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Biomedical subjects

D G Green

Publications and source records attributed to D G Green.

At least 19 recordsLinked to original sources
PubMed2001

Constitutive "light" adaptation in rods from G90D rhodopsin: a mechanism for human congenital nightblindness without rod cell loss.

A dominant form of human congenital nightblindness is caused by a gly90-->asp (G90D) mutation in rhodopsin. G90D has been shown to activate the phototransduction cascade in the absence of light in vitro. Such constitutive activity of G90D rhodopsin in vivo would desensitize rod photoreceptors and lead to nightblindness. In contrast, other rhodopsin mutations typically give rise to nightblindness by causing rod cell death. Thus, the proposed desensitization without rod degeneration would be a novel mechanism for this disorder. To explore this possibility, we induced mice to express G90D opsin in their rods and then examined rod function and morphology, after first crossing the transgenic animals with rhodopsin knock-out mice to obtain appropriate levels of opsin expression. The G90D mouse opsin bound the chromophore and formed a bleachable visual pigment with lambda(max) of 492 nm that supported rod photoresponses. (G+/-, R+/-) retinas, heterozygous for both G90D and wild-type (WT) rhodopsin, possessed normal numbers of photoreceptors and had a normal rhodopsin complement but exhibited considerable loss of rod sensitivity as measured electroretinographically. The rod photoresponses were desensitized, and the response time to peak was faster than in (R+/-) animals. An equivalent desensitization resulted by exposing WT retinas to a background light producing 82 photoisomerizations rod(-1) sec(-1), suggesting that G90D rods in darkness act as if they are partially "light-adapted." Adding a second G90D allele gave (G+/+, R+/-) animals that exhibited a further increase of equivalent background light level but had no rod cell loss by 24 weeks of age. (G+/+, R-/-) retinas that express only the mutant rhodopsin develop normal rod outer segments and show minimal rod cell loss even at 1 year of age. We conclude that G90D is constitutively active in mouse rods in vivo but that it does not cause significant rod degeneration. Instead, G90D desensitizes rods by a process equivalent to light adaptation.

Adaptation, Ocular↗
PubMed2000

Partial rescue of the ocular retardation phenotype by genetic modifiers.

The or(J) allele of the murine ocular retardation mutation is caused by a premature stop codon in the homeodomain of the Chx10 gene. When expressed on an inbred 129/Sv strain, the or(J) phenotype is characterized by microphthalmia and a thin, poorly differentiated retina in which the peripheral portion is affected to a greater extent than the central portion. Such mutant retinae lack differentiated bipolar cells and the optic nerve typically fails to form, leading to blindness. Here, we show that progeny from an outcrossed backcross between 129/Sv-or(J) /or(J) and Mus musculus castaneus produce animals that are homozygous for the or(J) mutation and exhibit a much ameliorated eye phenotype. Although not of normal size, such modified or(J) eyes are significantly larger than those in 129/Sv-or(J) /or(J) mice, and contain a better organized retina which includes bipolar cells. Furthermore, optic nerves are frequently present, and the eyes show a degree of function as reflected by electroretinogram and pupillary response. As in 129/Sv-or(J) /or(J) mice, however, modified or(J) eyes show incomplete growth and a lack of cell differentiation in the periphery of the retina. The selective, and apparently nonmodifiable, effect of the ocular retardation phenotype on the periphery of the retina indicates that Chx10 plays an important role in the central-to-peripheral gradient of retinal development. These findings demonstrate that the ocular retardation phenotype can be greatly modified by the genetic background, and help to define a role for Chx10 in ocular development.

Animals↗
PubMed2000

Open problems in artificial life.

This article lists fourteen open problems in artificial life, each of which is a grand challenge requiring a major advance on a fundamental issue for its solution. Each problem is briefly explained, and, where deemed helpful, some promising paths to its solution are indicated.

Artificial Intelligence↗
PubMed1999

Effects of dystrophin isoforms on signal transduction through neural retina: genotype-phenotype analysis of duchenne muscular dystrophy mouse mutants.

Duchenne and Becker muscular dystrophy patients have mutations in the dystrophin gene. Most show reduced b-wave amplitudes in the dark-adapted electroretinogram (ERG). We studied normal C57BL/6J mice and five X-linked muscular dystrophy strains with different dystrophin mutations to determine whether the location of the mutation within the gene affects the mouse ERG and to correlate such effects with dystrophin isoform expression. Amplitudes and implicit times were measured for a-waves, b-waves, and digitally filtered oscillatory potentials. mdx and mdxCv5 mice, with mutations near the amino terminus and lacking expression of Dp427, had ERGs similar to those of C57BL/6J mice. mdxCv2 and mdxCv4 mice, with mutations in the center of dystrophin and who do not express isoforms Dp427, Dp260, or Dp140 (mdxCv4), had increased b-wave and oscillatory potential implicit times. mdxCv3 mice, with a mutation near the carboxy terminus resulting in deficiency of all dystrophin isoforms, had increased b-wave and oscillatory potential implicit times and reduced scotopic b-wave amplitudes. Fitting the a-wave data to a transduction activation phase mathematical model showed normal responses for all phenotypes, suggesting that the b-wave delays are due to defects beyond the rod outer segment, most likely at the rod to on-bipolar cell synapse. The variation in the ERG phenotype with the position of the dystrophin gene mutation suggests that there are different contributions by each isoform to retinal electrophysiology. Although Dp427 and Dp140 isoforms do not appear to be important contributors to the ERG, lack of Dp260 and possibly Dp71 isoforms is associated with an abnormal ERG.

Animals↗
PubMed1999

Electrophysiological properties of a new isolated rat retina preparation.

A piece of rat retina was mounted in an open chamber and perfused with a Ringer solution at 37 degrees C. The electroretinogram (ERG) was recorded between an extracellular microelectrode in contact with the rod outer segments and a reference electrode under the retina. The addition of 250-500 microM of glutamate to the media prevented the b-wave from decaying in amplitude with time. Minor components of the ERG, the scotopic threshold response (STR) and oscillatory potentials (OPs), were well maintained with glutamate in the media. Experiments on the spatial properties of the recordings indicated that a small area immediately around the microelectrode contributes most strongly to the response. The similarity of ERGs recorded in vivo from the cornea to the transretinal ERGs from the isolated retina of the same animal indicated that the functional integrity of the isolated retina was well preserved in the media with glutamate.

Action Potentials↗
PubMed1998

Serum-induced changes in the physiology of mammalian retinal glial cells: role of lysophosphatidic acid.

1. With a breakdown of the vascular-CNS barrier, serum enters the nervous system. Although this is a frequent pathophysiological event, knowledge of the effects of serum on the function of the nervous system is limited. In this study, we examined the effects of serum on the activity of ion channels in Müller cells: the principal glia of the retina. 2. Freshly dissociated Müller cells from the bovine and human retina were studied with the perforated-patch configuration of the patch clamp technique. In other experiments, electroretinograms (ERGs) were recorded from isolated rat retinas. 3. Perforated-patch recordings revealed that serum induced a calcium-permeable, non-specific cation (NSC) current. Approximately 40 s after induction of this current, an outwardly rectifying K+ current was also detected. Sensitivity to charybdotoxin and margatoxin indicated that this K+ current was due to the activation of Kv1.3 channels. This increase in the Kv1.3 current was dependent on extracellular calcium. 4. The NSC and Kv1.3 currents were activated by serum in 100% and 95% of the sampled Müller cells, respectively. Also, in a minority (21%) of the cells, the inwardly rectifying K+ current was inhibited slightly. These changes in ion channel activity were associated with depolarization of the Müller cells. 5. We hypothesized that activation of NSC channels would reduce the siphoning of K+ via the Müller cells. Consistent with this idea, ERGs from isolated retinas showed serum-induced reductions in the slow PIII component, which is generated by Müller cells responding to light-evoked changes in the extracellular K+ concentration. 6. Lysophosphatidic acid (LPA), a component of serum, had effects on Müller cells that were qualitatively similar to those induced by serum. 7. Our observations demonstrate that exposure to serum alters the activity of multiple types of ion channels in Müller glial cells of the mammalian retina. When there is a breakdown of the blood-retina barrier, LPA may be one of the serum-derived molecules which regulates the physiology of Müller cells.

Animals↗
PubMed1997

Electrophysiology and density of retinal neurons in mice with a mutation that includes the Pax2 locus.

PURPOSE: The Krd mouse has a deletion in chromosome 19 that includes the Pax2 gene locus. The aim of this study was to characterize in detail how these retinas differ from normal. METHODS: Both electroretinographic and anatomic methods were used to assess visual function. Full-field flash electroretinograms (ERGs) and planimetric densities were obtained from Krd and control animals. RESULTS: Measurements of the ERG show that in the Krd mice, both a- and b-wave amplitudes are attenuated relative to control by amounts that vary from animal to animal. The b-wave of the ERG generally is affected more severely than the a-wave. However, there is little or no shift of the curves relating the b-wave and a-wave amplitude to the intensity of the stimulus. Also, no change in the response kinetics seems to be associated with the attenuated responses. Estimates of planimetric cell density in the outer nuclear, inner nuclear, and ganglion cell layers show significant cell losses in affected animals that are more pronounced proportionally in the inner layers. Comparisons between electrophysiological and histologic measurements made on each eye show good correlation between the reduction in the ERG components and the magnitude of cell losses. CONCLUSIONS: These experiments show that the eyes of Krd mice have reduced ERGs and reduced cellular density. There is a loss of cells in all layers of the retina, but the inner layers are affected more severely. Consistent with this, the b-wave is reduced more than the a-wave. The normal functional dependency of the ERG on stimulus intensity and the normal response kinetics suggest the cellular losses are not associated with changes in cellular function.

Animals↗
PubMed1996

Optical modulation transfer and contrast sensitivity with decentered small pupils in the human eye.

Human observers experience a large decrement in visual acuity when a small artificial pupil is displaced from the center to the edge of the dilated natural pupil. This decrement in visual resolution, called the Campbell effect, has been attributed to the retina, the ocular optics, or a combination of the two. Given the uncertainty about the relative magnitudes of these two components over the range of spatial frequencies used in normal vision, we have obtained objective measurements of the retinal image quality and psychophysical measurements of visual performance, with decentered pupils. The contributions of monochromatic aberrations were determined by using double pass measurements of the modulus of the optical transfer function (MTF). For all of the observers, there was a substantial decrement in the MTF with decentering, showing that even when using a 1.5 mm pupil and appropriate spherical/cylindrical refractive corrections, there is a considerable contribution of monochromatic aberrations to the effect. We have compared these optical MTFs with the psychophysical contrast sensitivity functions (CSFs) measured under exactly the same conditions using green gratings generated on the screen of a color monitor. At the low and intermediate spatial frequencies considered (2-16 c/deg), we find the fall in the CSF is much greater than the fall in the monochromatic MTF, with the difference becoming greater as the spatial frequency increases. We show that this discrepancy can be mostly attributed to the effect of transverse chromatic aberration due to the bandwidth of the green stimulus used for the CSF measurements. In conclusion, the combination of the ocular transverse chromatic aberration and monochromatic aberrations accounts for the loss in visual sensitivity found with a decentered small pupil at low and intermediate spatial frequencies.

Color↗
PubMed1995

Double-pass measurements of the retinal-image quality with unequal entrance and exit pupil sizes and the reversibility of the eye's optical system.

We have used a modified double-pass apparatus with unequal entrance and exit pupil sizes to measure the optical transfer function in the human eye and have applied the technique to three different problems. First, we confirm that in the eye the double-pass spread function is the cross correlation of the input spread function with the output spread function [J. Opt. Soc. Am. A 12, 195 (1995)]. Consequently, when entrance and exit pupil sizes are equal, phase information is lost from the double-pass images. Second, we show that in double-pass measurements the eye behaves like a reversible optical system. That is, when entrance and exit pupils are equal, the double-pass image results from two passes through an optical system having a transfer function that is the same in both directions. To test for reversibility in the living eye we have used a double-pass apparatus with different exit and entrance pupil sizes (one of them small enough to consider the eye diffraction limited), so that the ingoing and the outgoing transfer functions are different. The measured image quality was unchanged when the pupils were interchanged, i.e., when the first-pass entrance pupil size becomes the second-pass exit pupil size, and vice versa. Third, the technique provides a means for inferring the complete optical transfer function of the eye, including the phase transfer function, and the shape of the point-spread function.

Accommodation, Ocular↗
PubMed1995

mdxCv3 mouse is a model for electroretinography of Duchenne/Becker muscular dystrophy.

PURPOSE: To identify an animal model for the abnormal scotopic electroretinogram found in a majority of Duchenne and Becker muscular dystrophy patients. METHODS: Ganzfeld electroretinograms were recorded in dark-adapted normal C57BL/6 mice, and two strains of mice with different X-linked muscular dystrophy mutations (mdx and mdxCv3). Responses for the right eye were averaged and the amplitudes and implicit times of the a-wave and b-wave were measured. The electroretinogram was digitally filtered to extract the oscillatory potentials. Statistical analyses included one-way analysis of variance and the Scheffé S test. RESULTS: While the electroretinogram in mdx was normal, in mdxCv3 the scotopic b-wave was markedly reduced and the oscillatory potentials were delayed, similar to changes observed in Duchenne and Becker muscular dystrophy patients. Some of the mdxCv3 animals demonstrated negative configuration electroretinograms, with the b-wave amplitude reduced compared to that of the a-wave. CONCLUSIONS: Abnormalities found in the electroretinograms of Duchenne and Becker muscular dystrophy patients led to the identification of dystrophin in human retina and the discovery that dystrophin is required for normal retinal electrophysiology. These results indicate that mdxCv3 is a model for elucidating the role of dystrophin in retina and suggest that dystrophin isoforms, consisting of only the C-terminal domains of the full-length protein, may be important to the development of normal retinal electrical potentials.

Animals↗
PubMed1994

Kidney and retinal defects (Krd), a transgene-induced mutation with a deletion of mouse chromosome 19 that includes the Pax2 locus.

The semidominant mutation Krd (kidney and retinal defects) was identified in transgenic line Tg8052. Krd/+ mice have a high incidence of kidney defects including aplastic, hypoplastic, and cystic kidneys. Retinal defects in Krd/+ mice include abnormal electroretinograms and a reduction of cell numbers that is most extreme in the inner cell and ganglion layers. Viability of Krd/+ mice is strongly influenced by genetic background, and growth retardation is observed in young animals. Homozygosity results in early embryonic lethality. Fluorescence in situ hybridization of a transgene-specific probe localized the insertion site to the distal region of mouse Chromosome 19. The sequence of the insertion site revealed transgene insertion into a LINE element with deletion of a single nucleotide from the 3' terminus of the transgene. A polymorphic microsatellite, D19Umi1, was identified in a junction clone and mapped in several large crosses. D19Umi1 is located 1.7 +/- 1.0 cM distal to Pax2, which encodes a paired type transcription factor expressed in embryonic kidney and eye. Deletion of Pax2 from the transgenic chromosome was demonstrated by Southern analysis of genomic DNA from (Krd/+ x SPRET/Ei)F1 mice. Additional genetic and molecular data are consistent with an approximately 7-cM deletion that includes the loci stearoyl CoA desaturase (Scd1), pale ear (ep), D19Mit17, D19Mit24, D19Mit27, D19Mit11, and Pax2. This deletion, Del(19)TgN8052Mm, will be useful for genetic and functional studies of this region of mouse Chromosome 19.

Abnormalities, Multiple↗
PubMed1994

Synaptic targeting domains of synapsin I revealed by transgenic expression in photoreceptor cells.

Synapsins are abundant nerve terminal proteins present at all synapses except for ribbon synapses, e.g. photoreceptor cell synapses. Multiple functions have been proposed for synapsins, including clustering of synaptic vesicles and regulation of synaptic vesicle exocytosis. To investigate the physiological functions of synapsin and to ascertain which domains of synapsin are involved in synaptic targeting in vivo, we expressed synapsin Ib and its N- and C-terminal domains in the photoreceptor cells of transgenic mice. In these cells synapsin Ib is targeted efficiently to synaptic vesicles but has no significant effect on the development, structure or physiology of the synapses. This suggests that synapsin I does not have dominant physiological or morphoregulatory functions at these synapses. Full-length synapsin Ib and the N-terminal domains of synapsin Ib but not its C-terminal domains are transported to synapses, revealing that the molecular apparatus for synaptic targeting of synapsins is also present in cells which form ribbon synapses that normally lack synapsins. This apparatus appears to utilize the conserved N-terminal domains that are shared between all synapsins.

Animals↗
PubMed1994

Extrinsic current and flash sensitivity in turtle cones.

The effects of hyperpolarizing current and background light on intracellular responses of red cones in turtle were compared. Even though a background light always reduced response amplitude, hyperpolarizing current did so in only 25% of the cells studied. When hyperpolarizing current reduced response amplitude it also produced changes in response kinetics and the intensity-response relationships, but these changes differed from those produced by background light. Considerably greater hyperpolarization was required with current than with light to produce equivalent reductions in amplitude. The results suggest that current reduces amplitude by activating a membrane conductance, while background light acts through a different mechanism.

Action Potentials↗
PubMed1992

Effect of contact lens correction of sine wave contrast sensitivity in keratoconus patients after penetrating keratoplasty.

Contrast sensitivity testing reveals visual deficits not detected by standard acuity tests, providing a more sensitive measure of visual performance. Sine wave contrast sensitivity functions were examined for spectacle and contract lens correction of keratoconic eyes after penetrating keratoplasty (PKP). Contrast sensitivity was significantly higher for middle and high spatial frequencies with a rigid gas permeable contact lens than with glasses, even when Snellen acuity was identical for both forms of correction. Contrast performance correlated with subjective assessments of vision quality. This may be related to the quality of the keratometry mire images and suggests that mild irregularities in graft contour may be affecting contrast sensitivity. When considering the options for optical correction of grafted eyes, an evaluation of contrast sensitivity may help determine whether contact lens correction would optimize visual performance.

Adult↗