Neurobiology 2 - A - Cellular Properties of Neurons

Lecture “Neurobiology”
SK.Bio-NF.7001: Neurobiology
Prof. Dr. André Fiala

Molecular Neurobiology of Behavior, University of Göttingen
Part Two A: Cellular Properties of Neurons

Crash-Course in Basic Cell Biology for Non-Biology Students
Not all students of this lecture study Biology or a related topic of Life Science.
Some study Physics or Neuroinformatics. It will be difficult for them to follow the
lecture if basic knowledge about Cell Biology is lacking.
The next slides will provide a crash-course for those non-Biologists. All students
who have heard that already multiple times are encouraged to see whether they
would have known everything already.
For Students of Physics or Informatics and for self-studying it is

recommended to consults the respective chapters of a general textbook of
Biology, e.g.:
Neurons are cells that contain the same organelles as all cells
The Soma (Perikaryon)
Nucleus
rough and smooth endoplasmatic reticulum
Ribosomes und Polyribosomes
Mitochondria
Golgi-apparatus
Cytoskeleton
Gene transcription in the nucleus
Protein synthesis in the cytosol; can be precisely localized, e.g. at synapses!
Mitochondria produce ATP:
Present in axones, dendrites, synaptic structures. Cellular respiration, Krebs-Martin cycle;
Electron microscopic section: presynaptic terminal in the cerebellum. Please note the cell
membranes and the synaptic vesicles!
The Golgi apparatus: many functions!
- production of vesicles
- synthesis and modification of plasma membrane elements
- production of primary lysosomes
- post-translations modifications of proteins, e.g, cleavage;
- delivery of proteins and membranes to specific destinations („trafficking“)
The cytoskeleton: 3 „bones“
Cytoskeleton proteins are not static! They determime the plastic shape of the neuron!
3 main types:
a) microtubules
b) neurofilaments
c) microfilaments
Microtubules
Blue arrow: longitudinal Red arrow: transverse section

section across dendrite. across dendrite.
- Microtubules consist of two different monomers

- They form a polar strand (+ and – end)
- 13 monomers per „ring“
- polymerisation requires energyx: GTP-hydrolysis
Electron microscopic section: microtubules in dendrites. - In the mammlian brain > 20 isoformes!
- The stability is maintaines by microtubules-
associated proteins (MAPs). In Aaxons different MAPs
than in dendrites!
Microtubules
A large number of regulatory proteins are associated with microtubules.
- Microtubules consist of two different monomers

- They form a polar strand (+ and – end)
- 13 monomers per „ring“
- polymerisation requires energyx: GTP-hydrolysis
- In the mammlian brain > 20 isoformes!
- The stability is maintaines by microtubules-
associated proteins (MAPs). In axons different MAPs
than in dendrites!
Excursion: a MAP (microtubule-associated protein) called Tau plays a role in
Alzheimer‘s disease.
Tau becomes excessively phosphorlyted by an enzyme called Glykogen-Synthase-Kinase GSK3. This enzyme gets
activated by b-Amyloid.
Excursion: a MAP (microtubule-associated protein) called Tau plays a role in
Alzheimer‘s disease.
After cells have degenerated and die, clumps of Tau can be detected.
Staining of neurofilaments Staining of Tau Co-localization in dead cells.

Neurofilaments
- 3-10 x more than microtubules

- belong to the protein family of cytokeratins
- very stable, almost completely polymerized
- determine lerngthz and thickness of axons
Neurofilaments (Arrow)
P = Perikaryon of a Purkinje-Cell of the cerebellum
N = Nucleus
Neurofilaments
Intermediate filaments to which neurofilaments belong are a large family of

proteins.
- 3-10 x more than microtubules

- belong to the protein family of cytokeratins
- very stable, almost completely polymerized
- determine lerngthz and thickness of axons
Actin filaments (microfilaments)
- polar polymers of actin monomers

- GTP or GDP-binding
- several genes for actin
- dense mesh below the membrane
- fast polymerization and degradation
Actin filaments ina dendritic spine, a plastic extrusion of the dendrites of some neurons.
A: Axon
S: Spine
Actin filaments (microfilaments)
A large number of regulatory proteins are associated with microfilaments or

constitute different types of microfilaments.
- polar polymers of actin monomers

- GTP or GDP-binding
- several genes for actin
- dense mesh below the membrane
- fast polymerization and degradation
Dendritic spines
- small protrusions on dendrites of glutamatergic

central brain neurons in mammals
- variable shape, modifiable
- provide thousands of synaptic contacts to one cell
- plastic in shape
- important for learning and memory (plasticity!)
- Deseases can affect spine morphology
- Shape determines electrical properties of

synapses (resistance!) and biochemical properties.
Dendritic spines
Penzes, P., Woolfrey, K., & Srivastava, D. (2011). Epac2-mediated dendritic spine remodeling: Implications for disease. Molecular
and Cellular Neuroscience, 46, 368-380 .

Development and Regulation of Dendritic Spine Synapses
Barbara Calabrese, Margaret S. Wilson, and Shelley Halpain
Physiology 2006 21:1, 38-47 Dendritic spines
Fast anterograde axonal transport
Transports organelles, vesicles, mitohondria, elements of sER, …
- Kinesin and Microtubuli!
- ATP-dependent
- independent from soma
1 Cyclus = 16 nm, 2 ATP!

Fast anterograde axonal transport
Transports organelles, vesicles, mitohondria, elements of sER, …
There are many different forms of kinesins!

Fast retrograde axonal transport
Transports organelles, vesicles, mitohondria, endosomes, etc. towards the soma.
- MAP-1C (Microtubule-associated ATPase)

und Microtubuli!
- MAP-1C simlar to Dynein of Ciliates!

Dynein-Dynactin complex acts as motor!
- ATP-dependent
- independent of soma
Comparison between fast anterograde and retrograde transport
Slow axonal transport, only anterograde:
delivery of cytoskeleton and cytoplasmic elements to periphery.
- Delivery of cytoskeletal and cytoplasmic constituents to the

periphery.
- Studies suggest that cytoplasmic dynein may move
microtubules with their plus ends leading.
- Neurofilaments may move on their own or may hitchhike
on microtubules.
- Once cytoplasmic structures reach their destinations,
they are degraded by local proteases at a rate that allows
either growth (in the case of growth cones) or
maintenance of steady-state levels.
- The different composition and organization of cytoplasmic
elements in dendrites suggest that different pathways
may be involved in delivery of cytoskeletal and
cytoplasmic materials to dendrites. In addition, some
mRNAs are transported into dendrites, but not into axons
The cell membranes of all cells including neurons consist of phospholipid bilayers.
Important for neurons: these bilayers are insulators (charged ions cannot pass easily).
Biological bilayers are usually composed of amphiphilic phospholipids that have a hydrophilic phosphate head and a
hydrophobic tail consisting of two fatty acid chains.
The “fluid membrane mosaic model” states that membrane proteins are embedded in
the phosopholipid bilayer.
These membrane proteins are important to regulate transport across the membrane.
Carbohydrates are covalently linked to proteins (glycoproteins) or lipids (glycolipids) and also an important part of cell
membranes. Membrane carbohydrates perform two main functions: participate in cell recognition and adhesion, either cell-
cell signaling or cell-pathogen interactions, and they have a structural role as a physical barrier.
Also cell organelles are often composed of phospholipid membranes
Polar „head“
with phosphate
group
Unpolar „tail“
of carbohydrate
chains
extracellular
Phospho
lipid-
bilayer
Miljushli
intracellular pidss
General structure of an amino acid
Sequences of hydrophobic amino acids can form

transmembrane domains
Sequences of hydrophilic amino acids can form

intracellular and extracellular domains
Transport across membranes: passive (no energy consumption, driving force = concentration gradient; approaches equilibrium) and
active (energy required, usually ATP, results in non-equilibrium).
https://www.sciencedirect.com/topics/medicine-and-dentistry/membrane-transport
There are several principal types of transport across membranes:
- simple diffusion, e.g. gases (O2, CO2, NO, CO, etc.), small non-charged molecules, also H2O.
- channel-mediated (facilitated diffusion), e.g., ions, H2O through aquaporin channels, small molecules;
- carrier-mediated (facilitated diffusion), e.g., ions, glucose, small molecules;
- active transport, e.g., ions, small molecules; against concentration gradient;
Transport across membranes

amino acid residue
peptide bond
amino acids
Tertiary structure
Quartary structure
subunits
Primary structure: amino acid sequence
Secondary structure
Ion channels are transmembrane proteins

Ion channels consist of subunits
a) central, water-filled pore ,

Heterooligomer Homooligomer Homomer
b) transmembrane-domains of
glyco-proteins
c) 2 oder more subunits that can
be the same or different amino
acid sequences.
Associated proteins can be attached for regulation of channel

properties
Ion channels consist of subunits
Ion channels
1. Ion channels conduct ions

2. Recognize and select
specific ions
3. Open and close upon
electrical, chemical or
mechanical signals.
- Ion selectivity (1:10 – 1:100)

- Mechanisms for regulation
Ion channels
Patch-Clamp-technique: ACh-Receptor-Kanal, 25 pS
Properties of ion channels: opening and closing
a) Voltage-dependent
channels
b) Ligand-dependent
channels
c) Mechanically gated
channels
d) „resting channels“
e) Second-messenger-
regulated channels
Properties of ion channels: ligands, antagonists, agonists
Exogeneous factors can influece

opening and closing (z.B. toxins,
drugs).
Reversibel (Curare) or irreversibel

(e.g., a-Bungarotoxin).
A ligand-gated ion channel is an ionotropic receptor

Ion channels: there are different, structural families
Example: 5 subunits
Connexins
One polypetide
Ion channels: the opening propabilty can be modulated
Ion channels can desensitize, i.e.,

opening decresaes their opening
probability.
Modulation of channels:
Examples:
a) Temporally restricted inactivation

(refractory period!)
b) The conducted ion itself

modulates the opening state.
c) Phosphorylation (covalent
bonds) or dephosphorylation
modulates the opening
probability.
Models, how the S4 domain could transduce voltage into
conformational changes
Different splice variants of a channel protein .
Example: a potassium channel; differential expression of splice variants in the brain

Ion channels: there are different, structural families
Satiation of  Binding of individual ions onto channel pore.
Ionic current is passiv (no ATP

required),
Anions and cations along the

elctrochemical gradient.
+
Example for modulation of opening poperties through phosphorylation: Na -channel
Phosphorylation sites in
the amino acid sequence mediated
by protein kinases.
PK-A: protein kinase A
PK-C: protein kinase C
TTX: site of action of tetrodotoxin

(blocks channel).
How selective are ion channels?
Mesaurements of permeability (P)
potassium-channel: sodium-channel
PX/PK X PX/PNa
< 0,02 Lithium 0,93

< 0,01 Sodium 1,00
1,00 Potassium 0,09
2,3 Thallium 0,33
0,91 Rubidium
0,13 Ammonium 0,16
< 0,03 Hydroxylamin 0,94
< 0,03 Hydrazin 0,59
< 0,02 Guanidin 0,13
Selectivity filter: electrostatic interaction with carbonyl groups in the channel pore.
Pharmacological drugs that can affect ion channels
• Local anesthetics: lidocain, benzocain, procain (sodium channels)

• Peptide toxins (scorpion toxins, conotoxins, sea anemone toxins)
(block inactivation of sodium channels)
• Alkaloid-biotoxins (Veratridin, Batrachotoxin, Aconitin)
(open sodium channels, change ion selectivity)
• Tetrodotoxin (TTX), saxitoxin (block of sodium channels)
1 mg lethal, TTX in several unrelated species
(buffer fish, californian newt)  „convergent evolution“
• Ligand binding 1:1  Counting of sodium channels:

thin axon: ~ 3/µm2, giant axon: ~ 500/µm2,
node of ranvier: 3.000 – 10.000/µm2
• Measuring opening time (patch clamp): ~ 20 ms, ~100 ions pass.
Ion channels: ions flow not only along a concentration gradient but also along
an electrical potential
Direction of ion flux
Ion channels: ionic flow can be skewed in one direction.
Ohmic channel and rectifying channel: ions are more readily conducted in one direction
Voltage-gated ion channels: super-families
Some types of ligand-gated ion channels

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Lecture “Neurobiology”

Prof. Dr. André Fiala

Part Two A: Cellular Properties of Neurons

For Students of Physics or Informatics and for self-studying it is

rough and smooth endoplasmatic reticulum

Ribosomes und Polyribosomes

Present in axones, dendrites, synaptic structures. Cellular respiration, Krebs-Martin cycle;

Blue arrow: longitudinal Red arrow: transverse section

- Microtubules consist of two different monomers

A large number of regulatory proteins are associated with microtubules.

- Microtubules consist of two different monomers

Staining of neurofilaments Staining of Tau Co-localization in dead cells.

- 3-10 x more than microtubules

Intermediate filaments to which neurofilaments belong are a large family of

- 3-10 x more than microtubules

Actin filaments (microfilaments)

- polar polymers of actin monomers

Actin filaments (microfilaments)

A large number of regulatory proteins are associated with microfilaments or

- polar polymers of actin monomers

- small protrusions on dendrites of glutamatergic

- variable shape, modifiable

- provide thousands of synaptic contacts to one cell

- important for learning and memory (plasticity!)

- Deseases can affect spine morphology

- Shape determines electrical properties of

and Cellular Neuroscience, 46, 368-380 .

1 Cyclus = 16 nm, 2 ATP!

There are many different forms of kinesins!

- MAP-1C (Microtubule-associated ATPase)

- MAP-1C simlar to Dynein of Ciliates!

- Delivery of cytoskeletal and cytoplasmic constituents to the

General structure of an amino acid

Sequences of hydrophobic amino acids can form

Sequences of hydrophilic amino acids can form

There are several principal types of transport across membranes:

Transport across membranes

Primary structure: amino acid sequence

Ion channels are transmembrane proteins

a) central, water-filled pore ,

Associated proteins can be attached for regulation of channel

1. Ion channels conduct ions

- Ion selectivity (1:10 – 1:100)

Exogeneous factors can influece

Reversibel (Curare) or irreversibel

A ligand-gated ion channel is an ionotropic receptor

Ion channels can desensitize, i.e.,

a) Temporally restricted inactivation

b) The conducted ion itself

Example: a potassium channel; differential expression of splice variants in the brain

Ionic current is passiv (no ATP

Anions and cations along the

PK-A: protein kinase A

PK-C: protein kinase C

TTX: site of action of tetrodotoxin

< 0,02 Lithium 0,93

• Local anesthetics: lidocain, benzocain, procain (sodium channels)

• Ligand binding 1:1  Counting of sodium channels:

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