The thyroid and photoperiodic control of seasonal reproduction in American tree sparrows (Spizella arborea)

To explore the role of the thyroid gland in the control of seasonal reproduction in obligately photoperiodic American tree sparrows (Spizella arborea), the effects of (1) thyroxine administered in drinking water to thyroid-intact photosensitive or photorefractory birds, and (2) radiothyroidectomy before and after photostimulation and during photorefractoriness were examined. Chronic administration of pharmacological doses of thyroxine induced testicular growth and usually regression in initially photosensitive birds held on short or intermediate daylengths. Some thyroxine-treated birds with regressed testes were absolutely photorefractory, but most remained photosensitive. Exogenous thyroxine never induced testicular growth in photorefractory birds moved to short days, though it often impeded, and sometimes even blocked, the recovery of photosensitivity. Although circumstantial, these effects of exogenous thyroxine are consistent with an hypothesis that assigns to thyroid hormones two roles — one stimulatory and the other inhibitory — in the control of seasonal reproduction. Radiothyroidectomy before photostimulation inhibited (but did not prevent) photoinduced testicular growth, blocked spontaneous testicular regression, suppressed molt, and prevented photorefractoriness. Moreover, as demonstrated by testicular growth after thyroxine replacemnt therapy, radiothyroidectomy during photorefractoriness later restored photosensitivity despite continued photostimulation. Thus, euthyroidism is an essential condition for maximizing (but not for initiating) photoinduced testicular growth and for triggering and maintaining photorefractoriness in photostimulated tree sparrows. However, when performed early during photostimulation, radiothyroidectomy neither immediately induced nor later blocked spontaneous testicular regression. Thus, endogenous thyroid hormones and long days may interact during a critical period to program a sequence of physiological events that plays out as photorefractoriness in chronically photostimulated birds. Such an organizational event cannot be permanent, for seasonal reproduction is episodic and its control mechanism necessarily cyclic. Because thyroidectomy simulated the well-known restorative effect of short days (and exogenous thyroxine impeded it), short days may dissipate photorefractoriness by creating a milieu wherein thyroid hormones are deficient or inactive.Abbreviations ANOVA analysis of variance - bTSH bovine thyroid stimulating hormone - GnRH gonadotropin-releasing hormone - LH luteinizing hormone - nL: nD daily light: dark regime (n is duration in hours) - SEM standard error of the mean - SNK Student-Newman-Keuls test - T₄ thyroxine - TH thyroid hormone - TR thyroid hormone receptor     

Corticosterone treatment has no effect on reproductive hormones or aggressive behavior in free-living male tree sparrows, Spizella arborea

We examined the effect of corticosterone on plasma levels of reproductive hormones (testosterone, dihydrotestosterone, and luteinizing hormone) and territorial defense behavior in male tree sparrows, Spizella arborea. Birds receiving Silastic implants filled with corticosterone (B) had significantly higher plasma levels of B than control birds, which received empty implants, and exhibited pectoral muscle wastage and a decrease in body mass. We evaluated the hormonal and agonistic responses of the two implanted groups of birds using a simulated territorial intrusion (STI) 2 to 4 days after they were implanted. Corticosterone-treated and control birds did not differ in their circulating levels of reproductive hormones or in their behavioral responses to STI (latency to respond to intrusion, number of songs, and closest approach to a decoy and tape recording). Unlike previous studies of north temperate passerines, high physiological levels of exogenous B had no effect either on circulating levels of reproductive hormones or on territorial behaviors associated with breeding. Nonetheless, untreated tree sparrows do mount a robust adrenocortical response, having a two- to fourfold increase in plasma B levels during a 1-h period of capture. Thus, adrenocortical responsiveness is maintained in these birds, but elevated levels of glucocorticoids do not suppress reproductive hormones or associated behaviors. We believe that this hormonal and behavioral refractoriness to glucocorticoids-or uncoupling of the stress response from the reproductive axis-may be advantageous for species having extreme temporal constraints on their breeding schedules.     

Hypothalamic neurosecretion and photoinduced testicular growth in the tree sparrow,Spizella arborea

Summary To assess the role of the hypothalamic neurosecretory system in the mechanism of photoperiodic control of testicular growth in Spizella arborea, we have examined the effect of (1) photostimulation on the density of paraldehyde-fuchsin-stainable (PAF+) material in the neurovascular zone of the median eminence, and (2) interruption of the hypothalamo-hypophysial (neurosecretory) tract on the photoperiodic testicular response. Birds were captured during midwinter and retained on 8-hr daily photoperiods several months prior to the experiment. Interruption of the neurosecretory tract was effected by a small knife fashioned from stainless steel wire; radiography was employed to facilitate stereotaxic placement of the knife assembly.Extending the daily photoperiod from 8 to 20 hrs induced rapid testicular growth in photosensitive Spizella arborea, but had no consistent effect on the density of PAF+ material in the palisade layer of the anterior median eminence. Whereas some terminal controls showed moderate depletion after 21 days' exposure to 20-hr daily photoperiods, others retained as much or more stainable material than initial controls in which testicular growth was not induced. Moreover, testicular growth was induced at essentially identical rates whether the PAF+ innervation of the anterior median eminence was intact or eliminated to varying degrees. The finding that the rate of photoperiodically induced testicular growth is independent of the density or pattern of accumulation of PAF+material in the neurovascular zone of the median eminence, coupled with the inconstant effect of photostimulation on the density of stainable material therein, strongly suggests that the eminential component of the hypothalamic neurosecretory system is not a part of the mechanism that controls photoperiodic testicular growth in S. arborea.Contribution No. 980, Division of Biology, Kansas Agricultural Experiment Station, Manhattan 66502. This investigation was supported in part by the Undergraduate Research Participation Program (GY-771) of the National Science Foundation. We are grateful to French's Pet Bird Laboratory, Rochester, N.Y., for the parakeet foods, to Richard S. Donham, Michael D. Ruff, Thomas G. Shane, and Philip G. Watt for assisting with field operations, to Mr. Gary R. Dierking for preparing the drawing, and to Mr. Don Pihlaja for assisting with the photomicrography.     

A test of the hypothesis that T3 is the "seasonality" thyroid hormone in American tree sparrows (Spizella arborea): intracerebroventricular infusion of iopanoic acid, an inhibitor of T3 synthesis and degradation

This study tested the hypothesis that L-3,5,3'-triiodothyronine (T3) is the bioactive "seasonality" thyroid hormone in American tree sparrows (Spizella (arborea). The experimental approach coupled thyroid hormone replacement therapy after radiothyroidectomy with photostimulation and intracerebroventricular infusion of iopanoic acid, an inhibitor of L-3,5,3'-triiodothyronine synthesis and degradation. Endpoints were testis length, molt score, and hypothalamic content of chicken gonadotropin-releasing hormone 1. The hypothesis predicts that thyroidectomized male tree sparrows moved to long days and given thyroxine in combination with iopanoic acid will lack L-3,5,3'-triiodothyronine and so will not express thyroid hormone-dependent photoperiodic testicular growth (a vernal component of seasonality) and photorefractoriness or postnuptial molt (autumnal components of seasonality). It further predicts that iopanoic acid will enhance the efficacy of L-3,5,3'-triiodothyronine and so will facilitate the expression of seasonality in thyroidectomized males given L-3,5,3'-triiodothyronine replacement therapy. Iopanoic acid had no significant effect on any component of seasonality in thyroid-intact males given vehicle, or in thyroidectomized males given thyroxine or L-3,5,3'-triiodothyronine. Thyroid-intact males, as well as thyroidectomized males infused with thyroxine alone, commonly expressed all components of seasonality. Thyroidectomized males given L-3,5,3'-triiodothyronine alone exhibited photoperiodic testicular growth, but did not become photorefractory or initiate molt. While these results confirm that thyroid hormone acts centrally to program American tree sparrows for seasonality, they do not support the hypothesis that L-3,5,3'-triiodothyronine is the bioactive "seasonality" thyroid hormone, and they challenge the view that thyroxine is merely a prohormone.     

Seasonal acclimatization of metabolism in Eurasian tree sparrows (Passer montanus)

Acclimatization to winter conditions is an essential prerequisite for survival of small passerines of the northern temperate zone. Changes in photoperiod, ambient temperature and food availability trigger seasonal acclimatization in physiology and behavior of many birds. In the present study, seasonal adjustments in several physiological, hormonal, and biochemical markers were examined in wild-captured Eurasian tree sparrows (Passer montanus) from the Heilongjiang Province in China. In winter sparrows had higher body mass and basal metabolic rate (BMR). Consistently, the dry mass of liver, heart, gizzard, small intestine, large intestine and total digestive tract were higher in winter than in that in summer. The contents of mitochondrial protein in liver, and state-4 respiration and cytochrome c oxidase (COX) activity in liver and muscle increased significantly in winter. Circulating level of serum triiodothyronine (T₃) was significantly higher in winter than in summer. Together, these data suggest that tree sparrows mainly coped with cold by enhancing thermogenic capacities through increased organ masses and heightened activity of respiratory enzymes activities. The results support the view that prominent winter increases in BMR are manifestations of winter acclimatization in tree sparrows and that seasonal variation in metabolism in sparrows is similar to that in other small temperate-wintering birds.     

Fitness consequences of hybridization between house sparrows (Passer domesticus) and tree sparrows (P. montanus)

Gene transfer may occur following hybridization between closely related species if hybrids are viable and able to breed with individuals of one or both of the parental species. House (Passer domesticus) and tree sparrows (P. montanus) occasionally hybridize and produce viable offspring. Previously, we concluded that male tree × house sparrow hybrids are most probably fertile based on the observation of a male F1 hybrid feeding the nestlings with a female house sparrow in two consecutive clutches. However, recent DNA analyses based on blood samples revealed that all nestlings (4) in the first clutch were sired by a neighbouring house sparrow male, whereas nestlings in the second clutch (2) were not blood sampled and most probably died before fledging. This indicates that extensive extra-pair fertilization confounded our previous conclusion, and indicates that social partnership and attending behaviour can be imprecise measures of paternity.     

Habitat fragmentation causes bottlenecks and inbreeding in the European tree frog (Hyla arborea)

A genetic study of the European tree frog, Hyla arborea, in Denmark was undertaken to examine the population structure on mainland Jutland and the island of Lolland after a period of reduction in suitable habitat and population sizes. The two regions have experienced the same rate of habitat loss but fragmentation has been more severe on Lolland. Genetic variation based on 12 polymorphic DNA microsatellites was analysed in 494 tree frogs sampled from two ponds in Jutland and 10 ponds on Lolland. A significant overall deviation from Hardy-Weinberg expectations could be attributed to three ponds, all on Lolland. This was most probably caused by an inbreeding effect reducing fitness, which was supported by the observed significant negative correlation between larva survival and mean F(IS) value and mean individual inbreeding coefficient. A significant reduction in genetic variation (bottleneck) was detected in most of the ponds on Lolland. Population-structure analysis suggested the existence of at least 11 genetically different populations, corresponding to most of the sampled population units. The results indicated that the populations were unique genetic units and could be used to illustrate the migration pattern between newly established ponds arisen either by natural colonization of tree frogs or by artificial introduction. A high degree of pond fidelity in the tree frogs was suggested. A severe fragmentation process reducing population size and fitness within some of the populations probably caused the significant reduction in genetic variation of tree frog populations on Lolland.     

Isolation and characterization of microsatellite loci for the European tree frog (Hyla arborea)

We developed 11 new microsatellite markers for the European tree frog (Hyla arborea), and tested patterns of polymorphism in 54 adults (27 males and 27 females) from two ponds close to Lausanne (Western Switzerland). One marker was sex linked and two pairs displayed linkage disequilibrium. Comparisons of allele numbers with heterozygosity values support a stepwise-mutation model at neutral equilibrium, with mutation rates spanning nearly two orders of magnitude. These markers will prove useful for population genetic studies and fine-scale investigations requiring genetic assignment techniques.     

Corticosterone-induced gonadosuppression in photostimulated tree sparrows.

To explore the possibility that adrenal hormones play a role in the natural termination of testicular function in chronically photo-stimulated tree sparrows, corticosterone, the principal adrenal steroid of birds, was implanted intracerebrally in photoresponsive tree sparrows 3 weeks after they were transferred to a gonadostimulatory photoregime. Three weeks later, plasmas were assayed for immunoreactive luteinizing hormone (LH), testes were weighed, and implantation sites were determined. When implanted in a region of the basal hypothalamus known to be androgen sensitive, corticosterone reduced the concentration of plasma LH and blocked testicular growth. Control implants in the optic chiasma or tracts were not gonadoinhibitory. These results confirm the potential for inhibition of gonadotropin secretion by an effect of corticosterone on the hypothalamohypophyseal axis, but the physiological significance and mechanism of corticosterone's antigonadotropic effect remain to be established.     

Molecular evidence for phylogenetic relationships among buntings and American sparrows (Emberizidae)

To help clarify controversial phylogenetic relationships within the family Emberizidae, we sequenced 1238 bp of mitochondrial DNA from the cytochrome b gene and a flanking portion of ND5. Although the longspurs ( Calcarius ) and the snow buntings ( Plectrophenax ) have been grouped with the Old World buntings ( Emberiza ) in traditional classifications, our molecular phylogenies constructed with maximum likelihood and maximum parsimony place these genera basal to a clade in which the Old World buntings and North American sparrows are sister groups. Contrary to the hypothesis that the radiation within Emberiza is recent following a westward expansion of emberizid stock into Eurasia from North America, we found that the level of genetic divergence among Old World buntings approximates those among different genera in North American sparrows. Thus the radiation of the Emberizidae seems to have occurred at roughly the same time in the Palaearctic and Nearctic. Our results are consistent with earlier analyses of allozymes, but sequences from multiple genes and new morphological analyses are required to fully resolve phylogenetic relationships within the Emberizidae.     

Unilateral nervous control of the syrinx in java sparrows (Padda oryzivora)

Summary The syrinx of song birds contains two sound sources, the internal tympaniform membranes located one in each bronchus, that are controlled by the syringeal musculature. These muscles are innervated by the cervicalis descendens superior (CDS) branch of the hypoglossal nerve. Unilateral sections of the left CDS nerve in Java sparrows markedly disrupted tonal quality of the songs, although temporal parameters were unaltered. Bilateral CDS nerve sections caused greater disruption in frequency characteristics and temporal parameters were altered. Most birds died due to asphyxia soon after the operation. Right CDS nerve sections had much less effect, except on long whistles where extra low frequency sounds appeared, probably from the denervated right bronchus. Intact song pattern was restored within four months without re-innervation of the right syringeal musculature. This unilateral dominance in the control of the syrinx agrees with previous work and adds to the support for the two voice theory of sound production in birds (Greenewalt, 1968; Stein, 1968).Abbreviation CDS Nervus cervicalis descendens superior     

Heterozygosity-fitness correlations among wild populations of European tree frog (Hyla arborea) detect fixation load

Quantifying the impacts of inbreeding and genetic drift on fitness traits in fragmented populations is becoming a major goal in conservation biology. Such impacts occur at different levels and involve different sets of loci. Genetic drift randomly fixes slightly deleterious alleles leading to different fixation load among populations. By contrast, inbreeding depression arises from highly deleterious alleles in segregation within a population and creates variation among individuals. A popular approach is to measure correlations between molecular variation and phenotypic performances. This approach has been mainly used at the individual level to detect inbreeding depression within populations and sometimes at the population level but without consideration about the genetic processes measured. For the first time, we used in this study a molecular approach considering both the interpopulation and intrapopulation level to discriminate the relative importance of inbreeding depression vs. fixation load in isolated and non-fragmented populations of European tree frog (Hyla arborea), complemented with interpopulational crosses. We demonstrated that the positive correlations observed between genetic heterozygosity and larval performances on merged data were mainly caused by co-variations in genetic diversity and fixation load among populations rather than by inbreeding depression and segregating deleterious alleles within populations. Such a method is highly relevant in a conservation perspective because, depending on how populations lose fitness (inbreeding vs. fixation load), specific management actions may be designed to improve the persistence of populations.     

Sex‐chromosome differentiation parallels postglacial range expansion in European tree frogs (Hyla arborea)

Occasional XY recombination is a proposed explanation for the sex‐chromosome homomorphy in European tree frogs. Numerous laboratory crosses, however, failed to detect any event of male recombination, and a detailed survey of NW‐European Hyla arborea populations identified male‐specific alleles at sex‐linked loci, pointing to the absence of XY recombination in their recent history. Here, we address this paradox in a phylogeographic framework by genotyping sex‐linked microsatellite markers in populations and sibships from the entire species range. Contrasting with postglacial populations of NW Europe, which display complete absence of XY recombination and strong sex‐chromosome differentiation, refugial populations of the southern Balkans and Adriatic coast show limited XY recombination and large overlaps in allele frequencies. Geographically and historically intermediate populations of the Pannonian Basin show intermediate patterns of XY differentiation. Even in populations where X and Y occasionally recombine, the genetic diversity of Y haplotypes is reduced below the levels expected from the fourfold drop in copy numbers. This study is the first in which X and Y haplotypes could be phased over the distribution range in a species with homomorphic sex chromosomes; it shows that XY‐recombination patterns may differ strikingly between conspecific populations, and that recombination arrest may evolve rapidly (<5000 generations).     

Consequences of genetic erosion on fitness and phenotypic plasticity in European tree frog populations (Hyla arborea)

The detrimental effects of genetic erosion on small isolated populations are widely recognized contrary to their interactions with environmental changes. The ability of genotypes to plastically respond to variability is probably essential for the persistence of these populations. Genetic erosion impact may be exacerbated if inbreeding affects plastic responses or if their maintenance were at higher phenotypic costs. To understand the interplay 'genetic erosion-fitness-phenotypic plasticity', we experimentally compared, in different environments, the larval performances and plastic responses to predation of European tree frogs (Hyla arborea) from isolated and connected populations. Tadpoles from isolated populations were less performant, but the traits affected were environmental dependant. Heterosis observed in crosses between isolated populations allowed attributing their low fitness to inbreeding. Phenotypic plasticity can be maintained in the face of genetic erosion as inducible defences in response to predator were identical in all populations. However, the higher survival and developmental costs for isolated populations in harsh conditions may lead to an additional fitness loss for isolated populations.     

Embryonic development of the tree frog, Hyla arborea

A staged series with time data and original photographs of the embryonic development of the tree frog, Hyla arborea are presented, following Gosner’s generalized table. To contribute to species specific embryological studies of anurans, the external features of development of this species were described. Duration of embryonic development, from fertilization to 25^th stage, is about 9 days at 20 ± 1 °C. Cleavage is holoblastic and unequal. Unlike the typical anuran development, neurular rotation was not observed in stage 15 or in any other stages. Other developmental stages resulted in a normal-looking. Embryos of H. arborea hatched at stage 20 or 21, which varied individually. Hatching occurred in the 4^th or 5^th day after fertilization. The mean total length of hatchlings was calculated as 6.38 mm. The mean size of ovum was estimated 1.4 mm in diameter.     

Genetic differentiation of the Iberian tree frogs Hyla arborea molleri and Hyla meridionalis (Amphibia: Anura)

Genetic differentiation of Hyla arborea molleri and H. meridionalis was studied through allozyme electrophoresis and through essays of artificial hybridization. Samples were taken at several localities in Portugal, where these species occur either in sympatry or allopatry, and at one locality in France. Of the 21 presumptive loci analyzed, 6 were monoallelic, while 9 loci were discriminative between both species. A relatively high level of genetic differentiation was found between the two species (NEI's D = 0.725), contrasting with their morphological similarity. Remarkably low levels of genetic variability were found within species, with greater differentiation found among samples of H. meridionalis than of H. a. molleri. Although the capacity for interspecific hybridization was low, hybrid embryos resulted from both reciprocal crosses, but with much greater success in the cross ♂H. a. molleri x δH. meridionalis. Hybrids showed greater similarity with the male parent, and showed heterozygote allozymic phenotypes for discriminative loci. Results are compared with those of similar studies on the same and different species of Hyla, and differentiation of H. a. molleri and H. meridionalis is discussed in relation to their taxonomic status and origin.     

Multiscale determinants of tree frog (Hyla arborea L.) calling ponds in western Switzerland

A tree frog (Hyla arborea L., 1758) metapopulation in western Switzerland was studied during spring 2001. All potential calling ponds in an area of 350km² were searched for tree frog calling males. Twenty-nine out of 111 ponds sheltered between 1 and 250 callers. Most ponds were occupied by less than 12 males. Pond parameters were measured at three different levels using field analysis and a Geographical Information System (GIS). The first level was water chemistry and pond-associated measures. The second level was the surrounding land use in a 30m buffer around the pond. The third level consisted of landscape indices on a broader scale (up to 2km). Logistic regression was applied to identify parameters that can predict the presence of calling males in a pond. Response variable was the presence or absence of callers. Four significant parameters allowed us to explain about 40% of the total deviance of the observed occupational pattern. Urbanization around the pond had a highly negative impact on the probability of presence of calling males. Hours of direct sunlight on the pond was positively correlated with callers. Higher water conductivity was associated with a lesser probability of species presence. Finally, the further the closest two-lane road, the higher the probability of callers presence. Our results show that presence or absence of callers is influenced by factors acting at various geographical scales.