  • DNA is transcribed to mRNA (messenger RNA)
  • mRNA three-part code is translated to an amino acid by ribosomes
  • Amino acids are assembled into chains to form proteins

(basically DNA to RNA, RNA to protein)

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2
Q

What goes a Nissl stain show?

A
  • Nissl was found to be selective for RNA and so Nissl granules (dark patches) represents large stacks of rough endoplasmic reticulum.
  • These stacks are close to the soma of the cell therefore Nissl cells are good for visualizing cell bodies
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3
Q

What is meant by the term phospholipid bilayer?

A
  • This is what the cell membrane is
  • Hyrophobic ends in middle, Hyrophilic ends inside/outside
  • Not permeable to ions therefore need proteins to allow function (transport/ channels allow ions to cross)
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4
Q

What is the structure of the proteins embedded in the cell membrane?

A
  • May span membrane, or be inserted partway
  • Complex folded 3D structure
  • Composed of subunits (same/different)
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5
Q

Are Synaptic vesicles (SSV and LDCV) simple phospholipid spheres?

A

No have multiple membrane associated proteins

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6
Q

What is an essential feature of a neuron’s membrane in allowing it to be part of a communication network?

A
  • Needs to be polarized (different charges indifferent parts)
  • Proteins associated with the neuronal membrane are essential for this as allow specificity
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7
Q

What is the result of a neuron having a large membrane area?

A
  • 200x greater membrane area than the diameter of a neurons soma
  • Therefore need to make lots of proteins to maintain this area and so need lots of RER and specialized vesicles
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8
Q

What is the cytoskeleton made of?

A

Proteins

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9
Q

What can changes to membrane proteins cause?

A
  • Proteins are essential in neuronal function and need to be in the ‘right’ place
  • Changes to the location/ shape of these proteins causes dysfunction
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10
Q

How do kinases and phosphates work to create cellular energy?

A
  • Protein Kinases add a phosphate group from ATP activating the target and forming ADP (byproduct)
  • Protein Phosphatases remove a phosphate group deactivating target
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11
Q

What is the general method for identifying and locating proteins?

A
  • Called immunohistochemistry
  • Have protein of interest (antigen: receptor, vesicle etc.) which we manufacture a primary antibody for in a species like a goat or rabbit. The secondary antibody is what then binds to the primary antibody (often called an anti-goat or anti-rabbit depending on the original species the antibody was made in).
  • It’s a marker that binds to the secondary antibody that can be visualized and allows us to locate the protein of interest in electron microscopy. The marker can either be a fluorescent molecule (shine light in order to see) or enzyme (creates a coloured by product via a substrate)
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12
Q

What can the location of proteins (obtained via immunohistochemistry tell us)?

A
  • About their function/ precision of function and structure

- Also can help us tell the difference between axons and dendrites

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13
Q

What is ankG and where is it found?

A
  • scaffolding protein
  • proximal axon
  • nodes
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14
Q

What is Caspr and where is it found?

A
  • transmembrane protein
  • member of neurexin family
  • cell adhesion
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15
Q

What is MAP2 and where is it found?

A
  • microtubule associated protein

- absent in most astrocytes

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16
Q

What is PO and where is it found according to immunohistochemistry?

A

Myelin protein

17
Q

What does immunohistochemistry show in regards to dendrites?

A
  • Dendrites come up as green and F-actin as red
  • When merge/ both present get yellow
  • Picture shows yellow only in the spines
  • This matches what we know about the function of actin as it helps to grow/ stabilize the dendritic spines
18
Q

What is important when using immunohistochemistry?

A

Need to have some idea of a proteins function/ location before you then go and stain for it as you can’t stain everything only specific things. Not an exploratory thing. Need to use it to confirm hypotheses.

19
Q

Is a synaptic clef just a space?

A
  • No, its a very dense fibrous matrix of associated proteins (Appears dark in TM)
  • Bunch of proteins stick through both membranes (the pre and post synaptic membranes) to hold them together preventing widening of the synapse/ gap
20
Q

What is the difference in specialization of the pre and post-synaptic membranes?

A

-Pre= specialized for neurotransmitter release (contains active zones).
Contains: Calcium channels, Docking proteins for vesicles (SNAREs etc), Neurotransmitter autoreceptors, Reuptake channels, Vesicle accessory proteins

-Post= specialized for receiving neurotransmitters (contains receptors).
Contains: Voltage gated ion channels. G-Protein coupled receptors.

21
Q

Why is does the soma use so much energy/ ATP?

A

Because of the major requirement to make many diverse synaptic proteins

22
Q

What is PSD-95? What does it illustrate?

A

-Scaffold protein, under the plasma membrane for the clustering of
receptors, ion channels etc.
-Shows the importance of synapse proteins remain in the correct location

23
Q

Complex neuron shapes need to…

How is this accomplished?

A
  • Maintain their shape
  • Stay connected to next neuron (at synaptic cleft)

Accomplished via:
-Clef proteins (trans-synaptic) which are secreted from the presynaptic membrane and adhere/ interact with molecules on the surface of adjacent cells.

24
Q

How do neurons utilize myosin?

A

Associate with actin, similar to function in muscle. Help movement/ trafficking.

25
Q

What is the purpose of synaptic tagging?

A

-To let the actin/ trafficking mechanism know which synapse to drop cargo at (hypothesized mechanism)

26
Q

What is local protein synthesis?

A
  • mRNA transported to synapse and only transcribed into protein on synaptic activation
  • Location of polyribosomes at the post synaptic membrane supports this idea
27
Q

What is FISH?

A
  • Fluorescence in situ hybridization
  • Have target RNA. A fluorescent tag tells us where the RNA is as turns on when bound to target.
  • This can be useful as it can indicate what proteins are being formed.
28
Q

What does puromycin labelling tell us?

A
  • Allows the identification of newly synthesized proteins (contain puromycin)
  • Antibodies against puromycin + target (means we identify the proteins we are interested in not just all proteins).
  • Cause a PLA signal
29
Q

Why does there need to be local protein synthesis?

A

Because there is so many dendritic branches and neurons are big. If all proteins were made in the soma and were trafficked would take too long + too much energy.

30
Q

What is a case where protein synthesis goes wrong?

A
  • Alzheimer’s disease (Tau, amyloid beta)

- Treatments could target this

ANAT242 (25 decks)

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