Functional analysis of pax2/5/8 genes and their genetic interactions in zebrafish ear development

The vertebrate inner ear is a sensory organ responsible for auditory and vestibular function. Since its complex structure and cell types arise from a simply structured group of ectodermal cells, called the otic placode, the development of the inner ear has been a popular subject in embryology and developmental biology for decades. To date, many regulatory molecules and their functions have been identified in inner ear development showing considerable conservation among vertebrates. In vertebrates, Fgfs (fibroblast growth factors) from surrounding tissues are the main otic inducer and regulate various otic genes expression. Under the control of Fgf signals, pax2/5/8 genes are expressed in the otic region in the critical stages of otic development suggesting their function in otic development. In order to understand the function of pax2/5/8 genes and their interactions in the developing ear, we utilize zebrafish as a model system. Among zebrafish pax2/5/8 genes, pax8 is the earliest gene expressed in the preotic region while pax2a and pax2b are expressed slightly later. We found that pax8 is initially required for normal otic induction. Subsequently, pax8, pax2a and pax2b function redundantly to maintain otic fate. After otic placodes are induced by an Fgf signaling network, expression of Fgf3, one of otic inducers in zebrafish, persists in the hindbrain rhombomere 4. To investigate the function of the persistant Fgf3 expression, we examined a mutant with expanded Fgf3 expression in the hindbrain. Together with fgf3 knockdown results, we discovered that Fgf3 has later roles in specifying the anteroposterior (A-P) axis in the otic vesicle and regulating hair cell formation. We further identified pax5 as one of the genes regulated by the hindbrain Fgf3 activity, and pax5 function to be required for utricular hair cell survival

RNA-Guided Genome Editing in Drosophila with the Purified Cas9 Protein

We report a method for generating Drosophila germline mutants effectively via injection of the complex of the purified Cas9 protein, tracrRNA, and gene-specific crRNAs, which may reduce delayed mutations because of the transient activity of the Cas9 protein, combined with the simple mutation detection in GO founders by the T7E1 assay.

Cold shock domain proteins and glycine-rich RNA-binding proteins from Arabidopsis thaliana can promote the cold adaptation process in Escherichia coli

Despite the fact that cold shock domain proteins (CSDPs) and glycine-rich RNA-binding proteins (GRPs) have been implicated to play a role during the cold adaptation process, their importance and function in eukaryotes, including plants, are largely unknown. To understand the functional role of plant CSDPs and GRPs in the cold response, two CSDPs (CSDP1 and CSDP2) and three GRPs (GRP2, GRP4 and GRP7) from Arabidopsis thaliana were investigated. Heterologous expression of CSDP1 or GRP7 complemented the cold sensitivity of BX04 mutant Escherichia coli that lack four cold shock proteins (CSPs) and is highly sensitive to cold stress, and resulted in better survival rate than control cells during incubation at low temperature. In contrast, CSDP2 and GRP4 had very little ability. Selective evolution of ligand by exponential enrichment (SELEX) revealed that GRP7 does not recognize specific RNAs but binds preferentially to G-rich RNA sequences. CSDP1 and GRP7 had DNA melting activity, and enhanced RNase activity. In contrast, CSDP2 and GRP4 had no DNA melting activity and did not enhance RNAase activity. Together, these results indicate that CSDPs and GRPs help E.coli grow and survive better during cold shock, and strongly imply that CSDP1 and GRP7 exhibit RNA chaperone activity during the cold adaptation process

Acute Kidney Injury due to Menstruation-related Disseminated Intravascular Coagulation in an Adenomyosis Patient: A Case Report

The authors report a case of acute kidney injury (AKI) resulting from menstruation-related disseminated intravascular coagulation (DIC) in an adenomyosis patient. A 40-yr-old woman who had received gonadotropin for ovulation induction therapy presented with anuria and an elevated serum creatinine level. Her medical history showed primary infertility with diffuse adenomyosis. On admission, her pregnancy test was negative and her menstrual cycle had started 1 day previously. Laboratory data were consistent with DIC, and it was believed to be related to myometrial injury resulting from heavy intramyometrial menstrual flow. Gonadotropin is considered to play an important role in the development of fulminant DIC. This rare case suggests that physicians should be aware that gonadotropin may provoke fulminant DIC in women with adenomyosis

Effects of 4-week Training Using Laboratory Index on Competition Record of Elite Female Middle-distance Runner: A Case Report

PURPOSE To investigate the effect of a 4-week training using a laboratory index on the competition record of elite female middle-distance runners. METHODS A female, middle-distance, trained runner with the following characteristics: age, 20 years; height, 168.7 cm; weight, 64.3 kg; 27.2% fat; and VO2max, 56.4 VO2 mL/kg/min; volunteered to participate in this study. Before the training program, the participant took part in a 1,500-m track and field national event in April and the onset of blood lactate threshold was analyzed using (OBVA/ vOBLA), VO2max/vVO2max, and tVO2max tests. After completing the 4-week training program, the participant again took part in the 1,500-m track and field national event in May and her parameters were reassessed using the OBVA/vOBLA, VO2max/vVO2max, and tVO2max tests. RESULTS Pre- and post-training indicate that vVO2max did not improve; however, tVO2max (206 seconds pre-training vs. 251.51 seconds post-training) and VO2max (56.4 vs. 59.3 VO2 mL/kg/min)significantly. However, this improvement in relative VO2 was due to weight loss, and the absolute value of VO2 (3.63 vs. 3.62 L/min) did not change. The 1,500-m race record in track and field events decreased significantly from a pre-training value of 5 minutes 03 seconds to a post-training value of 4 minutes 52 seconds. CONCLUSIONS The results of this study indicate that utilizing laboratory indicators including vOBLA, vVO2max, and tVO2max may be extremely valuable when prescribing training programs for middle-distance runners

Effects of Acute Exercise on Mitochondrial Function, Dynamics, and Mitophagy in Rat Cardiac and Skeletal Muscles

Purpose This study aimed to investigate the effects of single-bout exercise on mitochondrial function, dynamics (fusion, fission), and mitophagy in cardiac and skeletal muscles. Methods Fischer 344 rats (4 months old) were randomly divided into the control (CON) or acute exercise (EX) group (n=10 each). The rats performed a single bout of treadmill exercise for 60 minutes. Mitochondrial function (e.g., O2 respiration, H2O2 emission, Ca2+ retention capacity), mitochondrial fusion (e.g., Mfn1, Mfn2, Opa1), mitochondrial fission (e.g., Drp1, Fis1), and mitophagy (e.g., Parkin, Pink1, LC3II, Bnip3) were measured in permeabilized cardiac (e.g., left ventricle) and skeletal (e.g., soleus, white gastrocnemius) muscles. Results Mitochondrial O2 respiration and Ca2+ retention capacity were significantly increased in all tissues of the EX group compared with the CON group. Mitochondrial H2O2 emissions showed tissue-specific results; the emissions showed no significant differences in the left ventricle or soleus (type I fibers) but was significantly increased in the white gastrocnemius (type II fibers) after acute exercise. Mitochondrial fusion and fission were not altered in any tissues of the EX group. Mitophagy showed tissue-specific differences: It was not changed in the left ventricle or white gastrocnemius, whereas Parkin and LC3II were significantly elevated in the soleus muscle. Conclusions A single bout of aerobic exercise may improve mitochondrial function (e.g., O2 respiration and Ca2+ retention capacity) in the heart and skeletal muscles without changes in mitochondrial dynamics or mitophagy

Structural determinants crucial to the RNA chaperone activity of glycine-rich RNA-binding proteins 4 and 7 in Arabidopsis thaliana during the cold adaptation process

Although glycine-rich RNA-binding proteins (GRPs) have been determined to function as RNA chaperones during the cold adaptation process, the structural features relevant to this RNA chaperone activity remain largely unknown. To uncover which structural determinants are necessary for RNA chaperone activity of GRPs, the importance of the N-terminal RNA recognition motif (RRM) and the C-terminal glycine-rich domains of two Arabidopsis thaliana GRPs (AtGRP4 harbouring no RNA chaperone activity and AtGRP7 harbouring RNA chaperone activity) was assessed via domain swapping and mutation analyses. The results of domain swapping and deletion experiments showed that the domain sequences encompassing the N-terminal RRM of GRPs were found to be crucial to the ability to complement cold-sensitive Escherichia coli mutant cells under cold stress, RNA melting ability, and freezing tolerance ability in the grp7 loss-of-function Arabidopsis mutant. In particular, the N-terminal 24 amino acid extension of AtGRP4 impedes the RNA chaperone activity. Collectively, these results reveal that domain sequences and overall folding of GRPs governed by a specific modular arrangement of RRM and glycine-rich sequences are critical to the RNA chaperone activity of GRPs during the cold adaptation process in cells