Classic Experiment 18.

Looking at Muscle
Contraction
he contraction and relaxation of striated muscles allow us to perform
all of our daily tasks. How does this happen? Scientist have long
looked to see how fused muscles cells, called myofibrils, differ from other
cells that cannot perform powerful movement. In 1954, Jean Hanson and
Hugh Huxley published their microscopy studies on muscle contraction,
which demonstrated the mechanism by which it occurs.

Background

The Experiment

The ability of muscles to perform work has long been a fascinating process. Voluntary muscle contraction is performed
by striated muscles, which are named for their appearance
when viewed under the microscope. By the 1950s, biologists
studying myofibrils, the cells that make up muscles, had
named many of the structures they had observed under the
microscope. One contracting unit, called a sarcomere, is
made up of two main regions called the A band, and the I
band. The A band contains two darkly colored thick striations and one thin striation. The I band is made up primarily
of light-colored striations, which are divided by a darkly colored line known as the Z disk. Although these structures
had been characterized, their role in muscle contraction remained unclear. At the same time, biochemists also tried to
tackle this problem by looking for proteins that are more
abundant in myofibrils than in other non-muscle cells. They
found muscles to contain large amounts of the structural
proteins actin and myosin in a complex with each other.
Actin and myosin form polymers that can shorten when
treated with adenosine triphosphate (ATP).
With these observations in mind, Hanson and Huxley
began their study of cross striations in muscle. In a few short
years, they united the biochemical data with the microscopy
observations and developed a model for muscle contraction
that holds true today.

Hanson and Huxley primarily used phase-contrast microscopy in their studies of striated muscles that they isolated from rabbits. The technique allowed them to obtain
clear pictures of the sarcomere, and to take careful measurements of the A and the I bands. By treating the muscles
with a variety of chemicals, then studying them under the
phase-contrast microscope, they were able to successfully
combine biochemistry with microscopy to describe muscle
structure as well as the mechanism of contraction.
In their first set of studies, Hanson and Huxley employed
chemicals that are known to specifically extract either
myosin or actin from myofibrils. First, they treated myofibrils with a chemical that specifically removes myosin from
muscle. They used phase-contrast microscopy to compare
untreated myofibrils to myosin-extracted myofibrils. In the
untreated muscle, they observed the previously identified
sarcometic structure, including the darkly colored A band.
When they looked at the myosin-extracted cells, however,
the darkly colored A band was not observed. Next, they extracted actin from the myosin-extracted muscle cells. When
they extracted both myosin and actin from the myofibril,
they could see no identifiable structure to the cell under
phase-contrast microscopy. From these experiments, they
concluded that myosin was located primarily in the A band,
whereas actin is found throughout the myofibril.

With a better understanding of the biochemical nature

of muscle structures, Huxley and Hanson went on to study
the mechanism of muscle contraction. They isolated individual myofibrils from muscle tissue and treated them with
ATP, causing them to contract at a slow rate. Using this technique, they could take pictures of various stages of muscle
contraction by using phase-contrast microscopy. They could
also mechanically induce stretching by manipulating the
coverslip, which allowed them to also observe the relaxation
process. With these techniques in hand, they examined how
the structure of the myofibril changes during contraction
and stretch.
First, Huxley and Hanson treated myofibrils with ATP,
then photographed the images they observed under phasecontrast microscopy. These pictures allowed them to measure the lengths of both the A band and the I band at various stages of contraction. When they looked at myofibrils
freely contracting, they noticed a consistent shortening of
the lightly colored I band, whereas the length of the A band
remained constant (see Figure 18.1). Within the A band, they
Z disk

I bands

S 2.8
A 1.5
I 1.3

Stretched
120%
A band
Relaxed
100%

S 2.3
A 1.5
I 0.8

Contracted
90%

S 2.0
A 1.5
I 0.5

Contracted
80%

S 1.8
A 1.5
I 0.3

Contracted
60%

S 1.5

FIGURE 18.1 Schematic diagram of muscle contraction

and stretch observed by Hanson and Huxley. The lengths of
the sarcomere (S), the A band (A), and the I band (I) were measured from 60 percent contraction (bottom) to 120 percent
stretch (top). The lengths of the sacromere, the I band, and A
band are noted on the left. Notice that from 120 percent stretch
to 70 percent contraction the A band does not change in the
length, whereas the length of the I band can stretch to 1.3 microns, then contract to 0.3 microns. At 60 percent contraction,
the I band disappears, and the A band shortens to the overall
length of the sarcomere. [Adapted from J. Hanson and H. E.
Huxley, 1955, Symp. Soc. Exp. Biol. Fibrous Proteins and their
Biological Significance 9:249.]

observed the formation of an increasingly dense area

throughout the contraction.
Next, the two scientists examined how the myofibril
structure changes during a simulated muscle stretch. They
stretched isolated myofibrils mounted on glass slides by manipulating the coverslip. They again photographed phasecontrast microscopy images and measured the lengths of the
A and the I bands. During stretch the length of the I band
increased, rather than shortened, as it had in contraction.
Once again, the length of the A band remained unchanged.
The dense zone that formed in the A band during contraction, became less dense during stretch.
From their observations, Hanson and Huxley developed
a model for muscle contraction and stretch (see Figure 18.1).
In their model, the actin filaments in the I band are drawn
up into the A during contraction, and thus the I band becomes shorter. This allows for increased interaction between
the myosin located in the A band and the actin filaments.
As the muscle stretches, the actin filaments withdraw from
the A band. From these data, they proposed that muscle
contraction is driven by actin moving in and out of a mass
of stationary myosin molecules.

Discussion
By combining microscopic observations with known biochemical treatments of muscle fibers, Hanson and Huxley
were able to describe the biochemical nature of muscle structures and outline a mechanism for muscle contraction. A
large body of research continues to focus on understanding
the process of muscle contraction. Scientists now know that
muscles contract by ATP hydrolysis driving a conformational change in myosin that allows it to push actin along.
Researchers are continuing to uncover the molecular details
of this process, while the mechanism contraction proposed
by Hanson and Huxley remains in place.