9 results on '"Rumi Hasegawa"'
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2. Differentiation of stem progenitor CD9/SOX2-positive cells is promoted with increased prolactin-producing and endothelial cells in the pituitary
- Author
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Kotaro HORIGUCHI, Ken FUJIWARA, Takehiro TSUKADA, Takashi NAKAKURA, Saishu YOSHIDA, Rumi HASEGAWA, and Shu TAKIGAMI
- Subjects
cd9 ,lactotrophs ,pituitary gland ,pregnancy ,stem cells ,Reproduction ,QH471-489 ,Internal medicine ,RC31-1245 - Abstract
Sex-determining region Y-box 2 (SOX2)-positive cells are stem/progenitor cells in the adenohypophysis, comprising the anterior and intermediate lobes (AL and IL, respectively). The cells are located in the marginal cell layer (MCL) facing Rathke’s cleft (primary niche) and the parenchyma of the AL (secondary niche). We previously demonstrated in vitro that the tetraspanin superfamily CD9 and SOX2 double-positive (CD9/SOX2-positive) cells in the IL-side MCL migrate to the AL side and differentiate into hormone-producing and endothelial cells in the AL parenchyma. Here, we performed in vivo studies to evaluate the role of IL-side CD9/SOX2-positive cells in pregnancy, lactation, and treatment with diethylstilbestrol (DES; an estrogen analog) when an increased population of prolactin (PRL) cells was observed in the AL of the rat pituitary. The proportions of CD9/SOX2-, CD9/Ki67-, and PRL/TUNEL-positive cells decreased in the primary and secondary niches during pregnancy and DES treatment. In contrast, the number of CD9/PRL-positive cells increased in the AL-side MCL and AL parenchyma during pregnancy and during DES treatment. The proportion of PRL/Ki67-positive cells increased in the AL-side MCL and AL parenchyma in response to DES treatment. Next, we isolated CD9-positive cells from the IL-side MCL using an anti-CD9 antibody. During cell culture, the cells formed free-floating three-dimensional clusters (pituispheres). Furthermore, CD9-positive cells in the pituisphere differentiated into PRL cells, and their differentiation potential was promoted by DES. These findings suggest that CD9/SOX2-positive cells in the IL-side MCL may act as adult stem cells in the AL parenchyma that supply PRL cells under the influence of estrogen.
- Published
- 2022
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3. The multiciliated cells in Rathke’s cleft express CYP26A1 and respond to retinoic acid in the pituitary
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Kotaro Horiguchi, Ken Fujiwara, Takehiro Tsukada, Takashi Nakakura, Saishu Yoshida, Rumi Hasegawa, and Shu Takigami
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Male ,Histology ,Pituitary Gland, Anterior ,Pituitary Gland ,Animals ,Tretinoin ,Cell Biology ,Rats, Wistar ,Retinoic Acid 4-Hydroxylase ,Rats ,Pathology and Forensic Medicine - Abstract
The adenohypophysis consists of the anterior and intermediate lobes (AL and IL). The marginal cell layer (MCL), including the ventral region of the IL and the dorsal region of the AL lining the Rathke's cleft, acts as the primary stem/progenitor cell niches in adult adenohypophysis. The cells of the MCL on the IL side consisted of cluster of differentiation 9 (CD9)-positive stem/progenitor cells with or without motile cilia. However, any additional cellular properties of multiciliated CD9-positive cells are not known. The present study aimed to identify the character of the multiciliated cells in stem cell niche of the pituitary gland. We observed the fine structure of the multiciliated cells in the MCL of male Wistar rats at an early stage after birth and in adulthood (P60) using scanning electron microscopy. Since the previous study showed that the MCL cells of adult rats synthesize retinoic acid (RA), the present study determined whether the multiciliated cells are involved in RA regulation by the expression of retinal aldehyde dehydrogenase 1 (RALDH1) and CYP26A1, an enzyme synthesizing and degrading RA, respectively. Results showed that 96% of multiciliated cells in adult male rats expressed CYP26A1, while 60% expressed RALDH1. Furthermore, the isolated CD9-positive cells from the IL side MCL responded to RA and activated the degradation system of RA by increasing Cyp26a1 expression. These findings indicated that multiciliated cells are involved in RA metabolism in the MCL. Our observations provide novel insights regarding the stem cell niche of the adult pituitary.
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- 2022
4. CD9-positive cells in the intermediate lobe of the pituitary gland are important supplier for prolactin-producing cells in the anterior lobe
- Author
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Ken Fujiwara, Rumi Hasegawa, Saishu Yoshida, Shu Takigami, Yoshito Takeda, Kotaro Horiguchi, Takehiro Tsukada, Takashi Nakakura, and Shunji Ohsako
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Male ,0301 basic medicine ,Pituitary gland ,Histology ,Cell ,Population ,Biology ,Tetraspanin 29 ,Pathology and Forensic Medicine ,Prolactin cell ,Mice ,03 medical and health sciences ,0302 clinical medicine ,SOX2 ,medicine ,Animals ,Rats, Wistar ,Progenitor cell ,education ,education.field_of_study ,Cell Biology ,Prolactin ,Rats ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Pituitary Gland ,embryonic structures ,Stem cell ,030217 neurology & neurosurgery - Abstract
A supply of hormone-producing cells from stem/progenitor cells is critical to sustain the endocrine activity of the pituitary gland. In the adenohypophysis composing the anterior and intermediate lobe (AL and IL, respectively), stem/progenitor cells expressing sex-determining region Y-box 2 (SOX2) and S100β are located in the marginal cell layer (MCL) facing Rathke's cleft (primary niche) and the parenchyma of the AL (secondary niche). Our previous studies using mice and rats indicated that the tetraspanin superfamily CD9 and CD81 are expressed in S100β/SOX2-positive cells of primary and secondary niches (named CD9/CD81/S100β/SOX2-positive cell), and the cells located in the AL-side niches exhibit plasticity and multipotency. However, it is unclear whether CD9/CD81/S100β/SOX2-positive cells in the IL-side primary niche are stem/progenitor cells for the AL or IL. Here, we successfully isolated pure CD9/CD81/S100β/SOX2-positive cells from the IL-side primary niche. They had a higher level of S100β and SOX2 mRNA and a greater pituisphere forming capacity than those of CD9/CD81/S100β/SOX2-positive cells isolated from the AL. They also had capacity to differentiate into all types of adenohypophyseal hormone-producing cells, concomitantly with the loss of CD9 expression. Loss of CD9 and CD81 function in CD9/CD81/S100β/SOX2-positive cells by siRNA treatment impaired prolactin cell differentiation. Consistently, in the pituitary gland of CD9/CD81 double knockout mice, dysgenesis of the MCL and a lower population of prolactin cells were observed. These results suggest that the CD9/CD81/S100β/SOX2-positive cells in the MCL of the IL-side are potential suppliers of adult core stem cells in the AL.
- Published
- 2021
5. CX3CL1/CX3CR1-signalling in the CD9/S100β/SOX2-positive adult pituitary stem/progenitor cells modulates differentiation into endothelial cells
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Ken Fujiwara, Takehiro Tsukada, Yukio Kato, Shunji Ohsako, Rumi Hasegawa, Kotaro Horiguchi, Saishu Yoshida, Takako Kato, Takashi Yashiro, and Shu Takigami
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Male ,0301 basic medicine ,Chemokine ,Histology ,CX3C Chemokine Receptor 1 ,S100 Calcium Binding Protein beta Subunit ,Tetraspanin 29 ,Vascular remodelling in the embryo ,03 medical and health sciences ,SOX2 ,Precursor cell ,CX3CR1 ,Animals ,Rats, Wistar ,Progenitor cell ,Receptor ,CX3CL1 ,Molecular Biology ,030102 biochemistry & molecular biology ,biology ,Chemokine CX3CL1 ,SOXB1 Transcription Factors ,Stem Cells ,Endothelial Cells ,Cell Differentiation ,Cell Biology ,Rats, Inbred F344 ,Rats ,Cell biology ,Medical Laboratory Technology ,030104 developmental biology ,Pituitary Gland ,embryonic structures ,biology.protein ,Signal Transduction - Abstract
Approximately 8% of CD9-, S100β- and SOX2-triple positive (CD9/S100β/SOX2-positive) stem/progenitor cells in the anterior lobe of the rat pituitary gland have previously been shown to differentiate into endothelial cells in vitro, suggesting that they play a role in vascularisation as tissue-resident vascular precursor cells. In the present study, we focused on chemokine ligands to further characterise the CD9/S100β/SOX2-positive cells and found that they distinctively express CX3C chemokine ligand 1 (Cx3cl1). Immunohistochemical analysis of the anterior lobe showed that CX3CL1-positive cells comprised 7.8% in CD9-positive cells. By cultivation of the CD9-positive cells on laminin-coated plates, we observed that the expression levels of Cx3cl1 decreased, while those of Sox18, an endothelial cell-progenitor marker, and Cx3cr1, a CX3CL1 receptor, increased. Furthermore, in a rat model of prolactinoma, the most common pituitary tumour, which is accompanied by frequent neo-vasculogenesis in the anterior lobe, we have confirmed a decrease in Cx3cl1 expression and an increase in Cx3cr1 expression, as well as a prominent increase in Sox18 expression. These findings suggest that CX3CL1/CX3CR1 signalling in CD9/S100β/SOX2-positive cells plays an important role in resupplying endothelial cells for vascular remodelling in the anterior lobe.
- Published
- 2020
6. Cluster of differentiation (CD) 9-positive mouse pituitary cells are adult stem/progenitor cells
- Author
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Rumi Hasegawa, Takashi Nakakura, Takehiro Tsukada, Saishu Yoshida, Shu Takigami, Shunji Ohsako, Kotaro Horiguchi, and Ken Fujiwara
- Subjects
0301 basic medicine ,Male ,Pituitary gland ,Histology ,Cell ,Biology ,Antibodies ,Tetraspanin 29 ,03 medical and health sciences ,Mice ,SOX2 ,medicine ,Animals ,Progenitor cell ,Molecular Biology ,Mice, Inbred ICR ,030102 biochemistry & molecular biology ,Cluster of differentiation ,Cell growth ,Stem Cells ,Cell Differentiation ,Cell Biology ,Immunohistochemistry ,Cell biology ,Medical Laboratory Technology ,030104 developmental biology ,medicine.anatomical_structure ,Pituitary Gland ,embryonic structures ,Stem cell ,Developmental biology - Abstract
SOX2-positive cells are stem/progenitor cells that supply hormone-producing cells; they are found in the anterior lobe of the rodent pituitary gland. However, they are likely composed of several subpopulations. In rats, a SOX2-positive cell populations can be distinguished by the presence of S100β. We identified the novel markers cluster of differentiation (CD) CD9 and CD81, members of the tetraspanin superfamily, for the identification of S100β/SOX2-positive cells. Recently, CD9/CD81 double-knockout mice were generated. Although they grew normally until 3 weeks after birth, they exhibited atrophy of the pituitary gland. These findings suggested that CD9/CD81/S100β/SOX2-positive cells in the mouse pituitary are adult stem/progenitor cells. To substantiate this hypothesis, we examined CD9 and CD81 expression in the adult and developing anterior lobe. Immunohistochemistry showed that CD9/CD81-positive cells began appearing from postnatal day 0 and settled in the stem cell niches (marginal cell layer and parenchyma) of the adult anterior lobe while expressing S100β. We next isolated CD9 -positive cells from the adult anterior lobe, using the anti-CD9 antibody for cell characterisation. The cells in culture formed free-floating three-dimensional clusters (pituispheres); moreover, induction into all types of hormone-producing cells was successful. Furthermore, reduction of CD9 and CD81 mRNAs by siRNAs inhibited cell proliferation. These findings indicate that CD9/CD81/S100β/SOX2-positive cells may play a role as adult stem/progenitor cells in SOX2-positive subpopulations, thus supplying hormone-producing cells in the postnatal anterior lobe. Furthermore, CD9 and CD81 are implicated in cell proliferation. The current findings provide novel insights into adult pituitary stem/progenitor cells.
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- 2020
7. Identification of THY1 as a novel thyrotrope marker and THY1 antibody-mediated thyrotrope isolation in the rat anterior pituitary gland
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Shu Takigami, Takako Kato, Rumi Hasegawa, Takehiro Tsukada, Shunji Ohsako, Takashi Nakakura, Naoko Kanno, Yukio Kato, Kotaro Horiguchi, and Saishu Yoshida
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Male ,0301 basic medicine ,medicine.medical_specialty ,Pituitary gland ,Biophysics ,Thyrotropin ,Cell Separation ,In situ hybridization ,urologic and male genital diseases ,Biochemistry ,03 medical and health sciences ,Anterior pituitary ,Antigen ,Pituitary Gland, Anterior ,Thyrotropic cell ,Internal medicine ,medicine ,Animals ,CD90 ,Rats, Wistar ,Molecular Biology ,Thymocytes ,biology ,Cell Biology ,Juxtacrine signalling ,Molecular biology ,030104 developmental biology ,Endocrinology ,medicine.anatomical_structure ,CD18 Antigens ,biology.protein ,Thy-1 Antigens ,Antibody ,Biomarkers - Abstract
Contact-dependent (juxtacrine) signaling is important for local cell-to-cell interaction and has received attention in recent years regarding its role in pituitary function, differentiation, and development. This study investigated one of the juxtacrine-related molecules, thymocyte differentiation antigen 1 (THY1), in the anterior lobe of the rat pituitary gland. Western blot analysis revealed expression of the THY1 protein in the adult rat anterior lobe. We also found that the THY1 ligand, integrin-β2 (ITGB2), is also expressed in the pituitary gland. In situ hybridization and immunohistochemical analyses showed that both THY1 mRNA and protein were present in almost, if not all, thyroid-stimulating hormone (TSH)-immunopositive cells (thyrotropes) and that ITGB2 was co-expressed in these cells. As THY1 appeared to represent a novel marker for thyrotropes, we then attempted to isolate these cells from various anterior lobe cells by the use of a THY1 antibody and a pluriBead-cascade cell isolation system. This technology allowed the isolation of thyrotropes with 83% purity at about 17-fold enrichment. Furthermore, the isolated THY1-immunopositive cells had higher Tsh mRNA levels compared with THY1-immunonegative cells and released TSH in response to thyrotropin-releasing hormone. These findings indicated that THY1 represents a potent thyrotrope marker and that the thyrotrope isolation method using the THY1 antibody may serve as a powerful tool to analyze their function including juxtacrine regulation through THY1/ITGB2 interaction.
- Published
- 2016
8. Isolation and characterisation of CD9-positive pituitary adult stem/progenitor cells in rats
- Author
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Shu Takigami, Yukio Kato, Takehiro Tsukada, Rumi Hasegawa, Ken Fujiwara, Shunji Ohsako, Takashi Nakakura, Takako Kato, Saishu Yoshida, Kotaro Horiguchi, Takashi Yashiro, and Ken Arae
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0301 basic medicine ,Male ,Pituitary gland ,Endothelium ,Cellular differentiation ,lcsh:Medicine ,S100 Calcium Binding Protein beta Subunit ,Bone morphogenetic protein ,Tetraspanin 29 ,Article ,03 medical and health sciences ,Pituitary Gland, Anterior ,medicine ,Animals ,Prolactinoma ,Progenitor cell ,Rats, Wistar ,lcsh:Science ,Cells, Cultured ,Cell Proliferation ,Multidisciplinary ,Cluster of differentiation ,biology ,lcsh:R ,Cell Differentiation ,In vitro ,Cell biology ,Rats ,Adult Stem Cells ,030104 developmental biology ,medicine.anatomical_structure ,embryonic structures ,biology.protein ,lcsh:Q ,Endothelium, Vascular ,Antibody - Abstract
S100β protein and SOX2-double positive (S100β/SOX2-positive) cells have been suggested to be adult pituitary stem/progenitor cells exhibiting plasticity and multipotency. The aim of the present study was to isolate S100β/SOX2-positive cells from the adult anterior lobes of rats using a specific antibody against a novel membrane marker and to study their characteristics in vitro. We found that cluster of differentiation (CD) 9 is expressed in the majority of adult rat S100β/SOX2-positive cells, and we succeeded in isolating CD9-positive cells using an anti-CD9 antibody with a pluriBead-cascade cell isolation system. Cultivation of these cells showed their capacity to differentiate into endothelial cells via bone morphogenetic protein signalling. By using the anterior lobes of prolactinoma model rats, the localisation of CD9-positive cells was confirmed in the tumour-induced neovascularisation region. Thus, the present study provides novel insights into adult pituitary stem/progenitor cells involved in the vascularisation of the anterior lobe.
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- 2017
9. Expression of chemokine CXCL10 in dendritic-cell-like S100β-positive cells in rat anterior pituitary gland
- Author
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Takehiro Tsukada, Ken Fujiwara, Hiroki Ueharu, Masashi Higuchi, Takako Kato, Rumi Hasegawa, Kotaro Horiguchi, Shu Takigami, Takashi Yashiro, Yukio Kato, Mo Chen, Shunji Ohsako, and Saishu Yoshida
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Male ,Pituitary gland ,medicine.medical_specialty ,Histology ,S100 Calcium Binding Protein beta Subunit ,CXCR3 ,Ligands ,Pathology and Forensic Medicine ,Anterior pituitary ,Pituitary Gland, Anterior ,Internal medicine ,medicine ,CXCL10 ,Animals ,Rats, Wistar ,CXCL14 ,CXCL16 ,Cells, Cultured ,Chemistry ,Cell Biology ,Dendritic cell ,Dendritic Cells ,Chemokine CXCL12 ,Cell biology ,Chemokine CXCL10 ,Protein Transport ,medicine.anatomical_structure ,Endocrinology ,XCL2 ,Receptors, Chemokine ,Rats, Transgenic - Abstract
Chemokines are mostly small secreted polypeptides whose signals are mediated by seven trans-membrane G-protein-coupled receptors. Their functions include the control of leukocytes and the intercellular mediation of cell migration, proliferation, and adhesion in several tissues. We have previously revealed that the CXC chemokine ligand 12 (CXCL12) and its receptor 4 (CXCR4) are expressed in the anterior pituitary gland, and that the CXCL12/CXCR4 axis evokes the migration and interconnection of S100β-protein-positive cells (S100β-positive cells), which do not produce classical anterior pituitary hormones. However, little is known of the cells producing the other CXCLs and CXCRs or of their characteristics in the anterior pituitary. We therefore examined whether CXCLs and CXCRs occurred in the rat anterior pituitary lobe. We used reverse transcription plus the polymerase chain reaction to analyze the expression of Cxcl and Cxcr and identified the cells that expressed Cxcl by in situ hybridization. Transcripts of Cxcl10 and its receptor (Cxcr3 and toll-like receptor 4, Tlr4) were clearly detected: cells expressing Cxcl10 and Tlr4 were identified amongst S100β-positive cells and those expressing Cxcr3 amongst adrenocorticotropic hormone (ACTH)-producing cells. We also investigated Cxcl10 expression in subpopulations of S100β-positive cells. We separated cultured S100β-positive cells into the round-type (dendritic-cell-like) and process-type (astrocyte- or epithelial-cell-like) by their adherent activity to laminin, a component of the extracellular matrix; CXCL10 was expressed only in round-type S100β-positive cells. Thus, CXCL10 produced by a subpopulation of S100β-positive cells probably exerts an autocrine/paracrine effect on S100β-positive cells and ACTH-producing cells in the anterior lobe.
- Published
- 2014
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