Gonadal development and diandric protogyny in two populations of Dascyllus reticulatus from Madang, Papua New Guinea

Two Dascyllus reticulatus populations from Madang, Papua New Guinea exhibited diandric protogyny. In both populations, gonads began as undifferentiated, and then developed oocytes in the primary growth stage and an ovarian lumen. From this ovarian state or from more developed ovaries containing oocytes beyond the primary‐growth stage, some gonads developed into testes. The first sign of testicular development was degeneration of oocytes, degeneration of oocytes in the primary growth stage in ovarian gonads and degeneration of oocytes of all growth stages present including the primary growth stage in ovaries, which was then followed by development of spermatogenic tissue. In both populations, most of the fish that had gonads with degenerating oocytes were smaller than the smallest mature females, indicating that development towards testes was mostly initiated in immature gonads containing only pre‐vitellogenic oocytes. On some occasions, however, females as large as other mature females also had gonads with degenerating oocytes, suggesting that development towards testes may have occurred in mature ovaries as well. This latter notion is further strengthened by the discovery of a fish having a gonad that contained both degenerating vitellogenic oocytes and developing spermatogenic tissue. Taken together, these results suggest that D. reticulatus can exhibit diandric protogyny, because testes in D. reticulatus developed from juvenile gonads as well as from mature ovaries.     

Annual changes in the ovarian activity of the freshwater teleost, Barbus luteus (Heckel) from Southern Iraq

The present paper describes the morphology, histology, and annual cyclical changes in the ovaries of Barbus luteus (Heckel). The ovaries are paired structures, joined by short oviducts. The ovarian structure consists of a number of lamellae, which contain the germ cells in various phases of maturation—immature, maturing and mature. Two types of yolk have been reported in the oocyte. The endocrine function of the gonads has been attributed to the follicular cells forming the 'pre-ovulatory corpus luteum' and 'postovulatory corpus luteum'. The ovaries undergo a regular annual cycle which has been divided into five stages, based on various criteria, viz, ovarian size, colour, gonosomatic index, average ova-diameter, histological composition, and the spawning activity.     

Dysgenesic rat ovaries subjected to gonadotrope stimulation]

Dysgenesia of the ovaries was produced by the destruction of the germinal cells during the foetal life by injecting the pregnant rat with Misulban. In the absence of ovocytes, follicular organization did not take place and cordal structure were observed in the ovary. The post-puberal evolution of these gonads led to polymorphous structures according to age and individual cases. Study of dysgenesic ovaries of rats treated by stilboestrol before puberty, showed mainly that the ovarian stroma was affected in response to the inhibition of the pituitary gonadotropic activity. It is concluded that the development of these ovaries is under the control of the hypothalamo=pituitary axis.     

Meiosis-inducing and meiosis-preventing effects of sex steroid hormones on hamster fetal ovaries in organ culture

Day 11 to day 15 p.c. female gonads were cultured for 6-8 days in chemically-defined media. In day 11 and day 12 p.c. ovaries grown in a non-hormonal medium, the germ cells were unable to enter meiosis; they were retained at a stage of oogonia or more frequently at a preleptotene stage. Ovaries of the same ages cultured in an estradiol-containing medium showed germ cells progressing through meiotic prophase in a way close to that in ovaries of equivalent age in vivo. That was the case of the germ cells in day 13 to day 15 p.c. ovaries maintained in a non-hormonal medium. In a testosterone-containing medium, the germ cells in day 13 and day 14 p.c. ovaries were prevented from entering meiosis; by contrast, those in day 15 p.c. ovaries underwent meiotic prophase normally. These results indicated that each of both hormones was able to exert its corresponding (meiosis-inducing or meiosis-preventing) effect before a definite critical time of ovarian development. The possibility is suggested that the germ cell differentiation in the female and male gonads in vivo would also depend on estrogens or androgens precociously synthesized in the gonads or supplied from other organs via the fetal blood.     

Differentiation of ambisexual gonads and immunohistochemical localization of P450 cholesterol side-chain cleavage enzyme during gonadal sex differentiation in the protandrous anemonefish, Amphiprion clarkii

To clarify the relationship between steroid hormones and sex differentiation of the protandrous anemonefish Amphiprion clarkii, we histologically examined its gonadal differentiation. From hatching to 30 days post hatching (dph), all of the gonads surveyed were sexually undifferentiated. The gonads of all fish first differentiated into ovaries at 60 dph, and the oocytes gradually developed and increased in number as the ovaries grew up until 183 dph. Some cysts of differentiated spermatogenic germ cells appeared in the ovaries at 214 dph, and ambisexual gonads with both ovarian and testicular tissues formed by 273 dph. Using immunohistochemistry, we then investigated the expression of cytochrome P450 cholesterol side-chain cleavage enzyme (P450scc), during gonadal sex differentiation. P450scc-immunopositive reactions first appeared in sexually undifferentiated gonads at 30 dph. Beginning at 60 dph, the number of strongly positive cells increased throughout the differentiation of the ovaries and continued to increase during the testicular differentiation until 210 dph. Immunopositive cells were observed more frequently in ovarian tissue than in testicular tissue in the ambisexual gonads at 270 dph. These results suggest that endogenous steroid hormones are important for the sex differentiation, including the primary sex differentiation and subsequent testicular differentiation, of the anemonefish.     

Embryological study on gynogenesis in onion (Allium cepa L.)

Haploid induction in onion can, to date, be induced only via gynogenesis by culturing unfertilized flowers, ovaries or ovules. The process of haploid embryo induction has been macroscopically well studied, but only limited data exist from microscopic examination of ovule development status at the inoculation stage and of the origin of gynogenic embryos. Microscopic studies were carried out using individual donor plants with relatively high embryo induction frequencies (45.9 embryos formed per 100 flowers, on average, for 2 years). Ovaries from flower bud culture were fixed at 1 week intervals up to the 7th week of culture. These were compared with pollinated ovaries at 1 or 2 weeks after pollination. In total, 1428 unfertilized embryo sacs were examined. The results indicate that, at the time of inoculation, ovules within ovaries 2.0–3.0 mm in diameter contained two- or four-nucleate embryo sacs in the smallest ovaries to mature embryo sacs in the largest ovaries. It seems likely that the embryos are actually induced from ovaries cultured at the immature stage. After 1 or 2 weeks in culture, the egg apparatus primarily consisted of distinctly enlarged synergids and the egg cell, which was often detached from the micropylar pole. But free nuclear endosperm was also formed. From the 2nd to 7th week in culture, formation of haploid embryos (from globular to the almost mature cylindrical stage) was detected in 5.7% of the ovules. Their origin, for several reasons, was most likely the egg cell. In addition, ovules containing endosperm only (3.6%) and ovules containing the egg apparatus (0.5%) or both endosperm and embryo (0.4%) were detected. This observation is probably unique and has not yet been reported in other species studied. Received: February 2001 / Revision accepted: 20 April 2001     

莴苣胚囊细胞分离

用酶解和解剖方法分离了莴苣的卵细胞,助细胞,中央细胞和合子。莴苣子房先在酶液中酶解40～50min,然后在不含酶的分离液中用解剖针解剖子房。在解剖出的胚囊中,可看到卵细胞,两个助细胞和中央细胞的轮廓。将胚囊的合点端切破,轻轻挤压胚囊的珠孔端,四个细胞即可逸出。在最佳条件下,90min可从40个子房中分离出29个胚囊,进一步从中分离出11个卵细胞。分离出的胚囊细胞用显微操作仪收集备用。莴苣卵细胞的成功分离为进行离体受精探索创造了条件。     

Characterization of Sus scrofa Small Non-Coding RNAs Present in Both Female and Male Gonads

Small non-coding RNAs (sncRNAs) are indispensable for proper germ cell development, emphasizing the need for greater elucidation of the mechanisms of germline development and regulation of this process by sncRNAs. We used deep sequencing to characterize three families of small non-coding RNAs (piRNAs, miRNAs, and tRFs) present in Sus scrofa gonads and focused on the small RNA fraction present in both male and female gonads. Although similar numbers of reads were obtained from both types of gonads, the number of unique RNA sequences in the ovaries was several times lower. Of the sequences detected in the testes, 2.6% of piRNAs, 9% of miRNAs, and 10% of tRFs were also present in the ovaries. Notably, the majority of the shared piRNAs mapped to ribosomal RNAs and were derived from clustered loci. In addition, the most abundant miRNAs present in the ovaries and testes are conserved and are involved in many biological processes such as the regulation of homeobox genes, the control of cell proliferation, and carcinogenesis. Unexpectedly, we detected a novel sncRNA type, the tRFs, which are 30–36-nt RNA fragments derived from tRNA molecules, in gonads. Analysis of S. scrofa piRNAs show that testes specific piRNAs are biased for 5′ uracil but both testes and ovaries specific piRNAs are not biased for adenine at the 10^th nucleotide position. These observations indicate that adult porcine piRNAs are predominantly produced by a primary processing pathway or other mechanisms and secondary piRNAs generated by ping-pong mechanism are absent.     

Development of 7--11-week-old human embryonic gonads in organ culture]

The author suggests a simplified method of organic culture of the human gonads. Gonads (160) from 7--11-week embryos were cultivated on millipore filters in penicillin bottles. During the first week of cultivation the central necroses were observed both in the male and the female gonads, but later they disappeared in some of the explants. In the ovaries from the 9--11-week embryos the germ cells underwent development which continued to the pachytene stage, while in the ovaries from the 7--9-week embryos the development of the germ cells failed to reach the leptotene stage. In the case of testis germ cells they remained at the stage of spermatogonia.     

The problem of sexual inversion in the minnow, Phoxinus phoxinus (L.)

The incidence of sexual inversion in Phoxinus phoxinus has been studied by examining histologically (a) the gonads of 406 adult specimens caught from the wild during a period of 16 months, (b) the gonads of 168 adults kept in captivity in groups with an experimentally altered sex-ratio, and (c) the differentiation of the gonads in the fry.
All the adults had either testes or ovaries and no case of intersexuality was observed. For both testis and ovary, we recognized two phases, depending on the seasonal cycle: an active spring-summer phase, corresponding with the period of reproduction, and a quiescent autumn-winter phase. The ovaries showed an asynchrony in oocyte development, which is typical of species that spawn many times during the breeding season.
In P. phoxinus the ovaries are easily recognizable just one month after hatching and testes are well differentiated in 2-month-old fingerlings. Differentiation is, therefore, precocious and follows a pattern typical of gonochoristic species in which hermaphroditism never occurs.     

Lymphoid aggregates in gonads of embryos, hatchlings, and young of turtles with temperature-dependent sex determination

Cellular infiltrations forming lymphoid-like aggregates were previously observed in gonads of two turtle species exhibiting temperature-dependent sex determination (TSD): at hatching in Chelydra serpentina; at and after hatching in Emys orbicularis. We show here that such aggregates are also present in gonads of Testudo graeca by the end of embryonic development, suggesting that their occurrence is general in turtles. Since in C. serpentina, infiltrations were observed mainly in testes exhibiting remnants of the germinal epithelium, it was assumed that their occurrence was an expression of maleness leading to rejection of this epithelium. The generality of this hypothesis was tested in E. orbicularis by looking for lymphoid-like aggregates in three types of gonads (testes, ovotestes, and ovaries) and for the stages at which they occur. Gonads were from embryos, hatchlings, and young incubated at various temperatures. Ovotestes obtained by treatment with an aromatase inhibitor of eggs incubated at female-producing temperature were also examined. In these gonads, the differentiation of Sertoli cells in testicular cords/tubes was ascertained by expression of SOX9. Moreover, the cell composition of aggregates was determined on electron micrographs. Aggregates appear in ovaries and ovotestes by the end of embryonic development and are present in the majority of these gonads at hatching, and at least up to one year after hatching. They are composed mainly of lymphocytes and fibroblasts. Aggregates are not present in typical testes. Since they occur in most ovaries, they cannot be seen as an expression of maleness. Rather, lymphocytic infiltration and formation of lymphoid aggregates in turtle gonads can be seen as components of the immune system, and can be under the control of gonadal endogenous sex steroids.     

The gonads of Aegla platensis Schmitt (Decapoda, Anomura, Aeglidae): a macroscopic and histological perspective

The aim of this study is to characterize the gonads of Aegla platensis, relating aspects of colour and size of ovaries, testes and vasa deferentia (VD) to histological observations. The ovaries are H‐shaped, extending from behind the stomach and ending at the 1st or 2nd abdominal somite. The male has a pair of testes from which the VD issue and extend over the pereon. For males and females, macroscopic categories of gonad development are defined. Stages of the male gonad are defined as types 1, 2 and 3. Type 1 includes the less developed gonads, and type 3 includes extremely developed testes and completely differentiated VD. The ovaries are classified into four stages according to their colour: I (white), II (yellow), III (orange) and IV (red), beginning with the least developed gonads. The number and size of oogonia and oocytes depend on the stage of the ovary and coincide with the different degrees of development as indicated by ovary colour. In males, the presence or absence of spermatozoa in the testes and VD is determined only for the more developed gonads. Neither spermathecae (female) nor spermatophores (male) were found. The lack of spermatophore in the present species is a rare characteristic among the anomurans.     

Morphological and histological features of the gonad structure in hermaphroditic walleye pollock <Emphasis Type="Italic">Theragra chalcogramma</Emphasis> (Gadidae)

Morphology of hermaphroditic gonads is studied in the individuals of walleye pollock Theragra chalcogramma collected in different regions of the seas of the North Pacific from 1987 to 2014. Based on the external morphological characteristics of sexual glands of 17 fishes, the most frequent types of the structure of bisexual gonads are described. Histological analysis shows the absence of differences in the composition and condition of oocytes in the left and right-paired ovaries, and the testes and ovaries of each individual are at the same maturity stage. In the ovigerous lamellae of ovaries in the areas adjacent to testes the zones of oocyte resorption are observed. These zones are located exclusively near the joining of the female and male parts of the gonads. The development of ovaries and testes of bisexual gonads is similar to that in the majority of normally maturing males and females collected during the same period. The occurrence of hermaphrodites is 0.0003–0.0016% from the total number of investigated exemplars.     

Detection of VHSV IVb within the gonads of Great Lakes fish using in situ hybridization

Viral haemorrhagic septicaemia virus (VHSV) genotype IVb was recently detected as the cause of numerous mortality events in Great Lakes fish. In situ hybridization was used to examine the gonads from 13 fish, including freshwater drum Aplodinotus grunniens and muskellunge Esox masquinongy that were infected naturally, as well as rainbow trout Oncorhynchus mykiss and fathead minnows Pimphales promelas, which were experimentally infected. Although the ovaries and testes of fish infected by VHSV IVb had few lesions, viral RNA was present in the ovaries of the rainbow trout and fathead minnow and was abundant in the gonads of muskellunge and in the ovaries of freshwater drum. Viral RNA was present mainly surrounding yolk vacuoles/granules or adjacent to the germinal vesicle, with lesser amounts found within the germinal vesicle, in the mesovarium and/or tunica albuginea and blood vessels of the ovary. Viral RNA was also found in and surrounding primary and secondary spermatocytes of the muskellunge.     

Immunolocalization of cytochrome P450 17alpha-hydroxylase/c17-20 lyase in the ovary of pregnant pigs and fetal gonads

The study was designed to localize P450 17alpha-hydroxylase/c17-20 lyase (P450c17) in the ovaries of pregnant pigs and fetal gonads. Immunoexpression of P450c17 was investigated in the porcine ovaries (follicles and corpora lutea; CL) collected on days 10, 18, 32, 50, 70 and 90 post coitum (p.c.), and fetal gonads (testes and ovaries) on days 50, 70 and 90 p.c. The presence of P450c17 in ovarian follicles was demonstrated on all examined days of pregnancy but was restricted to theca interna cells. In CL, P450c17 was detected on all examined days of pregnancy but only in small luteal cells. In the female porcine fetuses, P450c17 immunostaining was found in oocyte nests and granulosa cells of primary ovarian follicles, while in the male fetuses in fetal Leydig cells. In conclusion, the immunolocalization of P450c17, detected in the ovaries of pregnant pigs and fetal porcine gonads, indicates the potential sites of androgen synthesis. We suggest that androgens may play a role in the maintenance of pregnancy and in the development of prenatal gonads in pigs.     

Gender differences in transcriptional signature of developing rat testes and ovaries following embryonic exposure to 2,3,7,8-TCDD

Dioxins are persistent organic pollutants interfering with endocrine systems and causing reproductive and developmental disorders. The objective of our project was to determine the impact of an in utero exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on reproductive function of male and female offspring in the rat with a special emphasis on the immature period. We used a low dose of TCDD (unique exposure by oral gavage of 200 ng/kg at 15.5 days of gestation) in order to mirror a response to an environmental dose of TCDD not altering fertility of the progeny. We choose a global gene expression approach using Affymetrix microarray analysis, and testes of 5 days and ovaries of 14 days of age. Less than 1% of the expressed genes in gonads were altered following embryonic TCDD exposure; specifically, 113 genes in ovaries and 56 in testes with 7 genes common to both sex gonads. It included the repressor of the aryl hydrocarbon receptor (Ahrr), the chemokines Ccl5 and Cxcl4 previously shown to be regulated by dioxin in testis, Pgds2/Hpgds and 3 others uncharacterized. To validate and extend the microarray data we realized real-time PCR on gonads at various developmental periods of interest (from 3 to 25 days for ovaries, from 5 to the adult age for testes). Overall, our results evidenced that both sex gonads responded differently to TCDD exposure. For example, we observed induction of the canonic battery of TCDD-induced genes coding enzymes of the detoxifying machinery in ovaries aged of 3-14 days of age (except Cyp1a1 induced at 3-10 days) but not in testes of 5 days (except Ahrr). We also illustrated that inflammatory pathway is one pathway activated by TCDD in gonads. Finally, we identified several new genes targeted by TCDD including Fgf13 in testis and one gene, Ptgds2/Hpgds regulated in the two sex gonads.     

Juvenile bisexuality in the red sea bream,Pagrus major

The histology of the gonad of the red sea bream,Pagrus major, was examined in order to study the early gonadal development, sexual maturation and sex ratio in a natural population. A total of 1,117 fish between the ages of 4 months and 8 years were examined. Gonads of 4-month-old fish were either sexually undifferentiated with a central cavity, or ovarian in form. Gonads of 12- and 18-month-old fish were ovaries or bisexual gonads, while those of 2-year-old fish were ovaries, bisexual gonads or testes. Fish aged between 3 and 8 years had ovaries or testes, except for a few bisexual gonads found in 3- and 4-year-old fish. The chronological appearance of females, hermaphrodites and males in that order, and histological evidence, suggested that the testis originates from the ovary via a bisexual gonad in the juvenile stage. The sex ratio of females to males at the age of 2 years and over was about 1:1, suggesting that hermaphroditic red sea bream appear in about 50% of the juvenile population. The sexual pattern in this species, therefore, is concluded to be gonochorism with a bisexual juvenile stage.