Neurons and Muscles

Neurons and Muscles

A. Malcolm Campbell, PhD

Christopher J. Paradise, PhD
Neurons and Muscles
Copyright A. Malcolm Campbell and Christopher J. Paradise. 2016.

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 Abstract
Whenever a dancer or an athlete performs amazing feats, it is the con-
sequence of two very interesting cell types: neurons and muscles. When
the two of these cell types work together, animals can move in complex
ways with surprising control. Not only do they work together to pro-
duce movement, they have many traits in common. They both convert
chemical signals into electrical information, and then back into chemical
information again. This book will examine how neurons process informa-
tion and communicate to adjacent cells. This book presents how muscle
cells know when to contract and how contraction leads to bigger muscles.
Finally, the last chapter presents how long-term memories are formed. In
all three chapters, some of the original data that have contributed to our
understanding of these two fascinating cell types are reproduced to pro-
vide supporting evidence for the function of these two cell types.

Keywords
allosterically modulated, transmembrane, covalent modulation, ligand-
gated, ion channels, depolarized, voltage-gated, refractory period, patch
clamp, myelin, nodes of Ranvier, secretory vesicles, skeletal muscle, motor
neurons, acetylcholine, sarcomere, striated, longitudinal sections, cross
sections, actin, myosin, troponin, tropomyosin, T-tubules, hypertrophy,
hyperplasia, performance-enhancing drugs, short-term memory, long-
term memory, sensoryneurons, synapse, cAMP, PKA, MAPK
 Contents
Preface...................................................................................................ix
Acknowledgments....................................................................................xi
Introduction.........................................................................................xiii
Chapter 1 Neurons Receive and Send Information.............................1
Chapter 2 Muscles Contract and Grow Bigger..................................21
Ethical, Legal, and Social Implications: Use of
Performance Enhancing Drugs.....................................29
Chapter 3 Memories Require New Proteins in Neurons....................33
Ethical, Legal, and Social Implications: Concerns
About Memory Research..............................................41
Conclusion............................................................................................45
Glossary................................................................................................47
Index....................................................................................................49
 Preface
This book about neurons and muscles is part of a thirty book series that
collectively surveys all of the major themes in biology. Rather than just
present information as a collection of facts, the reader is treated more like
a scientist, which means the data behind the major themes are presented.
Reading any of the thirty books by Campbell and Paradise provides read-
ers with biological context and comprehensive perspective so that readers
can learn important information from a single book with the potential to
see how the major themes span all size scales: molecular, cellular, organ-
ismal, population and ecologic systems. The major themes of biology en-
capsulate the entire discipline: information, evolution, cells, homeostasis
and emergent properties.
In the twentieth century, biology was taught with a heavy emphasis
on long lists of terms and many specific details. All of these details were
presented in a way that obscured a more comprehensive understanding.
In this book, readers will learn how neurons and muscles work and some
of the supporting evidence behind our understanding. Instead of believ-
ing or simply accepting information, readers of this book will learn about
the science behind the production of proteins the way professional scien-
tists dowith experimentation and data analysis. In short, data are put
back into the teaching of biological sciences.
Readers of this book who wish to see the textbook version of this
content can go to www.bio.davidson.edu/icb where they will find
pedagogically-designed and interactive Integrating Concepts in Biology for
introductory biology college courses or a high school AP Biology course.
 Acknowledgments
Publishing this book would not have been possible without the generous
gift of Dr. David Botstein who shared some of his Breakthrough Prize
with AMC. Davids gift allowed us to hire talented artists (Tom Webster
and his staff at Lineworks, Inc.) and copyeditor Laura Loveall. Thanks go
to Kristen Mandava for project management and guidance on the pub-
lishing process. In particular, we are indebted to Katie Noble and Melissa
Hayban for their many hours of help and attention to detail.
Kristen Eshleman, Paul Brantley, Bill Hatfield and Olivia Booker
helped us with technology at Davidson College. We are grateful to ad-
ministrators Tom Ross, Clark Ross, Carol Quillen, Wendy Raymond,
Verna Case, and Barbara Lom who had confidence in us and encouraged
us to persist despite setbacks along the way.
These books were the product of the shared labor of my two vision-
ary coauthors Laurie Heyer and Chris Paradise. We shared the dream
and the hardships and developed this book from scratch. My family has
been very supportive and I thank Susan, Celeste and Paulina for their
support and patience. I also want to thank Jan Serie, my pedagogical
mentor, who taught me so much about the art and science of helping stu-
dents learn. I benefited from the support of the Howard Hughes Medi-
cal Institute grant 52006292, the James G. Martin Genomics Program,
and Davidson College. This book would not have survived its first draft
without my students who endured the typos and the early versions of this
book. These undergraduates participated in a bold experiment to see if
beginners could construct their own knowledge, retain what they learned,
and transform the way they see themselves and the discipline of biology.
While many people said that beginning students were not up to the task,
my students proved them wrong.
 Introduction
This book focuses on neurons and muscles. In many ways, cells are similar
to computers. Cells and computers both work from a set of directions and
respond to inputs. Both store information, come in a variety of shapes,
consume energy, use electricity to function, and both can be infected by
viruses. Over time, humans have engineered smaller and smaller comput-
ers to the point that cells have been built to function as computers. The
next generation of computers will include human cells that have been
programmed with DNA and can be implanted into humans to aug-
ment our current functions. This chapter looks at two major functions
of neuronscommunication to another cell and memory formation.
A muscle contracts when it receives the appropriate signal from a neuron.
Neurons can communicate information over long distances very quickly.
Muscles process the output from a neuron and convert that informa-
tion into a contraction that can lead to larger muscles. Neurons encode
memory by depositing new proteins that alter the cells function to benefit
the organism.
 CHAPTER 1

Neurons Receive and Send

Information

At birth, all newborns know how to move their muscles (Figure 1). They
kick, scream, and nurse instinctively. Although they can move their mus-
cles from the first minute of life, they do not have to consciously think
about the cellular actions required to move their muscles. All they have
to do is tell their body to respond, and it does. How fast can nerve cells
work? In the blink of an eye. The desire to blink tells the muscles that
control the eyelids to contract and relax in a matter of milliseconds. Since
diffusion alone is too slow to transmit the blink command from the
brain to the eyelid muscles, cells must use a different mechanism for com-
munication. The desire to blink starts as a chemical signal that stimulates
a nerve whichextends to the eyelid muscles. The neuron, or nerve cell,
receives the chemical input and converts the signal to an electrical cur-
rent that races down the axon to its terminal adjacent to the muscle cell.
At the neuromuscular junction, the neuron converts the electrical signal
back into a chemical message that is transmitted a very short distance
to the muscle, which interprets the information and contracts. The pro-
duction of cellular electricity begins with a modified version of signal
transduction.
Before understanding the neurons signal transduction, consider the
resting state of a neuron (Table 1). Cells do not maintain equal con-
centrations of ions inside and out. Because ions are charged particles,
they cannot pass through phospholipid bilayer membranes unassisted. In
most animal cells, more sodium ions (Na+) are excluded from the cyto-
plasm and accumulate in the extracellular environment. Conversely, cells
sequester more potassium ions (K+) in their cytoplasm than are typically
found outside the cells. Cells expend an enormous amount of energy to
2 NEURONS AND MUSCLES

Figure 1 Neurons use chemical and electrical signals to communicate

information. Neurons receive chemical inputs and convert them to
electrical signals that quickly convert the information to chemical
outputs. Chemical information is indicated by boxes and atomic
symbols; lightning bolts symbolize e lectrical signals.
Source: Original art.

maintain this unbalanced ionic state with a higher concentration of Na+

outside and a higher concentration of K+ inside.
Every cell on the planet has to regulate ions in order to live. Every
animal cell has in its plasma membrane Na+/K+ pumps to help regulate
sodium and potassium ions (Figure 2). As the name implies, a pump
is an active transporter, meaning that it requires energy in the form of
adenosine triphosphate (ATP) to move the ions to establish concentra-
tion gradients. If diffusion were the only force, then sodium gradually
 Neurons Receive and Send Information 3

Table 1 Concentration of ions inside and outside animal cells.

ion intracellular concentration extracellular concentration
K+ 155 mM 4 mM
Na+ 12 mM 145 mM
Ca2+ 0.0001 mM 1.5 mM
Cl 4 mM 120 mM
Source: Common knowledge. Additional negatively charged ions are also involved but
this is beyond the scope of this book.

phosphate group

2 potassium ions

cytoplasm
membrane

gamma beta
subunit subunit
A B extracellular

Figure 2 Primary and quaternary structure of a Na+/K+ pump. A,

Primary structure of one of three human Na+/K+ pumps encoded
bythree genes. The protein contains about 1000 amino acids. B,
Tertiary structure of the same Na+/K+ pump in panel A, along with
two additional subunits, gamma and beta.
Source: Panel A is public domain. Panel B is public domain from PDB file 3B8E.

would diffuse into cells and potassium out of cells. The Na+/K+ pump
helps maintain the ion concentration gradients shown in Table 1. As with
many ion pumps, ATP is converted to adenosine diphosphate (ADP),
and the terminal phosphate is covalently added to the pump protein
on an aspartic acid, which is abbreviated D. The phosphate covalently
modulates the proteins shape and thus changes its function. The discov-
ery of the Na+/K+ pump was rewarded with a Nobel Prize, but it took
many more years to discover how the pump moves charged particles in
opposite directions across plasma membranes. Physiologists in the 1960s
slowly uncovered how the pump functioned. First, they added each ion
to the pump in the presence of radioactive ATP to determine which ion
stimulates the pumps phosphorylation. Only when sodium was added to
4 NEURONS AND MUSCLES

the cytoplasmic side of the pump did the pump bind to ATP and phos-
phorylate itself on the aspartic acid. Additional research over the next
20 years allowed physiologists and biochemists to completely diagram
how the Na+/K+ pump worked.
Neurons use electrical communication to move information down
their lengths as rapidly as possible. Chemical communication informs a
neuron what action it should take and what action it should pass on, such
as telling a muscle cell to contract. When neurons and muscles are resting,
they have thirty times more potassium in their cytoplasm than on the out-
side, and twelve times more sodium outside compared to inside. Na+/K+
pumps help maintain these ion gradients as a form of potential energy
that can be used later. For every ATP consumed, the Na+/K+ pump
moves three sodium ions out of the cell and two potassium ions into the
cell. This 3:2 ratio is a vital ion ratio because it contributes to the electrical
potential every animal cell on the planet needs to perform a wide range
of functions. By exporting one more Na+ than K+ it imports with each
Na+/K+ pump cycle, the cytoplasm gradually becomes more negative
compared to the extracellular world. One ion at a time, cells accumulate
an overall membrane potential, which is a separation of charged particles
in the form of Na+ and K+ ions. Animal cells have a resting membrane
potential in the range of 20 to 200 millivolts (mV), although the
precise number varies by cell type and species. The membrane potential
is also substantially maintained by a series of K+ ion leak channels not
discussed in this book.
Neurons that communicate to muscle cells at the neuromuscular junc-
tion have a resting membrane potential of approximately 70 mV, but
neurons are not resting to maintain these ion gradients. The membrane
potential is caused by a polarization of ions. All animal cells spend about
half of their energy maintaining ion gradients, so it may seem like an oxy-
moron to call these resting membrane potentials. When neurons are not
using electrical communication, they are described as resting, and the lack
of electrical activity is what is meant by resting membrane potential.
The Na+/K+ pump changes its shape in response to covalent modula-
tion when it is phosphorylated or dephosphorylated. Figure 3 illustrates
the steps. The pump (1) is allosterically modulated when it binds three so-
dium ions (2), which leads to ATP binding and phosphorylation (3). Once
 Neurons Receive and Send Information 5

Adenosine-PPP
1 P
ion pump
no ions 2 3
bound ion pump Adenosine-PP ion pump
3 Na+ 3 Na+ 3 Na+
(inside) bound bound
2 K+ (inside) 4
6 ion pump
ion pump 5 no ions
2 K+ bound bound
ion pump
2 K+ P 3 Na+
bound (outside)
P 2 K+ (outside)

Figure 3 Chemical activity of the Na+/K+ pump. Diagram of the

Na+/K+ pump cycle. Circles represent modulated states of the enzyme
and subsequent shape changes.
Source: Original art.

phosphorylated, the covalently modulated pump has a shape change in the

transmembrane domains and develops a low affinity for Na+. The pump
opens toward the extracellular world and lets go of the three sodium ions
toward the outside of the cell (4), which causes another shape change. Now
the pump has a high affinity for two potassium ions (5), which bind from
the extracellular environment to allosterically modulate the pump again.
Upon binding two potassium ions, the pump breaks its covalent bond to
the phosphate (6), which reverses the covalent modulation from step 3.
The transmembrane domains move again, close the opening on the extra-
cellular side, produce an opening on the cytoplasmic side, and the pump
develops a low affinity for K+. The pump releases the two K+ ions into the
cytoplasm, which allosterically modulates the pump again, and the cycle re-
peats (1). In completing the cycle, the Na+/K+ pump regains its high affin-
ity for sodium ions inside the cell, and the pump resumes its never-ending
cycle. The heart medication, digitalis, works by stalling a dose-dependent
subset of cardiac Na+/K+ pumps so that they cannot transport ions any
more. The net result is a reduction of membrane potential closer to 50 mV
with more sodium inside the cells than normal.
Neurons use the potential energy in the form of a membrane poten-
tial to produce an electrical current, which is the movement of charged
6 NEURONS AND MUSCLES

particles. In Figure 1, the upper chemical communication box highlights

the area of neuron activation. A neurotransmitter ligand binds to its re-
ceptor on the neurons plasma membrane, which initiates a new form of
signal transduction (Figure 4). This neurotransmitter receptor is a ligand-
gated sodium channel. Ion channels bind to ligands and gate open
or closed because the structure of the ion channel functions as if it has a
door that can open or close. Once the ion channel gate is open, cells can
use the potential energy of ion gradients generated in part by the Na+/K+
pump. When the ligand binds, the ion channel allows only sodium ions

pore
beta delta
alpha subunit subunit
subunit
alpha alpha
beta subunit subunit
subunit
extracellular

gamma
subunit pore
membrane

delta cytoplasm
subunit
alpha gamma
subunit subunit

A top view B side view

open
extracellular closed extracellular ligand

cytoplasm cytoplasm

C D

Figure 4 Gated sodium ion channel structure and function. A, Channel

viewed from the extracellular side is composed of five subunits. B, Side
view of the same sodium channel but illustrated to show secondary
structures. Diagram of the gating mechanism that blocks (C) or
permits (D) Na+ to flow down its concentration gradient.
Source: Panels A and B are from PDB file 2bg9. Panels C and D are original art.
 Neurons Receive and Send Information 7

to flow down their chemical and electrical concentration gradients into

the cytoplasm. Ion channels are not as specific as enzymes, but they
are selective. Only ions of a particular charge and atomic radius can fit
through the pore of any given ion channel. Ion channels span the plasma
membrane, with ligand binding on one side and the transmembrane pore
within the phospholipid bilayer.
Once a Na+ ion channel opens, it allows positively charged Na+ ions
to move, producing an electrical current. When the polarization of ions
is reduced (extracellular Na+ ions enter the cell), then the membrane is at
least partially depolarized. Cellular current can be measured by placing
an electrode into a neuron and quantifying movement of ions across the
membrane (Figure 5). Rather than relying on a chemical signal to initi-
ate the current, investigators typically start their experiments by applying
electricity directly to the cell and then measuring ions rushing into the
neuron down the length of the axon. One benefit of using electricity to
stimulate neurons during experiments is that the level of input stimulus

input output

measure

neuron

A
applied

applied
current

current

+50 +50
potential (mV)

potential (mV)
membrane

membrane

time time
0 0

50 50
70 70
0 1 2 0 1 2
B time (ms) time (ms)

Figure 5 Stimulation and measurement of electrical current in

neuron. A, Stimulating electrode delivers known input current, and a
measuring electrode detects output current (membrane potential) at
its location along the axon. B, Two different stimulating inputs (10 or
20 mV) are applied to a neuron, and the resulting output membrane
potential is measured in the axon.
Source: Panel A is original art. Panel B is modified from Phillips et al., figure 17.2.
8 NEURONS AND MUSCLES

is easily controlled. Figure 5B shows the averaged results from repeated

experiments of applying each of two different levels of stimulating current
to neurons. When a cell was depolarized by only 10 mV (left side inset),
the measuring electrode (left side lower graph) detected exactly what was
applied to the cell, and the voltage decayed over time. However, when the
neuron was depolarized by 20 mV, the output current was much greater
than the input current, and the membrane potential increased by 120 mV
to a maximum of +50 mV. Through experiments similar to Figure 5,
physiologists had discovered that neurons require a threshold level of
stimulation before they will propagate their own electrical impulse
down their axons. When a neuron is stimulated at least as high as the
threshold, then the cell produces a complete wave of depolarization. In
further experiments, the physiologists found that no matter how much
above thethreshold their input stimulation, the neuron always pro-
duced thesame sized wave of depolarization in what has been called an
all-or-none response. Either the neuron is stimulated and it produces a
full depolarization (all), or threshold is not reached and the neuron fails
to produce its own depolarization wave (none).
Once biologists realized that neurons could be stimulated fully with
a threshold depolarization, they designed many more experiments to un-
derstand how cells propagate a wave of current (Figure 6). Investigators
stimulated a neuron once and simultaneously measured the membrane
potential at three locations along the axon. Electrodes further from the
activating electrode detected less depolarization. The investigators rede-
signed their experiment to measure membrane potential over a longer
time period (Figure 7). When they allowed enough time for the cells de-
polarization wave to travel down the axon, they detected the all-or-none
depolarization at every point down the length of the neuron. They called
this traveling wave of depolarization the action potential.
Ligand-gated ion channels densely populate a small portion of the
neurons plasma membrane where the chemical signal is first delivered.
Surrounding these ion channels and covering the vast majority of the
surface area on a neuron are a different type of gated ion channel called a
voltage-gated ion channel. As the name implies, voltage-gated ion chan-
nels open and close in response to depolarization of the membrane instead
of ligands binding receptors. At resting membrane potential, the sodium
 Neurons Receive and Send Information 9

applied
current
time

current generator

input

axon of neuron

measure

output
applied

applied

applied
current

current

current
time time time

Figure 6 Depolarization at one point in time. A neuron was

stimulated (top left) and the depolarization was measured (bottom
three probes) simultaneously at three distances from the site of
activation.
Source: Modified from Alberts et al., 1989, figure 19.12.

ions are more abundant outside a neuron, but once the membrane is lo-
cally depolarized to the threshold level or more, then the voltage-gated
sodium channels open. Like all proteins, these voltage-gated ion channels
do not know what is happening outside of their tiny local environment.
All they can sense is their structure as determined by the local charged
ions interacting with the side chains of their amino acids. Like ligand-
gated ion channels, voltage-gated ion channels behave like a drinking
fountain in that the fountain is always ready to let the water flow but the
fountain must be gated first. The voltage-gated ion channels are selective,
and the first wave of ion channels open to permit only Na+ ions to rush
into the neuron.
In Figure 5, threshold was approximately 50 mV, or a change of
+20 mV. At threshold depolarization, the voltage-gated sodium chan-
nels open, and they contribute further to the local depolarization until
the membrane potential is inverted to about +45 mV. Positive 45 mV
10 NEURONS AND MUSCLES

applied
current time

current generator

propogation
input

axon of neuron

measure #1 measure #2 measure #3

measure #1

measure #2

measure #3

0 1 2 3
time (milliseconds)

Figure 7 Depolarization over time. A neuron was stimulated (top

left) and the depolarization was measured over 3 milliseconds at three
distances (below neuron) from the site of activation.
Source: Modified from Alberts et al., 1989, figure 19.12.

represents the full depolarization for the cell in Figure 5. In Figure 7, the
depolarization wave moved down the length of the axon, and at each
position, the membrane potential reached +45 mV. The magnitude of
the action potential exceeded the initial stimulating voltage (+20 mV)
because once threshold was reached in a local area, all of the neighbor-
ing voltage-gated sodium channels opened and the membrane potential
was made less negative, causing more voltage-gated sodium channels to
 Neurons Receive and Send Information 11

open. As with any form of diffusion, intracellular sodium ions spread to

more voltage-gated sodium channels, and localized depolarization reaches
threshold again but in a new area. This process produces a full action
potential that will spread the entire length of the neuron. The duration of
a wave of depolarization is longer than the stimulating impulse because
many more ion channels open over time and allow more sodium ions to
diffuse to the next zone of voltage-gated sodium channels, which also
open in response to threshold depolarization.
To prevent an echo of the action potential from bouncing back to
the site of its origin, all voltage-gated sodium channels have a refractory
period of time when they cannot open even if the local threshold depo-
larization is exceeded (Figure 8). The area of depolarization has time to
repolarize (about 70 mV again) until the channels have rested long
enough to become responsive again to local depolarization. Neuronal ac-
tion potentials are one-way phenomena, and they do not move backward
to the area of the original stimulation.
Each ion channel permits about 1 million ions per second to move
from one side of the membrane to the other, which results in a rapid
rise in local ion concentration. Unlike ion channels, one Na+/K+ pump
completes its pumping cycle about 33 times per second, which results
in approximately 100 Na+ ions moved out every second. Sodium ions
rush into a neuron about 10,000 times faster than they can be removed
by pumps, yet a neuron can repolarize in milliseconds and not minutes.

cannot
open closed
extracellular Na+ channel extracellular K+ channel extracellular K+ channel

cytoplasm cytoplasm cytoplasm

A after depolarization B depolarized C shortly after depolarization

Figure 8 Neuron repolarization. A, Once closed, a voltage-gated

sodium channel cannot open again for a few milliseconds. B and C,
Voltage-gated potassium channels require threshold depolarization,
but their gating is time-delayed.
Source: Original Art.
12 NEURONS AND MUSCLES

Therefore, the neuron needs an equally fast mechanism to return the

membrane to the resting potential. It would be difficult to move Na+
ions back out quickly because the cell would be moving Na+ ions into a
Na+-rich area. Na+ ions would not rush back out via passive diffusion as
fast as they rushed in. Scattered among the voltage-gated sodium chan-
nels are counteracting voltage-gated potassium channels. With the cyto-
plasm now positively charged, compared to the outside, and the chemical
gradient of K+ ions shown in Table 1, K+ ions experience electrical and
chemical gradients to leave the cell (Figures 8B and 8C). Voltage-gated
sodium channels opened immediately upon threshold depolarization,
but voltage-gated potassium channels opened a few milliseconds after the
threshold depolarization has been reached. Because K+ ions pass through
their voltage-gated K+ channels at about 1 million ions per second, the
membrane repolarizes by the exodus of K+ ions as quickly as it depolar-
ized but with a slight time delay due to the time difference of opening for
Na+ and K+ ion channels.
Depolarization and repolarization occur within milliseconds of each
other, but this presents a new problem. Before a neuron can generate a
new action potential, it must reestablish the ion gradients in Table 1.
Neurons are ready to fire again in less than a second. The rate of blink-
ing is controlled by the rate of action potentials moving through neurons
and communicating to muscles. The only protein available to pump K+
ions back into the nerve cell and Na+ ions out again is the same Na+/K+
pump that contributed to the resting membrane potential in the first
place. Millions of ions have to be pumped at the expense of ATP which
helps explain why nearly half of a cells energy is spent maintaining the
ion gradients in Table 1.
Neurons need hundreds of more Na+/K+ pumps than voltage-gated
sodium channels to match ion transport rates. Different membrane pro-
teins must be produced in different amounts, and their densities in the
plasma membrane can vary over several orders of magnitude. Action
potentials do not echo backward because the Na+ ion channels have
a refractory period, so they cannot reopen immediately toward the ac-
tion potentials origin. Furthermore, mixed among the voltage-gated
Na+ channels are voltage-gated K+ channels which quickly regenerate a
net negative charge in the cytoplasm. The density of voltage-gated Na+
 Neurons Receive and Send Information 13

electrode
measures
current

electrode clamp

membrane patch

axon plasma membrane

Figure 9 Patch clamp method measures single channel activity. Pipet

removes a patch of membrane containing one ion channel.
Source: Original Art.

channels and voltage-gated K+ channels are approximately equal. They

both facilitate ion movement at about the same rate. The much more
abundant but slower Na+/K+ pumps help reset a neuron before it can be
activated again by a new ligand binding to the ligand-gated ion channels
that can initiate a new action potential. These four types of proteins (a
ligand-gated ion channel, two voltage-gated channels, and an ion pump)
play important roles in the electrical communication in neurons.
A neurons function is the cumulative response of hundreds of thou-
sands of individual proteins working independently. Physiologists discov-
ered that a very small pipet tip placed onto the plasma membrane of
a neuron can pull off a patch of membrane and hold it in the narrow
opening (Figure 9). The patch clamp method allows scientists to watch
the activity of one or a few ion channels at a time instead of the averaged
response of an entire cell. By isolating only one channel in a patch of
membrane, they could watch the current fluctuate as the lone gate opens
and closes. When one ion channel opens, the current goes from zero to
2 picoamps (pA). If a second channel were present, the current could rise
to 4 pA. An ion channel does not stay open for a constant amount of
time. The duration of an open channel ranges from about 1 millisecond
to over 10 milliseconds. This variability of open times indicates that ion
channels are inherently noisy and that random movements of amino acid
side chains play a role in the channels behavior.
14 NEURONS AND MUSCLES

If three ion channels were independently stimulated by a defined elec-

trical input, they would not open and close synchronously. One channel
might close very quickly, another might partially closed three times, and
the third one might opened once but stayed open consistently. An ac-
tion potential is the summation of many individual ion channels from
a population of voltage-gated sodium channels. Eventually, all of the
channels will close and enter their obligatory refractory period before the
stimulating input has ended. Action potentials in neurons result from
many voltage-gated sodium channels followed by many voltage-gated po-
tassium channels and many more Na+/K+ pumps to help the ions return
to the gradients in Table 1.
A neuron conducts electricity down its length as a consequence of
thousands of ion channels and pumps. Neurons communicate electrical
information like a copper wire carries information in the form of electri-
cal current. However, vertebrates and some other species have evolved
a cellular insulation that enhances the rate of electrical communication
down axons (Figure 10). Nerve cells that appear white in animals are
covered by insulating cells, called Schwann cells, that form a lipid-rich
myelin sheath surrounding the axons. The myelin sheath is more than
just insulation, because myelinated axons conduct electricity faster than
non-myelinated axons of the same diameter.
Electron micrographs reveal that myelin is composed of a Schwann
cells extended membrane wrapping around the axon 20 to 30 times to
produce a hydrophobic, fatty insulation. The Schwann cells multi-layered

axon of nerve cell

myelin from Schwann cell

Figure 10 Myelinated axons conduct electricity faster than non-

myelinated axons. Diagram of two axons and how myelin accelerates
the conductance of electricity in myelinated nerves.
 Neurons Receive and Send Information 15

plasma membrane wrapping produces an insulator of sufficient thickness

to ensure that the ions do not leak from the axon but not so thick as to
waste cellular resources. Myelin insulation is not continuous for the entire
length of the axon. There are gaps between adjacent Schwann cells called
nodes of Ranvier, named for the man who discovered them in 1867.
Holes in the insulation are not harmful to its function. Instead, neurons
use a saltatory movement of ions across the plasma membrane to propa-
gate the action potential. Without extracellular Na+ ions near sodium
channels, a neuron could not use electricity to communicate down its
axon. The nodes of Ranvier allow neurons to transmit their electrical cur-
rent faster skipping every other step when racing up stairs. Slower, non-
myelinated neurons conduct electricity by small steps of Na+ and K+ ion
flow. Myelinated axons can skip the intermediate steps and jump faster
from one node of Ranvier to the next. The optimal distance between
nodes is determined by the rate of Na+ ion diffusion in the cytoplasm
of axons, and the threshold of voltage-gated ion channels clustered at the
nodes and absent from areas covered by Schwann cells.
Ion channels flick open and closed with random timing making it im-
possible to predict when a particular channel will open next or how long
it will stay open. However, while the channel is open, the current always
reaches the same magnitude because ions are rushing through the channel
at maximum velocity and cannot go any faster. The summation membrane
protential response of an action potential will terminate even if stimulus
persisted. The reason the current ends is that gradually, the collection of ion
channels enter their refractory period and cannot open again until later. By
the time the refractory period is over, the original stimulus has been termi-
nated and only a single action potential will travel the length of the neuron.
The benefit of myelin is only realized if nodes of Ranvier are appropri-
ately spaced. The spacing of the nodes allows waves of sodium to flood the
axon cytoplasm and spread to the next node where more voltage-gated
sodium channels are available to open. The myelin covers most of the
membrane, so ions cannot enter the cytoplasm except at the nodes. In
axons of the same diameter but without myelin, all of the ion channels
can open, and it takes longer for the action potential to gradually move
down the axon because incremental movement would be much shorter
than the bigger steps between Nodes.
16 NEURONS AND MUSCLES

The nerve impulse began with a chemical signal that activated

ligand-gated sodium channels to initiate the electrical communication.
Voltage-gated sodium channels propagate the signal down the axon,
while voltage-gated potassium channels reverse the depolarization.
With the benefit of myelin, axons send the action potential faster to
their terminal ends where the cells produce a new chemical signal to
communicate with the muscle cell. At the end of the action potential,
depolarization reaches the terminal plasma membrane. Located at the
terminal ends of axons are yet another type of ion channelvoltage-
gated calcium channels. These calcium channels are required to con-
vert the electrical signal back into a chemical signal that will reach the
neighboring cell.
The movement of Ca2+ ions through calcium channels at the nerve
terminus can be measured in a way that does not disturb the cell, unlike
the patch clamp method that kills the cell from which the patch was
taken. It is possible to load the cytoplasm of live cells with a fluorescent
dye that changes color depending on the concentration of Ca2+ ions.
Computers can covert the Ca2+ ion concentration to a rainbow color
scale from dark blue, which represents no detectable Ca2+, all the way
to red, which represents the highest levels of Ca2+ ions. While watch-
ing these neurons in real time through a microscope, investigators can
stimulate a neuron on one end and view the opposite end to see what
happens when the action potential arrives at the terminus. The nerve ter-
minus at the neuromuscular junction is flooded by calcium ions (turn-
ing the microscopic image red), because voltage-gated calcium channels
in the plasma membrane opened in response to the action potential and
the Ca2+ ions rushed down their electrical and chemical concentration
gradients (Table 1).
The action potential has been converted from a wave of sodium ions
to a terminal wave of Ca2+ ions. Both ion waves represent electrical cur-
rent rather than a chemical message that can be passed to the next cell.
To secrete its neurotransmitter neurons must perform a process called
exocytosis. Exocytosis causes cells to secrete molecules to the extracellular
environment because the membranes that form vesicles which contain
neurotransmitters fuse with the plasma membrane. During exocytosis,
the secretory vesicles release their contents to the outside the cell. At
 Neurons Receive and Send Information 17

the termini of axons, neurons accumulate secretory vesicles that contain

neurotransmitters similar to the chemical messengers that started their
own electrical signal. Exocytosis must be a tightly regulated process or
neurons would tell muscles to contract at times other than when they
were supposed to contract. Regulated exocytosis is initiated by a rise
in cytoplasmic Ca2+ ions, which is produced in response to the action
potential. The trigger for regulated exocytosis is a Ca2+-binding protein
located in the membranes of secretory vesicles. To determine the loca-
tion of the Ca2+-binding protein, investigators performed an experiment
where they applied antibodies tagged with tiny gold beads to thin slices of
the termini of neurons and then visualized the experimental results with
an electron microscope. The Ca2+-binding protein is located wherever
they saw multiple black dotson the outside of the secretory vesicles.
Once this protein binds Ca2+, it changes its shape through allosteric
modulation. The new protein shape allows it to interact with comple-
mentary proteins on the plasma membrane, so they adhere to each other.
As long as the nerve terminus has elevated Ca2+ ions, it will continue to
secrete neurotransmitter to the neighboring cell. Once the action poten-
tial stops, calcium is pumped out of the cytoplasm, exocytosis stops, and
the adjacent cell is no longer bombarded with neurotransmitters.
Voltage-gated Ca2+ channels fill the small volume of neurons termi-
nus with enough Ca2+ to initiate regulated exocytosis. In order to stop
secreting neurotransmitter, the neuron must be able to rapidly reduce the
concentration of Ca2+ ions. The Ca2+ ion gradient in Table 1 had been
established by Ca2+ pumps located in the plasma membrane. Therefore,
all of the Ca2+ that flooded the cytoplasm from outside the cell must be
returned rapidly back to the extracellular environment in order stop exo-
cytosis. Because pumps move ions more slowly than channels, there must
be many pumps for every one voltage-gated Ca2+ channel.
Embedded in the membrane of a neurons secretory vesicles are pro-
teins that can bind Ca2+. Their Ca2+ binding sites are located in the cyto-
plasm and not inside the lumen of the vesicles because the increased Ca2+
ions are located in the cytoplasm. A rise in Ca2+ leads to a new protein
shape and the fusion of secretory vesicles to the plasma membrane. Upon
fusion, a tunnel or opening forms between the membrane of the vesicle
and the plasma membrane of the cell, and the opening grows in size until
18 NEURONS AND MUSCLES

the vesicle becomes part of the plasma membrane and its contents are
completely outside the cell.
Every activated neuron goes through the same three steps: 1) receive
a chemical message; 2) produce an electrical current that causes an action
potential to move down the axon; and 3) produce a new chemical mes-
sage to secrete via exocytosis. Action potentials are waves of Na+ ions that
jump from one node of Ranvier to the next in myelinated neurons. At
the end of the axon, calcium floods the cytoplasm, which triggers regu-
lated exocytosis of the neurotransmitter chemical messenger. Nerve to
muscle communication is very complex, but every animal performs the
same process for every movement. The next chapter describes how similar
neurons and muscles are with regard to their shared use of action poten-
tials and ion concentration gradients.

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 Index
Actinpolymers, 24
Action potential, 8
in neurons, 14
prevent an echo of, 11
protential response of, 15
Active transporter, 2
Adenosine triphosphate (ATP), 2
to adenosine diphosphate (ADP), 3
binds to myosin, 25
myosin consumption, 2425
Aplysia, 3334
electrical shocks, purpose of, 34
gill retraction in, 35
neuron L7, stimulation of, 3435
neurons responsibility, 34
C/EBP, 38
Ca2+-binding protein, 17
Cardiac muscle, 21
Cellular current, measurement of, 7
Chemical communication, 4
Conscious contraction versus reflexive
muscle movement, 35
Covalent modulation, 5
CREB-1, 3738
CREB-2, 3738
Cyclic adenosine monophosphate
(cAMP), 36
Depolarization, 12
Exocytosis, 1617
of neurotransmitter, 38
G-protein, 36
Gastrocnemius, 28, 29
Gill retractions, 35
neural circuit connecting tail
shocks, 36
Homodimer, 38
Hyperplasia, 2728
Hypertrophy. See Hyperplasia
International Olympic Committee
(IOC), 30
Kandel, Eric, 33
Ligand-gated ion channels, 8
to initiate electrical
communication,16
Lighter protein rods, 2324
Long-term memory, 34
formation of, 38
key aspect of, 37
steps to, 41
MAPK, 37
Memories
people with photographic, 3839
require new proteins in neurons,
3343
Motor neurons, 22
versus sensory neurons, 34
mRNA, translation of, 41
Muscles
anatomy and structure, 21
ATP in, 2526
cell, longitudinal view of, 23
contraction and growth, 2131
experiments for bigger muscle
growth, 28
microscopic images of, 2223
neuron communication, 22
number of nuclei in, 22
performance-enhancing drugs
(PEDs), use of, 2931
sarcomeres, 22
T-tubules in, 2627
vertebrates, types of, 2122
Myelin sheath, 14
electron micrographs of, 1415
Myelinated axons, 14
Myosinpolymers, 2425
cycle, 25
rod polymers, 25
50 INDEX
Na+/K+ pump, 34
chemical activity of, 45
Neurons
benefit of myelin, 15
use electrical communication, 4, 14
function of, 13
membrane proteins for, 1213
memories require new proteins
in,3343
movement of Ca2+ ions, 1617
to muscle cells, 4
Na+/K+ pump, 34
at neuromuscular junction, 1
neurotransmitter receptor, 67
potential energy, 56
receive and send information, 118
repolarization of, 11
resting state of a neuron, 12
secretory vesicles, 1718
stimulation and measurement of
electrical current in, 78
with threshold depolarization, 8
Nodes of Ranvier, 15
Patch clamp method, 13
Performance-enhancing drugs
(PEDs), 2931
Phosphorylation by PKA,
3738
Plantaris, 28
Potent transcription factor, 38
Protease, 38
Protein kinase A (PKA), 3637
phosphorylation by, 3738
Proteins
four types of, 13
memory research, 4143
in muscles, 28
in neurons, memories require
new,3341
Refractory period, 11
Regulatory protein, 37
Repolarization, 12
Resting membrane potential, 4
Sarcomere, 22
composition of, 2324
membrane network surrounding,27
Sarcoplasmic reticulum (SR), 2627
Schwann cells, 14
Secretory vesicles, 1617
Sensory neurons
versus motor neurons, 34
serotonin neuron
communication,35
Serotonin
neuron communication, 35
neuron from tail and siphons
sensory neuron experiment,
3940
Short-term memory, 34Short-term
memory, 34, 3536, 4041
Skeletal muscle, 21
contraction of, 22, 26, 35
Smooth muscle, 21
Sodium ion channel, structure and
function, 6
Steroid injections, 30
Testosterone, 2930
Thicker dark protein rods, 23
Transmembrane domains, 5
Tropomyosin, 26
Troponin, 25, 26
T-tubules, 26
Vertebrates, types of, 21
Vesicle proteins, 37
Voltage-gated ion channel, 89
propagate signal down, 16
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