BME 401

MOLECULAR BIOENG- 1

MRS DARAMOLA
BELLSTECH
Specialized Mechanosensory
Organs and Cells
Biomechanics of Hearing
Hair Cells
StereoCilia

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Hearing and Mechanotransduction
• Outer and Middle ear convert pressure waves
in air to fluid movement. (Ear also converts
gravitational orientation and movement to
fluid movement.)
• The inner ear has specialized cells called hair
cells that detect fluid movement.
• Hair cells contain specialized mechanosensory
organelles called stereocilia
• Ion channels on stereocilia open to start a
calcium signal.
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How Sound Signals are Transmitted
fluid transmission &
electrical transmission

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mechanical
transmission

tympanic membrane (ear drum) transmits pressure

waves in air to movement of Auditory ossicles (bones)
Middle Ear

Ossicles transmit movement of tympanic membrane to

movement of oval window. 1.3-fold force, 1/18.6 area.
Inner Ear (head rotation)

(hearing)

(gravity)
 Fluid waves in the Cochlea
cross-section of Cochlea
scala vestibuli
scala = chamber
(perilymph)

pressure waves travel from base to apex of cochlea in the scala vestibuli
Reissner’s and Basilar membranes are soft and elastic
pressure depresses these membranes into scala tympani, which dissipates into round window.
 So pressure distorts basilar membrane
 Cochlear Structures
hair cells are attached to basilar membrane
Stereocilia bundles on hair cells face the tectorial membrane in the Organ of Corti
Inner hair cells don’t touch, but outer are embedded into membrane.
When basilar membrane moves, the contact between them and the Tectorial
membrane is sheared
 Stereocilia Structure and Function
stereocilia are composed of actin filaments and myosin
(motor proteins).
rows of stereocilia are organized so each is taller than
the next.
a tip link connects the neighboring stereocilia.
When the stereocilia tilt, the tip links stretch under
tensile force
The tip link opens an ion channel called TrpA on the
taller stereocilia, which leads to electrical signaling of
nerves.
 How the Ear Distinguishes Pitches

The basilar membrane oscillates the most

near the base for high frequencies, and near
the apex for low frequencies

http://www.cns.nyu.edu/~david/courses/perception/lecturenotes/pitch/pitch.html
Models for How Force Opens TRP Channels
ankyrin repeats and motor
proteins activate

membrane tension activates

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Barritt and Rychkov Nature Cell Biology 7, 105 - 107 (2005)
Specialized Mechanosensory
Organelles in Cells
True Cilia

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 True Cilia
• Found on: Endothelial cells (line blood vessels),
epithelial cells (line skin, urinary tract, intestines,
mouth, etc) and osteocytes (mechanosensors in
bone)
• Bundle of microtubules called Axenome inside
plasma membrane.
• Brushes of Motile Cilia move fluid past cells in
trachea and fallopian tubes. Same as single
flagella in sperm cells
• Nonmotile cilia sense fluid movement.
• Anchored via Basal body (a solid support)
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True Cilia
• 9 pairs of
microtubules.
• Two more in
center for
motile cilia
• Dynein motor
protein slides
microtubules
along each
other for
motile cilia.
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 Mechanosensor in Cilia
• Polycystic kidney disease arises from mutations
in ion channels near base of nonmotile cilia.
• But, it is not known how bending of cilia opens
these ion channels.
• Patients with this disease also have problems
with cardiovascular disease, suggesting that
cilia in endothelial cells regulate their function,
but endothelial cells lose cilia at high flow, so it
is not the only flow sensor.
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Mechanosensors in Other Cells

Biological Significance
Examples of Mechanosensors

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 Mechanosensors in Other Cells
• Cells without specialized mechanosensory
organelles can still sense mechanical forces.
• Cells sense the mechanical and chemical
environment. They require the proper signals to
thrive or differentiate.
• Cells undergo apoptosis if they are in the wrong
environment.
• Cancer Cells lose the ability to control when they
divide. Metastasized cancer cells spread throughout
the body because they no longer require the right
chemical and mechanical signals.
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 Other Cells Sense:
• Stiffness of their environment
• Fluid shear stress
• Localized tensile forces; pulling by the
environment
• Topology, Nanostructure and Microstructure.

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 Endothelial Cells
• Endothelial cells (EC) line the blood vessel walls,
creating a barrier between blood and tissues.
• A healthy endothelium emits antiinflammatory
signals that modulate the function of blood cells
to keep them passivated, or in a “resting state”.
• However, if endothelial cells become activated,
they can have the opposite effect, emitting pro-
inflammatory signals that activate blood cells.

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 Endothelial Cells sense strain and fluidic
shear stress

• High laminar steady or pulsatile flow keeps EC passivated,

while low or disturbed flow is pro-inflammatory. Because
of this, atherosclerotic lesions nearly always occur at
branch points in arteries, where flow is disturbed.

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Conway and Schwartz, 2013, Qiagen
 Endothelial Cell Response to Flow
• Long-term flow: Secrete nitric oxide and prostacyclin,
which cause smooth muscle cells to relax and are anti-
inflammatory. They also align with flow.l
• Onset of shear transiently activates inflammatory
pathways like reactive oxygen species.
• Tight junctions where neighboring cells bind each
other are what appears to mediate
mechanotransduction: this involves Cadherin, PECAM-
1 (both are adhesion and signaling molecules), and
binds Src.
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 Atheroschlerosis and Thrombosis
• Activated blood cells called platelets initiate thrombosis
through several pathways, leading to blood clots, which can
occlude arteries.
• Activated immune cells called monocytes cross the
endothelium, emit cytokines, and differentiate into foam
cells, which convert HDL to fatty deposits in the wall and
stimulate migration of muscle cells, to form an
atherosclerotic plaque that thickens and hardens the artery
walls. These can rupture, leading to occlusion of arteries.
• Occlusion of arteries in the brain causes strokes, while
occlusion in arteries feeding the heart causes heart attacks,
or myocardial infarction.
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 Differentiation of Stem Cells
• They differentiate according to stiffness of
substratum.

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Engler et al, 2006 Cell 126 p 677
 Focal Adhesion Complexes
• adhesive proteins (integrin, cadherin) and adaptors (talin,
vinculin, a-actinin) are part of mechanical pathway
• and are coupled to signaling proteins (Src, FAK, Pax)

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 Talin as Mechanosensor
• Talin unfolds under tensile force
• Partially unfolded Talin binds vinculin head domain
vinculin head
bound to H12 of
talin (X-ray)

native talin (X-ray) vinculin head

unfolded talin
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 Testing for Mechanosensing
Pull on Talin with magnetic Detect number of bound vinculin by
bead number of photobleaching events

Detect binding of fluorescent

Vinculin
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More Vinculin bind at Higher force
Talin Mechanosensing Conclusions
• Talin tail binds to actin filaments and head to
integrins, so it is in mechanical pathway.
• Talin native state is folded but unfolded state can
be induced by force.
• Two states have different activity by number of
vinculin they bind.
• How does vinculin induce signal transduction? It
is known to be activated when it binds talin, and
leads to reorganization of the cytoskeleton.
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 Other suggested Mechanosensors
• Integrins initiate signaling when
they bind, so are linked to signal
transduction pathways. They
are in the line of force, and the
active conformation is longer.
• G-protein coupled receptors
have been implicated.
• Some matrix proteins have been
implicated.
• Nuclei have been implicated
(gene expression controlled
mechanically?)
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 Summary 1: Molecular Biomechanics
• Mechanotransduction allows cells to have a
microscopic sense of touch.
• Mechanotransduction requires a mechanosensor, a
mechanical pathway, and a biochemical pathway.
• A mechanosensor must be have force induce an
alternative conformation with a different activity than
the native state.
• Talin and TRP channels are probably mechanosensors,
and many other molecules may be.
• The activity is determined by p1 and p2 using the
now-familiar molecular biophysics principles.
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 Summary 2: Function and Significance
• Hearing is enabled by TRP channels on stereocilia
on hair cells in the inner ear.
• Kidney processes are regulated flow sensing by
true cilia.
• Endothelial Cells sense fluid flow to control
inflammatory state.
• Stem Cells sense Mechanical force to control
differentiation
• Most tissue cells sense forces to control
apoptosis and prevent metastasis.
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