Biomedical subjects

Dong-Gen Luo

Publications and source records attributed to Dong-Gen Luo.

6 recordsLinked to original sources

PubMed2006

Parietal-eye phototransduction components and their potential evolutionary implications.

The parietal-eye photoreceptor is unique because it has two antagonistic light signaling pathways in the same cell-a hyperpolarizing pathway maximally sensitive to blue light and a depolarizing pathway maximally sensitive to green light. Here, we report the molecular components of these two pathways. We found two opsins in the same cell: the blue-sensitive pinopsin and a previously unidentified green-sensitive opsin, which we name parietopsin. Signaling components included gustducin-alpha and Galphao, but not rod or cone transducin-alpha. Single-cell recordings demonstrated that Go mediates the depolarizing response. Gustducin-alpha resembles transducin-alpha functionally and likely mediates the hyperpolarizing response. The parietopsin-Go signaling pair provides clues about how rod and cone phototransduction might have evolved.

3',5'-Cyclic-GMP Phosphodiesterases↗

PubMed2005

Intrinsically photosensitive retinal ganglion cells detect light with a vitamin A-based photopigment, melanopsin.

In mammals, intrinsically photosensitive retinal ganglion cells (ipRGCs) mediate non-image-forming visual functions such as pupillary light reflex (PLR) and circadian photoentrainment. This photosensitivity requires melanopsin, an invertebrate opsin-like protein expressed by the ipRGCs. The precise role of melanopsin remains uncertain. One suggestion has been that melanopsin may be a photoisomerase, serving to regenerate an unidentified pigment in ipRGCs. This possibility was echoed by a recent report that melanopsin is expressed also in the mouse retinal pigment epithelium (RPE), a key center for regeneration of rod and cone pigments. To address this question, we studied mice lacking RPE65, a protein essential for the regeneration of rod and cone pigments. Rpe65-/- ipRGCs were approximately 20- to 40-fold less photosensitive than normal at both single-cell and behavioral (PLR) levels but were rescued by exogenous 9-cis-retinal (an 11-cis-retinal analog), indicating the requirement of a vitamin A-based chromophore for ipRGC photosensitivity. In contrast, 9-cis-retinal was unable to restore intrinsic photosensitivity to melanopsin-ablated ipRGCs, arguing against melanopsin functioning merely in photopigment regeneration. Interestingly, exogenous all-trans-retinal was also able to rescue the low sensitivity of rpe65-/- ipRGCs, suggesting that melanopsin could be a bistable pigment. Finally, we detected no melanopsin in the RPE and no changes in rod and cone sensitivities due to melanopsin ablation. Together, these results strongly suggest that melanopsin is the photopigment in the ipRGCs.

Animals↗

PubMed2005

Impaired channel targeting and retinal degeneration in mice lacking the cyclic nucleotide-gated channel subunit CNGB1.

Cyclic nucleotide-gated (CNG) channels are important mediators in the transduction pathways of rod and cone photoreceptors. Native CNG channels are heterotetramers composed of homologous A and B subunits. In heterologous expression systems, B subunits alone cannot form functional CNG channels, but they confer a number of channel properties when coexpressed with A subunits. To investigate the importance of the CNGB subunits in vivo, we deleted the CNGB1 gene in mice. In the absence of CNGB1, only trace amounts of the CNGA1 subunit were found on the rod outer segment. As a consequence, the vast majority of isolated rod photoreceptors in mice lacking CNGB1 (CNGB1-/-) failed to respond to light. In electroretinograms (ERGs), CNGB1-/- mice showed no rod-mediated responses. The rods also showed a slow-progressing degeneration caused by apoptotic death and concurred by retinal gliosis. Cones were primarily unaffected and showed normal ERG responses up to 6 months, but they started to degenerate in later stages. At the age of approximately 1 year, CNGB1-/- animals were devoid of both rods and cones. Our results show that CNGB1 is a crucial determinant of native CNG channel targeting. As a result of the lack of rod CNG channels, CNGB1-/- mice develop a retinal degeneration that resembles human retinitis pigmentosa.

Animals↗

PubMed2005

Rod sensitivity of neonatal mouse and rat.

We have measured the sensitivity of rod photoreceptors isolated from overnight dark-adapted mice of age P12 (neonate) through P45 (adult) with suction-pipette recording. During this age period, the dark current increased roughly in direct proportion to the length of the rod outer segment. In the same period, the flash sensitivity of rods (reciprocal of the half-saturating flash intensity) increased by approximately 1.5-fold. This slight developmental change in sensitivity was not accentuated by dark adapting the animal for just 1 h or by increasing the ambient luminance by sixfold during the prior light exposure. The same small, age-dependent change in rod sensitivity was found with rat. After preincubation of the isolated retina with 9-cis-retinal, neonatal mouse rods showed the same sensitivity as adult rods, suggesting the presence of a small amount of free opsin being responsible for their lower sensitivity. The sensitivity of neonate rods could also be increased to the adult level by dark adapting the animal continuously for several days. By comparing the sensitivity of neonate rods in darkness to that of adult rods after light bleaches, we estimated that approximately 1% of rod opsin in neonatal mouse was devoid of chromophore even after overnight dark adaptation. Overall, we were unable to confirm a previous report that a 50-fold difference in rod sensitivity existed between neonatal and adult rats.

Aging↗

PubMed2004

AMPA receptor is involved in transmission of cone signal to ON bipolar cells in carp retina.

The present work focuses on characterization of glutamate receptor subtypes mediating cone signal transmission to ON bipolar cells (BCs) in the carp retina, using intracellular recording techniques. Glutamate (5 mM) hyperpolarized cone-dominant ON BCs, which was associated with a suppression of light responses, whereas Co(2+) (1 mM) depolarized these cells and suppressed their light responses. On the other hand, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) of 20 microM caused a membrane depolarization and blocked depolarizing light responses, L-2-amino-4-phosphonobutryic acid (l-AP4) was without effect. The effects of AMPA were reversed by coapplication of GYKI 52466, an AMPA receptor selective non-competitive antagonist, but persisted in the presence of picrotoxin and strychnine. For rod-dominant ON BCs, both l-AP4 and AMPA reversibly blocked depolarizing light responses, but with membrane potential changes of opposite polarities (hyperpolarization for l-AP4 and depolarization for AMPA). In the inner retina, AMPA depolarized transient ON-OFF amacrine cells and blocked both ON and OFF cone-driven depolarizing responses, but l-AP4 did not. These results suggest that AMPA receptors, but not l-AP4 receptors, are involved in synaptic transmission of cone signal to ON bipolar cells in carp retina.

Animals↗

PubMed2002

Zn(2+) modulates light responses of color-opponent bipolar and amacrine cells in the carp retina.

The effects of Zn(2+) on color-opponent bipolar cells (BCs) and amacrine cells (ACs) were studied in the isolated superfused carp retina using intracellular recording techniques. Bath-applied Zn(2+) (25 micro M) depolarized R(+)G(-)-type BCs and suppressed both depolarizing responses of these cells to red (680 nm) flashes and hyperpolarizing ones to green (500 nm) flashes. Following Zn(2+) application, G(+)R(-)-type BCs were hyperpolarized, which was accompanied by a potentiation of their depolarizing responses to green flashes and a suppression of hyperpolarizing ones to red flashes. Similar Zn(2+) effects were observed in R(+)G(-)- and G(+)R(-)-type ACs. The Zn(2+) effects persisted in the presence of picrotoxin and strychnine, suggesting that modulation by Zn(2+) of GABA and glycine receptors was unlikely involved. Using whole-cell recording techniques, it was found Ca(2+) currents in cone terminals were dose-dependently suppressed by Zn(2+), suggesting that Zn(2+) may reduce glutamate release from cone photoreceptors. Furthermore, lowering extracellular Ca(2+), a procedure that increases glutamate release from photoreceptors, exerted actions on R(+)G(-)- and G(+)R(-)-type BCs, almost opposite to the Zn(2+) effects on these two types of BCs. It is therefore postulated that the Zn(2+) effects reported in the present work may reflect a consequence of the changes in input resistances of color-opponent BCs and driving forces for their light responses resulted from the reduced glutamate release by Zn(2+).

Action Potentials↗