Karen L. Gunther

Adaptive optics retinal imaging reveals S-cone dystrophy in tritan color-vision deficiency

Tritan color-vision deficiency is an autosomal dominant disorder associated with mutations in the short-wavelength-sensitive- (S-) cone-pigment gene. An unexplained feature of the disorder is that individuals with the same mutation manifest different degrees of deficiency. To date, it has not been possible to examine whether any loss of S-cone function is accompanied by physical disruption in the cone mosaic. Two related tritan subjects with the same novel mutation in their S-cone-opsin gene, but different degrees of deficiency, were examined. Adaptive optics was used to obtain high-resolution retinal images, which revealed distinctly different S-cone mosaics consistent with their discrepant phenotypes. In addition, a significant disruption in the regularity of the overall cone mosaic was observed in the subject completely lacking S-cone function. These results taken together with other recent findings from molecular genetics indicate that, with rare exceptions, tritan deficiency is progressive in nature.

Structure-activity studies related to ABT-594, a potent nonopioid analgesic agent: Effect of pyridine and azetidine ring substitutions on nicotinic acetylcholine receptor binding affinity and analgesic activity in mice

Analogs of A-98593 (1) and its enantiomer ABT-594 (2) with diverse substituents on the pyridine ring were prepared and tested for affinity to nicotinic acetylcholine receptor binding sites in rat brain and for analgesic activity in the mouse hot plate assay. Numerous types of modifications were consistent with high affinity for [3H]cytisine binding sites. By contrast, only selected modifications resulted in retention of analgesic potency in the same range as 1 and 2. Analogs of 2 with one or two methyl substituents at the 3-position of the azetidine ring also were prepared and found to be substantially less active in both assays.

Individual differences in chromatic (red/green) contrast sensitivity are constrained by the relative number of L- versus M-cones in the eye

Many previous studies have shown that the relative number of long-wavelength-selective (L) versus medium-wavelength-selective (M) cones in the eye influences spectral sensitivity revealed perceptually. Here, we hypothesize that the L:M cone ratio should also influence red/green chromatic contrast sensitivity. To test this, in each subject we derived an estimate of L:M ratio based on her red/green equiluminance settings (obtained with heterochromatic flicker photometry), and measured both red/green chromatic and luminance contrast sensitivity at different spatial and temporal frequencies. Factor analysis was applied to the data in order to reveal covariance between conditions. As expected, chromatic and luminance contrast sensitivity were found to be independent of one another, and no relationship was observed between L:M ratio and luminance contrast sensitivity. However, a significant relationship was observed between L:M ratio and chromatic contrast sensitivity, wherein subjects possessing the most symmetrical L:M cone ratios (i.e., near 1:1) appear to possess the relatively greatest chromatic contrast sensitivity. This relationship can be accounted for by a simple model based on the notion of random L- and M-cone inputs to the center and surround receptive fields of chromatic (L-M) mechanisms.

What covariance mechanisms underlie green:red equiluminance, luminance contrast sensitivity and chromatic (green:red) contrast sensitivity?

In order to investigate the mechanisms underlying green/red equiluminance matches in human observers and their relationship to mechanisms subserving luminance and/or chromatic (green/red) contrast sensitivity, we tested 21 human subjects along these dimensions at 16 different spatial and temporal frequencies (spatial frequency, 0.25-2 c/deg; temporal frequency, 2-16 Hz) and applied factor analysis to extract mechanisms underlying the data set. The results from our factor analysis revealed separate sources of variability for green/red equiluminance, luminance sensitivity and chromatic sensitivity, thus suggesting separate mechanisms underlying each of the three main conditions. When factor analysis was applied separately to green/red equiluminance data, two temporally-tuned factors were revealed (factor 1, 2-4 Hz; factor 2, 8-16 Hz), suggesting the existence of separate mechanisms underlying equiluminance settings at low versus high temporal frequencies. In addition, although the three main conditions remained separate in our factor analysis of the entire data set, our correlation matrix nonetheless revealed systematic correlations between equiluminance settings and luminance sensitivity at high temporal frequencies, and between equiluminance settings and chromatic sensitivity at low temporal frequencies. Taken together, these data suggest that the high temporal frequency factor underlying green/red equiluminance is governed predominantly by luminance mechanisms, while the low temporal frequency factor receives contribution from chromatic mechanisms.

Is epibatidine really analgesic? dissociation of the activity, temperature, and analgesic effects of (±)-epibatidine

The experiments in the present study were designed to determine if the activity, temperature, and analgesic effects of (+/-)-epibatidine treatment could be dissociated. Initially (i.e., 15 min) (+/-)-epibatidine treatment (0.1 mumol/kg = 28 micrograms/kg, IP) impaired rotorod performance, decreased activity, decreased temperature, and increased jump latency (e.g., analgesic effect). For the remaining time points measured (i.e., 30, 60, and 120 min), activity and temperature remained significantly reduced. In contrast, by 120 min (+/-)-epibatidines effects on rotorod performance and analgesia (jump latency) were not observed. When administered after (+/-)-epibatidine (0.05 mumol/kg, IP), mecamylamine treatment (5 mumol/kg = 1 mg/kg, IP) produced a potentiation of analgesia. This potentiation effect was not observed on activity and temperature measures. The effect of (+/-)-epibatidine treatment (0.1 mumol/kg, IP) was also determined in mice with central nicotinic receptor blockade induced by treatment with chlorisondamine (23 mumol/kg = 10 mg/kg, IP). An (+/-)-epibatidine-induced reduction in activity was not attenuated in chlorisondamine-treated mice and only a minimal effect was observed on (+/-)-epibatidine-induced hypothermia in chlorisondamine-treated mice. In contrast, in chlorisondamine-treated mice (+/-)-epibatidines analgesic effect was attenuated. Taken together, these data suggest that various centrally mediated effects of (+/-)-epibatidine can be dissociated.

Effects of intraseptal injection of 192-IgG-saporin in mature and aged Long-Evans rats

In this study, the effects intraseptal injections of the selective cholinergic immunotoxin, 192-IgG-saporin, were investigated in mature (6-month-old) and aged (24-26-month-old) male Long-Evans rats. Ten days following intraseptal injection of either 192-IgG-saporin or saline, testing began in a battery of behavioral tests modulated by the septohippocampal system including two versions of the Morris water maze (i.e. submerged platform task, and 2-platform spatial discrimination), inhibitory avoidance, and pre-pulse inhibition of acoustic startle. In both mature and aged rats, intraseptal injection of 192-IgG-saporin selectively reduced ChAT activity in the hippocampus and posterior cingulate cortex, without affecting ChAT activity of amygdala or parietal cortex. In general, in all of the behavioral tests analyzed, intraseptal 192-IgG-saporin treatment had no effect in mature animals. Age-related deficits were observed in the spatial memory tasks, however this impairment was largely a function of the poor performance of aged rats treated with the toxin. In addition, an increase in the response to an acoustic startle was found in aged rats treated with 192-IgG-saporin. Thus, although intraseptal injection of 192-IgG-saporin produced similar reductions of ChAT activity, performance of mature and aged rats in tasks believed to be modulated by the septohippocampal pathway tended to be differentially affected in mature and aged rats.

2-(Aryloxymethyl) azacyclic analogues as novel nicotinic acetylcholine receptor (nAChR) ligands

Abstract A series of 2-(aryloxymethyl) azetidine and pyrrolidine nAChR ligands in which the 3-pyridyl moiety of a previously described series 1 was replaced by a substituted phenyl group was explored. Aromatic substitution afforded analogues with K i values ranging from 3 to >10,000 nM. Generally, substitution at the ortho - and para -position was unfavorable, whereas electron-withdrawing groups at the meta -position improved the K i values.

A novel mutation in the short-wavelength-sensitive cone pigment gene associated with a tritan color vision defect

Inherited tritan color vision deficiency is caused by defects in the function of the short-wavelength-sensitive (S) cones. This heterozygous group of disorders has an autosomal dominant pattern of inheritance. Amino acid variations of the S cone opsin are rare and all that have been identified thus far are associated with inherited tritan color vision defects. Here we report the identification of a 30-year-old male who made errors on standard color vision tests consistent with the presence of a mild tritan color vision deficiency. We tested the hypothesis that his color vision impairment was due to a mutation in the S cone photopigment gene. He was found to be heterozygous for a mutation that caused the amino acid proline to be substituted in place of a highly conserved leucine at amino acid position 56 in the S cone opsin. This mutation was absent in 564 S cone photopigment genes from 282 subjects who did not make tritan errors. Thus, we conclude that this mutation disrupts the normal function of S cones.

The influence of Bay K 8644 treatment on (±)-epibatidine-induced analgesia

The purpose of this investigation was to determine if analogous to (-)-nicotines analgesic effect, the analgesic effect of the recently characterized potent nicotinic acetylcholine receptor (nAChR) agonist (+/-)-epibatidine was altered in response to treatment with the calcium channel agonist (+/-)-Bay K 8644. In addition, the effects of the enantiomers, (+)-Bay K 8644, reported to be a calcium channel antagonist, and (-)-Bay K 8644, reported to be a calcium channel agonist were examined. (+/-)-Bay K 8644 (2.8 mumol/kg; i.p.) produced a large analgesic response in mice in the hot-plate paradigm that rapidly dissipated by 30 min after treatment. This analgesic effect of (+/-)-Bay K 8644 was not prevented by pre-treatment with the nicotinic antagonist mecamylamine (5 mumol/kg; i.p.). Treatment with non-analgesic doses of the calcium channel agonists (+/-)- and (-)-Bay K 8644 (1.4 mumol/kg; i.p.) significantly potentiated the analgesic effect of (+/-)-epibatidine (0.05 mumol/kg; i.p.). Potentiation of (+/-)-epibatidines analgesic effect occurred when the agonists were administered prior to (+/-)-epibatidine or after (+/-)-epibatidine as long as analgesia testing was conducted 15 to 30 min after Bay K 8644 treatment. Pre-treatment with the calcium channel antagonist (+)-Bay K 8644 was found to attenuate (+/-)-epibatidine-induced analgesia. When given after (+/-)-epibatidine, (+)-Bay K 8644 had no effect on (+/-)-epibatidines analgesic effect. These data provide additional in vivo evidence that altering calcium dynamics can modulate neuronal nAChR function.

Nucleotide polymorphisms upstream of the X-chromosome opsin gene array tune L:M cone ratio.

In support of the long-held idea that cone ratio is genetically determined by variation linked to the X-chromosome opsin gene locus, the present study identified nucleotide differences in DNA segments containing regulatory regions of the L and M opsin genes that are associated with significant differences in the relative number of L versus M cones. Specific haplotypes (combinations of genetic differences) were identified that correlated with high versus low L:M cone ratio. These findings are consistent with the biological principle that DNA sequence variations affect binding affinities for protein components of complexes that influence the relative probability that an L versus M opsin gene will be silenced during development, and in turn, produce variation in the proportion of L to M cones.