OVARY DEVELOPMENT, FSH AND LH GENES EXPRESSION OF INDONESIAN LEAFFISH, Pristolepis grootii (Bleeker, 1852), INJECTED WITH LUTEINIZING HORMONE-RELEASING HORMONE ANALOG

Indonesian leaffish, Pristolepis grootii (Bleeker, 1852), is an undomesticated freshwater fish species native to the rivers, flooded swamps, and tributaries of Indonesia. The fish is mainly captured for consumption. In order to prevent its extinction and supply its growing demands, the artificial breeding of the fish should be developed. The purpose of this study was to determine the optimum dose of luteinizing hormone-releasing hormone analog (LHRHa) for stimulating the female P. grootii gonadal development at a dosage of 0, 1, 10, and 50 µg kg-1 of fish. Female fish (20.0 ± 0.6 g) were adapted for 30 days in the rearing environment and then separated into 12 aquariums with six fish per aquarium. Fish were then reared for another 21 days and fed with Tubifex sp. The LHRHa injection was conducted twice on day-7 and 14. Fish bodyweight, gonadosomatic index, gonad histology, blood estradiol-17â, and FSH-â and LH-â gene expression were evaluated at day 0, 7, 14, and 21. The results showed that the injection of the LHRHa hormone stimulated the development of fish gonads and was better achieved with a higher concentration of LHRHa. The best treatment was observed by the administration of 50 µg kg-1 of LHRHa that produced the fastest development among all treatments. This study demonstrated that the LHRHa induction could potentially stimulate the gonadal development of the newly domesticated fish. To our knowledge, this is the first study that reported the success of the induction of female gonad development in the Indonesian leaffish P. grooti.KEYWORDS:

Redescription of Ektaphelenchoides compsi Baujard, 1984 (Tylenchina: Aphelenchoididae) isolated from Pinus massoniana in Fujian Province, China

Summary Ektaphelenchoides compsi is redescribed morphologically with new molecular characterisation. It was isolated from a dead Pinus massoniana tree in Ningde City, Fujian Province, China. Detailed morphology of the spicule, female gonad, hemizonid position, arrangement of male caudal papillae and female tail terminus shape are documented. It is characterised by a lateral field with three lines (forming two bands), tripartite stylet 17.8 (17.0-19.4) μm long without basal thickenings, metacorpus rectangular with anterior 40% granular and posterior part weakly muscular, metacorpal valve slightly posterior to middle of metacorpus, excretory pore at level of nerve ring, vagina with thickened walls and strongly developed muscular bundles, vulval lips slightly protuberant, vulval flap absent, distal region of post-vulval uterine sac appearing as a weakly developed oogonia, anus and rectum indistinct, female posterior part (‘tail’) dorsally convex, conical, terminal region contracted into a bluntly pointed tip. The spicules are arcuate, 15.6 (14.3-16.3) μm along the chord, lamina smoothly curved to distal end, capitulum slightly concave, condylus well-developed with broadly rounded tip and slightly depressed at dorsal end, rostrum triangular with finely rounded tip, cucullus absent, and with seven caudal papillae present. The near full length 18S and 28S D2-D3 regions of rRNA genes sequences were characterised. The phylogenetic analyses revealed that the Fujian population of E. compsi grouped with the Zhejiang population of E. compsi, both being morphologically identical.

Effects of fish meal replacement with protein mixtures on growth, gonad development and amino acid composition of pre-adult red swamp crayfish, Procambarus clarkii (Girard, 1852) (Decapoda, Cambaridae)

A 100-day growth trial was conducted to evaluate the effects of dietary protein mixtures (PM) adding proportion on both sexes of the red swamp crayfish, Procambarus clarkii (Girard, 1852). Five diets were prepared by replacing 0%, 16.67%, 33.33%, 50%, and 66.67% of fish meal (FM) protein with protein mixtures in feed (designed Diet 1, Diet 2, Diet 3, Diet 4 and Diet 5). The results showed that growth performance presented a decreasing trend with an increase in dietary protein mixture levels. The GSI of female crayfish and the HSI of male crayfish initially increased and then decreased with increasing dietary protein mixtures. Interestingly, the GSI of males fed Diet 5 was higher than that in other groups (). Diet 2 was the best feed for female gonad development and male muscle quality. Females fed Diet 3 had the highest muscle quality among the groups; males had improved gonad development when fed Diet 5.

Evaluation of Probiotic-Fermented Feed Addition and LaserFiring to Accelerate Mature Broodstocks and Seed Productions of African Catfish (Clarias gariepinus)

This study aimed to determine the effects of fermented feed using probiotics and laser-firing to accelerate the mature broodstocks and seed productions of African catfish (Clarias gariepinus). Fish has used male and female broodstocks of catfish. The method was used a completely randomized design with three treatments: unfermented feed as control, probiotic-fermented feed (PFF), and probioticfermented feed+laser firing (PFF+Li). In the first study, a laser-firing dose of 1.125 Joule was performed on fish broodstocks every 15 days. The gonadal maturity time of male and female broodstocks was observed. The second study, treated female broodstocks, was mated with mature male broodstocks without any prior treatments (control). Fertilization rate, hatching rate, and seed production performances such as survival rate and total length were measured. The results showed that treatment of PFF+Li has a significant effect (P<0.05) on the gonadal maturity time of males and females. This treatment reaches the fastest time to mature of the female gonad (31-41 days) and the male gonad (32-37 days) than other treatments (P<0.05). This treatment also produced the highest fertilization, hatching, and survival rates of more than 90%, respectively, and the highest seed yield of 2.1-3.0 cm size compared to other treatments (P<0.05) in African catfish.

Self-organising human gonads generated by a Matrigel-based gradient system

Background Advances in three-dimensional culture technologies have led to progression in systems used to model the gonadal microenvironment in vitro. Despite demonstrating basic functionality, tissue organisation is often limited. We have previously detailed a three-dimensional culture model termed the three-layer gradient system to generate rat testicular organoids in vitro. Here we extend the model to human first-trimester embryonic gonadal tissue. Results Testicular cell suspensions reorganised into testis-like organoids with distinct seminiferous-like cords situated within an interstitial environment after 7 days. In contrast, tissue reorganisation failed to occur when mesonephros, which promotes testicular development in vivo, was included in the tissue digest. Organoids generated from dissociated female gonad cell suspensions formed loosely organised cords after 7 days. In addition to displaying testis-specific architecture, testis-like organoids demonstrated evidence of somatic cell differentiation. Within the 3-LGS, we observed the onset of AMH expression in the cytoplasm of SOX9-positive Sertoli cells within reorganised testicular cords. Leydig cell differentiation and onset of steroidogenic capacity was also revealed in the 3-LGS through the expression of key steroidogenic enzymes StAR and CYP17A1 within the interstitial compartment. While the 3-LGS generates a somatic cell environment capable of supporting germ cell survival in ovarian organoids germ cell loss was observed in testicular organoids. Conclusion The 3-LGS can be used to generate organised whole gonadal organoids within 7 days. The 3-LGS brings a new opportunity to explore gonadal organogenesis and contributes to the development of more complex in vitro models in the field of developmental and regenerative medicine.

The male and female gonad transcriptome of the edible sea urchin, Paracentrotus lividus: identification of sex-related and lipid biosynthesis genes

Paracentrotus lividus is the most abundant, distributed and desirable echinoid species in Europe. Although, economically important, this species has scarce genomic resources available. Here, we produced and comprehensively characterized the male and female gonad transcriptome of P. lividus. The P. lividus transcriptome assembly has 53,865 transcripts, an N50 transcript length of 1,842 bp and an estimated gene completeness of 97.4% and 95.6% in Eukaryota and Metazoa BUSCO databases, respectively. Differential gene expression analyses yielded a total of 3371 and 3351 up regulated genes in P. lividus male and female gonad tissues, respectively. Additionally, we analysed and validated a catalogue of pivotal transcripts involved in sexual development and determination (206 transcripts) as well as in biosynthesis and storage of lipids (119 transcripts) in male and female specimens. This study provides a valuable transcriptomic resource and will contribute for the future conservation of the species as well as the exploitation in aquaculture settings.HighlightsAssembly of a reference transcriptome of Paracentrotus lividus gonads.Differential gene expression between males and female gonads of Paracentrotus lividus.Identification and validation of pivotal genes involved in biosynthesis and storage of lipids.

Perivascular cells support folliculogenesis in the developing ovary

Granulosa cells, supporting cells of the ovary, are essential for ovarian differentiation by providing a nurturing environment for oogenesis. Sufficient numbers of granulosa cells are vital for establishment of follicles and the oocyte reserve; therefore, identifying the cellular source from which granulosa cells are derived is critical for understanding basic ovarian biology. One cell type that has received little attention in this field is the perivascular cell. Here we use lineage tracing and organ culture techniques in mice to identify ovarian Nestin+ perivascular cells as multipotent progenitors that contribute to granulosa, thecal, and pericyte lineages. Maintenance of these progenitors was dependent on vascular-mesenchymal Notch signaling. Ablation of postnatal Nestin+ cells resulted in a disruption of granulosa cell specification and an increased incidence of polyovular ovarian follicles, thus uncovering key roles for vasculature in ovarian differentiation. These findings may provide new insights into the origins of female gonad dysgenesis and infertility.

Adaptation of codon and amino acid use for translational functions in highly expressed cricket genes

Background For multicellular organisms, much remains unknown about the dynamics of synonymous codon and amino acid use in highly expressed genes, including whether their use varies with expression in different tissue types and sexes. Moreover, specific codons and amino acids may have translational functions in highly transcribed genes, that largely depend on their relationships to tRNA gene copies in the genome. However, these relationships and putative functions are poorly understood, particularly in multicellular systems. Results Here, we studied codon and amino acid use in highly expressed genes from reproductive and nervous system tissues (male and female gonad, somatic reproductive system, brain and ventral nerve cord, and male accessory glands) in the cricket Gryllus bimaculatus. We report an optimal codon, defined as the codon preferentially used in highly expressed genes, for each of the 18 amino acids with synonymous codons in this organism. The optimal codons were mostly shared among tissue types and both sexes. However, the frequency of optimal codons was highest in gonadal genes. Concordant with translational selection, a majority of the optimal codons had abundant matching tRNA gene copies in the genome, but sometimes obligately required wobble tRNAs. We suggest the latter may comprise a mechanism for slowing translation of abundant transcripts, particularly for cell-cycle genes. Non-optimal codons, defined as those least commonly used in highly transcribed genes, intriguingly often had abundant tRNAs, and had elevated use in a subset of genes with specialized functions (gametic and apoptosis genes), suggesting their use promotes the translational upregulation of particular mRNAs. In terms of amino acids, we found evidence suggesting that amino acid frequency, tRNA gene copy number, and amino acid biosynthetic costs (size/complexity) had all interdependently evolved in this insect model, potentially for translational optimization. Conclusions Collectively, the results suggest a model whereby codon use in highly expressed genes, including optimal, wobble, and non-optimal codons, and their tRNA abundances, as well as amino acid use, have been influenced by adaptation for various functional roles in translation within this cricket. The effects of expression in different tissue types and the two sexes are discussed.