Neuregulin1が雌妊孕性に果たす役割に関する研究

内容の要約広島大学(Hiroshima University)博士(農学)Doctor of Agriculturedoctora

Activation of Toll-like receptor 7/8 encoded by the X chromosome alters sperm motility and provides a novel simple technology for sexing sperm

In most mammals, the male to female sex ratio of offspring is about 50% because half of the sperm contain either the Y chromosome or X chromosome. In mice, the Y chromosome encodes fewer than 700 genes, whereas the X chromosome encodes over 3,000 genes. Although overall gene expression is lower in sperm than in somatic cells, transcription is activated selectively in round spermatids. By regulating the expression of specific genes, we hypothesized that the X chromosome might exert functional differences in sperm that are usually masked during fertilization. In this study, we found that Toll-like receptors 7/8 (TLR7/8) coding the X chromosome were expressed by approximately 50% of the round spermatids in testis and in approximately 50% of the epididymal sperm. Especially, TLR7 was localized to the tail, and TLR8 was localized to the midpiece. Ligand activation of TLR7/8 selectively suppressed the mobility of the X chromosome–bearing sperm (X-sperm) but not the Y-sperm without altering sperm viability or acrosome formation. The difference in sperm motility allowed for the separation of Y-sperm from X-sperm. Following in vitro fertilization using the ligand-selected high-mobility sperm, 90% of the embryos were XY male. Likewise, 83% of the pups obtained following embryo transfer were XY males. Conversely, the TLR7/8-activated, slow mobility sperm produced embryos and pups that were 81% XX females. Therefore, the functional differences between Y-sperm and X-sperm motility were revealed and related to different gene expression patterns, specifically TLR7/8 on X-sperm.This work was supported in part by Livestock Promotional Funds of Japan Racing Association (JRA) and by Hiroshima Cryopreservation Service Co. (to MS)

Agreement in rotator cuff muscles measurement between ultrasonography and magnetic resonance imaging

[Background/objective] It is important to assess the atrophy of the rotator cuff to better understand shoulder function and pain. Previously, magnetic resonance imaging has been used for the evaluation of atrophy of rotator cuff muscles, which is time consuming. Therefore, a measurement tool requiring little time and easy accessibility is clinically desirable to be used frequently in rehabilitation. Recently, rotator cuff muscles have been evaluated using ultrasonography. However, little is known about the agreement of evaluation in rotator cuff muscles between magnetic resonance imaging and ultrasonography. The purpose of this study was to demonstrate the agreement between the muscle thickness measurements of supraspinatus, infraspinatus, and teres minor muscles by ultrasonography and the cross-sectional area measured by magnetic resonance imaging in the patient with rotator cuff tears. [Methods] A total of 47 patients with rotator cuff tears were enrolled. There were the 37 small tears, four medium tears, and six large tears, and the involved rotator cuff muscles were the supraspinatus in 37 shoulders, and the supraspinatus and infraspinatus in 10 shoulders. The measuring variables were muscle thickness and cross-sectional area of supraspinatus, infraspinatus, and teres minor muscles by using magnetic resonance imaging. Further, the muscle thickness of the rotator cuff were assessed using ultrasonography. A single regression model was used for demonstrating the agreement between the cross-sectional area measurement by magnetic resonance imaging and the muscle thickness measured using ultrasonography and magnetic resonance imaging of rotator cuff muscles. Additionally, the Bland-Altman plots between magnetic resonance imaging and ultrasonography was analyzed. [Results] The cross-sectional area were correlated with the muscle thickness measurement of rotator cuff muscles by magnetic resonance imaging, significantly (supraspinatus: r = 0.84, infraspinatus: ρ = 0.63, teres minor: ρ = 0.61, all p < 0.001). There were significant agreements between the cross-sectional area measured by magnetic resonance imaging and muscle thickness measured by ultrasonography (supraspinatus: r = 0.80, infraspinatus: ρ = 0.78, teres minor: ρ = 0.74, all p < 0.001). Bland-Altman plots revealed significant correlations between the average and the difference of the two measurements in supraspinatus (r = 0.36, p = 0.012), infraspinatus (r = 0.38, p < 0.001), and teres minor (r = 0.42, p < 0.001). These results clarified the proportional bias between MRI and US. [Conclusion] This study showed that, similar to magnetic resonance imaging, ultrasonography is a useful tool for assessing muscle atrophy of supraspinatus, infraspinatus, and teres minor muscles

Compensation strategy of shoulder synergist muscles is not stereotypical in patients with rotator cuff repair

Rotator cuff tear is a common shoulder injury that causes shoulder dysfunction and pain. Although surgical repair is the primary treatment for rotator cuff tear, it is well recognized that impaired force exertion of muscles connecting to the involved tendon and subsequent complemental change in the force exertion of synergist muscles persist even after repair. This study aimed to identify the compensation strategy of shoulder abductors by examining how synergist muscles respond to supraspinatus (SSP) muscle force deficit in patients with rotator cuff repair. Muscle shear modulus, an index of muscle force, was assessed for SSP, infraspinatus, upper trapezius, and middle deltoid muscles in repaired and contralateral control shoulders of 15 patients with unilateral tendon repair of the SSP muscle using ultrasound shear wave elastography while the patients passively or actively held their arm in shoulder abduction. In the repaired shoulder, the shear modulus of the SSP muscle declined, whereas that of other synergist muscles did not differ relative to that of the control. To find the association between the affected SSP and each of the synergist muscles, a regression analysis was used to assess the shear moduli at the population level. However, no association was observed between them. At the individual level, there was a tendency of variation among patients with regard to a specific muscle whose shear modulus complementarily increased. These results suggest that the compensation strategy for SSP muscle force deficit varies among individuals, being nonstereotypical in patients with rotator cuff injury

The acceleration of reproductive aging in Nrg1flox/flox;Cyp19-Cre female mice

Irregular menstrual cycles, reduced responses to exogenous hormonal treatments, and altered endocrine profiles (high FSH/high LH/low AMH) are observed in women with increasing age before menopause. In this study, because the granulosa cell-specific Nrg1 knockout mice (gcNrg1KO) presented ovarian and endocrine phenotypes similar to older women, we sought to understand the mechanisms of ovarian aging and to develop anewstrategy for improving fertility in older women prior to menopause. In the ovary of 6-month-old gcNrg1KO mice, follicular development was blocked in bilayer secondary follicles and heterogeneous cells accumulated in ovarian stroma. The heterogeneous cells in ovarian stroma were distinguished as two different types: (i) the LH receptor-positive endocrine cells and (ii) actin-rich fibrotic cells expressing collagen. Both the endocrine and fibrotic cells disappeared following long-term treatment with a GnRH antagonist, indicating that the high levels of serum LH induced the survival of both cell types and the abnormal endocrine profile to reduce fertility. Moreover, follicular development to the antral stages was observed with reduced LH and the disappearance of the abnormal stromal cells. Mice treated with the GnRH antagonist regained normal, recurrent estrous cycles and continuously delivered pups for at least for 3 months. We conclude that endocrine and matrix alternations occur within the ovarian stroma with increasing age and that abolishing these alternations resets the cyclical release of LH. Thus, GnRH antagonist treatments might provide a new, noninvasive strategy for improving fertility in a subset of aging women before menopause.This work was supported in part by The Japan Society for the Promotion of Science (JSPS) KAKENHI, JP24688028, JP 16H05017 (to MS) and JP15J05331 (to TU), by Japan Agency for Medical Research and Development (AMED) 16gk0110015 h0001 (to MS), and by National Institute of Health (NIH)-HD-076980 (to JSR)

Gene Expression and Protein Synthesis in Mitochondria Enhance the Duration of High-Speed Linear Motility in Boar Sperm

Sperm motility patterns are continuously changed after ejaculation to fertilization in the female tract. Hyperactivated motility is induced with high glucose medium in vitro or the oviduct fluids in vivo, whereas sperm maintain linear motility in the seminal plasma or the uterine fluids containing low glucose. Therefore, it is estimated that sperm motility patterns are dependent on the energy sources, and the mitochondrial oxidative phosphorylation is activated to produce ATP in low glucose condition. To elucidate these hypotheses, boar sperm was incubated in different energy conditions with the transcription and translation inhibitors in vitro. Sperm motility parameters, mitochondrial activity, ATP level, gene expression and protein synthesis were analyzed. Sperm progressive motility and straight-line velocity were significantly increased with decreasing glucose level in the incubation medium. Moreover, the mitochondrial protein turnover meaning transcription and translation from mitochondrial genome in sperm is activated during incubation. Incubation of sperm with mitochondrial translation inhibitor (D-chloramphenicol) suppressed mitochondrial protein synthesis, mitochondrial activity and ATP level in sperm and consequently reduced the linear motility speed, but not the motility. Thus, it is revealed that the mitochondrial central dogma is active in sperm, and the high-speed linear motility is induced in low glucose condition via activating the mitochondrial activity for ATP generation.This work was supported in part by Livestock Promotional Funds of Japan Racing Association (JRA) grant no. H30-279 and by Hiroshima Cryopreservation Service Co., grant no. H30-1 (to MS). ZZ was supported by China Scholarship Council during a visit of “Zhendong Zhu” to Hiroshima University (#CSC201706300110).The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fphys.2019.00252/full#supplementary-materia

Femoral neck fracture with osteoporosis in Tokushima Prefecture

The numbers and features of occurrence, causes, treatments and prognosis of femoral neck fracture with osteoporosis in Tokushima prefecture were investigated in the 10th fiscal year of Heisei. 634 patients (154 males and 480 females) suffered from the femoral neck fracture. Females were 3.1 times as many as males. In females, the occurrence of the fracture had a tendency to increase to 85-years-old population. In fracture types, 253 cases were intracapsular type and 381 cases extracapsular type. Extracapsular type of the femoral neck fracture increased in proportion to aging. 384 cases were treated with osteosynthesis, 207 cases with femoral prosthesis and 43 cases with othermethods. The main cause of the fracture was trivial fall (79%). 288 cases returned to home and 258 cases still admitted in the secondary hospitals. 69 cases entered to the nursing home. Half of the patients who could walk with or without crutch before fracture were able to return to home, on the other hand, the ratio of patients who could return to home among the patients with no ability of walk before injury was less than 10%. Mortality rate was 2.1 % at the discharge

Class IA Phosphatidylinositol 3-Kinase in Pancreatic β Cells Controls Insulin Secretion by Multiple Mechanisms

SummaryType 2 diabetes is characterized by insulin resistance and pancreatic β cell dysfunction, the latter possibly caused by a defect in insulin signaling in β cells. Inhibition of class IA phosphatidylinositol 3-kinase (PI3K), using a mouse model lacking the pik3r1 gene specifically in β cells and the pik3r2 gene systemically (βDKO mouse), results in glucose intolerance and reduced insulin secretion in response to glucose. β cells of βDKO mice had defective exocytosis machinery due to decreased expression of soluble N-ethylmaleimide attachment protein receptor (SNARE) complex proteins and loss of cell-cell synchronization in terms of Ca2+ influx. These defects were normalized by expression of a constitutively active form of Akt in the islets of βDKO mice, preserving insulin secretion in response to glucose. The class IA PI3K pathway in β cells in vivo is important in the regulation of insulin secretion and may be a therapeutic target for type 2 diabetes

Ameloblastin induces tumor suppressive phenotype and enhances chemosensitivity to doxorubicin via Src-Stat3 inactivation in osteosarcoma

Ameloblastin (AMBN), the most abundant non-amelogenin enamel matrix protein, plays a role in ameloblast differentiation. Previously, we found that AMBN promoted osteogenic differentiation via the interaction between CD63 and integrin β1, leading to the inactivation of Src; however, how AMBN affects the malignant behavior of osteosarcoma is still unclear. Osteosarcoma affects the bone and is associated with poor prognosis because of the high rate of pulmonary metastases and drug resistance. Here we demonstrated that stable overexpression of AMBN induced apoptosis and suppressed colony formation and cell migration via the inactivation of Src-Stat3 pathway in human osteosarcoma cells. Moreover, AMBN induced chemosensitivity to doxorubicin. Thus, AMBN induced a tumor suppressive phenotype and chemosensitivity to doxorubicin via the AMBN-Src-Stat3 axis in osteosarcoma. Indeed, immunohistochemical expression of AMBN was significantly correlated with better outcome of osteosarcoma patients. Our findings suggest that AMBN can be a new prognostic marker and therapeutic target for osteosarcoma combined with conventional doxorubicin treatment