Transcriptomic analysis reveal novel genes with sexually dimorphic expression in the zebrafish gonad and brain

Our knowledge on zebrafish reproduction is very limited. We generated a gonad-derived cDNA microarray from zebrafish and used it to analyze large-scale gene expression profiles in adult gonads and other organs. We have identified 116638 gonad-derived zebrafish expressed sequence tags (ESTs), 21% of which were isolated in our lab. Following in silico normalization, we constructed a gonad-derived microarray comprising 6370 unique, full-length cDNAs from differentiating and adult gonads. Labeled targets from adult gonad, brain, kidney and ‘rest-of-body’ from both sexes were hybridized onto the microarray. Our analyses revealed 1366, 881 and 656 differentially expressed transcripts (34.7% novel) that showed highest expression in ovary, testis and both gonads respectively. Hierarchical clustering showed correlation of the two gonadal transcriptomes and their similarities to those of the brains. In addition, we have identified 276 genes showing sexually dimorphic expression both between the brains and between the gonads. By in situ hybridization, we showed that the gonadal transcripts with the strongest array signal intensities were germline-expressed. We found that five members of the GTP-binding septin gene family, from which only one member (septin 4) has previously been implicated in reproduction in mice, were all strongly expressed in the gonads. We have generated a gonad-derived zebrafish cDNA microarray and demonstrated its usefulness in identifying genes with sexually dimorphic co-expression in both the gonads and the brains. We have also provided the first evidence of large-scale differential gene expression between female and male brains of a teleost. Our microarray would be useful for studying gonad development, differentiation and function not only in zebrafish but also in related teleosts via cross-species hybridizations. Since several genes have been shown to play similar roles in gonadogenesis in zebrafish and other vertebrates, our array may even provide information on genetic disorders affecting gonadal phenotypes and fertility in mammals.Fellowships from Temasek Life Science Laboratory Internal research grants from Temasek Life Science LaboratoryWeb of Scienc

Potential Vorticity Evolution of a Protoplanetary Disk with An Embedded Protoplanet

We present two-dimensional inviscid hydrodynamic simulations of a protoplanetary disk with an embedded planet, emphasizing the evolution of potential vorticity (the ratio of vorticity to density) and its dependence on numerical resolutions. By analyzing the structure of spiral shocks made by the planet, we show that progressive changes of the potential vorticity caused by spiral shocks ultimately lead to the excitation of a secondary instability. We also demonstrate that very high numerical resolution is required to both follow the potential vorticity changes and identify the location where the secondary instability is first excited. Low-resolution results are shown to give the wrong location. We establish the robustness of a secondary instability and its impact on the torque onto the planet. After the saturation of the instability, the disk shows large-scale non-axisymmetry, causing the torque on the planet to oscillate with large amplitude. The impact of the oscillating torque on the protoplanet's migration remains to be investigated.Comment: 17 pages total with 9 figures (Fig.4,5,9 are in .jpg), accepted to Ap

Polygenic Sex Determination System in Zebrafish

Contains fulltext : 94011.pdf (publisher's version ) (Open Access

Gonad differentiation in zebrafish is regulated by the canonical Wnt signalling pathway

Zebrafish males undergo a ‘‘juvenile ovary-to-testis’’ gonadal transformation process. Several genes, including nuclear receptor subfamily 5, group A (nr5a) and anti-Mu¨ llerian hormone (amh), and pathways such as Tp53-mediated germ-cell apoptosis have been implicated in zebrafish testis formation. However, our knowledge of the regulation of this complex process is incomplete, and much remains to be investigated about the molecular pathways and network of genes that control it. Using a microarray-based analysis of transforming zebrafish male gonads, we demonstrated that their transcriptomes undergo transition from an ovary-like pattern to an ovotestis to a testislike profile. Microarray results also validated the previous histological and immunohistochemical observation that there is high variation in the duration and extent of commitment to the juvenile ovary phase among individuals. Interestingly, global gene expression profiling of diverging zebrafish juvenile ovaries and transforming ovotestes revealed that some members of the canonical Wnt/beta-catenin signaling pathway were differentially expressed between these two phases. To investigate whether Wnt/beta-catenin signaling plays a role in zebrafish gonad differentiation, we used the Tg (hsp70l:dkk1b-GFP)w32 line to inhibit Wnt/beta-catenin signaling during gonad differentiation. Activation of dkk1b-GFP expression by heat shock resulted in an increased proportion of males and corresponding decrease in gonadal aromatase gene (cyp19a1a) expression. The Wnt target gene, lymphocyte enhancer binding factor 1 (lef1), was also down-regulated in the process. Together, these results provide the first functional evidence that, similarly to mammals, Wnt/beta-catenin signaling is a ‘‘pro-female’’ pathway that regulates gonad differentiation in zebrafish.Web of Scienc

High dose of 8-OH-DPAT decreases maximal dentate gyrus activation and facilitates granular cell plasticity in vivo

Although several studies have emphasized a crucial role for the serotonergic system in the control of hippocampal excitability, the role of serotonin (5-HT) and its receptors in normal and pathologic conditions, such as temporal lobe epilepsy (TLE), is still unclear. The present study was therefore designed firstly to investigate the acute effect of 8-OH-DPAT, a mixed 5-HT1A/7 receptor agonist, at a high dose (1 mg/kg, i.p.) known to have antiepileptic properties, in a model of acute partial epilepsy in rats. For this purpose, a maximal dentate activation (MDA) protocol was used to measure electrographic seizure onset and duration. In addition, the effect of 8-OH-DPAT on in vivo dentate gyrus cell reactivity and short- and long-term plasticity was studied. Rats injected with 8-OH-DPAT exhibited a significant reduction in MDA and epileptic discharges, a decrease in paired-pulse facilitation and an increase in long-term potentiation. This study suggests that 8-OH-DPAT or in general 5-HT1A/7 agonists might be useful for the treatment of TLE and also have some beneficial effects on the comorbid cognitive disorders seen in epileptic patients.peer-reviewe

Selective Ablation of the Androgen Receptor in Mouse Sertoli Cells Affects Sertoli Cell Maturation, Barrier Formation and Cytoskeletal Development

The observation that mice with a selective ablation of the androgen receptor (AR) in Sertoli cells (SC) (SCARKO mice) display a complete block in meiosis supports the contention that SC play a pivotal role in the control of germ cell development by androgens. To delineate the physiological and molecular mechanism responsible for this control, we compared tubular development in pubertal SCARKO mice and littermate controls. Particular attention was paid to differences in SC maturation, SC barrier formation and cytoskeletal organization and to the molecular mediators potentially involved. Functional analysis of SC barrier development by hypertonic perfusion and lanthanum permeation techniques and immunohistochemical analysis of junction formation showed that SCARKO mice still attempt to produce a barrier separating basal and adluminal compartment but that barrier formation is delayed and defective. Defective barrier formation was accompanied by disturbances in SC nuclear maturation (immature shape, absence of prominent, tripartite nucleoli) and SC polarization (aberrant positioning of SC nuclei and cytoskeletal elements such as vimentin). Quantitative RT-PCR was used to study the transcript levels of genes potentially related to the described phenomena between day 8 and 35. Differences in the expression of SC genes known to play a role in junction formation could be shown from day 8 for Cldn11, from day 15 for Cldn3 and Espn, from day 20 for Cdh2 and Jam3 and from day 35 for ZO-1. Marked differences were also noted in the transcript levels of several genes that are also related to cell adhesion and cytoskeletal dynamics but that have not yet been studied in SC (Actn3, Ank3, Anxa9, Scin, Emb, Mpzl2). It is concluded that absence of a functional AR in SC impedes the remodeling of testicular tubules expected at the onset of spermatogenesis and interferes with the creation of the specific environment needed for germ cell development

LLAMA : stellar populations in the nuclei of ultra-hard X-ray-selected AGN and matched inactive galaxies

The relation between nuclear (.50 pc) star formation and nuclear galactic activity is still elusive; theoretical models predict a link between the two, but it is unclear whether active galactic nuclei (AGNs) should appear at the same time, before, or after nuclear star formation activity. We present a study of this relation in a complete, volume-limited sample of nine of the most luminous (log L14−195 keV > 1042.5 erg s−1 ) local AGNs (the LLAMA sample), including a sample of 18 inactive control galaxies (six star-forming; 12 passive) that are matched by Hubble type, stellar mass (9.5 . log M?/M . 10.5), inclination, and distance. This allows us to calibrate our methods on the control sample and perform a differential analysis between the AGN and control samples. We performed stellar population synthesis on VLT/X-shooter spectra in an aperture corresponding to a physical radius of ≈150 pc. We find young (.30 Myr) stellar populations in seven out of nine AGNs and in four out of six star-forming control galaxies. In the non-star-forming control population, in contrast, only two out of 12 galaxies show such a population. We further show that these young populations are not indicative of ongoing star formation, providing evidence for models that see AGN activity as a consequence of nuclear star formation. Based on the similar nuclear star formation histories of AGNs and star-forming control galaxies, we speculate that the latter may turn into the former for some fraction of their time. Under this assumption, and making use of the volume completeness of our sample, we infer that the AGN phase lasts for about 5% of the nuclear starburst phase