The Opitz syndrome gene MID1 is essential for establishing asymmetric gene expression in Hensen's node

Patterning the avian left-right (L/R) body axis involves the establishment of asymmetric molecular signals on the left and right sides of Hensen's node. We have examined the role of the chick Midline 1 gene, cMid1, in generating asymmetric gene expression in the node. cMid1 is initially expressed bilaterally, but its expression is then confined to the right side of the node. We show that this restriction of cMid1 expression is a result of repression by Shh on the left side of the node. Misexpression of cMid1 on the left side of the node results in bilateral Bmp4 expression and a loss of Shh expression. Correspondingly, downstream left pathway genes are repressed while right pathway genes are ectopically activated. Conversely, knocking down endogenous right-sided cMid1 results in a loss of Bmp4 expression and bilateral Shh expression. This results in an absence of right pathway genes and the ectopic activation of the left pathway on the right. Here, we present a revised model for the establishment of asymmetric gene expression in Hensen's node based on the epistatic interactions observed between Shh, cMid1, and Bmp4.     

An anatomic and genetic study of canine cryptorchidism.

Twelve cases of cryptorchidism were found in a colony of Minature Schnauzer purebred and crossbred dogs. At least nine affected dogs were derived from the same sire directly or indirectly. Of 12 affected dogs, five cases were unilateral and seven were bilateral. Eight of the 12 cases were subjected to anatomic study. Fixation of affected organs was by vascular perfusion or immersion. Testes were separated from epididymides and both were weighed. All unilateral retained testicles were on the right side, and right sided bilaterally retained testes were always smaller than their left sided counterparts. With only one exception, ectopic testes were in the abdominal positon. Developmentally, the morphologic appearance of the epididymis of abdominal testes was very primitive in bilateral cases but nearly normal in the unilateral cases. Degree of inbreeding was greater for bilateral cases than unilateral cases. High incidence of cryptorchidism in this colony provided good evidence for hereditary nature of the condition in the Miniature Schnauzer dog. Morphologic observations were suggestive of a multiple gene defect.     

The role of an endogenous PKA inhibitor, PKIalpha, in organizing left-right axis formation

Protein kinase inhibitor (PKI) is an endogenous inhibitor of cAMP-dependent protein kinase A (PKA). We have found that the alpha-isoform of PKI (PKIalpha) is asymmetrically expressed along the left-right (L-R) axis in chick embryos. At stage 6, PKIalpha is expressed on the right side of the node, and this asymmetric expression continues until stage 7+. After stage 8, PKIalpha expression returns symmetric. Treatment of embryos with antisense PKIalpha oligonucleotides increased the incidence of reversed heart looping. Antisense oligonucleotides also induced ectopic expression of the left-specific genes Nodal and Pitx2, and suppressed the expression of the right-specific gene SnR in the right lateral plate mesoderm. Similarly, treatment with PKA activators forskolin and Sp-cAMPs resulted in both reversed heart looping and bilateral expression of NODAL: Ectopic activin induced PKIalpha on the left side of the node, while ectopic Shh and anti-Shh antibody had no effect on PKIalpha expression. Taken together, these data suggest that PKIalpha induced by an activin-like molecule, through the inhibition of PKA activity, suppresses the Nodal-Pitx2 pathway on the right side of the body.     

Characteristics of cryptic/ectopic and contralateral scrotal testes in dogs between 1 and 2 years of age

Testicular malposition represents a common developmental genital defect in dogs and can affect one or both testes. In both humans and dogs, unilateral cryptorchism is more frequently detected and thought to be the expression of a genetic abnormality affecting both the undescended and scrotal testis. In the dog, there is evidence of degenerative processes affecting the maldescended testis. However, the histologic and functional changes that occur in the scrotal testis of unilateral cryptorchid or ectopic individuals remain a source of debate. Because the bilateral surgical removal of the testes leads to some undesirable side effects, the aim of this study was to evaluate the necessity for performing bilateral orchiectomy in young unilateral cryptorchid dogs. A morphologic study of both cryptic/ectopic and scrotal testes in young dogs affected by unilateral testicular maldescent was therefore conducted. The study was conducted on 10 dogs aged 1 to 2 yr and affected by unilateral testicular maldescent. We found that, in young dogs, even if no neoplastic lesions were observed, morphologic abnormalities are detectable between 1 and 2 yr of age in the maldescended testes with severity dependent on testicular position. In contrast, in the scrotal testes, the histologic and immunohistochemical exam failed to find signs of incorrect development or morphologic abnormalities. The results seem to suggest that, though the early removal of the undescended testis is recommended, continuous monitoring of the scrotal testis for the life of the dog is preferable to removing it considering the undesirable side effects related to castration.     

Generation of robust left-right asymmetry in the mouse embryo requires a self-enhancement and lateral-inhibition system

The bilateral symmetry of the mouse embryo is broken by leftward fluid flow in the node. However, it is unclear how this directional flow is then translated into the robust, left side-specific Nodal gene expression that determines and coordinates left-right situs throughout the embryo. While manipulating Nodal and Lefty gene expression, we have observed phenomena that are indicative of the involvement of a self-enhancement and lateral-inhibition (SELI) system. We constructed a mathematical SELI model that not only simulates, but also predicts, experimental data. As predicted by the model, Nodal expression initiates even on the right side. These results indicate that directional flow represents an initial small difference between the left and right sides of the embryo, but is insufficient to determine embryonic situs. Nodal and Lefty are deployed as a SELI system required to amplify this initial bias and convert it into robust asymmetry.     

Left and right contributions to the<Emphasis Type="Italic"> Xenopus</Emphasis> heart: implications for asymmetric morphogenesis

The left-right asymmetry of the vertebrate heart is evident in the topology of the heart loop, and in the dissimilar morphology of the left and right chambers. How left-right asymmetric gene expression patterns influence the development of these features is not understood, since the individual roles of the left and right sides of the embryo in heart looping or chamber morphogenesis have not been specifically defined. To this end, we have constructed a bilateral heart-specific fate map of the left and right contributions to the developing heart in the Xenopus embryo. Both the left and right sides contribute to the conoventricular segment of the heart loop; however, the left side contributes to the inner curvature and ventral face of the loop while the right side contributes to the outer curvature and dorsal aspect. In contrast, the left atrium is derived mainly from the original left side of the embryo, while the right atrium is derived primarily from the right side. A comparison of our fate map with the domain of expression of the left-right gene, Pitx2, in the left lateral plate mesoderm, reveals that this Pitx2-expressing region is fated to form the inner curvature of the heart loop, the left atrioventricular canal, and the dorsal aspect of the left atrium. We discuss the implications of these results for the role of left-right asymmetric gene expression in heart looping and chamber morphogenesis.     

Lateralization of syringeal function during song production in the canary

The canary (Serinus canaria) vocal organ, the syrinx, has two separate sound sources, one in the cranial end of each bronchus. Previous investigations of whether song syllables are produced unilaterally or bilaterally have provided two contradictory results, as one researcher suggested that almost all syllables are produced by the left side of the syrinx alone, whereas another researcher suggested that both sides contribute similarly to all syllables. Our experiments, which involved unilateral bronchus plugging followed later by denervation of the ipsilateral syringeal muscles, attempted to resolve this disagreement. The males with right bronchus plugs, singing on the left side of the syrinx alone, produced nearly normal songs, whereas the birds with left bronchus plugs, singing on the right side, sang quite poorly. Interpretation of these data is difficult because it is not clear how syringeal function would be affected if the airflow rate through the intact side is increased above normal, nor is it known if the bird can compensate for bronchus occlusion. Nonetheless, we suggest that in male canaries most syllables are normally sung by the left side alone, with some syllables being produced by the right side alone and some being sung by both sides together. Right nerve section had little effect on the right-bronchus-plugged males' ability to sing, but the repertoires of the left-plugged males were altered after left nerve section, indicating the possibility that signals carried by the left nerve exert an influence on the contralateral side.     

Left-right asymmetry in the sea urchin embryo is regulated by nodal signaling on the right side

The asymmetric positioning of internal organs on the left or right side of the body is highly conserved in vertebrates and relies on a Nodal signaling pathway acting on the left side of the embryo. Whether the same pathway also regulates left-right asymmetry in invertebrates and what is the evolutionary origin of the mechanisms controlling left-right determination are not known. Here, we show that nodal regulates left-right asymmetry in the sea urchin but that, intriguingly, its expression is reversed compared to vertebrates. Nodal signals emitted from the right side of the larva prevent the right coelomic pouch from forming the imaginal rudiment. Inhibition of Nodal signaling after gastrulation causes formation of an ectopic rudiment on the right side, leading to twinned urchins after metamorphosis. In contrast, ectopic activation of the pathway prevents formation of the rudiment. Our results show that the mechanisms responsible for left-right determination are conserved within basal deuterostomes.     

Behavioral and morphological asymmetries in chukar Alectoris chukar copulation

Birds often exhibit greater reproductive tract development on the left side than right side. Behavioral evidence from the three species for which data has been published indicates that these species copulate more frequently on the left side of females than on the right side. Missing from the literature are studies that compare asymmetry in copulation behavior to asymmetry in reproductive tract morphology of the same individuals of both sexes within a single species. To better understand the potential for cryptic sexual selection to influence avian copulation, we measured asymmetries in chukar Alectoris chukar copulation using 24 male and 29 female chukar brought into captivity from the wild. Chukar copulated (n=37) more from the left side (n=30) of females than the right side (n=7). The left testis of males was consistently greater in size, mass and volume than the right testis. The left ovary and oviduct of females was consistently functional with no observable development of the right ovary or oviduct. Left-side bias in direction of copulation, larger left testes, and functional left vaginal openings may act in concert to deliver spermatozoa to the oviduct, promoting fertilization.     

Morphologic study of the testes from spontaneous unilateral and bilateral abdominal cryptorchid boars

Macroscopical and histological characteristics were examined in both testes from three healthy boars, three boars with unilateral abdominal cryptorchidism on the right side, and three boars with bilateral abdominal cryptorchidism. Abdominal cryptorchidism, unilateral and bilateral, provoked a significant decrease of the weight and volume of the ectopic testes. The scrotal testis of the unilateral cryptorchid boars showed an increase in its volume and weight. Cryptorchidism also induced abnormalities in the histological structure of seminiferous tubules, lamina propria, and interstitial tissue of the abdominal testes. The number of seminiferous tubules decreased; the seminiferous epithelium was constituted by few spermatogonia with an atypical pattern and by abnormal Sertoli cells. The lamina propria showed a variable degree of thickening and collagenization. The interstitial tissue was very developed but displayed a decrease in the Leydig cell population. These abnormalities were more critical in bilateral cryptorchidism than in unilateral cryptorchidism. The scrotal testis of the unilateral cryptorchid boars showed normal appearance, but a decrease of the number of seminiferous tubules was observed. Moreover, the seminiferous tubules showed impaired spermatid maturation. The alterations observed in the abdominal testes of the unilateral and bilateral cryptorchid boars were attributed to defective proliferation and differentiation of Sertoli cells and Leydig cells. The anomalies in the scrotal testis of the unilateral cryptorchid boars were due to disturbances in the Sertoli cell activity.     

Structural and functional abnormality of ectopic testes in rats

Some males of a mutant strain of King-Holtzman rats exhibit an anomalous heritable defect manifested as either unilateral or bilateral ectopic testes. In the adult, these testes contain seemingly immature Sertoli and Leydig cells, seminiferous tubules greatly reduced in diameter, and exhibit arrested spermatogenesis. Thus, the affected testis is essentially sterile. An inability to produce normal amounts of testosterone and androstenedione by these gonads is probably a reflection of changes that have been effected in their Leydig cells. Thus, this study suggests that abnormal function of the Leydig and Sertoli cells and seminiferous tubule failure in these mutant animals result from the physiologically cryptorchid condition.     

Regulation of neuronal lineage decisions by the HES-related bHLH protein REF-1

Members of the HES subfamily of bHLH proteins play crucial roles in neural patterning via repression of neurogenesis. In C. elegans, loss-of-function mutations in ref-1, a distant nematode-specific member of this subfamily, were previously shown to cause ectopic neurogenesis from postembryonic lineages. However, while the vast majority of the nervous system in C. elegans is generated embryonically, the role of REF-1 in regulating these neural lineage decisions is unknown. Here, we show that mutations in ref-1 result in the generation of multiple ectopic neuron types derived from an embryonic neuroblast. In wild-type animals, neurons derived from this sublineage are present in a left/right symmetrical manner. However, in ref-1 mutants, while the ectopically generated neurons exhibit gene expression profiles characteristic of neurons on the left, they are present only on the right side. REF-1 functions in a Notch-independent manner to regulate this ectopic lineage decision. We also demonstrate that loss of REF-1 function results in defective differentiation of an embryonically generated serotonergic neuron type. These results indicate that REF-1 functions in both Notch-dependent and independent pathways to regulate multiple developmental decisions in different neuronal sublineages.     

Sex and age differences of relationships among stepping parameters for evaluating dynamic balance in the elderly

This study aimed to examine the relationships among various stepping parameters, sex, and age in the elderly. Healthy elderly Japanese individuals 60-85 years old (50 males and 61 females) performed 4 types of stepping motions for 20 s. Stepping motions included bilateral stepping (back/forth and right/left) and unilateral stepping (back/forth and right/left). The number of steps, the average connecting time of a foot during one step, and the average time of both feet touching the floor at the same time (bilateral connecting time) were measured with a foot switch sheet. The trial-to-trial reliability was very high (above 0.86) except for the bilateral connecting time in the bilateral stepping back/forth test for 70-85 year olds (males: 0.67, females: 0.68). With age, the number of steps was significantly smaller, and the average connecting time and the bilateral connecting time were shorter in all stepping tests. There were significant sex differences in bilateral connecting time for bilateral stepping right and left and the number of steps for the bilateral stepping back and forth and the unilateral stepping right and left tests. The number of steps and average connecting time showed high correlations between bilateral stepping right/left and back/forth (r=0.71-0.94) and between unilateral stepping back/forth and right/left (r=0.87-0.99). There were significant correlations of the average connecting time between bilateral and unilateral stepping motions (r=0.51-0.83), but both stepping motions are considered to have different motion properties from the viewpoint of center of gravity sway. The correlations between the bilateral connecting time and the number of steps in bilateral stepping were relatively low (males: /r/<0.70, females: /r/<0.57). The bilateral connecting time was near 0 s in many males; thus, it may depend greatly on individual or sex differences in stepping strategy. These results suggest that the stepping motions used in this study can evaluate dynamic balance ability, and that the unilateral test may be useful for the elderly who cannot walk independently with ease.     

The Incidence of Double Hypoglossal Canal in Japanese: Evaluation with Multislice Computed Tomography

ResultsDouble hypoglossal canal was identified in 16.9% of the patients, and was bilateral in 2.2%. Double hypoglossal canal was significantly more frequent on the left side than right (P = 0.004, odds ratio = 1.79) and in males than females (P = 0.011, odds ratio = 1.67). A larger left or right-sided canal was found in 31.6% and 12.2% of the patients, respectively, following the same side preference as that of double hypoglossal canal. Almost perfect agreement was achieved between the two readers (k = 0.975).ConclusionsIn this Japanese population, the prevalence of a double hypoglossal canal was 16.9%, of which 2.2% were bilateral. Double hypoglossal canal was more frequent in males than females, and on the left side than right.     

Case of unilateral wing formation in the female of the glowworm Lampyris noctiluca

On July 27, 1999, the first author found a unilaterally winged adult glowworm in a park in the city of Mainz. Except for the wings on the left side, the specimen exhibited female characteristics that extended to external sexual appendages, the lantern and the gonads. The internal organization showed some remarkable differences between right (wingless) and left (winged) side. The right ovary contained three times more mature eggs than the left side and the volume of the corpus allatum of this side was about one-third larger than that of the left side. This suggests that aptery and egg maturation are affected by corpus allatum activity, i.e., juvenile hormone production. The findings do not support the hypothesis of Naisse ([1966] Arch Biol Liège 77:139-201) that wing formation, as a secondary male characteristic, is controlled by an androgenic hormone from the testes in the glowworm. Thus, the observations on this exceptional specimen have implications for the current hypotheses concerning the control of sexual wing dimorphism in Lampyris noctiluca.     

Effect of tamoxifen on H-Y antigen expression and gonadal development in chicken embryos

Avian species follow the ZW/ZZ system of sex determination, which the female is heterogametic and expresses H-Y (or, more appropriately, 'H-W') antigen. We present the results of an investigation into the effects of the antiestrogen, tamoxifen, on gonadal differentiation and H-Y antigen expression in chickens. When given at doses of 0.25-2 mg per egg immediately before incubation, tamoxifen blocked regression of the right gonad in a significant number of 14-day-old female embryos. The nonregressed right gonad had a testis-like external appearance and, in some cases, contained what appeared to be spermatogenic tubules. Tamoxifen had no histologically detectable effect on the differentiation of the left ovary or the testes. In spite of tamoxifen's histological effects on right female gonads, it did not masculinize the steroidogenic capabilities of these gonads. Whether obtained from drug- or vehicle-treated embryos, the left and right female gonads always contained appreciable amounts of estrogen. In contrast, testes obtained from either drug- or vehicle-treated embryos did not contain detectable amounts of estrogen. Tamoxifen reduced the H-Y antigen levels in female liver and gonads. In both left and right female gonads, the reduction was to male levels. In female livers, tamoxifen reduced H-Y antigen to levels intermediate between those of normal males and females. Thus, the expression of H-Y antigen in both gonadal and nongonadal tissue is estrogen dependent, but the dependency appears to be more stringent for gonadal tissue.(ABSTRACT TRUNCATED AT 250 WORDS)