Homozygous Mutation of gsdf Causes Infertility in Female Nile Tilapia (Oreochromis niloticus).

Gonadal somatic cell-derived factor (Gsdf) is a member of the TGF-ß superfamily, which exists mainly in fishes. Homozygous gsdf mutations in Japanese medaka and zebrafish resulted in infertile females, and the reasons for their infertility remain unknown. This study presents functional studies of Gsdf in ovary development using CRISPR/Cas9 in Nile tilapia (Oreochromis niloticus). The XX wild type (WT) female fish regularly reproduced from 12 months after hatching (mah), while the XX gsdf-/- female fish never reproduced and were infertile. Histological observation showed that at 24 mah, number of phase IV oocyte in the XX gsdf-/- female fish was significantly lower than that of the WT fish, although their gonadosomatic index (GSI) was similar. However, the GSI of the XX gsdf-/- female at 6 mah was higher than that of the WT. The mutated ovaries were hyperplastic with more phase I oocytes. Transcriptome analysis identified 344 and 51 up- and down-regulated genes in mutants compared with the WT ovaries at 6 mah. Some TGF-ß signaling genes that are critical for ovary development in fish were differentially expressed. Genes such as amh and amhr2 were up-regulated, while inhbb and acvr2a were down-regulated in mutant ovaries. The cyp19a1a, the key gene for estrogen synthesis, was not differentially expressed. Moreover, the serum 17ß-estradiol (E2) concentrations between XX gsdf-/- and WT were similar at 6 and 24 mah. Results from real-time PCR and immunofluorescence experiments were similar and validated the transcriptome data. Furthermore, Yeast-two-hybrid assays showed that Gsdf interacts with TGF-ß type II receptors (Amhr2 and Bmpr2a). Altogether, these results suggest that Gsdf functions together with TGF-ß signaling pathway to control ovary development and fertility. This study contributes to knowledge on the function of Gsdf in fish oogenesis.

High Polymorphism in the Dmrt2a Gene Is Incompletely Sex-Linked in Spotted Scat, Scatophagus argus.

Unlike mammals and birds, many fishes have young sex chromosomes, providing excellent models to study sex chromosome differentiation at early stages. Previous studies showed that spotted scat possesses an XX-XY sex determination system. The X has a complete Dmrt3 copy (termed normal) and a truncated copy of Dmrt1 (called Dmrt1b), while the Y has the opposite (normal Dmrt1, which is male-specific, and a truncated Dmrt3 called Dmrt3△-Y). Dmrt1 is the candidate sex determination gene, while the differentiation of other sex-linked genes remains unknown. The spotted scat has proven to be a good model to study the evolution of sex chromosomes in vertebrates. Herein, we sequenced a neighbor gene of this family, Dmrt2, positioned farther from Dmrt1 and closer to Dmrt3 in the spotted scat, and analyzed its sequence variation and expression profiles. The physical locations of the three genes span across an estimated size of >40 kb. The open reading frames of Dmrt2a and its paralog Dmrt2b are 1578 bp and 1311 bp, encoding peptides of 525 and 436 amino acid residues, respectively. Dmrt2a is positioned close to Dmrt3 but farther from Dmrt1 on the same chromosome, while Dmrt2b is not. Sequence analysis revealed several mutations; insertions, and deletions (indels) on Dmrt2a non-coding regions and single-nucleotide polymorphisms (SNPs) on the Dmrt2a transcript. These indels and SNPs are sex-linked and showed high male heterogeneity but do not affect gene translation. The markers designed to span the mutation sites tested on four different populations showed varied concordance with the genetic sexes. Dmrt2a is transcribed solely in the gonads and gills, while Dmrt2b exists in the gonads, hypothalamus, gills, heart, and spleen. The Dmrt2a and Dmrt2b transcripts are profoundly expressed in the male gonads. Analyses of the transcriptome data from five other fish species (Hainan medaka (Oryzias curvinotus), silver sillago (Sillago sihama), Nile tilapia (Oreochromis niloticus), Hong Kong catfish (Clarias fuscus), and spot-fin porcupine fish (Diodon hystrix)) revealed testes-biased expression of Dmrt1 in all, similar to spotted scat. Additionally, the expression of Dmrt2a is higher in the testes than the ovaries in spotted scat and Hainan medaka. The Dmrt2a transcript was not altered in the coding regions as found in Dmrt1 and Dmrt3 in spotted scat. This could be due to the functional importance of Dmrt2a in development. Another possibility is that because Dmrt2a is positioned farther from Dmrt1 and the chromosome is still young, meaning it is only a matter of time before it differentiates. This study undeniably will aid in understanding the functional divergence of the sex-linked genes in fish.

Comparative transcriptome analysis of male and female gonads reveals sex-biased genes in spotted scat (Scatophagus argus).

Scatophagus argus is a new emerging aquaculture fish in East and Southeast Asia. To date, research on reproductive development and regulation in S. argus is lacking. Additionally, genetic and genomic information about reproduction, such as gonadal transcriptome data, is also lacking. Herein, we report the first gonadal transcriptomes of S. argus and identify genes potentially involved in reproduction and gonadal development. A total of 136,561 unigenes were obtained by sequencing of testes (n = 3) and ovaries (n = 3) at stage III. Genes upregulated in males and females known to be involved in gonadal development and gametogenesis were identified, including male-biased dmrt1, amh, gsdf, wt1a, sox9b, and nanos2, and female-biased foxl2, gdf9, bmp15, sox3, zar1, and figla. Serum estradiol-17ß and 11-ketotestosterone levels were biased in female and male fish, respectively. Sexual dimorphism of serum steroid hormone levels were interpreted after expression analysis of 20 steroidogenesis-related genes, including cyp19a1a and cyp11b2. This gonadal transcript dataset will help investigate functional genes related to reproduction in S. argus.

Expression and transcriptional regulation of gsdf in spotted scat (Scatophagus argus).

Gonadal soma-derived factor (Gsdf) is critical for testicular differentiation and early germ cell development in teleosts. The spotted scat (Scatophagus argus), with a stable XX-XY sex-determination system and the candidate sex determination gene dmrt1, provides a good model for understanding the mechanism of sex determination and differentiation in teleosts. In this study, we analyzed spotted scat gsdf tissue distribution and gene expression patterns in gonads, as well as further analysis of transcriptional regulation. Tissue distribution analysis showed that gsdf was only expressed in testis and ovary. Real-time PCR showed that both gsdf and dmrt1 were expressed significantly higher in testes at different phases (phase III, IV and V) compared to ovaries at phase II, III and IV, while gsdf was expressed significantly higher in phase II ovaries than those of phase III and IV. Western blot analysis also showed that Gsdf was more highly expressed in the testis than ovary. Immunohistochemistry analysis showed that Gsdf was expressed in Sertoli cells surrounding spermatogonia in the testis, while it was expressed in the somatic cells surrounding the oogonia of the ovary. Approximately 2.7 kb of the 5' upstream region of gsdf was cloned from the spotted scat genomic DNA and in silico promoter analysis revealed the putative transcription factor binding sites of Dmrt1 and Sf1. The luciferase reporter assay, using the human embryonic kidney cells, demonstrated that Dmrt1 activated gsdf expression in a dose-dependent manner in the presence of Sf1 in spotted scat. These results suggest that Gsdf could play a role in regulating the development of spermatogonia and oogonia, and also participate in male sex differentiation by acting as a downstream gene of Dmrt1 in spotted scat.

InAs/GaAs Quantum Dot Dual-Mode Distributed Feedback Laser Towards Large Tuning Range Continuous-Wave Terahertz Application.

In this paper, a laterally coupled distributed feedback (LC-DFB) laser based on modulation p-doped multiple InAs/GaAs quantum dot (QD) structures has been fabricated. The device exhibits a high side-mode suppression ratio (SMSR) of > 47 dB and a high thermal stability of dλ/dT = 0.092 nm/K under continuous-wave (CW) operation, which is mainly attributed to the high material gain prepared by modulation p-doping and rapid thermal annealing (RTA) process, and the significantly reduced waveguide losses by shallow-etched gratings and its close proximity to the laser ridge feature in the LC-DFB laser. With this superior performance of the DFB laser, the wide tunable dual-wavelength lasing operation has been obtained by delicately defining different periods for the grating structures on the two sides of the laser ridge or combining the reduced laser cavity length. The wavelength spacing between the two lasing modes can be flexibly tuned in a very wide range from 0.5 to 73.4 nm, corresponding to the frequency difference from 0.10 to 14 THz, which is the largest tuning range by the utilization of single device and hence providing a new opportunity towards the generation of CW THz radiation.

Near-infrared and mid-infrared semiconductor broadband light emitters.

Semiconductor broadband light emitters have emerged as ideal and vital light sources for a range of biomedical sensing/imaging applications, especially for optical coherence tomography systems. Although near-infrared broadband light emitters have found increasingly wide utilization in these imaging applications, the requirement to simultaneously achieve both a high spectral bandwidth and output power is still challenging for such devices. Owing to the relatively weak amplified spontaneous emission, as a consequence of the very short non-radiative carrier lifetime of the inter-subband transitions in quantum cascade structures, it is even more challenging to obtain desirable mid-infrared broadband light emitters. There have been great efforts in the past 20 years to pursue high-efficiency broadband optical gain and very low reflectivity in waveguide structures, which are two key factors determining the performance of broadband light emitters. Here we describe the realization of a high continuous wave light power of >20 mW and broadband width of >130 nm with near-infrared broadband light emitters and the first mid-infrared broadband light emitters operating under continuous wave mode at room temperature by employing a modulation p-doped InGaAs/GaAs quantum dot active region with a 'J'-shape ridge waveguide structure and a quantum cascade active region with a dual-end analogous monolithic integrated tapered waveguide structure, respectively. This work is of great importance to improve the performance of existing near-infrared optical coherence tomography systems and describes a major advance toward reliable and cost-effective mid-infrared imaging and sensing systems, which do not presently exist due to the lack of appropriate low-coherence mid-infrared semiconductor broadband light sources.

Realization of III-V Semiconductor Periodic Nanostructures by Laser Direct Writing Technique.

In this paper, we demonstrated the fabrication of one-dimensional (1D) and two-dimensional (2D) periodic nanostructures on III-V GaAs substrates utilizing laser direct writing (LDW) technique. Metal thin films (Ti) and phase change materials (Ge2Sb2Te5 (GST) and Ge2Sb1.8Bi0.2Te5 (GSBT)) were chosen as photoresists to achieve small feature sizes of semiconductor nanostructures. A minimum feature size of about 50 nm about a quarter of the optical diffraction limit was obtained on the photoresists, and 1D III-V semiconductor nanolines with a minimum width of 150 nm were successfully acquired on the GaAs substrate which was smaller than the best results acquired on Si substrate ever reported. 2D nanosquare holes were fabricated as well by using Ti thin film as the photoresist, with a side width of about 200 nm, but the square holes changed to a rectangle shape when GST or GSBT was employed as the photoresist, which mainly resulted from the interaction of two cross-temperature fields induced by two scanning laser beams. The interacting mechanism of different photoresists in preparing periodic nanostructures with the LDW technique was discussed in detail.

Core temperature changes in resuspended red blood cells (RBCs) and pediatric RBCs removed from refrigerated storage.

BACKGROUND: The 30-minute rule, whereby intact red blood cell (RBC) products may be returned to stock if returned to 4 degrees C storage within 30 minutes of issue, was established many years ago. It was based on observations that the core temperature of units of whole blood removed from storage temperatures of 1 to 6 degrees C, and left at room temperature, would reach 10 degrees C at between 45 minutes and 1 hour. STUDY DESIGN AND METHODS: Forty-one units of RBCs resuspended leukoreduced and 8 units of pediatric RBCs resuspended leukoreduced were exposed to ambient temperature for periods of time between 0 and 60 minutes. Core temperatures of all units were measured at 1-minute or 5-minute intervals. RESULTS: Resuspended RBCs units reached a mean core temperature of 10 degrees C at 15 minutes, 12.7 degrees C at 30 minutes, and 15 degrees C at 60 minutes. Pediatric RBCs reached a mean core temperature of 12.8 degrees C at 15 minutes, 15.5 degrees C at 30 minutes, and 17.8 degrees C at 60 minutes. CONCLUSION: In view of our results, and the range of RBC products now available, it may be timely for blood services to review and reduce the 30-minute rule.