Pineal melatonin rhythms in the lizard Anolis carolinensis: II. Photoreceptive inputs

The pineal organ of the lizard Anolis carolinensis acts as a transducer of photoperiodic information, since light can affect the pineal melatonin rhythm (PMR). The synthesis and secretion of melatonin may be a major mechanism whereby a circadian pacemaker within the pineal can control circadian clocks located elsewhere. An investigation into potential routes by which light could affect the PMR showed that (1) removal of the photosensory parietal eye did not affect the PMR as compared to controls under either a light-dark (LD) 12:12 cycle and a constant temperature (32 degrees C) or an LD 12:12 cycle and a daily temperature cycle (32 degrees C/20 degrees C); (2) removal of both the parietal eye and the lateral eyes did not affect the PRM of anoles held in LD 12:12 (constant 32 degrees C); (3) the PMR of blinded anoles re-entrained to a 10-hr shift in the phase of the LD cycle as rapidly as that of sighted anoles; (4) blocking light penetration to the brains of anoles, but leaving the lateral eyes exposed, blocked the ability of anoles to re-entrain to a 10-hr shift in the phase of an LD cycle. The data support the hypothesis that light directly affects the PMR in Anolis and that other potential photic inputs (parietal eye, lateral eyes) play little or no role. This conclusion is supported by previous neurophysiological and ultrastructural studies showing that the lizard pineal possesses functional photoreceptors.     

Pineal melatonin rhythms in the lizardAnolis carolinensis: effects of light and temperature cycles

Summary Pineal and ocular melatonin was assessed, over 24 h periods, in male lizards (Anolis carolinensis) entrained to 24 h light-dark (LD) cycles and a constant 32 C, and in lizards entrained to both 24 h LD cycles and 24 h temperature cycles (32 C/20 C). At a constant temperature, the duration of the photoperiod has a profound effect on the duration, amplitude, and phase of the pineal melatonin rhythm (Fig. 1). The pineal melatonin rhythm under cyclic temperature peaks during the cool (20 C) phase of the cycle regardless of whether or not the cool phase occurs during the light or dark phase of a LD 1212 cycle (Fig. 3). Under a temperature cycle and constant dim illumination, a pineal melatonin rhythm is observed which peaks during the cool phase of the temperature cycle, but the amplitude of the rhythm is depressed relative to that observed under LD (Fig. 2). Illumination up to 2 h in duration does not suppress the nocturnal melatonin peak in theAnolis pineal (Fig. 4). No melatonin rhythm was observed in the eyes ofAnolis under either 24 h LD cycles and a constant temperature (Fig. 1), or under simultaneous light and temperature cycles (Fig. 3). Ocular melatonin content was, in all cases, either very low or non-detectable.Abbreviations HIOMT hydroxyindole-O-methyltransferase - NAT N-acetyltransferase     

Incubation temperature modifies neonatal thermoregulation in the lizard Anolis carolinensis

The thermal environment experienced during embryonic development can profoundly affect the phenotype, and potentially the fitness, of ectothermic animals. We examined the effect of incubation temperature on the thermal preferences of juveniles in the oviparous lizard, Anolis carolinensis. Temperature preference trials were conducted in a laboratory thermal gradient within 48 hr of hatching and after 22-27 days of maintenance in a common laboratory environment. Incubation temperature had a significant effect on the upper limit of the interquartile range (IQR) of temperatures selected by A. carolinensis within the first 2 days after hatching. Between the first and second trials, the IQR of selected temperatures decreased significantly and both the lower limit of the IQR and the median selected temperature increased significantly. This, along with a significant incubation temperature by time interaction in the upper limit of the IQR, resulted in a pattern of convergence in thermoregulation among treatment groups. The initial differences in selected temperatures, as well as the shift in selected temperatures between first and second trials, demonstrate plasticity in temperature selection. As a previous study failed to find environmentally induced plasticity in temperature selection in adult A. carolinensis, this study suggests that this type of plasticity is exclusive to the period of neonatal development.     

Annual testicular cycle of the lizard Anolis carolinensis: effects of pinealectomy and melatonin

The effects of pinealectomy and exogenous melatonin treatment on the reproductive system of male anoles were examined at several different times of year. In September pinealectomy of anoles exposed to either a stimulatory LD 14:10 light cycle or a nonstimulatory LD 10:14 light cycle induced significant testicular growth and development over that observed in sham-operated anoles. At a nonphotosensitive time of year (December) pinealectomy also had a significant progonadal effect but no effect of pinealectomy was seen in February-March. Daily melatonin injections given either in the morning or afternoon (or both) failed to block gonadal growth either (1) in sham-operated or pinealectomized anoles exposed to LD 14:10 in the fall or (2) in pinealectomized lizards exposed to LD 10:14 in the fall. Continuous melatonin administration via subcutaneous silastic implants blocked the progonadal effects of pinealectomy in the winter (December). The results show that pinealectomy can have significant progonadal effects; these effects are seasonal but can encompass phases of the annual testicular cycle which are either photoperiod-dependent or temperature-dependent; and melatonin may be a reproductively active factor involved.     

Light at night cannot suppress pineal melatonin levels in the lizard Anolis carolinensis

Up to 2 hr exposure of anoles to high intensity natural or artificial illumination at mid-dark does not suppress pineal melatonin levels. The results support the hypothesis that the lizard pineal is completely insensitive to acute exposure to light at night which is in direct contrast to the effects of light in higher vertebrates.     

Compensatory testicular hypertrophy in the lizard Anolis carolinensis

In reproductively responsive, male Anolis carolinensis undergoing artificially induced testicular recrudescence, unilateral orchidectomy of the left testis produced compensatory hypertrophy of the remaining testis. Testosterone inhibited this compensatory testicular hypertrophy on a weight basis, but did not reduce the rate of spermatogenic development. These results suggest that there is a mechanism of testosterone feedback in Anolis carolinensis that controls gonadotropin secretion during the recrudescent phase. In reproductively thermorefractory lizards, unilateral orchidectomy had no effect on the remaining testis. Administration of exogenous follicle-stimulating hormone to refractory animals increased testicular weight and stage of spermatogenic development. Sensitivity to gonadotropin, as well as failure of unilateral orchidectomy to produce compensatory hypertrophy in refractory male anoles, suggests that the control of the refractory period in A. carolinensis results from physiological mechanisms in the pituitary gland or brain rather than in the testis.     

Latent effects of egg incubation temperature on growth in the lizard Anolis carolinensis

Varied egg incubation temperatures can result in immediate effects on the phenotype of reptiles, and also latent effects that can augment or contradict effects evident at egg hatching. I examined the effects of incubation temperature on embryonic development, hatching morphology, and subsequent growth in multiple populations of the lizard Anolis carolinensis. Eggs from wild-caught females in four populations were incubated at up to three temperatures, 23.5, 27, and 30 degrees C. Measures of body size were collected immediately after hatching and weekly thereafter, while juveniles were maintained in a common laboratory environment for 8 weeks. Cooler incubation temperatures resulted in longer incubation periods but did not affect conversion of egg mass to hatchling mass. Incubation temperature did not affect hatchling mass or snout vent length (SVL), but did affect subsequent growth in both mass and SVL, which varied by population. Cooler incubation temperatures generally resulted in greater overall growth over 8 weeks of housing all juveniles in a common environment. In A. carolinensis, egg incubation temperature had latent effects on juvenile growth despite the absence of any detected immediate effects on hatchling phenotype. Therefore, the total impact and evolutionary importance of developmental environment should not be assessed or assumed based solely on the phenotype of reptiles at birth or hatching.     

Sperm transfer and storage in the lizard,Anolis carolinensis

The relationship of the hemipenis to the cloaca in copula and sperm storage and transport in the female oviduct were studied in Anolis carolinensis using light and scanning electron microscopy. During copulation, the hemipenis does not penetrate beyond the cloaca, but the two apical openings of the bifurcate sulcus spermaticus appose the openings of the oviducts from the cloaca. Sperm enter the sperm storage tubules between 2 and 6 hr after insemination and small amounts of sperm reach the infundibulum 6 to 24 hr following mating. Sperm storage tubules are embedded in the wall of the utero-vaginal transition, and are formed by the folding and fusion of the oviducal epithelium. The importance of the hemipenile-cloacal relationship and the role of sperm storage in the life history of A. carolinensis are discussed.     

Chromatophores and color change in the lizard,Anolis carolinensis

Summary The skin of the lizard, Anolis carolinensis, changes rapidly from bright green to a dark brown color in response to melanophore stimulating hormone (MSH). Chromatophores responsible for color changes of the skin are xanthophores which lie just beneath the basal lamina containing pterinosomes and carotenoid vesicles. Iridophores lying immediately below the xanthophores contain regularly arranged rows of reflecting platelets. Melanophores containing melanosomes are present immediately below the iridophores. The ultrastructural features of these chromatophores and their pigmentary organelles are described. The color of Anolis skin is determined by the position of the melanosomes within the melanophores which is regulated by MSH and other hormones such as norepinephrine. Skins are green when melanosomes are located in a perinuclear position within melanophores. In response to MSH, they migrate into the terminal processes of the melanophores which overlie the xanthophores above, thus effectively preventing light penetration to the iridophores below, resulting in skins becoming brown. The structural and functional characteristics of Anolis chromatophores are compared to the dermal chromatophore unit of the frog.This study was supported in part by GB-8347 from the National Science Foundation.Contribution No. 244, Department of Biology, Wayne State University.The authors are indebted to Dr. Joseph T. Bagnara for his encouragement during the study and to Dr. Wayne Ferris for his advice and the use of his electron microscope laboratory.     

Glucosylceramide in the androgen-responsive kidney of the lizard Anolis carolinensis

During the spring breeding season of the American chameleon, Anolis carolinensis, elevated levels of glucosylceramides which contain hydroxy fatty acids are produced in the kidneys of males but not females. Hyperproduction of this glycolipid is also induced by testosterone. The testosterone-induced hypertrophy of epithelial cells in the proximal tubules of the mouse kidney seems an analogous phenomenon and an elevated concentration of specific glycolipids in the male mouse kidney has been previously demonstrated. Thus the formation of renal glycolipids in response to testosterone may be a widespread feature in vertebrates.     

Mast cell histamine and ovarian follicular growth in the lizard Anolis carolinensis.

The hypothesis is proposed that histamine released from mast cells in the theca of ovarian follicles increases thecal hyperemia and vessel permeability, and thus plays a role in follicular growth in Anolis carolinensis. Mast cells are present in the stroma and theca, and the number of thecal mast cells increases as follicles grow. The levels of histamine in follicular walls varies with follicular size. Histamine causes vasodilatation of thecal vessels. Antihistamine blocks the effects of histamine and, when given alone, stimulates vasoconstriction. Antihistamine also blocks estradiol-induced growth of large follicles. These findings are consistent with the above hypothesis.     

Thermoregulatory function of the parietal eye in the lizard Anolis carolinensis

Summary Animals were acclimitized to a daily cycle of 15–25°C and a photoperiod of LD 12:12. Parietalectomized animals in an experimental thermal gradient (15°C–40°C, LD 12:12) selected significantly higher temperatures than controls at all hours of the day except from 0800–1200. These results suggest that the lizard parapineal plays a direct role in thermoregulation in addition to the indirect function as an illuminometer.     

Embryology and cytodifferentiation of the pituitary gland in the lizard Anolis carolinensis

Anolis embryos have limb buds at the time eggs are laid and require about 39 days to complete development at 28°C. Rathke's pouch is present at five days, and the subdivisions of the adenohypophysis are differentiated by ten days after oviposition. The cells of the rostral half of the pars distalis (PD) are derived from the anterior face of Rathke's pouch; cells of the caudal half from the posterior face. Lateral lobe cells differentiate on the lateral margins of the developing caudal PD, and knob-like outgrowths of this tissue attach to the walls of the diencephalon to form the pars tuberalis (PT). Subsequently, the cells of the PT lose their connection with the PD and become a pair of flattened oblong plaques. They reach maximal size in midincubation, and are gradually invaded by nervous elements and incorporated into the walls of the hypothalamus. Electron micrographs demonstrate that the embryonic PT is secretory. Ultrastructurally the pars intermedia (PI) and PD are composed of parenchymous secretory cells in a framework of stellate cells. Stellate cells surround the lumen of Rathke's pouch and are connected laterally by complex junctions that exclude the secretory cells from the luminal surface. They extend in sheet-like processes among the secretory cells to the outer margin of the gland where they form a partial sheath within the basal lamina around the secretory tissue. As development proceeds, the lumen becomes subdivided and the resulting reduced lumina are recognizable as the forerunners of the follicles of the adult adenohypophysis. The cells of the PI are differentiated into secretory or stellate cells halfway through incubation. At this time only half of the cells of the PD can be so classified. Four of the five granulated cell types described in the adult are recognizable by mid-incubation; the fifth cell type (prolactin cell) becomes distinguishable within ten days thereafter, and at hatching appears to be actively synthesizing secretory products.