PHOTOTRANSDUCTION: Phase II

The Biochemical Phase

 The activated form metarhodopsin II, which is generated by light, diffused rapidly from the
photoreceptor disk membrane towards the plane of the plasma membrane to trigger an
enzymatic cascade.
 Metarhodopsin II recruits the inactive heterotrimeric G-protein transducing, triggering a
nucleotide exchange from the guanosine disphosphate (GDP) bound to its α-subunit, to
guanosine triphosphate (GTP).
 This subsequently liberates the complex the α-transducin-GTP complex from association with β-
γ dimer.
 The α-transducin-GTP complex then associates with either two γ subunits of the similarly
inactive heterotetrameric cGMP phosphodiesterase (PDE), freeing catalytically active α-β
subunit so that it could now actively effect the hydrolysis of 3’-5’ cyclic guanosine
monophosphate (cGMP) to 5’-guanosine monophosphate (5’-GMP).
 In some cases of retinitis pigmentosa
o Mutations in the TRANSDUCIN gene prevent it from disinhibiting PDE.
o The resulting excess in CGMP levels is regarded to be toxic to the retina.
The Biochemical Phase in Rods and Cones Compared
 A smilar cascade occurs within cone cells except that the PDE in cone cells consists of TWO α-
catalytic subunits (α2) instead of α-β catalytic subunits found in rods.

PHOTOTRANSDUCTION: Phase III

The Electrical Phase
 Most neural membranes have resting potentials of around -60mV, being dominated by the
relative distribution of K+ IONS in and out of the cell.
o However, the membranes of the rod and cones cells, which are quite “leaky” to Na +
permitting the reversible entry of the cation, and are thus able to attain a LESS
NEGATIVE resting potential of around -30mV.
 The electrophysiological response of the photoreceptors to light involves a change in the
permeability of the plasma membrane to certain cations.
o As photons are absorbed by photoreceptors, OPSIN is activated and the same cation
channels, which initially permit the entry of Na + and Ca2+ found in the outer segment ,
start to CLOSE, thus preventing further influx of Na +, without affecting the rate of its
efflux in the inner segment through the action of Na +-K+-ATPase.
 This event renders the membrane potential to be MORE NEGATIVE relative to
the resting state, HYPERPOLARIZATION thus occurs.
 What accomplishes the closure of these cation channels is the substantial
decrease in levels of cGMP = a compound that acts to keep these ligand-gated
cation channels open.
  It has been experimentally determined that the effect of cGMP on these
channels is DIRECT rather than mediated by covalent modification or by binding
of a cystosolic protein.
 The opening of these cation channels by cgmp is likewise highly cooperative,
requiring the binding of at least three molecules of cgmp per channel protein.
 The high degree of cooperativity is seen among cation channels makes them
MORE SENSITIVE to minute changes in CGMP concentration.
 It must be rememere that upon ILLUMINATION, levels of Cgmp dramatically
DECREASE upon the action of the UPREGULATED Cgmp-specific PDE, eventually
leading to the closure of these cation channels.
 HYPOTHESES RELATING HYPERPOLARIZATION of PHOTORECEPTOR CELLS TO VISUAL
EXCITATIONS:
1. Changes in the membrane potential of photoreceptors influence adjacent bipolar cells, thus
reducing intercellular communication electrotonic in nature.
2. In the dark, photoreceptors release an excitatory neurotransmitter (presumably glutamate
or aspartate).
a. However, upon hyperpolarization, less neurotransmitter gets released, thereby
decreasing the stimulation of post-junctional bipolar cells.
3. In the dark, rods and cons may be discharging an INHIBITORY (instead of an excitatory)
neurotransmitter to nearby bipolar cells.
a. Upon illumination, hyperpolarization decreases the releases of the putative
inhibitory neurotransmitter, thus evoking an opposite excitatory response- a
phenomenon called IMPULSE INVERSION
i. This excitatory response is then transmitted to ganglial neurons, which then
send action potentials to optic nerve.
 Regardless of the exact neurophysiologic mechanism, the foregoing hyperpolarization of
photoreceptor cells and in the response of bipolar cells are both considered GRADED
PHENOMENA.
o One photon of light absorbed INCREASES the membrane potential by 1 Mv, with effect
peaking about 1 second after excitation.
 This effect becomes LARGER and FASTER as more and more photons get
simultaneously ABSORBED, reaching the highest level once light energy in 100
photons have activated visual pigments in a single rod.
 Meanwhile, the reaction of the neighboring ganglion cells is an ALL or NONE phenomena i.e.,
whether this cell is triggered, it responds by sending an action potential to cortical centers.

THE ELECTRICAL PHASE IN RODS AND CONES COMPARED

RODS CONES
-12 -15
Sensitivity 1 – 3 x 10 Amps 10 x 10 Amps
(absorption of one photon)
Response time Cones are FOUR times faster than that of rods.
Rods are more sensitive than Cones are better adapted to
 cones even under low-light detect rapid changes in the
conditions. environment.
Cluster of rod cells connects to
each bipolar cell and multiple
bipolar cells connect to each
ganglion cell.

Rods have Cones

 Summation of signals  Lack of signal summation
 HIGHER PIGMENT  Provide BETTER VISUAL
content ACUITY with a one-to-one
 GREATER molar correspondence with
sensitivity bipolar cells
 Strong VISUAL RESPONSE  Exhibit lower sensitivity
OF RODS even to faint compared to rods.
light, especially at night,
alongside some
compromise on precise
spatial discrimination.
 Night Vision mainly
appears in shades of gray
and lacks clarity since the
brain uses combined
signals from rod cells of
the retina.

IMPORTANCE OF SIGNAL AMPLIFICATION IN PHOTOTRANSDUCTION

 Foregoing cascade occurring in visual phototransduction is considered so effective that a single
photon of light is able to prevent the influx of at least 1 million Na+ ions in the outer segment of
the photoreceptor cells.
 In vertebrates, including humans, absorption of a single photon by a rod is estimated to
decrease the current by 3%, roughly equivalent to closing 25 to 1000 Na+ channels , or a change
in membrane potential by 1 mV.
o This scheme highlights the presence and the amplifying effect of a diffusible molecule
that could traverse the disk membranes where rhodopsin is embedded towards plasma
membrane of the photoreceptor outer segment where Na + channels are found- a
characteristic aptly subserved by the second messenger, cGMP.
 One molecule of activated rhodopsin is able to recruit 500 transducin molecules and form
approximately 20 -transducin-GTP complexes, a number sufficient to fully capacitate a single
phosphodiesterase molecule.
 Each phosphodiesterase molecule then hydrolyzes many cGMP molecules at a rate of range of
4,200 molecules/second, before it decays.
REGULATION OF PHOTOTRANSDUCTION
 In order to maintain sensitivity to changing light intensities, phosphodiesterase activation must
be TRANSIENT.
o This property is made possible by hydrolysis of bound GTP to GDP and inorganic
phosphate by instrinsic GTPase activity residing in transducin-GTP complex.
o HYDROLYTIC PROCESS  takes place than a second when transducin is bound to PDE 
promotes reassociation of  subunit of transducin with  and  subunits  to reform
the heterotrimeric inactive state  PDE reforms into its heterotritrameric, resting,
inactive state
 In order to switch off the response to light, as what happens during continuous stimulation
o the enzyme rhodopsin kinase PHOSPHORYLATES metarhodopsin II.
o Rhodopsin kinase TRANSFER nine (9) phosphate groups per molecule of bleached
rhodopsin to selected serine and threonine residues in protein’s carboxyl terminus.
 PHOSPHORYLATED METARHODOPSIN II  becomes associated with sequestering molecule,
ARRESTIN 1.
o This mechanism renders metarhodopsin incapable of activating additional molecules of
transducin.
o Additionally, cGMP must be RESYNTHESIZED in order to reopen cation channels in the
outer segment membrane.
 This is brought about by the effector enzyme, guanylate cyclase, which
catalyzes the conversion of GTP to CGMP. This enzyme is potently INHIBITED by
Ca+ ions (at intercellular concentrations of at least 100nM)
 Ca+ ions enter photoreceptor cells via the same cGMP-gated cation channels in
the photoreceptor outer segment.
 Ca2+ ions is then EXPORTED out of the cell by another transport protein, in the
exchange for Na+.
 This combined Ca2+ influx and efflux mechanism produces intracellular
concentration about 500 nM.
o On illumination, however, it should be remembered, that these cation channel CLOSE
thereby preventing further entry of Ca2+ ions without affecting its efflux by the Na +/Ca2+-
K exchanger.
 This phenomenon thus results in substantial drop in Ca + ion concentration,
which then releases guanylate cyclase inhibition.
 This DISINHIBITION facilitate the conversion of GTP  cGMP, the ULTIMATE
LIGAND that keeps cation channels in the outer segment open for Na + influx,
thus restoring the membrane potential to its RESTING STATE.
 Going further, the Ca2+ binding protein recoverin inhibits rhodopsin kinase at
high [Ca2+], but the inhibition is relieved when [Ca 2+] drops after illumination.
 o More photons are needed to excite a constantly illuminated rod than to excite one in
the dark; a properly termed as photo-adaptation  enables the rod cell to perceive
light-dark contrast over a wide range of background light intensities.

The Biochemical Basis of Olfaction

 The sense of smell is subserved by the activity of olfactory neurons, which possess long thin
cilia that extend from one end of cell into the overlying mucous layer.
o These cilia allow for a large surface area for interaction with potential odorant.
o Chemically, odorants are relatively small organic compounds sufficiently volatile to be
carried as vapors into the nose.
 The ability to discriminate odors is a function of different olfactory receptors in the tongue and
nasal passages; and the ability to integrate sensory inputs from different types of olfactory
receptors in case of odorants that have a “hybrid” pattern.
 The SHAPE of an odorant largely determines its characteristic SMELL.
o It is the structure of the same odorant that allows it to interact specific binding surfaces
on receptor proteins present in the ciliary membrane of the olfactory neurons.
o These olfactory receptors are members of the SEVEN-TRANSMEMBRANE receptor
family, exhibiting marked variability in the central region between helices 4 and 5, thus
suggesting that this region serves as the SITE OF ODORANT BINDING.
o The human genome is believed to encode roughly 500-750 odorant receptors.
 However, more than 50% of these have been discovered to be pseudogenes containing
mutations that prevent the formation of a full-length, functional odorant receptor.
o This loss may allude to the loss of acuity in the sense of smell of higher mammals
including man, presumably due to the reduction in the dependence on the sense of
smell for survival.
o In humans, odorant-receptor proteins are approximately 30-60% identical with each
other.
o Certain anosmias, i.e. the inability to smell specific compounds, may be due to
mutations in the olfactory receptors , which are supposedly sensitive to the given
odorants.
 Odorants are detected mainly in the main olfactory epithelium- the region situated at the
superior portion of the nasal cavity, which contains roughly 1 million sensory neurons.
o The interaction between odorant and receptor triggers a change in receptor
conformation resulting DISPLACEMENT of bound GDP by GTP on Golf, a transmembrane
G protein analogous to transducing for vision.
o The activated Golf (Golf α-adenylyl cyclase in turn catalyzes the conversion of ATP to
cyclic adenosine monophosphate (cAMP).
o With the increase in intracellular cAMP concentration, cAMP-gated Na + and Ca2+
channels of the ciliary membrane OPEN, promoting the influx of Na+ and Ca2+, thus
producing a small depolarization called RECEPTOR POTENTIAL.
 o Ca2+ influx then triggers the openin of Cl- channels, which ultimately DEPOLARIZES the
cell.
o If a sufficient concentration of odorants is present, the receptor potential becomes
strong enough to cause specific olfactory neuron to fire an action potential,  then
relayed to the brain and registers as a specific smell.
o Each olfactory neuron expresses only a single olfactory receptor gene .
o These neurons are likewise linked with specific sites in the brain, thus producing a
spatial map of odorant-responsive neuronal activity.
o These events occur within a period of 100-200 milliseconds.
 It has been experimentally found that a simple 1:1 correspondence between odorants and
receptors does not exist.
o Almost every odorant activates a number of receptors, at varying extents, and almost
every receptor is activated by more than 1 odorant.
o In principle, the combinatorial mechanism allows the relatively small array receptors to
distinguish a large number possible odorants.
 As the olfactory stimulus is withdrawn, several mechanisms turn off the foregoing transduction
mechanism.
o A cAMP-specific phospodiesterase CATALYZES the conversion of cAMP to 5’-adenosine
monophosphate (5’-AMP), thus returning cAMP concentration to baseline levels.
o Similar to what occurs after photo-excitation, Golf via its intrinsic GTPase activity
hydrolyzes its bound GTP to GDP, thereby inactivating itself.
o Similarly, phosphorylation of the olfactory receptors by a specific kinase prevents its
interaction with Golf.
o Finaly, some odorants may get enzymatically broken down by oxidases present in the
system.

The Biochemical Basis of Gustation

 The perception of taste or gustation reflects the activity of gustatory receptors clustered within
taste buds on the tongue surface.
 The difference in specificity among the five taste (sweet, bitter, sour, salty or savory/umami)
reflects the fact that sense of taste is a number of independent sense all utilizing the same
organ- the tongue.
 Further analysis reveals that there are from 50 to 100 members of the family of gustatory
receptors in the entire human genome.
 In contrast to olfaction, each taste receptor cell expresses many different members of the
receptor protein family. Humans are able to characterize tastants to a limited repertoire of
tastes, since many of these chemicals stimulate the same neurons.
 Grossly, the tongue is characterized by structures called papillae, where taste buds are
concentrated. Each taste bud contains approximately 150 cells, containing fingerlike projections
called microvilli, which extend from on end of the sensory neuron to the surface of the tongue.
 Sweet receptors are considered to be part of seven-transmembrane receptor protein family.
 o As tastant molecules bind to specific receptors, the associated G-protein gustducin is
activated, leading to the exchange of bound GDP to GTP.
o Gustducin α-GTP then stimulates cAMP levels then activate protein kinase A, which is
responsible for phosphorylating and thus closing ligand-gated K + channels in the
basolateral portion of the plasma membrane.
 This phenomenon results in the REDUCTION of K + efflux, thue DEPOLARIZING the
cell.
 Salty tastants are detected NOT via 7-transmembrane receptors, but by their DIRECT PASSAGE
through ION CHANNELS expressed on the surface cells in the tongue.
o One such class of channels has first been characterized by its role in salt reabsorption
and its sensitivity to amiloride inhibition.
o Na+ ions, as derived from NaCl, are able to pass directly through channels and produce a
significant transmembrane current, inhibition of these channels mutes the taste of salt,
thus substantially lowering the activation of sensory neurons in response to the sodium
ion (Na+).
 Sour tastes are detected by direct interaction of hydrogen H +, with corresponding ion channels.
o The ligand, however, typically, consists of high concentrations of H +, rather than Na+
ions.
o This H+ influx induces substantial transmembrane current and changes in membrane
polarization, producing sensation of sour taste.
 Umami is a perception distinct from the other basic tastes, subserved by the receptor for the
amino acid and neurotransmitter glutamate.
o This gustatory receptor is believed to have evolved through changes in the expression of
an existing glutamate receptor gene.