Normal Gonads Research Articles

Diagnosis and treatment of rare complications of pelvic fractures

PurposeTo enhance the awareness of rare complications of pelvic fracture and describe the correct diagnosis and effective treatment. MethodsA total of 188 cases of pelvic fractures were retrospectively reviewed, and four patients who suffered from four types of rare pelvic fracture complications were described, namely ureteral obstruction caused by retroperitoneal hematoma-induced abdominal compartment syndrome (ACS), bowel entrapment, external iliac artery injury, and open scrotal sac injury. ResultsWe demonstrated that combined measures should be employed to prevent the occurrence of ACS following major pelvic fractures. Ureteral catheter support may be a good option at an early stage when ACS occurred. Contrasted computed tomography examination and sufficient awareness are keys to a correct diagnosis of bowel entrapment following pelvic fractures. Recognition of risk factors, early diagnosis, and prompt treatment of suspected injury of the external iliac artery are keys to patient survival and to avoid limb loss. Scrotal and/or testicular injury complicated by pelvic fractures should be carefully treated to maintain normal gonad function. Additionally, establishment of a sophisticated trauma care system and multi-disciplinary coordination are important for correct diagnosis and treatment of rare complications in pelvic fractures. ConclusionsRare complications of pelvic fractures are difficult to diagnose and negatively impact outcome. Recognition of risk factors and sufficient awareness are essential for correct diagnosis and prompt treatment.

Diagnosis and treatment of rare complications of pelvic fractures

PurposeTo enhance the awareness of rare complications of pelvic fracture and describe the correct diagnosis and effective treatment. MethodsA total of 188 cases of pelvic fractures were retrospectively reviewed, and four patients who suffered from four types of rare pelvic fracture complications were described, namely ureteral obstruction caused by retroperitoneal hematoma-induced abdominal compartment syndrome (ACS), bowel entrapment, external iliac artery injury, and open scrotal sac injury. ResultsWe demonstrated that combined measures should be employed to prevent the occurrence of ACS following major pelvic fractures. Ureteral catheter support may be a good option at an early stage when ACS occurred. Contrasted computed tomography examination and sufficient awareness are keys to a correct diagnosis of bowel entrapment following pelvic fractures. Recognition of risk factors, early diagnosis, and prompt treatment of suspected injury of the external iliac artery are keys to patient survival and to avoid limb loss. Scrotal and/or testicular injury complicated by pelvic fractures should be carefully treated to maintain normal gonad function. Additionally, establishment of a sophisticated trauma care system and multi-disciplinary coordination are important for correct diagnosis and treatment of rare complications in pelvic fractures. ConclusionsRare complications of pelvic fractures are difficult to diagnose and negatively impact outcome. Recognition of risk factors and sufficient awareness are essential for correct diagnosis and prompt treatment.

Interspecific hybridization between Crassostrea angulata and C. ariakensis

Interspecific hybridization can generate heterosis, which is proven to be a useful tool in selective breeding programs for oyster culture. Crassostrea angulata and C. ariakensis are two important economic shellfish species in China. We conducted 2 × 2 reciprocal crosses to determine whether these two species can cross-fertilize and their hybrids can hatch, survive and perform heterosis. Fertilization was found symmetrical without delay. The rate of fertilization success of C. angulata ♀ × C. ariakensis ♂ was lower than that of C. ariakensis ♀ × C. angulata ♂, and the success rate of both hybridizations was lower than that of two intraspecific crosses each. During the planktonic period, survival rate of the progeny was lower in the hybrid crosses than in the intraspecific crosses. On day 360, mean shell height of the progeny of C. angulata ♀ × C. angulata ♂ was highest, which was followed by that of C. angulata ♀ × C. ariakensis ♂, C. ariakensis ♀ × C. ariakensis ♂ and C. ariakensis ♀ × C. angulata ♂ in a descending order. Morphology of adults produced by the hybrid crosses was similar to that of C. angulata. Both hybrids underwent normal gonad development and produced mature gametes in the mating season. This study provided new insights into the quantitative traits in interspecific crosses of Crassostrea species, thus being of guidance value for selective breeding of oyster.

Genomic variation in the hybrids of white crucian carp and red crucian carp: evidence from ribosomal DNA.

In this study, we conducted a cross of white crucian carp (♀)×red crucian carp (♂) (WR), and characterized the morphology, reproduction and genetics of the progeny. Different from parents, WR with the gray color showed the hybrid morphological traits of both parents. WR possessed normal gonads producing mature eggs or sperm, and exhibited high fertilization rate (90.2%) and high hatchery rate (80.5%), which contributed to produce and enlarge the population. WR with the same DNA content as parents was a diploid fish with 100 chromosomes (2n=100). Amplified ITS of 45S rDNA, in WR the sequences consisting of 884 bp bases of the entire ITS-1 region, 5.8 S region, and entire ITS-2 region. The sequences showed high similarity between WR and its parents and leaned towards male inheritance. In WR, NTS of 5S rDNA consisted of three length types with total 654 bp bases. From sequence analysis of NTS, WR shared 94.2% and 95.1% similarities with their female and male parent, respectively. Sequence analysis of ITS and NTS revealed that there existed recombination and variation in the hybrid progeny, which was the genetic base for adaptation and speciation. In conclusion, we obtained WR from hybridization and it exhibited hybrid traits in morphology and variation in genetic composition showing essential difference with its parents. The obtainment of WR has important significance in fish genetic breeding.

Kiss and Tell: Deletion of Kisspeptins and Receptors Reveal Surprising Results

The well-known expression “Kiss and Tell” usually refers to revealing one’s sexual exploits or private matters. In the case of the transcription activator-like effector nucleases gene knockout studies by Tang et al (1), sequential removal of the 2 known kisspeptins and their receptors surprisingly reveals that this system is not required for reproduction. The authors rightly note that this challenges the idea that kisspeptin neurons are central regulators of vertebrate reproduction (eg, Refs. 2, 3 among many). To date, most observations leading to this conclusion are based on studies in mammals, including humans, rats, and mice, and thus may not reflect the diversity of neuroendocrine control mechanisms across the vertebrate groups. On the surface, this lack-of-effect study in zebrafish could be quite discouraging, except in the light of the evolution of neuropeptidergic systems controlling reproductive function. Zebrafish are members of a group of fish called teleosts. Teleosts are wonderfully diverse in their reproductive strategies, often beautiful, economically important, and there are nearly 30 000 species currently known (4). The evolutionof these ray-finned fishgoesbackapproximately 200 million years before present, after undergoing the third lineage-specific genome duplication (5). Complete or partial gene duplications in many teleosts have presented real challenges when trying to sort out the evolution and specific functions of reproductive neuropeptides. Many neuropeptides, neurotransmitter synthesis enzymes, and neurohormone receptors duplicated and/or mutated over time, resulting in an array of novel molecular substrates for the evolution of hormone systems. This may be one reason that teleosts are successful, being both widespread in distribution and fecund. These evolutionary duplication events in teleosts have generated multiple numbers of genes and peptides in comparison with their mammalian counterparts. For example, it has been observed that most fish species harbor 2 or 3 distinct GnRH genes and peptides (6, 7). Their location and functions are different, and early GnRH knockdown studies in trout revealed that reproduction may continue even with reduced GnRH levels (8). Subsequent studies revealed a more important role for the hypophysiotropic forms of GnRH in tilapia (9) and common carp (10). In both cases, reproduction was not inhibited in all animals expressing the GnRH antisense transgene. For the common carp, a close relative of zebrafish (family Cyprinidae, eg, carps, minnows, and goldfish), abnormal sexual development and infertility were observed in about 40% of the animals expressing the antisense transgene targeting the hypophysiotropic form of GnRH (10). On the other hand, the remaining 60% had normal gonads with sperm or eggs (10), indicating that reproduction was still possible in these carp. What about the kisspeptin knockout fish? Through rigorous analysis, the authors demonstrated that spermatogenesis, folliculogenesis, and reproductive capacity were not impaired in 6 mutant lines missing kisspeptins or kisspeptin receptors (1). It should be noted that thekisspeptin systemhasbeendescribed indetail in several fish, and there are significant differences between species, and in comparison with mammals. If one considers only zebrafish, Kah’s group (11), using specific antibodies, has clearly demonstrated that there are 2 independent kisspeptin systems (kiss1 and kiss2). The first is the habenular kiss1 system that is likely involved in nonreproductive activities, perhaps metabolic control. On the other hand, distribution of kiss2 neurons in hypophysiotropic areas

Generation of mouse functional oocytes in rat by xeno-ectopic transplantation of primordial germ cells.

Primordial germ cells (PGCs) are germ cell progenitors in the fetal genital ridge; female PGCs give rise to definitive oocytes that contribute to the next generation. Artificial PGCs have been induced in vitro from pluripotent stem cells and gonad-like tissue has been induced in vivo by cotransplantation of PGCs with PGC-free gonadal cells. To apply these technologies to human infertility treatment or conservation of rare species, PGC transplantation must be established in xenogenic animals. Here, we established a xenogeneic transplantation model by inducing ovary-like tissue from PGCs in xenogenic animals. We transplanted enzymatically dispersed PGCs with PGC-free gonadal cells under the kidney capsule of xenogenic immunodeficient animals. The transplanted cells formed ovary-like tissues under the kidney capsule. These tissues were histologically similar to the normal gonad and expressed the oocyte markers Vasa and Stella. In addition, mouse germinal vesicle-stage oocyte-like cells collected from ovary-like tissue in rats matured to metaphase II via in vitro maturation and gave rise to offspring by intracytoplasmic sperm injection. Our studies show that rat/mouse female PGCs and PGC-free gonadal cells can develop and reconstruct ovary-like tissue containing functional oocytes in an ectopic xenogenic microenvironment.

Metal levels in the liver, muscle, gill, intestine, and gonad of Lake Van fish (Chalcalburnus tarichi) with abnormal gonad.

Recently, an increasing number of studies have shown that Lake Van--the biggest soda lake in the world--is polluted due to an increasing population. Studies have shown abnormalities in the Lake Van fish (Chalcalburnus tarichi), the sole fish species that inhabits the lake. Unlike the vitellogenic and mature oocytes in normal gonads, abnormal gonads show large amounts of connective tissue and young oocytes. In this study, metal levels (nickel [Ni], copper [Cu], cobalt [Co], iron [Fe], zinc [Zn], cadmium [Cd], lead [Pb], and manganese [Mn]) in the muscle, liver, gill, intestine, and gonad of Lake Van fish with normal and abnormal gonads were assessed. Further, the metal contents in the wastewater from the wastewater treatment facility situated near Lake Van in Van City were assessed. All the metal levels, except that of Zn, were high in the Lake Van environment (P<0.05). The highest metal content in the tissues was for Fe, while the lowest level was for Co. The Pb level was found to be very high in both fish groups. Cd was not found in the tissues of both fish groups. The levels of Fe, Cu, Pb, and Mn were not significant in the tissues of both normal and abnormal fish groups. Zn level was significantly high in the livers and gonads of fish with abnormal gonads, and Co level was significantly high only in the livers (P<0.05). Consequently, high levels of Zn in the liver and gonads and high levels of Co in the liver may be factors causing the abnormal gonads in the Lake Van fish.

LIN28Expression in Malignant Germ Cell Tumors Downregulateslet-7and Increases Oncogene Levels

Despite their clinicopathologic heterogeneity, malignant germ cell tumors (GCT) share molecular abnormalities that are likely to be functionally important. In this study, we investigated the potential significance of downregulation of the let-7 family of tumor suppressor microRNAs in malignant GCTs. Microarray results from pediatric and adult samples (n = 45) showed that LIN28, the negative regulator of let-7 biogenesis, was abundant in malignant GCTs, regardless of patient age, tumor site, or histologic subtype. Indeed, a strong negative correlation existed between LIN28 and let-7 levels in specimens with matched datasets. Low let-7 levels were biologically significant, as the sequence complementary to the 2 to 7 nt common let-7 seed "GAGGUA" was enriched in the 3' untranslated regions of mRNAs upregulated in pediatric and adult malignant GCTs, compared with normal gonads (a mixture of germ cells and somatic cells). We identified 27 mRNA targets of let-7 that were upregulated in malignant GCT cells, confirming significant negative correlations with let-7 levels. Among 16 mRNAs examined in a largely independent set of specimens by quantitative reverse transcription PCR, we defined negative-associations with let-7e levels for six oncogenes, including MYCN, AURKB, CCNF, RRM2, MKI67, and C12orf5 (when including normal control tissues). Importantly, LIN28 depletion in malignant GCT cells restored let-7 levels and repressed all of these oncogenic let-7 mRNA targets, with LIN28 levels correlating with cell proliferation and MYCN levels. Conversely, ectopic expression of let-7e was sufficient to reduce proliferation and downregulate MYCN, AURKB, and LIN28, the latter via a double-negative feedback loop. We conclude that the LIN28/let-7 pathway has a critical pathobiologic role in malignant GCTs and therefore offers a promising target for therapeutic intervention.

Differential Expression of PRAMEL1, a Cancer/Testis Antigen, during Spermatogenesis in the Mouse

PRAME belongs to a group of cancer/testis antigens (CTAs) that are characterized by their restricted expression in normal gametogenic tissues and a variety of tumors. The PRAME family is one of the most amplified gene families in the mouse and other mammalian genomes. Members of the PRAME gene family encode leucine-rich repeat (LRR) proteins functioning as transcription regulators in cancer cells. However, the role of PRAME in normal gonads is unknown. The objective of this study is to characterize the temporal and spatial expression of the mouse Pramel1 gene, and to determine the cellular localization of the PRAMEL1 protein during the mouse spermatogenesis. Our results indicated that the mouse Pramel1 was expressed in testis only. The mRNA and protein expression level was low in the newborn testes, and gradually increased from 1- to 3-week-old testes, and then remained constant after three weeks of age. Immunofluorescent staining on testis sections with the mouse PRAMEL1 antibody revealed that PRAMEL1 was localized in the cytoplasm of spermatocytes and the acrosomal region of round, elongating and elongated spermatids. Further analyses on the testis squash preparation and spermatozoa at a subcellular level indicated that the protein localization patterns of PRAMEL1 were coordinated with morphological alterations during acrosome formation in spermatids, and were significantly different in connecting piece, middle piece and principal piece of the flagellum between testicular and epididymal spermatozoa. Collectively, our results suggest that PRAMEL1 may play a role in acrosome biogenesis and sperm motility.

Mayer–Rokitansky–Kuster–Hauser Type-B Anomaly with MURCS Association and Gonadal Dysgenesis

The Mayer–Rokitansky–Kuster–Hauser (MRKH) syndrome affects 1 out of 4,500 women [1]. It is a malformation of the female genitals due to interrupted embryonic development of the mullerian (paramesonephric) ducts in otherwise chromosomally, phenotypically, and endocrinologically normal female. It is second to Turner’s syndrome as a cause of primary amenorrhea and was described by Mayer (1829), Rokitansky (1838), Kuster (1910), and Hauser and Schreiner (1961) in various literature studies, which was later designated as MRKH syndrome. MRKH syndrome is broadly subdivided into type A (typical) having symmetric uterine remnants and normal fallopian tubes and type B (atypical) with asymmetric uterine buds and abnormally developed fallopian tubes and other organ system anomalies. Gonadal agenesis or dysgenesis is a chromosomal aberration with a separate spectrum of anomalies having an overall incidence of about 1:2,500 live birth females, half of which have a mosaic pattern. These patients have multiple somatic abnormalities like short stature, broad chest, webbed neck, low hair line, sometimes low I.Q., cardiac abnormalities (1/3 cases), and renal abnormalities (35–70 %). They usually have pre-pubertal female genitalia, bilateral streak gonads, but usually normal uterus and vagina (Figs ​(Figs1,1, ​,22). Fig. 1 Photograph showing webbing of neck Fig. 2 Photograph showing the absence of left thumb and rudimentary right thumb Case Report An 18-year-old girl with primary amenorrhea attended the O.P.D. with normal I.Q. (1st year college student) and without such family history. On examination, her height is 154 cm, weight—42 kg, with webbing of neck, telecanthus; and her left thumb was absent and right thumb rudimentary. Systemic examination was normal. Breasts were well developed and axillary and pubic hairs were present. On perineal examination, there was a hymenal fringe and blind vagina. Uterus was absent on per rectal examination. Routine examination of blood, ECG, and chest X-ray reported normal. X-ray cervical spine showed fusion of the cervical vertebrae (Klippel Feil Syndrome). Serum FSH, LH, TSH, and testosterone were normal. USG [02/10/07] showed right-sided crossed fused ectopic kidney with normal corticomedullary differentiation, uterus absent, and normal gonads. Diagnostic laparoscopic [04/10/07] finding was asymmetric mullerian nodule [rt > lt] with rudimentary fallopian tubes, and both ovaries were normal with signs of ovulation. Karyotyping was Turner’s mosaics pattern (46XX/45XO). Diagnosis of MRKH type-B with gonadal dysgenesis was made. Vaginoplasty was done on 12/10/07 after proper counseling as parents wanted to arrange marriage. Post-operative period was uneventful.

Localised spawning omission in snapper, Chrysophrys auratus (Sparidae)

Failure to spawn in a significant proportion of adult fish may greatly decrease the reproductive output of a population and lead to overestimates of recruitment to the fish stock. Reproductive output of the commercially important sparid, Chrysophrys auratus, around the Kaipara Harbour, New Zealand, is particularly important as this harbour is the primary source for the C. auratus population along the west coast of the North Island of New Zealand. We tested the hypothesis that C. auratus do not spawn inside the Kaipara Harbour by comparing monthly gonad and otolith samples from fish caught within the harbour with those of fish from surrounding coastal waters. Fish from coastal waters showed normal gonad development with peak spawning in spring. By comparison, almost all adult fish from the harbour had underdeveloped gonads, with mean gonadosomatic indices ≤1. Histological gonad analyses of C. auratus caught in the harbour showed vitellogenic oocytes were reabsorbed before spawning. Therefore, recruitment to the harbour must originate from spawning that occurs outside the harbour. This first record of spawning omission in C. auratus has important implications for fisheries management, as it may lead to overestimation of the spawning stock and increase the risk of broad-scale population depletion, through the targeted fishing of spawning aggregations.

RanBPM is essential for mouse spermatogenesis and oogenesis

RanBPM is a recently identified scaffold protein that links and modulates interactions between cell surface receptors and their intracellular signaling pathways. RanBPM has been shown to interact with a variety of functionally unrelated proteins; however, its function remains unclear. Here, we show that RanBPM is essential for normal gonad development as both male and female RanBPM(-/-) mice are sterile. In the mutant testis there was a marked decrease in spermatogonia proliferation during postnatal development. Strikingly, the first wave of spermatogenesis was totally compromised, as seminiferous tubules of homozygous mutant animals were devoid of post-meiotic germ cells. We determined that spermatogenesis was arrested around the late pachytene-diplotene stages of prophase I; surprisingly, without any obvious defect in chromosome synapsis. Interestingly, RanBPM deletion led to a remarkably quick disappearance of all germ cell types at around one month of age, suggesting that spermatogonia stem cells are also affected by the mutation. Moreover, in chimeric mice generated with RanBPM(-/-) embryonic stem cells all mutant germ cells disappeared by 3 weeks of age suggesting that RanBPM is acting in a cell-autonomous way in germ cells. RanBPM homozygous mutant females displayed a premature ovarian failure due to a depletion of the germ cell pool at the end of prophase I, as in males. Taken together, our results highlight a crucial role for RanBPM in mammalian gametogenesis in both genders.

Effects of exposure to the xenoestrogen octylphenol and subsequent transfer to clean water on liver and gonad ultrastructure during early development of Zoarces viviparus embryos

Female eelpouts (Zoarces viviparus L.) are exposed during early pregnancy to nominal concentrations of 100 microg/L of 4-tert-octylphenol (OP) or 0.5 microg/L of 17beta-estradiol (E2). Effects on maternal metabolism and on liver and gonad development in embryos were examined and compared with controls (C) during exposure and after transfer to clean water (depuration). In the mother fish, significantly higher concentrations of plasma vitellogenin (vtg) and calcium were found in the two exposed groups, when compared with the C group after exposure and depuration. When compared, however, with the respective values after exposure, vtg had decreased significantly after depuration. The hepatosomatic index was normalized after depuration. In both exposed groups, the hepatocytes were rounded and not distinctly polygonal as in the controls. The amount of glycogen was considerably less while the number of mitochondria increased, and the rER significantly proliferated after exposure as well as after depuration. The gonads of nine of more than 28 embryos in the group treated with OP exhibited a number of abnormalities as compared with the normal gonad development in both sexes. Feminization of the male gonads in the exposed specimens and a number of histopathological features were observed in all the abnormal gonads, whereas reliable male features, such as formation of seminiferous tubules or spermioduct, were not observed. This study showed that 4t-tert-OP and 17beta-estradiol exert estrogenic effects during very early development of the embryos and that depuration had a positive effect on the motherfish and her embryos.

Gonad Maturation Cycle of the Sea Urchin Strongylocentrotus nudus Population Inhabiting an Artificial Seaweed Forest, Samchuk, Korea

We determined the seasonal gonad maturation in Strongylocentrotus nudus sea urchins inhabiting an artificially enhanced seaweed forest along the Samchuk Coast of Korea from April 2006 to March 2007. A total of 30 sea urchins per month were collected from the study area, and gonadosomatic index (GSI), gonad index (GI), egg diameter, and RNA/DNA variation were measured for each specimen. GSI values of female and male urchins achieved maximums of 17.6 and 17.0, respectively, in June. Based on histological studies, maximum GI values occurred in July (4.6 for females and 4.8 for males). A mean ovarian egg diameter of <TEX>$73.7\;\pm\;14.2\;{\mu}m$</TEX> was measured in August; during the main spawning period in September, mean egg diameter reached a maximum of <TEX>$74.2\;\pm\;17.8\;{\mu}m$</TEX>. The RNA/DNA ratio and RNA content for both males and females showed a distinct peak during the ripe stage in July, but another peak occurred in the spring season from March to April, when urchins deposit protein into the nutritive phagocytes of immature gonads prior to gametogenesis. The reproductive cycle of S. nudus is divided into five stages: early active (December-May), late active (March-July), ripe (July-September), spent and degenerative (August-November), and inactive (October-February). Our continuous removal of sea urchins from the study area did not influence the reproductive cycle, as populations quickly recovered, and achieved normal gonad development cycle in the site.

An expression screen for RhoGEF genes involved in C. elegans gonadogenesis

The gonad in Caenorhabditis elegans is an important model system for understanding complex morphogenetic processes including cellular movement, cell fusion, cell invasion and cell polarity during development. One class of signaling proteins known to be critical for the cellular events underlying morphogenesis is the Rho family GTPases, particularly RhoA, Rac and Cdc42. In C. elegans orthologues of these genes have been shown to be important for gonad development. In our current study we have extended those findings by examining the patterns of 5′ cis-regulatory element (5′CRE) activity associated with nineteen putative guanine nucleotide exchange factors (GEFs) encoded by the C. elegans genome predicted to activate Rho family GTPases. Here we identify 13 RhoGEF genes that are expressed during gonadogenesis and characterize the cells in which their 5′CREs are active. These data provide the basis for designing experiments to examine Rho GTPase activation during morphogenetic processes central to normal gonad development.

Global Gene Expression in the Human Fetal Testis and Ovary1

This study describes a temporal profile of gene expression from normal human fetal testes and ovaries. Gonads from 34 fetuses between 9 wk and 20 wk of gestation were obtained from the Department of Pathology and the Birth Defects Research Laboratory at the University of Washington. Relative transcript levels were determined using the Affymetrix Human Genome U133A Plus 2.0 arrays. Sex determination occurs in the human gonad at approximately 6 wk of gestation with development of the testis driven by expression of SRY. In this study, SRY transcript was present and elevated at 9 wk of gestation in the testis but was absent in the ovary. The transcript levels of other testis-specific factors SOX9 and AMH and the steroidogenic genes CYP17A1, CYP11A1, STAR, and HSD17B3 were all significantly higher in the testis. In contrast, transcripts known to be involved in meiosis, including STRA8, SPO11, SYCP3, TEX11, TEX14, and STAG3, showed highest expression in the fetal ovary beginning at Week 12. These gene expression profiles will be a resource for understanding and defining normal gonad development and provide the opportunity to dissect abnormal development.

Unexplained gonad alterations in whitefish ( Coregonus spp.) from Lake Thun, Switzerland: Levels of persistent organic pollutants in different morphs

Since 2000, a surprisingly high number of macroscopical gonad alterations has been reported in whitefish ( Coregonus spp.) from Lake Thun, Switzerland. This unique phenomenon is still unexplained and has received much public attention. As one possible trigger for these effects, the presence of persistent, bioaccumulative and toxic compounds acting as endocrine disruptors in the lake has been discussed. In this study, concentrations of selected persistent organic pollutants were examined in two morphs of whitefish from Lake Thun and their link to the observed abnormalities was investigated. Analyzed compound classes included polychlorinated biphenyls, polychlorinated dibenzo- p-dioxins and dibenzofurans, polychlorinated naphthalenes, polybrominated diphenyl ethers and hexabromocyclododecanes. The target substances were identified in all samples and concentrations of the analyzed compounds were highly correlated among each other. These correlations show that the analyzed substances have the same distribution pattern throughout the lake and that uptake, accumulation and elimination processes are similar. Significant differences in contaminant levels within the samples existed between the two analyzed morphs of whitefish, most likely due to different age, food patterns and growth rate. No difference in contaminant levels was observed between fish with abnormal gonads and fish with normal gonads, suggesting no causal link between the investigated lipophilic organohalogen compounds present in fish and the observed gonad abnormalities in whitefish from Lake Thun. A comparison to existing data shows that concentrations in Lake Thun whitefish are at the lower bound of contaminant levels in whitefish from Swiss lakes or from European waters.

Formation and biological characterization of three new types of improved crucian carp

The improved tetraploids (G(1)xAT) were obtained by distant crossing and gynogenesis and the high-body individuals accounted for 2% among G(1)xAT. After mating with each other, the high-body individuals produced three kinds of bisexual fertile diploid fishes: high-body red crucian carp, high-body fork-like-tails goldfish and gray common carp. The high-body red crucian carp mating with each other formed three types of improved crucian carp (ICC) including improved red crucian carp (IRCC), improved color crucian carp (ICCC) and improved gray crucian carp (IGCC). The phenotypes, chromosome numbers, gonadal structure and fertility of the three kinds of ICC and their offspring were observed. All the three kinds of ICC possessed some improved phenotypes such as higher body, smaller head and shorter tail. The ratios of the body height to body length of IRCC, ICCC and IGCC were 0.54, 0.51 and 0.54, respectively. All of them were obviously higher than that of red crucian carp 0.41 (P<0.01). Three kinds of ICC had the same chromosome number as red crucian carp with 100 chromosomes. All the ICC possessed normal gonads producing mature eggs or sperm, which was important for the production of an improved diploid population. Compared with red crucian carp, all the ICC had stronger fertility such as higher gametes production, higher fertilization rate and higher hatchery rate. Three types of improved diploid fish population were generated from the three kinds of ICC by self-crossing, respectively. The ICC can serve as ornamental fish and edible fish. They are also ideal parents to produce triploids by mating with tetraploids. The new ICC plays an important role in biological evolution and fish genetic breeding.

FOXL2 and SOX9 as parameters of female and male gonadal differentiation in patients with various forms of disorders of sex development (DSD)

The transcription factors SOX9 and FOXL2 are required for male and female mammalian gonadal development. We have used specific antibodies to investigate the role of these key proteins in disorders of sex development (DSD), specifically inter-sex states. In normal gonads, SOX9 was found to be restricted to the presence of (pre-)Sertoli cells, while FOXL2 was found in granulosa cells, and in stromal cells interpreted as early ovarian stroma. Both proteins were found within a single patient, when testicular and ovarian development was present; and within the same gonad, when both differentiation lineages were identified, as in ovotesticular DSD (ie hermaphrodite). Especially SOX9 was informative to support the presence of early testicular development (ie seminiferous tubules), expected based on morphological criteria only. In a limited number of DSD cases, FOXL2 was found within reasonably well-developed seminiferous tubules, but double staining demonstrated that it was never strongly co-expressed with SOX9 in the same cell. All seminiferous tubules containing carcinoma in situ (CIS), the malignant counterpart of a primordial germ cell, ie the precursor of type II germ cell tumours of the testis, seminomas and non-seminomas, showed the presence of SOX9 and not FOXL2. In contrast, gonadoblastomas (GBs), the precursor of the same type of cancer, in a dysgenetic gonad, showed expression of FOXL2 and no, or only very low, SOX9 expression. These findings indicate that gonadal differentiation, ie testicular or ovarian, determines the morphology of the precursor of type II germ cell tumours, CIS or GB, respectively. We show that in DSD patients, the formation of either ovarian or/and testicular development can be visualized using FOXL2 and SOX9 expression, respectively. In addition, it initiates a novel way to study the role of the supportive cells in the development of either CIS or GB.

Ambiguous Genitalia in the Newborn

Efficient and accurate evaluation of the newborn who has ambiguous genitalia is required to provide appropriate medical therapy and assuage parental anxiety. Genital ambiguity usually is due to virilization of genetic females or undervirilization of genetic males who have normal gonads. Congenital adrenal hyperplasia is the most common condition leading to inappropriate virilization in females. Defects in testosterone production, metabolism, or peripheral action can lead to ambiguous genitalia in males. In any condition involving ambiguous genitalia or question of sex assignment, a karyotype should be obtained within 24 hours of delivery. Parents should be apprised of the situation in a professional manner with the appropriate level of detail; sex assignment should be withheld until sufficient data are gathered to make an accurate diagnosis. Families can be counseled with the latest available information and resources to make the best decisions for their individual situations.