A potential mate influences reproductive development in female, but not male, pine siskins.

The role of photoperiod in avian reproductive timing has been well studied, and we are increasingly recognizing the roles of other environmental cues such as social cues. However, few studies have evaluated the extent to which males and females of the same species respond similarly to the same type of cue. Moreover, previous studies have rarely examined how variation in the quality or nature of a given social cue might modulate its effect. Here, we examine the sensitivity of male and female pine siskins (Spinus pinus) to a potential mate as a stimulatory cue for gonadal recrudescence, and we investigate whether variation in the relationship between a bird and its potential mate modulates the effect of that potential mate. Birds were initially housed without opposite sex birds on a 12L:12D photoperiod with ad libitum food. After gonadal recrudescence had begun males and females were randomly paired with an opposite sex bird or housed alone. An additional group of males was paired with estradiol-implanted females. In males, these social treatments had no effect on testis length, cloacal protuberance length, luteinizing hormone (LH) levels, or testosterone levels. In females, presence of a potential mate had a significant and positive effect on ovary score, defeathering of the brood patch, and LH levels. Among paired birds, the degree of affiliation within a pair corresponded to the extent of reproductive development in females, but not males. Thus, reproductive timing in females appears to be sensitive to both the presence of a potential mate and her relationship with him.

Testosterone manipulation postcastration does not alter cloacal gland growth differences in male quail selected for divergent plasma corticosterone stress response.

Japanese quail selected for reduced (low-stress, LS) rather than exaggerated (high-stress, HS) plasma corticosterone response to brief restraint have consistently shown greater cloacal gland (CG) development, an androgen-dependent trait. In this study, the effects of testosterone implants on levels of plasma testosterone and CG development in castrated LS and HS quail were determined. Stress-line males were castrated and randomly allocated to 1 of 3 testosterone treatments: the empty testosterone (ET), low testosterone (LT), or high testosterone (HT) implant group. Cloacal gland volume was determined at 4 weekly intervals that represented ranges of 1 to 9 d, 8 to 17 d, 15 to 24 d, and 22 to 31 d after castration and testosterone implantation. Levels of plasma testosterone were also assessed at the end of the study. Development of the CG was affected by quail line (LS > HS), testosterone treatment (HT > LT > ET), and time of measurement (1 to 9 d < 8 to 17 d < 15 to 24 d = 22 to 31 d after castration and testosterone implantation). A significant interaction between testosterone treatment and time of measurement on CG volume was also detected (with CG volume generally increasing with time in LT- and HT-treated quail, but not in ET-treated quail). However, even though HT implant treatments induced higher CG development than did LT treatments beyond the first interval of CG volume measurement, and despite the finding of greater CG volumes in LS than HS quail during the last 2 measurement intervals within each of the LT and HT groups, no interaction was observed between testosterone implant dosages and quail stress line on CG volume. Thus, by the end of the study, regardless of testosterone dose, CG volume was consistently greater in LS quail than in their HS counterparts. In addition, although, as expected, the testosterone implant treatment significantly altered levels of plasma testosterone (HT > LT > ET), neither quail line nor its interaction with testosterone treatment affected plasma testosterone. The present findings suggest that the often-observed depressed CG development in the HS line may be independent of testosterone effects.

Temporal control of urate oxidase activity in Drosophila: evidence of an autonomous timer in malpighian tubules.

The appearance of urate oxidase activity in the Malpighian tubules of Drosophlia melanogaster is synchronized with the time of emergence of the imago from the puparium. A developmental clock within the Malpighian tubules specifies the time of appearances of urate oxidase activity.

Embryonic and posthatching treatments with sex steroids demasculinize the motivational aspects of crowing behavior in male Japanese quail.

Demasculinizing action of embryonic estrogen on crowing behavior in male Japanese quails was examined. Eggs were treated with either 20 microg of estradiol benzoate (EB) or vehicle on the 10th day of incubation. Chicks hatched from both groups of eggs were injected daily with either testosterone propionate (TP; 10 microg/g b.w.), 5alpha-dihydrotestosterone (DHT, a non-aromatizable androgen; 10 microg/g b.w.), or vehicle from 11 to 50 days after hatching, and during this period their calling behaviors were observed. Irrespective of embryonic treatments, all birds received posthatching treatment with either TP or DHT, but not with vehicle, emitted crows in place of distress calls in a stress (non-sexual) context of being isolated in a recording chamber. The posthatching TP, but not posthatching DHT, induced crowing in a sexual context (crowing in their home-cages) from much earlier age than posthatching vehicle in the birds received control embryonic treatment with vehicle. The same TP treatment, however, completely eliminated the crowing in a sexual context in the birds received EB during their embryonic life. In the birds treated with either posthatching DHT or posthatching vehicle, the crowing in a sexual context was only slightly decreased by embryonic EB treatment. These data suggest that posthatching estrogen, derived from testosterone aromatization, enhances the demasculinizing action of embryonic estrogen, and thus strongly reduces the sexual motivation for crowing behavior. This demasculinizing action, however, would not influence vocal control system which generates acoustic pattern of crowing in the presence of androgens allowing the birds to crow in a non-sexual context.

Cloacal gland growth differences in high and low plasma corticosterone stress response line male quail reared under short daylengths.

Mail quail selected (generation 32) for reduced [low stress (LS)] or exaggerated [high stress (HS)] plasma corticosterone stress response to brief mechanical restraint (5 min of immobilization) were studied for differences in the growth of their cloacal glands when reared essentially lifelong on short day lengths of 8L:16D. Post-brooding, at 4 wk of age, 96 quail (48 LS+48 HS) were housed in cages (1 LS and 1 HS male/cage), and the short day light treatment was instigated. Using a digital caliper, cloacal gland (CG) size measurements (length and width, mm) were made biweekly beginning at 42 d of age and ending at 196 d of age (28 wk old). The CG volume (CVOL) was calculated from these measurements using a literature-proposed formula. The CVOL in both lines increased similarly with increasing age from 6 to 12 wk of age. However, beginning at 14 wk of age and thereafter, LS males showed greater (P<0.05) CVOL than HS ones. No further increases in CVOL were observed in either line at 24 wk of age beyond that seen at 22 wk. This study demonstrated that although both lines show CG development under short days, eventually CG growth becomes comparatively stymied in the HS males. Furthermore, the maintenance of similarly lower CVOL in HS than LS males, CVOL that showed no further increase from 22 to 24 wk of age, suggests that holding these quail lifelong on short days results in, comparatively, yet another permanent negative reproductive consequence in quail selected for exaggerated plasma corticosterone stress responsiveness.

Rate of testicular maturation, in relation to gonadotrophin and testosterone levels, in quail exposed to various artificial photoperiods and to natural daylengths.

Rates of testicular growth and plasma levels of FSH, LH and testosterone were determined in Japanese quail exposed to various fixed photoperiods (number of hours of light: number of hours of darkness): 12L : 12D, 13L : 11D, 14L : 10D, 16L : 8D and 20L : 4D and to natural daylengths. All five artificial photoperiods stimulated spermatogenesis, with the testes reaching maturity after 30-40 days. Maximum rates of testicular growth occurred with 14L : 10D, 16L : 8D or 20L : 4D but the rate was reduced by 50% in birds exposed to 12L : 12D.. This reduction was due to decreased growth in the seminiferous tubule epithelium (and hence in tubule diameter); the duration of spermatogenesis hardly being affected. Near maximum growth rates occurred with 13L : 11D. The hormone profiles offer an explanation for the differential rates of testicular growth. In the three longest photoperiods, FSH rose from 20 ng/ml to peak levels of 300-400 ng/ml after 10 days. As the testes matured, so the level of FSH decreased to 50-100 ng/ml. This pattern was not seen under 12L : 12D ; the level of FSH rose slowly to about 100 ng/ml and showed no peak of secretion. With 13L : 11D a small peak was found, which decreased at maturity. In quail with testes is greater than 1500 mg, the level of FSH was invariably about 100 ng/ml. Patterns of LH secretion were rather similar with all treatments, but testosterone was affected by photoperiod; lower levels were found under 12L : 12D than 20L : 4D. The rate of photoperiodically induced testicular growth was proportional to the levels of FSH, and possibly also testosterone, in the circulation. Outdoors, testicular growth began when daylengths reached about 12 h. Maturity occurred within the next 40 days. The levels of FSH rose steadily but did not show a peak of secretion. In general, the highest levels of hormone were found in July just before gonadal regression which occurred when the daylengths were still quite long.

Relative potencies of testosterone and 5 alpha-dihydrotestosterone on crowing and cloacal gland growth in the Japanese quail (Coturnix coturnix japonica).

It has been suggested that testosterone is less effective at inducing crowing behaviour in young birds than in adults because of the presence of higher levels of steroid 5 beta-reductase in the young brain, which converts testosterone to inactive 5 beta-reduced metabolites. This hypothesis was tested indirectly by comparing the relative potencies of 5 alpha-dihydrotestosterone (5 alpha-DHT), which cannot be converted to 5 beta-metabolites, and testosterone at inducing crowing in young gonadectomized male and female quail. The promotion of cloacal gland growth by these treatments was also assessed since there are no age-related changes in 5 beta-reductase in this organ. Silicone elastomer implants (2 X 5, 5 and 10 mm) containing 5 alpha-DHT were more effective at stimulating crowing than similar implants of testosterone whilst there was little difference in their potency at inducing cloacal gland growth. These results are consistent with the hypothesis that brain steroid 5 beta-reductase regulates the behavioural activity of testosterone in the brain of young birds.

Cloacal gland of the Japanese quail: androgen dependence and metabolism of testosterone.

A cloacal gland complex whose growth and development is androgen-dependent exists in the Japanese quail. In-vitro incubation studies of the cloacal gland using 4-14C-labelled testosterone as substrate allowed the positive identification of five metabolites: androstenedione, 5 beta-dihydrotestosterone (5 beta-DHT), 5 beta-androstane-3 alpha, 17 beta-diol, 5 alpha-DHT and 5 alpha -androstane-3 alpha, 17 beta-diol. More polar metabolites, not yet chemically identified, were detected in trace amounts. Androstenedione appeared to be the main testosterone metabolite in immature birds while in mature birds on long daylengths testosterone was preferentially metabolized to 5 alpha-DHT. This change may have been in response to the higher levels of plasma steroids found in mature birds. When various testosterone metabolites, contained in silicone elastomer capsules, were implanted s.c. into castrated birds maintained on a photostimulatory light regime, 5 alpha-DHT, 5 alpha-androstane-3,17-dione, androstenedione and 5 alpha-androstan-3 alpha-ol-17-one were shown to be equipotent with testosterone in stimulating the development of the cloacal gland. 5 alpha-Androstane-3 alpha, 17 beta-diol and 5 alpha-androstan-3 beta-ol-17-one stimulated some growth while 5 beta-DHT, 5 alpha-androstane-3 beta, 17 beta-diol, 5 alpha-androstane-3 alpha, 17 beta-diol, 5 beta-androstane-3,17-dione, androst-5-en-3 beta-ol-17-one and androst-5-ene-3 beta, 17 beta-diol were completely ineffective.

Testosterone induces hypothalamic aromatase during early development in quail.

The effects of testosterone on androgen metabolizing enzymes were examined in the developing hypothalamus of male and female quail using an in vitro radiometric assay which measures metabolite formation in individual brain samples. Testosterone (T) administered by subcutaneous silastic implants to gonadectomized 4-day-old chicks increased formation of estradiol-17 beta (E2) in both preoptic area + anterior hypothalamus (PA) and posterior hypothalamus + tuberal area (HT) to adult levels. The T-induced increase in E2 formation occurred to the same degree in both sexes. The increase was very small in control non-target areas, neostriatum intermediale + hyperstriatum ventrale (VN), of either sex. Testosterone had no effect on formation of 5 alpha-dihydrotestosterone (5 alpha-DHT), 5 beta-dihydrotestosterone (5 beta-DHT) and 5 beta-androstane 3 alpha, 17 beta-diol (5 beta, 3 alpha-diol) in PA. Kinetic analysis of the rate of E2 production by hypothalamic tissue from castrated chicks (CX-chicks) and castrates treated with T (CX + T-chicks) indicates that the increase in hypothalamic aromatase activity by T corresponds to induction of the enzyme: the Vmax (maximum velocity) was increased by T (CX-chicks, 21; CX + T-chicks, 91 fmol/mg FW/h), whereas the Km was unaffected (CX-chicks, 5.5; CX + T-chicks, 4.7 X 10(-8) M). Testosterone treatment, effective for inducing PA and HT aromatase activity, also activated crowing and caused cloacal gland development; neither of these effects were sexually dimorphic. Our results indicate that: (1) T induces aromatase specifically in the hypothalamus during early post-hatching development, other pathways of T metabolism are not affected; and (2) the inducible aromatase is not sexually dimorphic in the developing brain. Since there are sex differences in adult brain aromatase, we conclude that capacity for induction of the hypothalamic aromatase becomes sexually differentiated after the post-hatching period.

Developmental sex differences in brain aromatase activity are related to androgen level.

Sex differences in the metabolism of testosterone (T) in the developing brain of quail were examined using an in vitro microassay. During each developmental stage (day before hatching, hatching and 2 days after hatching) aromatase activity was higher in hypothalamic areas than in a control neostriatal area. There was no sex difference in oestradiol-17 beta (E2) formation in the late embryonic brain or at hatching. But aromatase activity in the male preoptic-anterior hypothalamic area was 50% higher than in females by day 2. No regional differences in brain 5 beta-reductase activity were detected at any of the developmental stages sampled. There was a sex difference in production of catabolic 5 beta-reduced metabolites. Male 5 beta-reductase activity declined continuously from high embryonic levels in all areas, whereas female enzyme activity showed an increase at hatching. In contrast to plasma progesterone, levels of T were higher in the male than in the female by day 1 after hatching. We suggest that elevated circulating T in the male after hatching may account for the sexual dimorphism in brain aromatase activity.

Developmental changes in Malpighian tubule cell structure.

Structural changes which occur in the Malpighian tubule yellow region primary cells during larval-pupal-adult development of the skipper butterfly Calpodes ethlius are described. The developmental changes in cell structure are correlated with functional changes in fluid transport (Ryerse, 1978a) in a way which supports osmotic gradient models of fluid secretion. Larval tubules are specialized for fluid secretion with deep basal infolds and elongate mitochondria-containing apical microvilli which provide channels in which osmotic gradients could be set up. The Malpighian tubule cells are extensively remodelled at pupation when fluid transport is switched off, but they persist intact through metamorphosis. At this time, the basement membrane doubles in thickness, the mitochondria are retracted from the microvilli and are isolated for degradation in autophagic vacuoles, and both apical and basal plasma membranes are internalized via coated vesicles for degradation in multivesicular bodies, which results in the shortening of the microville and the disappearance of the basal infolds. Mitochondria are re-inserted into the microvilli, and the basal infolds re-form in pharate adult stage Malpighian tubules when fluid secretion resumes. Adult tubules are similar in general structure to larval tubules and contain mitochondria in the microvilli and basal infolds. However, they differ from larval tubules in that they are capable of very rapid fluid transport, have a reduced tubule diameter and tubule wall thickness, a much thicker basement membrane and peripherally associated tracheoles. Mineral concretions of calcium phosphate accumulate in larval tubules, persist through metamorphosis and decline in number in adults, suggesting they serve some anabolic role.

Relationship between throat feather-color pattern and photodependency of testicular function in the athens randombred control line of Japanese quail.

The relationship between throat feather-color and testicular regression and recrudescence under short photoperiods of 6 hr light (L):18 h dark (D) was examined in the Athens randombred control line of Japanese quail. In contrast to earlier studies, no differences were found in testicular regression when quail with brick red, mixed red and white, central red surrounded by white, or all white throat feathers were shifted from a photoperiod of 16L:8D to one of 6L:18D. The cloacal protuberance areas (CPA) of all groups ranged between 1.81 +/- .09 and 1.90 +/- .19 cm2 while birds were housed on the long photoperiod. Cloacal protuberance areas of all groups were markedly reduced after 3 wk of exposure to 6L:18D. Testes weights of all groups ranged between 46.7 and 54.8 mg after 3 wk of exposure to the short photoperiod. In a second experiment, cloacal foam gland development of males of the four throat feather patterns was examined during 26 wk after changing the photoperiod from 16L:8D to 6L:18D. Foam glands of all groups regressed within 3 wk of exposure to 6L:18D. Glands of birds with brick red throats or mixed red and white feathers recrudesced by 11 wk under the short photoperiod. Foam glands of birds with white throats or a band of white surrounding a patch of red did not show recrudescence at 11 or 14 wk but did so by 26 wk.

A circadian rhythm in photosensitivity as the basis for the testicular response of Japanese quail to intermittent light.

In order to examine the relationship between the circadian rhythm in photosensitivity and intermittent light in stimulating testicular development, Japanese quail were exposed to intermittent light providing a single dark period interruption varying from .5 to 10 hr in duration. The treatment lasted for 3 weeks. Changes in the cloacal gland size were monitored throughout the test and testis weights were taken at the end of the test (7 and 11 weeks of age). The results indicated that quail have a circadian rhythm in photosensitivity occurring approximately 12 hr after the onset of light (dark-light interface) which lasts for 4 to 6 hr. Testicular stimulation occurred only when light was given within the photosensitive phase. However, this rhythm is subject to phase shifting by night interrupting light flashes. This resulted in variable testicular responses depending upon the relationship between the position of the new photosensitive phase and the available light. Photostimulation of the testes was proportional to the duration of light within the photosensitive phase up to a maximum of 4 hr of light. Intermittent light may be as effective as a single, long photoperiod in stimulating testes development providing that an adequate amount of light is given during the photosensitive phase.

Cloacal gland and testes development in male Japanese quail selected for divergent adrenocortical responsiveness.

The time course of sexual development in male quail from lines selected for either a reduced (low stress, LS) or an exaggerated (high stress, HS) plasma corticosterone response to brief restraint was determined. Sexual development was assessed by examination of cloacal gland area (CAREA) and volume (CVOL), proportion of individuals that produced cloacal gland foam, and the intensity of cloacal gland foam production (CFP) at 4, 5, 6, 8, 10, 14, and 16 wk of age. These intervals encompass ages during which growing, photostimulated quail would be expected to evolve from being totally prepubescent to well into adulthood. Testes weight (TW) and the proportion of individuals that had achieved a combined TW of 500 mg (PI500) were also measured at 6 and 16 wk of age. Mean CAREA and CVOL were similar in LS and HS quail at 4 wk of age. Beginning at 5 wk of age and thereafter, CAREA was greater (P < 0.05) in LS than in HS quail. Similarly, beginning at 6 wk of age and thereafter, CVOL was greater (P < 0.05) in LS than in HS quail. More LS than HS birds also expressed foam at 4 (P < 0.05), 5 (P = 0.15), and 6 wk (P < 0.05) of age. From 8 wk on, all birds were in foam production. CFP results mimicked those found for CAREA and CVOL in that LS quail tended toward higher CFP values than HS quail at all intervals of measurement. TW were higher (P < 0.02) in LS than HS quail when data from the 2 ages at which testes were harvested were combined. At 6 wk of age, the LS and HS birds showed a PI500 line difference (P < 0.08) of 1.0 and 0.91, respectively; by 16 wk, all birds exhibited a maximum PI500. The results suggest that 1) selection for reduced adrenocortical responsiveness in Coturnix is accompanied by an accelerated onset of puberty in males, and 2) enhanced reproductive development in LS males remains extant throughout early to middle-age adulthood.

The cloacal gland--an external indicator of testicular development in coturnix.

The quantitative relationship between cloacal gland size and testes weight during the stages of initial growth, involution and recrudescence is described. Cloacal gland foam was found to be a reliable indicator of testes size and function. The mean testes weight associated with the first presence of cloacal gland foam was 377 mg. and this occurred at a mean age of 29 days. The physical measurement of the cloacal gland provides a valuable noninvasive method of predicting testicular activity especially suited for long term experiments involving multiple measurements where a few coturnix may be used repeatedly (as their own control) rather than periodic killing of a large group of animals.

Influence of different photoperiods on development of gonad, cloacal gland and circulating thyroid hormones in male Japanese quail Coturnix coturnix japonica.

One day old chicks of Japanese quail were exposed to different photoperiods (LD, 8:16, 13.5:10.5, 16:8 and LL) and observations (testes weight, cloacal gland size, body weight and circulating thyroxine and triiodothyronine) were taken at the age of 3, 5, 7, 9 and 16 weeks. Results indicate that immediate reproductive development occurred in birds exposed to long photoperiods (greater than 12 hr). Growth under LD 8:16, was not apparent till 7th week and by 16 weeks, degree of gonadal development was similar in all the birds, irrespective of photoperiodic treatment. Whereas body weight of the intermediate and long day (LD 13.5:10.5, 16:8 and LL) treated birds increased upto 5th week and remained constant thereafter. But the chicks maintained under short day length (LD 8:16), showed spontaneous increase till the end of the study and birds were much heavier compared to all other groups. Plasma T4 concentration increased with increasing age till 9th week and remained unaltered thereafter. On the other hand T3 level did not change till 7th week followed by a decline. It is suggested that the initiation and degree of gonadal growth in quail depends on the availability of daily photoperiod, until the achievement of full breeding condition. Peak level of T4 observed in 9 week old birds may be involved in the development of photorefractoriness at that age.

Disruption of the temporally regulated cloaca endodermal ß-catenin signaling causes anorectal malformations.

The cloaca is temporally formed and eventually divided by the urorectal septum (URS) during urogenital and anorectal organ development. Although congenital malformations, such as anorectal malformations (ARMs), are frequently observed during this process, the underlying pathogenic mechanisms remain unclear. ß-Catenin is a critical component of canonical Wnt signaling and is essential for the regulation of cell differentiation and morphogenesis during embryogenesis. The expression of ß-catenin is observed in endodermal epithelia, including URS epithelia. We modulated the ß-catenin gene conditionally in endodermal epithelia by utilizing tamoxifen-inducible Cre driver line (Shh(CreERT2)). Both ß-catenin loss- and gain-of-function (LOF and GOF) mutants displayed abnormal clefts in the perineal region and hypoplastic elongation of the URS. The mutants also displayed reduced cell proliferation in the URS mesenchyme. In addition, the ß-catenin GOF mutants displayed reduced apoptosis and subsequently increased apoptosis in the URS epithelium. This instability possibly resulted in reduced expression levels of differentiation markers, such as keratin 1 and filaggrin, in the perineal epithelia. The expression of bone morphogenetic protein (Bmp) genes, such as Bmp4 and Bmp7, was also ectopically induced in the epithelia of the URS in the ß-catenin GOF mutants. The expression of the Msx2 gene and phosphorylated-Smad1/5/8, possible readouts of Bmp signaling, was also increased in the mutants. Moreover, we introduced an additional mutation for a Bmp receptor gene: BmprIA. The Shh(CreERT2/+); ß-catenin(flox(ex3)/+); BmprIA(flox/-) mutants displayed partial restoration of URS elongation compared with the ß-catenin GOF mutants. These results indicate that some ARM phenotypes in the ß-catenin GOF mutants were caused by abnormal Bmp signaling. The current analysis revealed the close relation of endodermal ß-catenin signaling to the ARM phenotypes. These results are considered to shed light on the pathogenic mechanisms of human ARMs.

Dysregulation of Wnt inhibitory factor 1 (Wif1) expression resulted in aberrant Wnt-ß-catenin signaling and cell death of the cloaca endoderm, and anorectal malformations.

In mammalian urorectal development, the urorectal septum (urs) descends from the ventral body wall to the cloaca membrane (cm) to partition the cloaca into urogenital sinus and rectum. Defective urs growth results in human congenital anorectal malformations (ARMs), and their pathogenic mechanisms are unclear. Recent studies only focused on the importance of urs mesenchyme proliferation, which is induced by endoderm-derived Sonic Hedgehog (Shh). Here, we showed that the programmed cell death of the apical urs and proximal cm endoderm is particularly crucial for the growth of urs during septation. The apoptotic endoderm was closely associated with the tempo-spatial expression of Wnt inhibitory factor 1 (Wif1), which is an inhibitor of Wnt-ß-catenin signaling. In Wif1(lacZ/lacZ) mutant mice and cultured urorectum with exogenous Wif1, cloaca septation was defective with undescended urs and hypospadias-like phenotypes, and such septation defects were also observed in Shh(-/-) mutants and in endodermal ß-catenin gain-of-function (GOF) mutants. In addition, Wif1 and Shh were expressed in a complementary manner in the cloaca endoderm, and Wif1 was ectopically expressed in the urs and cm associated with excessive endodermal apoptosis and septation defects in Shh(-/-) mutants. Furthermore, apoptotic cells were markedly reduced in the endodermal ß-catenin GOF mutant embryos, which counteracted the inhibitory effects of Wif1. Taken altogether, these data suggest that regulated expression of Wif1 is critical for the growth of the urs during cloaca septation. Hence, Wif1 governs cell apoptosis of urs endoderm by repressing ß-catenin signal, which may facilitate the protrusion of the underlying proliferating mesenchymal cells towards the cm for cloaca septation. Dysregulation of this endodermal Shh-Wif1-ß-catenin signaling axis contributes to ARM pathogenesis.

Dietary phytoestrogens and photoperiodic response in a male songbird, the Dark-eyed Junco (Junco hyemalis).

Many commercial bird diets are made with soy products that contain phytoestrogens (i.e., plant compounds that have weak agonist activity at estrogen receptors), but the effects of these compounds on bird physiology and behavior are largely unknown. The primary phytoestrogens present in soy are the isoflavones genistin and diadzin, which have been shown to affect reproductive measures in many taxa. Two groups of wild-caught male Dark-eyed Juncos (Junco hyemalis) were fed a diet either made with water-washed soy protein (soy(+)) or made with soy protein that had been alcohol-washed to extract isoflavones (soy(-)). Both groups exhibited a photoperiodic response to long days. Plasma luteinizing hormone (LH) concentrations increased within the first week of long day (LD) exposure for both groups, and over the course of the experiment LH was higher in the soy(+) group, although concentrations for both groups were lower than have been reported in free-living juncos. The rate of cloacal protuberance (CP) growth was significantly affected by diet, with the soy(-) birds beginning to increase their CPs about a week faster than soy(+) birds after exposure to LD. There was no group difference in food intake, fat score, body mass, or behavioral measures during the study or in testis weight at the end of the study. Although effects of dietary phytoestrogens detected were subtle (i.e., rate of CP growth), those investigating subtle effects of hormonally active substances (e.g., endocrine disruptors) or environmental cues affecting the reproductive axis in songbirds may want to consider eliminating phytoestrogens from their experimental diets.

Variations in the photoperiodic cloacal response of Japanese quail: association with testes weight and feather color.

The size of the cloacal gland was found to be a reliable indicator of testicular activity of Japanese quail. Six experiments were performed to examine the effects of alternating long and short photoperiod on the size of the cloacal gland of male Japanese quail. Three types of photoperiodic cloacal responses were distinguished. Type I birds became refractory to short photoperiods after they had experienced 5 weeks or more of short days. They maintained large cloacal glands under subsequent condition of alternating long and short photoperiod. Type II birds were intermediate types I and III birds did not become refractory to short photoperiods after experiencing 5 weeks or more of short days. The cloacal glands responded to conditions of alternating long and short photoperiods with increases or decreases in size. Feather color on the throat was found to correspond to the type of cloacal response. Type I birds had brick-red throat feathers. Type II birds had white feathers intermingled with brick-red feathers. Type III had white throat feathers. The percentages of types I, II, and III observed in the experimental population was 67, 18, and 15%, respectively. Type III birds were used to study the effects of blinding on the cloacal response to short photoperiod. Five out of eight blinded type III birds did not lose the responsiveness to short photoperiod. These results are consistent with the view that extraocular photoreceptors participate in the photoperiodic gonadal response of Japanese quail.