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Spatially and temporally controlled gene expression, including transcription, several mRNA processing steps, and the export of mature mRNA to the cytoplasm, is essential for developmental processes. It is well known that RNA helicases of the DExD/H-box protein family are involved in these gene expression processes, including transcription, pre-mRNA splicing, and rRNA biogenesis. Although one DExD/H-box protein, Prp5, a homologue of vertebrate Ddx46, has been shown to play important roles in pre-mRNA splicing in yeast, the in vivo function of Ddx46 remains to be fully elucidated in metazoans. In this study, we isolated zebrafish morendo (mor), a mutant that shows developmental defects in the digestive organs and brain, and found that it encodes Ddx46. The Ddx46 transcript is maternally supplied, and as development proceeds in zebrafish larvae, its ubiquitous expression gradually becomes restricted to those organs. The results of whole-mount in situ hybridization showed that the expression of various molecular markers in these organs is considerably reduced in the Ddx46 mutant. Furthermore, splicing status analysis with RT-PCR revealed unspliced forms of mRNAs in the digestive organ and brain tissues of the Ddx46 mutant, suggesting that Ddx46 may be required for pre-mRNA splicing during zebrafish development. Therefore, our results suggest a model in which zebrafish Ddx46 is required for the development of the digestive organs and brain, possibly through the control of pre-mRNA splicing.

Balanced and precisely controlled processes between self-renewal and differentiation of hematopoietic stem cells (HSCs) into all blood lineages are critical for vertebrate definitive hematopoiesis. However, the molecular mechanisms underlying the maintenance and differentiation of HSCs have not been fully elucidated. Here, we show that zebrafish Ddx46, encoding a DEAD-box RNA helicase, is expressed in HSCs of the caudal hematopoietic tissue (CHT). The number of HSCs expressing the molecular markers cmyb or T-cell acute lymphocytic leukemia 1 (tal1) was markedly reduced in Ddx46 mutants. However, massive cell death of HSCs was not detected, and proliferation of HSCs was normal in the CHT of the mutants at 48 h postfertilization. We found that myelopoiesis occurred, but erythropoiesis and lymphopoiesis were suppressed, in Ddx46 mutants. Consistent with these results, the expression of spi1, encoding a regulator of myeloid development, was maintained, but the expression of gata1a, encoding a regulator of erythrocyte development, was downregulated in the mutants. Taken together, our results provide the first genetic evidence that zebrafish Ddx46 is required for the multilineage differentiation of HSCs during development, through the regulation of specific gene expressions.

BackgroundMedial tibial stress syndrome (MTSS) is one of the most common causes of exercise-related leg pain in runners. Because stopping training due to pain from MTSS could decrease the athlete’s competitiveness, it is necessary to construct MTSS prevention and treatment programs. However, the effect of running, which is believed to cause MTSS, on shear elastic modulus of the posterior lower leg is unclear. Therefore, the purpose of this study was to investigate the effect of 30 min of running on shear elastic modulus of the posterior lower leg in healthy subjects.MethodsTwenty healthy males volunteered to participate in this study (age, 20.9 ± 0.6 y; height, 169.6 ± 4.5 cm; weight, 62.6 ± 5.2 kg). The shear elastic modulus of the posterior lower leg was measured using ultrasonic shear wave elastography before and immediately after a 30-min running task.ResultsShear elastic moduli of the flexor digitorum longus and tibialis posterior were significantly increased after 30 min running task. However, there were no significant changes in shear elastic moduli of the lateral gastrocnemius, medial gastrocnemius, peroneus longus and peroneus brevis.ConclusionThe results suggested that the increases in shear elastic moduli of flexor digitorum longus and tibialis posterior after running could be a risk factor for running-related MTSS development.

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BackgroundIn the present study, CFLs harvested from cadavers were categorized according to the differences in the angle of the CFL with respect to the long axis of the fibula and their shape, and then three-dimensional reconstructions of the CFLs were used to simulate and examine the differences in the angles of the CFLs with respect to the long axis of the fibula and how they affect CFL function.MethodsThe study sample included 81 ft from 43 Japanese cadavers. CFLs were categorized according to their angle with respect to the long axis of the fibula and the number of fiber bundles. Five categories were subsequently established: CFL20° (angle of the CFL with respect to the long axis of the fibula from 20° to 29°); CFL30° (range 30–39°); CFL40° (range 40–49°); CFL50° (range 50–59°); and CFL2 (CLFs with two crossing fiber bundles). Three-dimensional reconstructions of a single specimen from each category were then created. These were used to simulate and calculate CFL strain during dorsiflexion (20°) and plantarflexion (30°) on the talocrural joint axis and inversion (20°) and eversion (20°) on the subtalar joint axis.ResultsIn terms of proportions for each category, CFL20° was observed in 14 ft (17.3%), with CFL30° in 22 ft (27.2%), CFL40° in 29 ft (35.8%), CFL50° in 15 ft (18.5%), and CFL2 in one foot (1.2%). Specimens in the CFL20° and CFL30° groups contracted with plantarflexion and stretched with dorsiflexion. In comparison, specimens in the CFL40°, CFL50°, and CFL2 groups stretched with plantarflexion and contracted with dorsiflexion. Specimens in the CFL20° and CFL2 groups stretched with inversion and contracted with eversion.ConclusionsCFL function changed according to the difference in the angles of the CFLs with respect to the long axis of the fibula.

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Background: Anterior cruciate ligament (ACL) injury has been reported to have a higher incidence in women than in men. Purpose/Hypothesis: The purpose was to examine the relationship of anterior knee laxity (AKL), stiffness, and generalized joint laxity (GJL) with respect to the menstrual cycle. It was hypothesized that AKL and GJL would increase during the ovulation phase, when estrogen levels are high. Study Design: Descriptive laboratory study. Methods: A total of 15 female university students aged >20 years and with normal menstrual cycles were evaluated. AKL was measured as anterior tibial displacement of the femur after application of 44-, 89-, and 133-N loads to the tibia. Stiffness was calculated as Δ force/Δ displacement at loads between 44 and 89 N and between 89 and 133 N. The University of Tokyo joint laxity test was used for evaluation of GJL. The participants’ menstrual cycle was divided into the early follicular, late follicular, ovulation, and luteal phases using the basal body temperature method and an ovulation kit; AKL and GJL were measured once during each phase. Participants were also stratified according to the presence or absence of genu recurvatum (GR). Results: There was no significant difference in AKL, stiffness, or GJL among the menstrual phases. In the GR group, AKL values at 89 N and 133 N were significantly higher in the ovulation phase than in the early follicular phase ( P = .025 and P =.018, respectively); there were no significant differences in AKL among the phases in the non-GR group. In addition, the GR group in the ovulation phase had significantly higher AKL values at 44 N, 89 N, and 133 N compared with the non-GR group ( P = .013, P = .005, and P = .010, respectively). There were no significant differences in GJL among the phases in the GR or non-GR groups. Conclusion: Women with GR may have increased AKL in the ovulation phase when compared with the early follicular phase, which may be a risk factor for ACL injury. Clinical Relevance: The results of this study suggest that the ovulation phase may be related to the greater incidence of ACL injuries in women.