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Impairment of spermatogenesis and enhancement of testicular germ cell apoptosis induced by exogenous all-trans-retinoic acid in adult lizard Podarcis sicula.

In mammals, retinoic acid is involved in the regulation of testicular function by interaction with two families of nuclear receptors, retinoic acid receptor (RAR) and retinoid X receptor (RXR). Among RAR isoforms, the testicular cells of the lizard were found to express only RARalpha (3.7 kb) and RARbeta (3.4 kb) mRNAs, as reported here. In this study, the effects of exogenous all-trans-retinoic acid (atRA) on spermatogenesis of a non-mammalian seasonal reproducer were investigated. Daily intraperitoneal injections of atRA or atRA plus testosterone (atRA+T) were given for 2 weeks to adult males of the lizard Podarcis sicula. In animals treated with atRA, the seminiferous tubules were markedly reduced in cross-area. The seminiferous epithelium collapse was responsible for a sensible reduction in the number of germ cells and disruption in normal epithelial organization. In comparison, in atRA+T-treated lizards the loss of germinal cells was significantly less. The loss of germ cells observed in both experimental groups results from an induction of apoptotic process, as revealed by TUNEL analysis. Although low in number, apoptotic germ cells were also observed in the control groups (saline- and T-treated lizard), where the main germ cells undergoing apoptosis are primary spermatocytes (most frequently) and some spermatogonia. In conclusion, it is shown here that retinoic acid has deleterious effects on lizard spermatogenesis, causing a severe depletion of seminiferous epithelium, probably via induction of apoptotic processes. These effects are not completely inhibited by simultaneous administration of testosterone, although this hormone, once injected, is able to stimulate spermatogenesis and protect germinal cells from apoptotic cell death.

Animals↗

Pregnant female lizards Iberolacerta cyreni adjust refuge use to decrease thermal costs for their body condition and cell-mediated immune response.

Lizards often respond to increased predation risk by increasing refuge use, but this strategy may entail a loss of thermoregulatory opportunities, which may lead to a loss of body condition. This may be especially important for pregnant oviparous female lizards, because they need to maintain optimal body temperatures as long as possible to maximize developmental embryos rate until laying. However, little is known about how increased time spent at low temperatures in refuges affects body condition and health state of pregnant female lizards. Furthermore, it is not clear how initial body condition affects refuge use. Female Iberian rock lizards forced to increase time spent at low temperatures showed lower body condition and tended to show lower cell-mediated immune responses than control females. Therefore, the loss of thermoregulatory opportunities seems to be an important cost for pregnant females. Nevertheless, thereafter, when we simulated two repeated predatory attacks, females modified refuge use in relation to their body condition, with females with worse condition decreasing time hidden after attacks. In conclusion, female lizards seemed able to compensate increased predation risk with flexible antipredatory strategies, thus minimizing costs for body condition and health state.

Analysis of Variance↗

Ultrastructural aspects of dental tissues and their behavior in xenoplastic association (lizard quail).

Ultrastructural characteristics of tooth buds of the polyphyodont adult lizards Liolaemus tenuis and Liolaemus gravenhorsti have been elucidated. Xenoplastic combinations of lizard whole tooth buds and neural crest cells from embryos of the quail Coturnix coturnix japonica have been cultured in vitro. Mesenchymal cells (preodontoblasts) of lizard teeth early develop filopodia that contact the basal lamina. Fragments of quail neural crest isolated by dissection were recombined with isolated lizard tooth buds and cultured for 84 hours in dishes kept in an incubator at 37.8 degrees C in air. Some identifiable quail cells in these recombinants developed a cytoplasmic extension like that of an odontoblastic process. These results suggest that lizard tooth rudiments already determined for tooth development produce some non-species specific transmissible constituents which are capable of inducing quail cranial neural crest cells to express certain dental characteristics (odontoblastogenesis) not expressed in their normal development in vivo.

Animals↗

Odontogenesis and amelogenesis in interacting lizard-quail tissue combinations.

In this study we examined the possible inductive role of the dental papilla from polyphyodont lizard tooth germs. Flank skin sheets of quail ectoderm enzymatically separated from dermal tissue were recombined with lizard tooth papillae and placed on semisolid medium and cultured for 2 days. Subsequently, the recombinants were removed and placed on the chorioallantoic membrane of chick hosts and incubated for 6 days. After this period of 8 days in explant, control tissues differentiated according to their own phenotypes. Lizard dental papilla alone differentiated as fibroblasts. Quail flank skin ectoderm differentiated into epithelial sheets. Intact lizard tooth buds developed into teeth with dentine and incipient enamel. In the best experimental recombinants, advanced and relatively well-constructed teeth were observed, with clear indications of hard tissue deposition in association with quail epithelium. The results show that mesenchyme of the adult lizard dental papilla and embryonic quail ectoderm of heterotopic origin are capable of carrying out the complex sequence of morphogenetic interactions involved in normal odontogenesis.

Amelogenesis↗

Stable social aggregations in an Australian lizard, Egernia stokesii.

Social groups in many animal species contain family members, and are maintained by parental care, and then tolerance of related individuals in the group. Lizards rarely show prolonged parental care, and rarely form stable social aggregations, although cohesive groups have been reported in the Australian skink genus, Egernia. A population of Egernia stokesii was surveyed for six seasons on Camel Hill, South Australia. At this site individuals occupied rock crevices for refuges on an isolated rocky outcrop of about 1.5 ha. The population was divided into 17 stable social groups, each containing 2-17 individual lizards. Surveyed juveniles took more than 5 years to reach mature size, and most juveniles and subadults remained in the social group of their parents for that period and longer. There were 2-8 permanent adult members of each group. Group members shared common crevice refuges, basked close together (and sometimes on top of each other) and defecated in common scat piles. There was low mortality after the first 2 years of life, and low dispersal either into or out of the population. Some individuals were "floaters" that did not belong to a social group. Over the study a number of these became established in groups. The social structure of these lizards resembles the family groups reported in many species of birds and mammals where group members help to raise the offspring of relatives. The low level of parental care in lizards suggests that the evolution of this form of social organisation in lizards has resulted from different processes than in other vertebrate taxa.

Aging↗

Tail loss and thermoregulation in the common lizard Zootoca vivipara.

Tail autotomy in lizards is an adaptive strategy that has evolved to reduce the risk of predation. Since tail loss reduces body mass and moving ability-which in turn are expected to influence thermal balance-there is potential for a trade-off between tail autotomy and thermoregulation. To test this hypothesis, we studied a common lizard (Zootoca vivipara) population at high latitude, inhabiting a high-cost thermal environment. Z. vivipara is a small, non-territorial lizard known as a very accurate thermoregulator. We made two predictions: (1) the reduced body weight due to tail loss results in faster heating rate (a benefit), and (2) the reduction in locomotor ability after tail loss induces a shift to the use of thermally poorer microhabitats (a cost), thus decreasing the field body temperatures of active lizards. We did not find any effect of tail loss on heating rate in laboratory experiments conducted under different thermal conditions. Likewise, no significant relationship between tail condition and field body temperatures, or between tail condition and thermal microhabitat use, were detected. Thus, our results suggest that tail autotomy does not influence the accuracy of thermoregulation in small-bodied lizards.

Animals↗

Annual variations of adrenal gland hormones in the lizard Podarcis sicula.

The adrenal gland regulates metabolism and maintains normal electrolyte balance. Adrenal hormones are equivalent in all vertebrates; the chromaffin tissue produces adrenaline and noradrenaline and the steroidogenic tissue produces most of the steroid hormones present in mammals. Podarcis sicula belongs to the Squamata family of lizards and it is the most abundant lizard species in southern Italy. This species shows a reproductive annual cycle and the presence of seasonal variations in the activity of the hypothalamus-hypophyseal-thyroid axis. To investigate the existence of an annual cycle of lizard adrenal gland, we have measured plasma concentrations of corticosterone, ACTH, noradrenaline and adrenaline. We have shown that corticosterone rapidly increased from January to March to reach a peak value that persisted until July, then, it slowly decreased until December. ACTH levels increased from January to May and slowly decreased from July to December. Noradrenaline levels were higher in March and then decreased until December. On the contrary, adrenaline levels increased from March to July and slowly decreased until December. Our results demonstrate the existence of an annual cycle of the lizard adrenal gland activity. This is very interesting because its activity is important to rightly regulate the reproductive status of the Podarcis sicula lizard.

Adrenal Glands↗

Partial tail loss has no severe effects on energy stores and locomotor performance in a lacertid lizard, Takydromus septentrionalis.

Many species of lizards use caudal autotomy as a defense strategy to avoid predation, but tail loss entails costs. These topics were studied experimentally in the northern grass lizard, Takydromus septentrionalis. We measured lipids in the three-tail segments removed from each of the 20 experimental lizards (adult females) initially having intact tails to evaluate the effect of tail loss on energy stores; we obtained data on locomotor performance (sprint speed, the maximal length traveled without stopping and the number of stops in the racetrack) for these lizards before and after the tail-removing treatments to evaluate the effect of tail loss on locomotor performance. An independent sample of 20 adult females that retained intact tails was measured for locomotor performance to serve as controls for successive measurements taken for the experimental lizards. The lipids stored in the removed tail was positively correlated with tailbase width when holding the tail length constant, indicating that thicker tails contained more lipids than did thinner tails of the same overall length. Most of the lipids stored in the tail were concentrated in the proximal portion of the tail. Locomotor performance was almost unaffected by tail loss until at least more than 71% of the tail (in length) was lost. Our data show that partial tail loss due to predatory encounters or other factors may not severely affect energy stores and locomotor performance in T. septentrionalis.

Animals↗

Transmission of the blood parasite Hemolivia mariae between its lizard and tick hosts.

The haemogregarine Hemolivia mariae is found in the erythrocytes of a natural population of the lizard Tiliqua rugosa. It infects two tick species, Amblyomma limbatum and Aponomma hydrosauri, which parasitise lizards. In laboratory experiments, engorged Amb. limbatum nymphs from infected lizards transmitted the haemogregarine to uninfected lizards significantly more often than engorged Ap. hydrosauri nymphs. Dissections of larvae and nymphs of both species fed on the same infected hosts showed that Amb. limbatum ticks were significantly more likely to become infected than Ap. hydrosauri ticks. In Amb. limbatum, oval cysts containing parasite stages thought to be infective to the lizard host had developed within 15 days of engorged nymphs detaching from an infected host. The chance of Ap. hydrosauri becoming infected and the intensity of infection in Amb. limbatum increased when ticks were fed on infected hosts as larvae and as nymphs compared with those fed on an infected host only as a nymph. This suggests that infections can accumulate over the tick life stages. Since the two tick species have broadly parapatric distributions, the boundary between the tick species may have implications for the distribution of H. mariae.

Animals↗

Energetics of bluetongue lizards (Tiliqua scincoides) in a seasonal tropical environment.

We studied the physiological ecology of bluetongue lizards (Tiliqua scincoides) on the Adelaide River floodplain in tropical Australia to determine the seasonal patterns of energy expenditure and to determine the mechanisms by which seasonal differences were achieved. Field metabolic rates (FMR) were significantly lower in the dry season (37.6 kJ kg(-1) day(-1); n=9) than in the wet (127.3 kJ kg(-1) day(-1); n=7). Water flux was also lower in the dry season (6.8 ml kg(-1) day(-1); n=9) than in the wet (39.4 ml kg(-1) day(-1); n=7). Measurements of body temperatures (T(b)) and movements of free-ranging animals, and standard metabolic rate (SMR) of recently caught animals, allowed a detailed analysis of energy budgets for wet and dry seasons. In the dry, bluetongue lizards expended 90 kJ kg(-1) day(-1) less energy than in the wet season. Unlike some other lizards of the wet-dry tropics, SMR did not differ between seasons. About 5% of the seasonal difference in FMR was due to lower night time T(b) during the dry season, and about 7% was due to lower diurnal T(b). The remaining 88% of the decrease in energy expended in the dry season was due to a substantial decrease in other costs that may include reproduction, growth, digestion and activity. If we assume the animals fed daily and the costs of digestion are taken into account, the estimates are: 14% of the savings result from lower T(b) at night, 20% from lower T(b) in the day, and 66% result from decreased activity. It is therefore apparent that, unlike some agamid and varanid lizards that use a combination of behavioural and physiological mechanisms to conserve energy when food and water are limited, bluetongue lizards primarily use behavioural mechanisms to achieve a dramatic reduction in energy expenditure in the dry season.

Adaptation, Physiological↗

Geographic variation in the life history of the sagebrush lizard: the role of thermal constraints on activity.

Thermal constraints on the time available for activity have been proposed as a proximate mechanism to explain variation in suites of life history traits. The longer that an ectotherm can maintain activity, the more time it has to forage and the greater chance that it will encounter a predator and be eaten. Thus, the thermal environment may produce a trade off between growth and survival when variation in the environment favors increased activity. I used mark-recapture data from a demographic study of three natural populations of the sagebrush lizard (Sceloporus graciosus) and estimates of thermal opportunity for each population to evaluate whether variation in the thermal environment can explain patterns of growth and survival that occur over an elevational gradient. Lizards from the highest elevation population exhibited higher individual growth rates than those of lizards from lower elevation, while mortality rates increased with elevation for these populations. The covariation of fast growth and high mortality with increased thermal opportunity is the opposite trend expected if the thermal environment alone is to explain patterns of life history in these lizards. Additional factors including thermal heterogeneity in the distribution of microhabitats of lizards, adaptation to local environmental conditions, and a potential trade-off between resource acquisition and predation risk need to be addressed to obtain a satisfactory explanation of the causative mechanisms producing life history variation.

Animals↗

Changes in GABA and parvalbumin immunoreactivities in the cerebral cortex of lizards after narine occlusion.

Olfactory deprivation produced by narine occlusion has been suggested to reduce the activity in the cerebral cortex of lizards. Here we analyzed the short-term changes in GABA and parvalbumin (PV) immunoreactivities in the cerebral cortex of lizards after narine occlusion. The number and distribution of GABA- and parvalbumin-immunoreactive (IR) cells have been studied by immunocytochemistry in the cerebral cortex of control and olfactory-deprived lizards. The distribution of GABA-IR cells as well as that of PV-IR cells was similar in control and deprived animals, and PV-IR cells were GABA-IR in all cases. However, significant changes were observed in the absolute number of GABA- and PV-IR cells. GABA-IR cells were more abundant in deprived animals than in control ones. In contrast, the number of PV-IR cells decreased significantly and PV immunoreactivity in dendrites and boutons was lower in deprived animals. These results suggest that the reduction in the number of PV-IR cells in olfactory-deprived lizards occurs without loss of GABA cells, and that PV expression is under the control of olfactory activity and remains plastic in the cerebral cortex of adult lizards.

Animals↗

Demonstration of common and stage-specific anti-Hepatozoon mocassini antibodies in experimentally infected unnatural lizard hosts.

This study examines the immunological response to Hepatozoon mocassini in lizards. Three lizard species were infected experimentally with H. mocassini. Baseline and post-infection (PI) sera were assayed for anti-H. mocassini meront and gametocyte antibody by immunohistochemistry and IFA. Seroconversion occurred at 38 d PI with endpoint IFA titers of 1:64. Antisera from non-parasitaemic and parisitaemic lizards exhibited similar affinities for merozite and gametocyte antigens. Antibody specific for the membranes of gametocyte-infected erythrocytes was demonstrated exclusively in parasitaemic lizards. The results demonstrate that lizards infected with snake haemogregarines mount an antibody response with common and stage-specific components.

Aedes↗

A comparison of skin-test responses using antigen from Leishmania donovani and a lizard trypanosome.

261 individuals were skin tested with leishmanin from Leishmania donovani and with an antigen from a lizard trypanosome isolated in an area endemic for kala-azar in Ethiopia. 83 of the 261 responded positively, with indurations of 5 mm or greater, to one antigen but negatively to the other, and 42 reacted positively to both. Among the Afar people of the Awash valley in north-eastern Ethiopia and the Nyangatom people of south-western Ethiopia, positive response to both antigens was high, though many individuals responded to one antigen but not the other. Both these groups live as cattle pasturalists, a predominantly male occupation, in areas where known vectors of kala-azar as well as lizard-feeding sandflies exist. Among the Anuak and Suri people, who live in areas of south-western Ethiopia infested with tsetse flies, the lizard trypanosome antigen gave a markedly higher skin test positivity than did the leishmanin antigen. This suggests that the lizard trypanosome may give artificially high results in other immunological tests of exposure to trypanosomiasis in areas where lizard trypanosomes and man-biting sandflies co-exist with mammalian trypanosomes transmitted by Glossina.

Animals↗

Immunohistochemical localization of gastrin C-terminus, gastric inhibitory peptide (GIP) and endorphin in the pancreas of lizards with special reference to the hibernation period.

Gastrin C-terminus and GIP immunoreactive cells were observed in the pancreas of the desert lizard (Uromastyx aegyptia) captured during the hibernation period, but not in those collected in the active period. These cell types were encountered among the exocrine parenchyma, especially around ducts and among the ductal epithelial cells. Occasionally a few GIP cells were seen to occupy the islet periphery. No gastrin C-terminus or GIP immunoreactive cells were observed in the pancreas of the grass lizard (Mabuya quinquetaeniata)--which does not hibernate--collected in winter and in summer. In both species of lizards endorphin-like immunoreactivity was localized in the pancreatic PP-cells in specimens collected in winter and summer. It was assumed that the presence of the gastrin C-terminus and GIP cells in the desert lizard pancreas represents a response to the peculiar physiological state through which these lizards pass in hibernation.

Animals↗

Characterization of beta-keratins in lizard epidermis: electrophoresis, immunocytochemical and in situ-hybridization study.

Lizard scales are composed of alpha-(cyto-) keratins and beta-keratins. The characterization of the molecular weight and isoelectric point (pI) of alpha- and beta-keratins of lizard epidermis (Podarcis sicula) has been done by using two-dimensional electrophoresis, immunoblotting, and immunocytochemistry. Antibodies against cytokeratins, against a chicken scale beta-keratin or against lizard beta-keratin bands of 15-16kDa, have been used to recognize alpha- and beta-keratins. Acid and basic cytokeratins of 42-67kDa show a pI from 5.0 to 8.9. This indicates the presence of specific keratins for the formation of the stratum corneum. Main protein spots of beta-keratin at 15-17kDa, and pI at 8.5, 8.2, and 6.7, and one spot at 10kDa and pI at 7.3 were recognized. Therefore, beta-keratins are mainly basic proteins, and are used for the formation of the hard corneous layer of the epidermis. Ultrastructural immunocytochemistry confirms that beta-keratin is packed into large and dense bundles of beta-keratin cells of lizard epidermis. The use of a probe against a lizard beta-keratin in situ-hybridization studies confirms that the mRNA for beta-keratins is present in beta-cells and is localized around or even associated with beta-keratin filaments.

Actin Cytoskeleton↗

Natriuretic peptides are negative modulators of adrenocortical cell function of the eastern fence lizard (Sceloporus undulatus).

Elucidation of the role of natriuretic peptides (NPs) in vertebrate adrenal steroidogenesis has been facilitated by the use of freshly dispersed adrenocortical cells. Our recent characterization of lizard adrenocortical cells [Carsia, R.V., John-Alder, H.B., 2003. Seasonal alterations in adrenocortical cell function associated with stress-responsiveness and sex in the Eastern Fence Lizard (Sceloporus undulatus). Horm. Behav. 43, 408-420] provided the opportunity to examine the influence of atrial natriuretic peptides (ANPs) and related NPs on reptilian adrenal steroidogenesis at the cellular level. In the present report, the action of NPs on lizard adrenal steroidogenesis was investigated using freshly dispersed adrenocortical cells derived from the Eastern Fence Lizard (Sceloporus undulatus). Basal production rates of aldosterone and corticosterone and maximal angiotensin II (ANG II)-induced production rates of these corticosteroids were inhibited with high efficacy (75-90%) by rat ANP at potencies of 0.4-0.7 nM. By contrast, rat ANP had no effect on maximal production rates of these corticosteroids in response to a maximal steroidogenic concentration of adrenocorticotropin (ACTH; 1 nM). However, rat ANP inhibited aldosterone and corticosterone production rates in response to a half-maximal steroidogenic concentration of ACTH (10 pM; approximately 50 pg/ml), albeit with less efficacy ( approximately 50%) and potency (approximately 6 nM) than for ANG II. Rat and eel ANP and rat and chicken brain natriuretic peptide (BNP) were equally efficacious at inhibiting maximal ANG II-induced aldosterone and corticosterone production but with different potencies. The order of inhibitory potency was rat ANP = chicken BNP > eel ANP > rat BNP. However, a specific peptide ligand for the NP clearance receptor was without effect. This study indicates that ANP and related NPs are efficacious inhibitors of lizard adrenal steroidogenesis by acting directly at the level of the adrenocortical cell.

Adrenal Cortex↗

Isolation of Trypanosoma brucei from the monitor lizard (Varanus niloticus) in an endemic focus of Rhodesian sleeping sickness in Kenya.

Monitor lizards were sampled along the shores of Lake Victoria to detect natural infections of potentially human-infective trypanosomes. In an area with endemic rhodesian sleeping sickness, one of 19 lizards was infected (Busia, Kenya). Six of ten lizards also showed indirect evidence of infection with Trypanosoma brucei (antibody ELISA). In an area with no recent history of human disease (Rusinga Island), no parasites were found and no antibodies to T. brucei were detected. The isolate was identified as T. brucei through xenodiagnosis (completion of the life cycle in the salivary glands of tsetse), and through molecular techniques (positive reactions with a PCR primer and a microsatellite DNA probe characteristic of the subgenus Trypanozoon). Experimental infections of monitor lizards were also attempted with a variety of parasites and tsetse species. It was possible to infect monitor lizards with T. brucei but not with forest or savannah genotypes of Trypanosoma congolense. Parasites reached low levels of parasitaemia for a short period without generating any pathology; they also remained infective to tsetse and laboratory rats. The implications of these findings are discussed in relation to the endemicity of sleeping sickness.

Animals↗