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J C Besharse

Publications and source records attributed to J C Besharse.

80 records · Page 5Linked to original sources
PubMed1976

Renewal of normal and degenerating photoreceptor outer segments in the Ozark cave salamander.

To determine whether photoreceptor degeneration in the Ozark cave salamander is associated with cessation or changes in the kinetics of outer segment (OS) renewal, an autoradiographic study of 3H-leucine incorporation in photoreceptors was carried out. Six days after isotope injection rods and cones showed labeling in both inner and outer segments. Cone OS were diffusely labeled whereas rods contained a band of radioactivity at the base of the OS. At 13 and 21 days the radioactive band in rods was located progressively nearer the distal tip of the OS. The rate of rod OS renewal ranged from 0.30 to 0.38 mu of OS length per day at 18 degrees C. L-thyroxin induced metamorphosis and light increased the renewal rate compared to larvae in darkness, and adults with photoreceptors in an early stage of degeneration had a slightly higher renewal rate than larvae. Light and electron microscope autoradiographs of degenerate photoreceptors revealed that even in the final stages of degeneration when OS are reduced to small, irregular whorls of membrane, 3H-leucine labeling was present in inner segments and OS membranes. These observations demonstrate that OS renewal occurs in both larvae and adults, and suggest that photoreceptor degeneration may be due to disruption of some aspect of the OS disposal process.

Animals↗
PubMed1976

Effects of continuous light and darkness on the eyes of the troglobitic salamander Typhlotriton spelaeus.

Larval Typhlotriton spelaeus collected from five caves in Pulaski Co., Missouri, were kept as larvae or induced to transform in darkness or continuous fluorescent illumination. Larvae maintained in darkness for 215 and 279 days had smaller eyes, smaller rod inner and outer segments, and fewer metaphase figures in the germinative zone of the neural retina than comparable larvae maintained in light (258 lux). Except for visual cell size, differences were small and for each characteristic exceptions were observed. One larva kept in light showed early retinal degeneration comparable to that in transformed adults to T. spelaeus. All larvae exhibited optomotor behavior both before and after the experiment. Among animals induced to transform by L-thyroxin and maintained in darkness 111 to 366 days, visual cell and pigment epithelium degeneration was more extensive and more frequent than in animals kept for the same length of time in light (237-298 lux). In darkness the frequency of animals with retinal degeneration increased between 111 and 366 days. In light some animals exhibited pigment epithelium reduction with normal visual cells, and others had free, pigmented cells in the subretinal space. These effects were not comparable to degeneration in darkness. Eyelids covered the eyes of only a few animals in both light and dark treatments. The extent of eyelid encroachment over the eye greater in darkness than in light. Most animals exhibited optomotor responses after experiments, but responses of animals kept in darkness were impaired in comparison to those of animals kept in light.

Animals↗
PubMedDate not supplied

Tryptophan hydroxylase is expressed by photoreceptors in Xenopus laevis retina.

Serotonin has important roles, both as a neurotransmitter and as a precursor for melatonin synthesis. In the vertebrate retina, the role and the localization of serotonin have been controversial. Studies examining serotonin immunoreactivity and uptake of radiolabeled serotonin have localized serotonin to inner retinal neurons, particularly populations of amacrine cells, and have proposed that these cells are the sites of serotonin synthesis. However, other reports identify other cells, such as bipolars and photoreceptors, as serotonergic neurons. Tryptophan hydroxylase (TPH), the rate-limiting enzyme in the serotonin synthetic pathway, was recently cloned from Xenopus laevis retina, providing a specific probe for localization of serotonin synthesis. Here we demonstrate that the majority of retinal mRNA encoding TPH is present in photoreceptor cells in Xenopus laevis retina. These cells also contain TPH enzyme activity. Therefore, in addition to being the site of melatonin synthesis, the photoreceptor cells also synthesize serotonin, providing a supply of the substrate needed for the production of melatonin.

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PubMedDate not supplied

Spectral sensitivity of melatonin synthesis suppression in Xenopus eyecups.

Melatonin synthesis in retinal photoreceptors is stimulated at night by a circadian oscillator and suppressed acutely by light. To identify photoreceptor mechanisms involved in the acute suppression of melatonin synthesis, an action spectrum was measured for dark-adapted Xenopus laevis eyecups at night. Intensity-response curves at six wavelengths from 400 to 650 nm were parallel, suggesting that a single photopigment predominates in melatonin suppression. Half-saturating intensities at 400, 440, 480, and 533 nm were not significantly different from one another, at 1-2 x 10(8) quanta cm(-2) s(-1). Significantly higher intensities of 580- and 650-nm light were required for melatonin suppression. These results indicate a predominant role for the principal green-absorbing rods in acute regulation of retinal melatonin synthesis in response to light, and argue against an important role for the red-absorbing cones. Higher than expected sensitivity at short wavelengths suggests that photoreceptors sensitive to blue and/or violet light may also contribute to melatonin suppression.

Animals↗
PubMedDate not supplied

Circadian clock in Xenopus eye controlling retinal serotonin N-acetyltransferase.

Circadian rhythms are controlled by endogenous oscillators or clocks. These clocks exhibit a persistent period of approximately 24 h in constant conditions, a specific phase relationship to a periodic cue (zeitgeber) in the external environment, and plasticity in that the phase of the clock may be altered in response to a phase change in the zeitgeber. Although many processes exhibit circadian rhythmicity, the nature and location of endogenous clocks remain poorly defined. Recent evidence in vertebrates suggests that the mammalian suprachiasmatic nucleus and the avian pineal gland contain clocks that affect the rhythmicity of indoleamine metabolism. The vertebrate retina also exhibits a circadian rhythm of serotonin N-acetytransferase activity (NAT, EC 2.1.1.4), a key enzyme controlling melatonin synthesis, and of photoreceptor disk shedding. The latter process may be regulated by melatonin, and the immediate cellular events seem to be controlled locally within the eye. Although sustained oscillation and entrainment were not demonstrated, data suggesting that an ocular circadian clock influences disk shedding have been reported. We sought evidence for an ocular clock by studying retinal NAT activity in Xenopus eye cups maintained in culture and report here both sustained oscillation and entrainment of the in vitro system. The data indicate that in addition to the suprachiasmatic nucleus and pineal gland, the eye itself must be regarded as the locus of a circadian clock in vertebrates.

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