Kunimasa Ohta

Kyushu University | Kyudai · Faculty of Arts and Science

Background: Although glioblastoma (GBM) stem like cells (GSCs), which exhibit chemo-radio resistance and recurrence, are key prognostic factors in GBM patients, molecular mechanisms of GSCs development are largely unknown. Recent studies revealed that extrinsic ribosome incorporation into somatic cells played key roles in cell reprogramming process...

bacterial ribosome stimuli promotes stem-cell like characteristics in mouse primary fibroblast

Tsukushi (TSK) proteoglycan dysfunction leads to hydrocephalus, a condition defined by excessive fluid collection in the ventricles and lateral ventricular enlargement. TSK injections into the LV at birth are effective at rescuing the lateral ventricle (LV). TSK regulates the activation of the Wnt signaling to facilitate the proper expansion of the...

Neurogenesis persists in selected regions of the adult mouse brain; among them, the ventricular-subventricular zone (V-SVZ) of the lateral ventricles represents a major experimental paradigm due to its conspicuous neurogenic output. Postnatal V-SVZ neurogenesis is maintained by a resident population of neural stem cells (NSCs). Although V-SVZ NSCs...

Glioblastoma multiforme (GBM), a lethal brain tumor, contains a small population of GBM stem cells (GSCs), which potentially cause chemotherapeutic resistance and tumor recurrence. Recent studies have reported that incorporation of ribosomes and ribosomal proteins into somatic cells promote lineage trans-differentiation toward multipotency. In this...

Tsukushi (TSK), a leucine-rich peptidoglycan in the extracellular compartment, mediates multiple signaling pathways that are critical for development and metabolism. TSK regulates signaling pathways that eventually control cellular communication, proliferation, and cell fate determination. Research on TSK has become more sophisticated in recent yea...

Although glioblastoma (GBM) stem-like cells (GSCs), which retain chemo-radio resistance and recurrence, are key prognostic factors in GBM patients, the molecular mechanisms of GSC development are largely unknown. Recently, several studies revealed that extrinsic ribosome incorporation into somatic cells resulted in stem cell properties and served a...

The ribosome, which is present in all three domains of life, plays a well-established, critical role in the translation process by decoding messenger RNA into protein. Ribosomal proteins, in contrast, appear to play non-translational roles in growth, differentiation, and disease. We recently discovered that ribosomes are involved in reverting cellu...

In the central nervous system (CNS), which comprises the eyes, spinal cord, and brain, neural cells are produced by the repeated division of neural stem cells (NSCs) during the development of the CNS. Contrary to the notion that the CNS is relatively static with a limited cell turnover, cells with stem cell-like properties have been isolated from m...

Previously we reported that, lactic acid bacteria (LAB) can induce human dermal fibroblast (HDF) cells to form multipotent cell clusters which are able to transdifferentiate into three germ layer derived cell lineages. Later on, we confirmed that ribosome is responsible for the LAB-induced transdifferentiation and ribosomes from diverse organisms c...

In 2006, Takahashi and Yamanaka reported the generation of mouse and human induced pluripotent stem cells (iPS cells) by retroviral gene transduction of four transcription factors, famously known as Yamanaka factors (OCT4, SOX2, Klf4, and c-Myc) (Takahashi et al., 2007; Takahashi and Yamanaka, 2006). iPS cells show stemness properties (self-renewal...

Tsukushi is a small, leucine-rich repeat proteoglycan that interacts with and regulates essential cellular signaling cascades in the chick retina and murine subventricular zone, hippocampus, dermal hair follicles, and the cochlea. However, its function in the vestibules of the inner ear remains unknown. Here, we investigated the function of Tsukush...

The lateral ventricle (LV) is flanked by the subventricular zone (SVZ), a neural stem cell (NSC) niche rich in extrinsic growth factors regulating NSC maintenance, proliferation, and neuronal differentiation. Dysregulation of the SVZ niche causes LV expansion, a condition known as hydrocephalus; however, the underlying pathological mechanisms are u...

Although ribosomes are generally known to be a translational machinery, some ribosomal proteins also have accessory functions involving early development and differentiation. Previously, we reported that ribosome incorporation into human dermal fibroblasts generated embryoid body-like cell clusters, altered cellular fate, and differentiated into ce...

In 2006, Takahashi and Yamanaka reported the generation of mouse and human induced pluripotent stem cells (iPS cells) by retroviral gene transduction of four transcription factors, famously known as Yamanaka factors (OCT4, SOX2, Klf4, and c-Myc) (Takahashi et al., 2007; Takahashi and Yamanaka, 2006). iPS cells show stemness properties (self-renewal...

Glioblastoma multiform (GBM), a lethal brain tumor developing in the white matter of the adult brain, contains a small population of GBM stem cells (GSCs), which potentially cause chemotherapeutic resistance and tumor recurrence. However, the mechanisms underlying the pathogenesis and maintenance of GSCs remain largely unknown. A recent study repor...

Tsukushi (TSK)-a small, secreted, leucine-rich-repeat proteoglycan-interacts with and regulates essential cellular signaling cascades. However, its functions in the mouse inner ear are unknown. In this study, measurement of auditory brainstem responses, fluorescence microscopy, and scanning electron microscopy revealed that TSK deficiency in mice r...

Secreted proteoglycan molecule Tsukushi (TSK) regulates various developmental processes, such as early body patterning and neural plate formation by interacting with major signaling pathways like Wnt, BMP, Notch etc. In central nervous system, TSK inhibits Wnt signaling to control chick retinal development. It also plays important roles for axon gu...

Specialized microenvironment, or neurogenic niche, in embryonic and postnatal mouse brain plays critical roles during neurogenesis throughout adulthood. The subventricular zone (SVZ) and the dentate gyrus (DG) of hippocampus in the mouse brain are two major neurogenic niches where neurogenesis is directed by numerous regulatory factors. Now, we rep...

Objectives: Tsukushi (TSK), a member of the small leucine-rich repeat proteoglycan (SLRP) family, plays multifunctional roles by interacting with signaling molecules during development. However, the role of TSK in cancer remains unknown. The aim of the present study was to investigate the biological significance of TSK in lung cancer. Materials a...

Thermogenesis is an important contributor to whole-body energy expenditure and metabolic homeostasis. Although circulating factors that promote energy expenditure are known, endocrine molecules that suppress energy expenditure have remained largely elusive. Here we found that Tsukushi (TSK) is a liver-enriched secreted factor that is highly inducib...

Extracellular molecules coordinate the multiple signaling pathways spatiotemporally to exchange information between cells during development. Understanding the regulation of these signal molecule-dependent pathways elucidates the mechanism of intercellular crosstalks. CCN2/CTGF is one of the CCN family members that binds BMP2, fibronectin, aggrecan...

Ribosomes are intracellular organelles ubiquitous in all organisms, which translate information from mRNAs to synthesize proteins. They are complex macromolecules composed of dozens of proteins and ribosomal RNAs. Other than translation, some ribosomal proteins also have side‐jobs called “Moonlighting” function. The majority of these moonlighting f...

Living organisms have been classified into three domains—archaea, eukaryota, and prokaryota—based on their cell structure and genetic evolution (Woese CR, Kandler O, Wheelis ML. Proc Natl Acad Sci USA 87:4576–4579, 1990). The eukaryotic cells have organelles that originated from prokaryotes living within these cells as endosymbionts (Martin W, Hoff...

Recently, we reported that bacterial incorporation induces cellular transdifferentiation of human fibroblasts. However, the bacterium-intrinsic cellular- transdifferentiation factor remained unknown. Here, we found that cellular transdifferentiation is caused by ribosomes. Ribosomes, isolated from both prokaryotic and eukaryotic cells, induce the f...

Tsukushi (TSK) is a small signaling molecule which takes part in different developmental processes of multiple vertebrate organisms. The diverse activity of TSK depends on its ability to bind various intermediate molecules from different major signaling pathways. Interactions of TSK with BMP, FGF, TGF-β and Wnt pathways have already been confirmed....

Sonic Hedgehog (Shh) is a ventrally enriched morphogen controlling dorsoventral patterning of the neural tube. In the dorsal spinal cord, Gli3 protein bound to suppressor-of-fused (Sufu) is converted into Gli3 repressor (Gli3R), which inhibits Shh-target genes. Activation of Shh signalling prevents Gli3R formation, promoting neural tube ventralizat...

Sirtuin 7 (SIRT7) is an NAD(+)-dependent deacetylase/deacylase, and is involved in a variety of biological processes relevant to the transcription of rRNA, the DNA damage response, tumorigenesis, and metabolism. SIRT7 mRNA is expressed ubiquitously, including in the brain, but there is no detailed information about the anatomical distribution and f...

Introduction Endochondral ossification is one of a key process for bone maturation. Tsukushi (TSK) is a novel member of the secreted small leucine-rich repeat proteoglycan (SLRP) family. SLRPs localize to skeletal regions and play significant roles during whole phases of bone development. Although prior evidence suggests that TSK may be involved in...

Though the adult central nervous system has been considered a comparatively static tissue with little turnover, it is well established today that new neural cells are generated throughout life. Neural stem/progenitor cells (NS/PCs) can self-renew and generate all types of neural cells. The proliferation of NS/PCs, and differentiation and fate deter...

The thalamocortical tract carries sensory information to the neocortex. It has long been recognized that the neocortical pioneer axons of subplate neurons are essential for thalamocortical development. Herein we report that an axon guidance cue, draxin, is expressed in early-born neocortical neurons, including subplate neurons, and is necessary for...

Supplementary Figures 1-8.

During somitogenesis, segmentation of the body axis occurs by epithelial somites budding off from the rostral end of the unsegmented presomitic mesoderm (PSM), and its molecular regulation is achieved by a molecular oscillator and signaling molecules. Tsukushi (TSK) is a unique secreted protein and involved in diverse biological cascades in vertebr...

Though the central nervous system is considered a comparatively static tissue with limited cell turnover, cells with stem-cell properties have been isolated from most neural tissues. The spinal cord ependymal cells show neural stem cell potential in vitro and in vivo in injured spinal cord. However, very little is known regarding the ependymal nich...

During the wound-healing process, macrophages, fibroblasts, and myofibroblasts play a leading role in shifting from the inflammation phase to the proliferation phase, although little is known about the cell differentiation and molecular control mechanisms underlying these processes. Previously, we reported that Tsukushi (TSK), a member of the small...

Since the days of Hans Spemann, the ocular lens has served as one of the most important developmental systems for elucidating the fundamental processes of induction and differentiation. Lens is an important source of signals that influence the eye development and a variety of genes expressed by the lens have been identified. The identification of a...

Temporal and spatial coordination of multiple signaling pathways is essential for finely tuned cellular regulation in development. Extracellular signaling molecules that function in a concentration-dependent manner, collectively called morphogens, and their inhibitors both play pivotal roles in the regulation of embryogenesis. Understanding concern...

Background: Adhesion molecules are known to be instructive for both development and differentiation. During lens differentiation, epithelial cells undergo vertical elongation, with the anterior and posterior tips of the elongating fiber cells sliding along the epithelium and capsule, respectively. These cellular processes are highly coordinated th...

Nestin is an intermediate filament found in neurogenic progenitors and non-neuronal cells. Nestin-immunoreactivity (IR) in the brain often increases after brain damage. Here we show that amygdala kindling, which mimics the epileptic seizures, also induces nestin expression in the brain. Nestin-IR was greatly enhanced in the leptomeninges (pia and a...

The human gastrointestinal tract is colonized by a vast community of symbionts and commensals. Lactic acid bacteria (LAB) form a group of related, low-GC-content, gram-positive bacteria that are considered to offer a number of probiotic benefits to general health. While the role of LAB in gastrointestinal microecology has been the subject of extens...

Typical LAB-incorporated cell clusters generated by HDFs and LAB. (A) The cell clusters were not generated from HDFs without LAB that were dissociated with trypsin/EDTA. (B) Lactobacillus acidophilus (JCM 1021) is able to generate LAB-incorporated cell clusters from HDFs that were dissociated with trypsin. (C) Lactococcus lactis subsp. lactis (JCM...

LAB-incorporated cell clusters are ALP positive. (A) LAB-incorporated cell clusters generated by Lactobacillus acidophilus (JCM 1021) from HDFs are ALP positive. (B) LAB-incorporated cell clusters generated by Lactococcus lactis subsp. lactis (JCM 20101), Streptococcus salivarius subsp. thermophilus (JCM 20026), or Lactobacillus sp. (JCM 20061) fro...

Double immunostaining. To examine the heterogeneous expression pattern of pluripotency markers in LAB-incorporated cell clusters at 14 days after incorporation, double staining was performed with rabbit anti-NANOG (ReproCELL) and mouse anti-OCT3/4 (Santa Cruz Biotechnology) antibodies. After three washes with PBS, the sections (10 µm) were incubate...

RT-PCR analysis. RT-PCR analysis of native HDFs, LAB-incorporated cell clusters, and LAB-incorporated cell clusters induced to differentiate in culture medium (DMEM/F12 medium containing 10% FCS) for 12 days. Note that LAB-incorporated cell clusters induced to differentiate express α-fetoprotein and GATA6. The PCR was performed with the following p...

Teratoma assay. The control testes and the testes injected with LAB-incorporated cell clusters at 14 days after incorporation were extracted, embedded in paraffin and sectioned in 6-µm intervals followed by deparaffinization in xylene and processing through a graded series of alcohol concentrations. The samples were stained with hematoxylin and eos...

Cell death. To examine cell death, LAB-incorporated cell clusters at 14 days after incorporation were immunostained with rabbit anti-single-stranded DNA antibody (DAKO). After three washes with PBS, the sections (10 µm) were incubated with anti-rabbit IgG antibody conjugated with FITC in the presence of DAPI in the antibody dilutions for 2 h at RT....

Comparison between HDF-1,2 and LAB-incorporated cell clusters-1,2. (PDF)

List of 7580 genes that shows the difference by factor of two or more between LAB-incorporated cell clusters-1 vs. HDF-1 and LAB-incorporated cell clusters-2 vs. HDF-2. (PDF)

Generation of GFP-MEF-derived LAB-incorporated cell clusters. Lactic acid bacteria (Lactobacillus acidophilus; JCM 1021) can generate cell clusters from LAB-incorporated GFP-MEFs. Lower (A, B) and higher (C) magnifications of LAB-incorporated GFP-MEF clusters after 6 days of incorporation. (D) LAB-incorporated GFP-MEF clusters are ALP positive. (A,...

Time course of the differentiation of LAB-incorporated cells. LAB-incorporated cell clusters after 14 days of incorporation were treated to the conditions that induced differentiation into adipocytes, osteocytes, or chondrocytes. The cell clusters were crumbled into the single cell and differentiate into adipocytes or osteocytes (attached condition...

Cell differentiation. LAB-incorporated GFP-MEF clusters were cultured on PLL-laminin-coated coverslips for 14 days in DMEM/F12 medium containing 1% FCS and 20 ng/ml bFGF (Sigma). The following primary antibodies were used for immunocytochemistry: rabbit anti-desmin (Thermo), mouse anti-Tuj1 (DSHB), rabbit anti-α-SMA (Thermo), and rabbit anti-GFAP (...

List of cluster genes that shows the difference by factor of two or more between LAB-incorporated cell clusters-1 vs. HDF-1 and LAB-incorporated cell clusters-2 vs. HDF-2. (PDF)

Selected 37 pluripotency maker genes. (PDF)

List of 50 genes that is decreased by a factor of fifteen or more at least between LAB-incorporated cell clusters-1 vs. HDF-1 and LAB-incorporated cell clusters-2 vs. HDF-2 and has a GO term [development]. (PDF)

We have shown that draxin is a repulsive axon guidance molecule for a variety of neuron classes and that genetic deletion of draxin in mice results in the absence of all forebrain commissures. Moreover, we also identified a secreted molecule, Tsukushi (TSK), that belongs to the small leucine-rich proteoglycan family (SLRP) and inhibits signaling mo...

The hair follicle contains stem/progenitor cells that supply progeny for skin development and the hair cycle. Several signaling molecules belonging to the Wnt, BMP, shh, and transforming growth factor β (TGF-β) signaling cascades are involved in the normal hair follicle cycle. However, the systemic mechanism of how these humoral factors are control...

Draxin, a recently identified axon guidance protein, is essential for the formation of forebrain commissures, and can mediate repulsion of netrin-stimulated spinal commissural axons. Here, we report that draxin binds multiple netrin receptors: DCC (deleted in colorectal cancer), Neogenin, UNC5s (H1, H2, H3), and DSCAM (Down's syndrome cell adhesion...

The Wnt signaling pathway is essential for the development of diverse tissues during embryogenesis. Signal transduction is activated by the binding of Wnt proteins to the type I receptor low-density lipoprotein receptor-related protein 5/6 and the seven-pass transmembrane protein Frizzled (Fzd), which contains a Wnt-binding site in the form of a cy...

The anterior commissure (AC) is one of the important commissure projections in the brain that conveys information from one side of the nervous system to the other. During development, the axons from the anterior AC (aAC) and the posterior AC (pAC) course in the same dorsoventral plane and converge into a common fascicle for midline crossing. Previo...

In tetrapod limbs, an anteriormost digit has common traits of small, short and less-phalange morphology. In this study, we focused on three genes, Mkp3, Sef and Tsukushi (TSK), which have anterior-specific or anterior-prominent expression patterns in the developing limb bud at the autopod-forming stage. The anterior expression is not fixed in the p...

PC3 is a member of the BTG/Tob family of antiproliferative genes. Here, we report the results of an analysis of PC3 protein expression in spiral ganglion neurons of the rat cochlea at embryonic days 16 (E16) and 20 (E20), and postnatal days 4 (P4) and 7 (P7). PC3 expression was observed in the cytoplasm of ganglion neurons at E16 and E20, and this...

Axon guidance proteins are critical for the correct wiring of the nervous system during development. Several axon guidance cues and their family members have been well characterized. More unidentified axon guidance cues are assumed to participate in the formation of the extremely complex nervous system. We identified a secreted protein, draxin, tha...

We acquire information from the outside world through our eyes which contain the retina, the photosensitive component of the central nervous system. Once the adult mammalian retina is damaged, the retinal neuronal death causes a severe loss of visual function. It has been believed that the adult mammalian retina had no regenerative capacity. Howeve...