Shedding light on photoperiodism

Ignacio Provencio1
Department of Biology, University of Virginia, Charlottesville, VA 22903

n mammals, photoreception is restricted to the eye. By contrast, nonmammalian vertebrates possess

photoreceptors in a wide array of
extraocular tissues, including the pineal
complex and brain (1). The identity of the
deep brain photoreceptors has remained
elusive. They continue to be the least studied
of all nonvisual photoreceptor classes, although their existence in the vertebrates has
been known for nearly a century. In this issue
of PNAS, the Yoshimura laboratory presents a study that represents a signicant
stride toward identifying and characterizing
encephalic photoreceptors (2).
History of Encephalic Photoreception
In 1911, von Frisch (3) recognized in the
European minnow that light-induced
changes in skin coloration occurred even
in the absence of eyes and the photoreceptive pineal gland. However, crude lesions of the diencephalon abolished this
photoresponse, leading him to propose the
existence of encephalic photoreceptors. In
birds, the presence of such photoreceptors
was conclusively shown in 1944 through
the work of Benoit and Ott (4), who
showed that direct illumination of the hypothalamus through ne quartz rods (the
ber optics of the day) could induce testicular growth in blinded ducks. It was not
until the late 1960s that the pioneering
research of Menaker (5) indicated that
light-driven reproductive responses in
birds could be mediated entirely by photoreceptors outside of the retina and pineal. His group showed that, in addition
to regulating avian reproduction, encephalic photoreceptors also synchronize the
phase of daily activity rhythms to the
day:night cycle (6, 7).
Precisely pinpointing the anatomical site
of the avian encephalic photoreceptors
and identifying the photopigments that
render these photoreceptors light-sensitive
have proven difcult. Several groups have
attempted to localize these cells immunohistochemically. Rhodopsin-like immunoreactivity was initially observed in the
tuberal hypothalamus and septal forebrain
regions of the pigeon (8, 9). Later studies
indicated that melanopsin and vertebrate
ancient opsin (VA-opsin) were found in
the avian brain (10, 11).
Role in Photoperiodism
Determining the photopigment responsible for a given physiological response to
light can serve as a guidepost by which
the cells harboring the photopigment can

be identied and characterized. Action

spectrum analysis (i.e., establishing which
wavelengths of light optimally elicit a
photoresponse) is one method by
which photopigments can be linked to particular physiology. Foster et al. (12) showed
in the quail that the light-dependent increase in serum luteinizing hormone (LH),
a prerequisite for testicular growth, was
greatest in response to blue-green wavelengths, suggesting the involvement of a
rhodopsin-like photopigment (12).

Nakane et al. aggregate

the ndings of the
past into a realistic
working model of
photoperiodism in birds.
LH is released into the circulation from
the anterior pituitary in response to
gonadotropin-releasing hormone (GnRH).
GnRH arises from a sparse cohort of
neurons that terminate in the median
eminence of the mediobasal hypothalamus. Here, the GnRH is secreted into the
hypophyseal portal system, ultimately
leading to the release of LH from the anterior pituitary. Importantly, the secretion
of GnRH from nerve terminals in the
median eminence is gated by glial cells
called tanycytes (13, 14). The Yoshimura
laboratory had suggested previously that
this gating is dependent on the light-induced activation of thyroid hormone
through the enzymatic conversion of the
inactive T4 form to the active T3 form
(15, 16). The enzyme responsible for this
conversion, type 2 deiodinase, is expressed
in tanycytes in response to thyroid stimulating hormone (TSH). TSH is sensed by
the endfeet of tanycytes after it has been
secreted from thyrotropes located in the
pars tuberalis.
The big question remained: how is light
regulating TSH and consequently initiating this complex hormonal signaling cascade that ultimately leads to an activated
reproductive axis? In PNAS, Nakane et al.
(2) describe cells in the paraventricular
organ of the hypothalamus that express
the photopigment Opn5. They show that
these presumed photoreceptor cells send
projections to the external zone of the
median eminence, which is juxtaposed to
the par tuberalis, the site of TSH release.

1566215663 | PNAS | September 7, 2010 | vol. 107 | no. 36

Consolidating these ndings, Nakane et al.

(2) propose a model of photoperiodism in
birds that encompasses the entire process
from photoreception to gonadal induction.
The high amino acid sequence homology of Opn5 to other opsin sequences
certainly suggests that it functions as a
photopigment protein (1, 17). Nakane
et al. (2) conrm that Opn5 is indeed lightsensitive by heterologously expressing it in
Xenopus oocytes and recording lightinduced ionic currents (2). Oocytes not
expressing Opn5 showed no photoresponses. They completed an action spectrum and determined the sensitivity of
Opn5 to peak in the violet wavelengths
(420 nm). This is signicantly more blueshifted than the sensitivity of the encephalic photoreceptors described by Foster
et al. (12) (492 nm). Nakane et al. (2)
measure testicular size in response to
various wavelength exposures, and their
analysis shows that UV (UVB and UVA)
and blue wavelengths stimulate gonadal
growth as effectively as white light, a nding consistent with the involvement of
Opn5 (2). These results, however, do not
rule out the possibility that a blue-sensitive
photopigment, such as melanopsin or
perhaps, VA-opsin, mediates the photoperiodic response. Opsins maximally sensitive to blue light still absorb signicantly
in the violet and UV wavelengths (18).
Therefore, it remains a possibility that the
UV and blue wavelengths tested by Nakane et al. (2) stimulated testis growth
through photopigments other than Opn5.
As previously mentioned, VA-opsin has
indeed been identied in the developing
(19) and adult avian brain (11); it has also
been suggested (albeit not tested) that
a TSH-mediated photoperiodic response,
virtually identical to the one presented by
Nakane et al. (2), is initiated by VA-opsin
activation (11).
Role of Opn5
Therefore, the historical pendulum has
swung from a time when no encephalic
photopigments were denitively identied
to the current state where at least three
photopigments are known to reside in the
brain. What are all of these deep-brain
photopigments doing? Those localized to
Author contributions: I.P. wrote the paper.
The author declares no conict of interest.
See companion article on page 15264 in issue 34 of volume
107.
1

E-mail: ip7m@virginia.edu.

www.pnas.org/cgi/doi/10.1073/pnas.1010370107

must be regenerated by replacing the alltrans isomer with another 11-cis chromophore. Photopigments of invertebrates are
capable of photoconverting all-trans-retinoids back to 11-cis in situ through the
absorption of another photon (21). The
photopigments of vertebrates are incapable
of such in situ regeneration and must
relinquish their all-trans-retinaldehyde
and incorporate a new 11-cis-retinaldehyde molecule.
Multiple mechanisms have evolved to
do this. A specialized class of opsin has
emerged whose sole function is to incorporate the spent all-trans chromophore
and use the energy of an absorbed photon
to photoisomerize the retinoid back to the
11-cis conguration (22). The 11-cis chromophore eventually is used to regenerate
the photosensory opsins, rendering them
capable of initiating another round of
phototransduction.
Peropsin (23) and retinal pigment
epithelium-retinal G protein receptor opsin (RGR-opsin) (24) are two examples
of such auxiliary opsins (referred to as
photoisomerases). Interestingly, several
published comparisons of opsin amino
acid sequences have grouped Opn5 among
the photoisomerases rather than the photosensory opsins (17, 22, 25). Furthermore, the intron:exon structure of the

Opn5 gene more closely resembles that of

peropsin and RGR-opsin (17). These
ndings raise the possibility that, in the
avian brain, Opn5 does not function as the
photopigment directly mediating the photoperiodic response but rather, maintains
the sensitivity of an attendant photosensory protein, perhaps VA-opsin, by
ensuring that sufcient 11-cis retinoid
is available.
The relative importance of the various
opsins in encephalic photoperiodic signaling remains to be established. In
the meantime, the photosensory/photoisomerase capacity of Opn5 can be determine empirically by analyzing its
chromophore isomer preference as has
been done for other opsins (23, 26). Nevertheless, the strides made by the Yoshimura lab (2) over the last decade to
understanding the molecular and cellular
basis for avian photoperiodism have been
signicant. Nakane et al. (2) aggregate the
ndings of the past into a realistic working
model of photoperiodism in birds. This
model is likely to shed light on photoperiodic responses of seasonally breeding
mammals (27, 28), which, despite lacking
encephalic photoreceptors, also respond
to changing day lengths.

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PNAS | September 7, 2010 | vol. 107 | no. 36 | 15663

COMMENTARY

distinct areas of the brain may mediate

particular responses to light. For example,
photopigments found in the tuberal hypothalamus may mediate the photoperiodic reproductive responses, whereas
those found in the septal forebrain mediate the photoentrainment of circadian activity rhythms. This argument, however,
cannot be applied to Opn5 and VA-opsin;
these opsins are expressed in the same
anatomical neighborhood in the bird brain
(2, 11) and may be coexpressed.
Possessing multiple photopigments
could broaden the spectral range to which
a photoreceptor cell is sensitive, thereby
increasing the probability of photon capture under diverse ambient-light conditions. Alternatively, one of the
photopigments may be playing an auxiliary
role in light reception. Opsin-based photosensory pigments are G protein-coupled
receptors that detect light, because they
possess a covalently bound 11-cis-retinaldehyde chromophore that functions as
a photolabile ligand (20). On absorption of
a photon, the 11-cis-retinaldehyde is photoisomerized to all-trans-retinaldehyde
(21). This conformational change activates
the photopigment and initiates a signaling
cascade. After this occurs, the photopigment is spent and rendered useless for
additional signaling. The chromophore