Kim, Eun-Joong;Ro, Hyun-Ju;Huh, Tae-Lin;Lee, Chang-Joong;Choi, Jin-Hee;Rhee, Myung-Chull
Animal cells and systems
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v.12
no.4
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pp.219-230
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2008
We are reporting the expression patterns and possible biological functions of a novel Kinesin-like protein, Surhe, in the zebrafish. Homology studies of derived amino acid sequences suggest that Surhe has an amino-terminal kinesin motor domain that is similar to that of the emerging MKLP-1 subfamily [Kim and Endow, 2000] and two coiledcoil domains in a central region. Cellular localization studies in mammalian cells revealed that Surhe protein is located in cytoplasm, suggesting that Surhe may be involved in the intracellular transport. During the developmental process, surhe transcripts are highly expressed in early embryonic stages. Overexpression of the dominant negative form of Surhe significantly down-regulates the dorsalization markers, such as goosecoid, bozozok, and chordin. Taken together, we postulate that Surhe may be involved in dorsalization process as a motor molecule.
Junction-mediating and regulatory protein (JMY) is a regulator of both transcription and actin filament assembly. The actin-regulatory activity of JMY is based on a cluster of three actin-binding Wiskott-Aldrich syndrome protein homology 2 (WH2) domains that nucleate actin filaments directly and promote nucleation of the Arp2/3 complex. In addition to these activities, we examined the activity of JMY generation in early embryo of mice carrying mutations in the JMY gene by CRISPR/Cas9 mediated genome engineering. We demonstrated that JMY protein shuttled expression between the cytoplasm and the nucleus. Knockout of exon 2, CA (central domain and Arp2/3-binding acidic domain) and NLS-2 (nuclear localization signal domain) on the JMY gene by CRISPR/Cas9 system was effective and markedly impeded embryonic development. Additionally, it impaired transcription and zygotic genome activation (ZGA)-related genes. These results suggest that JMY acts as a transcription factor, which is essential for the early embryonic development in mice.
The inner ear constitutes a complex organ responsible for auditory perception and equilibrium. It comprises diverse cellular entities operating collaboratively to perceive and transmit sensory information to the brain. Inner ear disease is a sophisticated and multifactorial scenario substantially impacting the quality of life of affected individuals. Gaining insights into the developmental process of the inner ear is crucial for diagnosing and treating inner ear diseases, which can lead to hearing loss and impaired balance. Recent research in inner ear development and associated pathophysiology has focused on several pivotal domains, including identifying new genes and signaling pathways involved in inner ear development, using stem cells for inner ear regeneration, and developing novel therapies for inner ear diseases. Recent advances in genetics research have shed new light on the fundamental etiologies of inner ear diseases, with a growing body of evidence suggesting that genetic mutations might exert a pivotal influence on the development and progression of this condition. In this review, we have delved into certain common genetic mutations linked to inner ear disorders. We also discussed ongoing research endeavors and future directions for understanding the genetic mechanisms underlying this condition and potential therapeutic avenues.
Fertilized egg, by successive cell divisions, differentiates into different tissues and organs with various structures and functions. Different cells and tissues contain different proteins, products of selective gene expression. Not all the genes in any genomes are equally active, temporal and spatial gene expression being the general rule. Present paper attempts to review the tanscriptional mechanisms or the initiations of transcription from several angles. In some of the organisms the genes in the process of transcription or the genes in the inactive state can be seen under the light microscope. Some bands of Drosophila polytene chromosomes may exhibit a swollen or puff appearance under certain conditions. A puff, unfolded or decondensed form of chromomere, represents sets of intense transcriptional activity or RNA synthesis. The heterochromatic X chromosome whose genes remain inactive in the female mammals can be visualized as a dark staining structure called Barr body, Configuration of chromatin differs between transcribed and nontranscribed chromatin. Modification to the chromatin facilitates RNA synthesis. The movement of large polymerase molecule along the DNA would probably be facilitated if some modifications of the chromatin configuration is effected. Methylation of cytosines in CG sequences is associated with inactive genes. Methylation can play a role in determination of mammalian cells during embryogenesis. Demethylation is necessary for the gene to be expressed during development A histone modification that is also known to be correlated with transcriptional capacity of chromatin is acetylation of the lysine residues of the core histones. Chromatin containing a high level of histone acetylation is very sensitive to DNase 1. For the transcription to occur TBP must first bind to the TATA box. Another TF, TF IIB, then binds to the promoter-TBP complex, facilitating the access of RNA polymerase to the transcription initiation site. As recently as eight years ago researchers assumed that histones were irrelevant to the regulation of gene expression. Histones combine with the DNA to form nucleosome of the chromatin. Histones are vital participant in gene regulation. Histone and basal factors compete for access to TATA box. When DNA is exposed to basal factors before histones are introduced, the basal factors assemble on TATA boxes preventing the access of histones, allowing transcription to occur, for transcription to begin, activator protein at the upstream activation sequence or enhancer must interact with the tail of histone H4 at TATA box and cause the histone role particle to dissociate from the TATA box leading to partial breakup of the histone core particle and allowing the basal factors to bind to the TATA box. New concept of genomic flux in contrast to the old concept of static genome has been developed based on the powerful new molecular techniques. Genomic changes such as repetitive DNAs and transposable elements, it is assumed but not yet proved, may affect some of the developmental patterns that characterize particular cells, tissues, organs, and organisms. In the last decade or so remarkable achievement have been made in the researches of the structures and functions of TFs and the specific target sequences located in promoters or enhancers where these TFs bind. TFs have independent domains that bind DNA and that activate transcription. DNA binding domain of TFs serves to bring the protein into the right location. There are many types of DNA binding domains. Common types of motifs can be found that are responsible for binding to DNA. The motifs are usually quite short and comprise only a small part of the protein structure. Steroid receptors have domains for hormone binding, DNA binding, and activating transcription. The zinc finger motif comprises a DNA binding domain. Leucine zipper consist of a stretch of amino acids with a leucine residue in every seventh position Two proteins form a dimer because they interact by means of leucine zippers on similar α-helical domain. This positions their DNA binding basic domains for interaction with the two halves of a DNA sequence with dyad symmetry of TGACTCA, ACTGAGT.
Ku, Bo Mi;Yune, Young Phil;Lee, Eun Shin;Hah, Young-Sool;Park, Jae Yong;Jeong, Joo Yeon;Lee, Dong Hoon;Cho, Gyeong Jae;Choi, Wan Sung;Kang, Sang Soo
Development and Reproduction
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v.17
no.4
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pp.299-309
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2013
Transforming growth factor (TGF) family is well known to induce the chondrogenic differentiation of mesenchymal stem cells (MSC). However, the precise signal transduction pathways and underlying factors are not well known. Thus the present study aims to evaluate the possible role of C2 domain in the chondrogenic differentiation of human mesenchymal stem cells. To this end, 145 C2 domains in the adenovirus were individually transfected to hMSC, and morphological changes were examined. Among 145 C2 domains, C2 domain of protein kinase C eta ($PKC{\eta}$) was selected as a possible chondrogenic differentiation factor for hMSC. To confirm this possibility, we treated $TGF{\beta}3$, a well known chondrogenic differentiation factor of hMSC, and examined the increased-expression of glycosaminoglycan (GAG), collagen type II (COL II) as well as $PKC{\eta}$ using PT-PCR, immunocytochemistry and Western blot analysis. To further evaluation of C2 domain of $PKC{\eta}$, we examined morphological changes, expressions of GAG and COL II after transfection of $PKC{\eta}$-C2 domain in hMSC. Overexpression of $PKC{\eta}$-C2 domain induced morphological change and increased GAG and COL II expressions. The present results demonstrate that $PKC{\eta}$ involves in the TGF-${\beta}3$-induced chondrogenic differentiation of hMSC, and C2 domain of $PKC{\eta}$ has important role in this process.
Local protein synthesis at subsynaptic sites plays a key role in the regulation of the protein composition in local domains. In this study, we carried out immunocytochemistry of cultured rat hippocampal neurons in various developmental stages to investigate the expression of eIF4E and its binding protein, eIF4EBP1. Both proteins were distributed in dendrites. In addition, eIF4EBP1 was highly expressed in the nucleus throughout the development, whereas eIF4E was not expressed in the nucleus. Punctate expression of eIF4E and eIF4EBP1 was evident in DIV 3. The colocalization rates of eIF4E or eIF4EBP1 puncta with PSD95 were higher in the dendrogenic than in the mature stages. In contrast, the colocalization rates of eIF4E and eIF4EBP1 puncta were higher in the mature than in the dendrogenic stages. As eIF4E is inactive when it is bound to eIF4EBP1, these data indicate that most dendritic eIF4E's are active during development but that they are mostly under inhibition in mature neurons.
Journal of Korean Academic Society of Home Health Care Nursing
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v.8
no.1
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pp.5-24
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2001
The purpose of this study was to develop a parenting intervention program and determine the efficacy of the program with low-birth weight infants and their mothers. Nine dyads for the experimental group and twelve dyads for the control group discharged from the Neonatal Intensive Care Unit of a University Hospital in Seoul were recruited for the study. For the intervention group, programmed education and support which focused on the maternal sensitivity of the infant's behavior. rearing environment. motherinfant interaction and infant care were given to each subject. Individual counseling and home visits were provided at discharge, one week after discharge. and one and three months of corrected age in every infant. Structured questionaires were administered and feeding interactions were videotaped and coded by a blinded certified observer. A Quasi-experimental design was conducted for this study. Postpartum depression, maternal self esteem. infant care burden, HOME. mother-infant interaction, and infant development were measured. Results were in favor of the intervention versus the control group. On the Beck depression inventory, intervention mothers showed decreasing trends in depressive symptom vs control mothers although, there were statistically no significant differences between the two groups at each time. The mean score of experimental group was 11.55(mild depression state) at discharge and became 8,6(normal state) at 1 month of corrected age. On the other hand, the mean score of the control group was 13.92(mild depression state) at discharge and became 14.0. Maternal self esteem in both groups improved over time. Infant care burden in both groups was also shown to increase over time. There was a significant difference between the two groups in HOME(p=.0340) at 3 months of corrected age. HOME scores of the experimental group and the control's were 31.10 and 25.58, respectively. Mothers' emotional and language responses were significantly high in the intervention group compared with the control group(p=.0155). Intervention group (53.33) showed a significantly high quality of motherinfant interaction compared with the in control group (42.80)(p =.0340). Intervention group mothers appeared have a better quality of mother-infant interaction behaviors. On the other hand, there was no statistical difference in the infant part between groups. Intervention group infants had higher trends in a general developmental quotient: although, there was no statistical difference between groups. The general developmental quotient of intervention infants was 102.56 and control's was 91.28. However, the developmental quotient of the domain of 'individuality-sociality' was higher in the intervention group infants compared with the control's(p=.0155). The concerns identified by parents revealed two domains of an infants' health management -knowledge and skills in caregiving of lowbirthweight-infants, characteristics of lowbirthweight infants, identifying a developmental milestone, coping with emergency situations and relaxation strategies of mothers from the infant care burden. Interview data with the mothers of low-birth weight infants can be used to develop intervention program contents. Limited intervention time and frequency due to time and cost limitations of this study should be modified. The intervention should be continuously implemented when low-birth weight infants become three years old. An NNNS demonstration appeared to be a very effective intervention for the mothers to improve the quality of mother-infant interactions. Therefore intervening in the mothers of low-birth weight infants as early after delivery as possible is desirable. This study has shown that home visit interventions are worthwhile for mothers only beyond the approach as an essential factor in ability of facilitating a growth fostering environment. In conclusion. the intervention program of this study was very effective in enhancing the parenting for the mothers of low-birth weight infants, resulting in health promotion of low-birth weight infants. The home-visit outreach intervention program of this study will contribute to the health delivery system in this country where there is a lack of continuous follow-up programs for low-birth weight infants after discharge from NICU, if it is activated as part of the home visit programs in community health systems.
Objective: This study aimed to review scales and tests developed for or validated with children and their families that were published in the Korean Journal of Child Studies from 1984 to 2016. Specifically, the purpose of the present study was to analyze the contents and format of the selected instruments, as well as to evaluate their psychometric properties. Methods: Using several databases and journal archives from the Korean Journal of Child Studies, searches were implemented using the key terms: instrument, scale, development, and validation. Instruments from 76 selected studies were reviewed based on several characteristics, such as assessment areas, contents, respondents, responding types, and psychometric properties. Results and Conclusion: First, a majority of the reviewed instruments were developed for infants and children, whereas only one was developed for adolescents. With regard to their specific measurement domains, many instruments focused on social emotional development among children. Second, with a few exceptions, the selected studies provided appropriate evidence for the reliability of the instrument, including its internal consistency, inter-rater reliability, and spilt-half reliability. Many studies also reported on the criterion-related or construct validity of the instrument to establish its validity. Future studies need to develop instruments across diverse developmental areas that collect information from multiple sources and raters. In addition, more evidence on the reliability and validity of the reviewed instruments should be provided to demonstrate their psychometric qualities.
Maeng, Sejung;Yoon, Sung Woo;Kim, Eun Jeong;Nam, Yoon Kwon;Sohn, Young Chang
Development and Reproduction
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v.23
no.4
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pp.333-344
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2019
In vertebrate reproductive system, estrogen receptor (ER) plays a pivotal role in mediation of estrogenic signaling pathways. In the present study, we report the cDNA cloning, expression analysis, and transcriptional activity of ERβ1 subtype from medaka Oryzias dancena. The deduced O. dancena ERβ1 (odERβ1; 519 amino acids) contained six characteristic A/B to E/F domains with very short activation function 2 region (called AF2). A phylogenetic analysis indicated that odERβ1 was highly conserved among teleost ERβ1 subgroup. A conventional RT-PCR revealed that the odERβ1 transcripts were widely distributed in the multiple tissues, the ovary, brain, gill, intestine, kidney, and muscle. Further, the relatively higher odERβ1 expressions in the ovary and brain were clearly reproduced in RT-qPCR assay. When HA-fused odERβ1 expression vector was transfected into HEK293 cells, an immunoreactivity for odERβ1 was mainly detected in the nucleus part. Finally, an estrogen responsive element driven luciferase reporter assays demonstrated that the transcriptional activity of odERβ1 significantly increased by estradiol-17β (E2) in a dose dependent manner (p<0.05). However, fold-activation of odERβ1 in the presence of E2 was markedly weak, when it compared with those of O. latipes ERβ1. Taken together, these data suggest that odERβ1 represents a functional variant of teleost ERβ subtype and provides a basic tool allowing future studies examining the function of F domain of ERβ1 subtype and expanding our knowledge of ERβ evolution.
Erythropoietin (EPO) is a glycoprotein hormone secreted from primarily cells of the peritubular capillary endothelium of the kidney, and is responsible for the regulation of red blood cell production. We constructed and expressed dimeric cDNAs in Chinease hamster ovary (CHO) cells encoding a fusion protein consisting of 2 complete human EPO domains linked by a 2-amino acid linker (Ile-Asp). We described the activity of dimeric hyperglycosylated EPO (dHGEPO) mutants containing additional oligosaccharide chains and characterized the function of glycosylation. No dimeric proteins with mutation at the $105^{th}$ amino acid were found in the cell medium. Growth and differentiation of the human EPO-dependent leukemiae cell line (F36E) were used to measure cytokine dependency and in vitro bioactivity of dHGEPO proteins. MIT assay at 24 h increased due to the survival of F36E cells. The dHGEPO protein migrated as a broad band with an average molecular mass of 75 kDa. The mutant, dHGEPO, was slightly higher than the wild-type (WT) dimeri-EPO band. Enzymatic N-deglycosylation resulted in the formation of a narrow band with a molecular mass twice of that of of monomeric EPO digested with an N-glycosylation enzyme. Hematocrit values were remarkably increased in all treatment groups. Pharmacokinetic analysis was also affected when 2.5 IU of dHGEPO were intravenously injected into the tails of the mice. The biological activity and half-life of dHGEPO mutants were enhanced as compared to the corresponding items associated the WT dimeric EPO. These results suggest that recombinant dHGEPO may be attractive biological and therapeutic targets.
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