33 results on '"Shu TAKIGAMI"'
Search Results
2. Fluctuation of CD9/SOX2-positive cell populations during the turnover of GH- and TSH-producing cells in the adult anterior pituitary gland
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Kotaro HORIGUCHI, Yuto TSUTSUI, Ken FUJIWARA, Takehiro TSUKADA, Takashi NAKAKURA, Saishu YOSHIDA, Rumi HASEGAWA, and Shu TAKIGAMI
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cluster of differentiation 9 (cd9) ,growth hormone (gh) ,pituitary ,sex-determining region y-box 2 (sox2) ,thyroid-stimulating hormone (tsh) ,Reproduction ,QH471-489 ,Internal medicine ,RC31-1245 - Abstract
The adenohypophysis is comprised of the anterior and intermediate lobes (AL and IL, respectively). Cluster of differentiation 9 (CD9)- and sex-determining region Y-box 2 (SOX2)-positive cells are stem/progenitor hormone-producing cells in the AL. They are located in the marginal cell layer (MCL) facing Rathke’s cleft between the AL and IL (primary niche) and the parenchyma of the AL (secondary niche). We previously showed that, in rats, CD9/SOX2-positive cells in the IL side of the MCL (IL-side MCL) migrate to the AL side (AL-side MCL) and differentiate into prolactin-producing cells (PRL cells) in the AL parenchyma during pregnancy, lactation, and diethylstilbestrol treatment, all of which increase PRL cell turnover. This study examined the changes in CD9/SOX2-positive stem/progenitor cell niches and their proportions by manipulating the turnover of growth hormone (GH)- and thyroid-stimulating hormone (TSH)-producing cells (GH and TSH cells, respectively), which are Pit1 lineage cells, as well as PRL cells. After induction, the isolated CD9/SOX2-positive cells from the IL-side MCL formed spheres and differentiated into GH and TSH cells. We also observed an increased GH cell proportion upon treatment with GH-releasing hormone and recovery from continuous stress and an increased TSH cell proportion upon propylthiouracil treatment, concomitant with alterations in the proportion of CD9/SOX2-positive cells in the primary and secondary niches. These findings suggest that CD9/SOX2-positive cells have the potential to supply GH and TSH when an increase in GH and TSH cell populations is required in the adult pituitary gland.
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- 2023
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3. Differentiation of stem progenitor CD9/SOX2-positive cells is promoted with increased prolactin-producing and endothelial cells 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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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.
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- 2022
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4. Expression and functions of cluster of differentiation 9 and 81 in rat mammary epithelial cells
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Kotaro HORIGUCHI, Saishu YOSHIDA, Takehiro TSUKADA, Takashi NAKAKURA, Ken FUJIWARA, Rumi HASEGAWA, Shu TAKIGAMI, and Shunji OHSAKO
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cluster of differentiation (cd) 9 ,cd81 ,lactation ,mammary gland ,proliferation ,Reproduction ,QH471-489 ,Internal medicine ,RC31-1245 - Abstract
Cluster of differentiation (CD) 9 and CD81 are closely-related members of the tetraspanin family that consist of four-transmembrane domain proteins. Cd9 and Cd81 are highly expressed in breast cancer cells; however, their expression in healthy mammary glands is unclear. In this study, we performed quantitative real-time PCR to analyze the expression levels of Cd9 and Cd81. Histological techniques were employed to identify Cd9- and Cd81-expressing cells in rat mammary glands during pregnancy and lactation. It was observed that Cd9 and Cd81 were expressed in the mammary glands, and their expression levels correlated with mammary gland development. To identify cells expressing Cd9 and Cd81 in the mammary glands, we performed double immunohistochemical staining for CD9 and CD81, prolactin receptor long form, estrogen receptor alpha, or Ki67. The results showed that CD9 and CD81 were co-expressed in proliferating mammary epithelial cells. Next, we attempted to isolate CD9-positive epithelial cells from the mammary gland using pluriBead cell-separation technology based on antibody-mediated binding of cells to beads of different sizes, followed by isolation using sieves with different mesh sizes. We successfully isolated CD9-positive epithelial cells with 96.8% purity. In addition, we observed that small-interfering RNAs against Cd9 and Cd81 inhibited estrogen-induced proliferation of CD9-positive mammary epithelial cells. Our current findings may provide novel insights into the proliferation of mammary epithelial cells during pregnancy and lactation as well as in pathological processes associated with breast cancer.
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- 2020
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5. 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
6. CD9-positive cells in the intermediate lobe of the pituitary gland are important supplier for prolactin-producing cells in the anterior lobe
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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
7. 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.
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- 2020
8. Expression and functions of cluster of differentiation 9 and 81 in rat mammary epithelial cells
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Takashi Nakakura, Rumi Hasegawa, Shunji Ohsako, Takehiro Tsukada, Ken Fujiwara, Kotaro Horiguchi, Shu Takigami, and Saishu Yoshida
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Mammary gland ,Proliferation ,Biology ,Real-Time Polymerase Chain Reaction ,Tetraspanin 29 ,Tetraspanin 28 ,Mammary Glands, Animal ,CD81 ,Tetraspanin ,Pregnancy ,Lactation ,medicine ,Animals ,RNA, Small Interfering ,Rats, Wistar ,Diethylstilbestrol ,Cell Proliferation ,Cluster of differentiation ,Gene Expression Profiling ,Prolactin receptor ,Estrogen Receptor alpha ,Cell Differentiation ,Epithelial Cells ,Rats ,Cell biology ,Ki-67 Antigen ,medicine.anatomical_structure ,embryonic structures ,Pregnancy, Animal ,Immunohistochemistry ,Female ,Original Article ,Cluster of differentiation (CD) 9 ,Animal Science and Zoology ,Estrogen receptor alpha - Abstract
Cluster of differentiation (CD) 9 and CD81 are closely-related members of the tetraspanin family that consist of four-transmembrane domain proteins. Cd9 and Cd81 are highly expressed in breast cancer cells; however, their expression in healthy mammary glands is unclear. In this study, we performed quantitative real-time PCR to analyze the expression levels of Cd9 and Cd81. Histological techniques were employed to identify Cd9- and Cd81-expressing cells in rat mammary glands during pregnancy and lactation. It was observed that Cd9 and Cd81 were expressed in the mammary glands, and their expression levels correlated with mammary gland development. To identify cells expressing Cd9 and Cd81 in the mammary glands, we performed double immunohistochemical staining for CD9 and CD81, prolactin receptor long form, estrogen receptor alpha, or Ki67. The results showed that CD9 and CD81 were co-expressed in proliferating mammary epithelial cells. Next, we attempted to isolate CD9-positive epithelial cells from the mammary gland using pluriBead cell-separation technology based on antibody-mediated binding of cells to beads of different sizes, followed by isolation using sieves with different mesh sizes. We successfully isolated CD9-positive epithelial cells with 96.8% purity. In addition, we observed that small-interfering RNAs against Cd9 and Cd81 inhibited estrogen-induced proliferation of CD9-positive mammary epithelial cells. Our current findings may provide novel insights into the proliferation of mammary epithelial cells during pregnancy and lactation as well as in pathological processes associated with breast cancer.
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- 2020
9. CD9-positive cells in the intermediate lobe migrate into the anterior lobe to supply endocrine cells
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Rumi Hasegawa, Shunji Ohsako, Takashi Nakakura, Takehiro Tsukada, Saishu Yoshida, Kotaro Horiguchi, Ken Fujiwara, and Shu Takigami
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0301 basic medicine ,Male ,Histology ,Enteroendocrine cell ,Tetraspanin 29 ,03 medical and health sciences ,Tissue culture ,Downregulation and upregulation ,Cell Movement ,Pituitary Gland, Anterior ,Animals ,Progenitor cell ,Rats, Wistar ,Molecular Biology ,030102 biochemistry & molecular biology ,Cluster of differentiation ,Chemistry ,Cell migration ,Cell Biology ,Cell biology ,Rats ,Medical Laboratory Technology ,030104 developmental biology ,embryonic structures ,Endocrine Cells ,Developmental biology ,CD81 - Abstract
The adenohypophysis is composed of the anterior and intermediate lobes (AL and IL), and secretes important hormones for growth, sexual development, metabolism, and reproduction. In the marginal cell layer (MCL) facing Rathke's cleft between the IL and AL, cluster of differentiation (CD) 9-, CD81-, S100β-, and SOX2-quadruple positive (CD9/CD81/S100β/SOX2-positive) cells in the adult IL are settled as tissue-resident stem/progenitor cells supplying hormone-producing cells to the AL. However, it is unclear how CD9/CD81/S100β/SOX2-positive cells in the IL-side MCL migrate into the AL across Rathke's cleft. In the present study, we performed chimeric pituitary tissue culture using S100β/GFP-transgenic rats and Wistar rats, and traced the footprint of S100β/GFP-expressing cells. We detected IL-side S100β/GFP-expressing cells in the AL tissue, demonstrating that these cells migrate from the IL to the AL. However, the cells failed to migrate in the opposite direction. Consistently, scanning electron microscopic analysis revealed well-developed cytoplasmic protrusions in the IL-side MCL, but not in the AL-side MCL, suggesting that IL-side CD9/CD81/S100β/SOX2-positive cells had higher migratory activity. We also searched for a specific marker for IL-side CD9/CD81/S100β/SOX2-positive cells and identified tetraspanin 1 (TSPAN1) from microarray analysis. Downregulation of Tspan1 by specific siRNA impaired cell migration and significantly reduced expression of snail family transcriptional repressor 2 (Slug), a marker of epithelial-mesenchymal transition (EMT). Therefore, CD9/CD81/S100β/SOX2-positive cells in the IL-side MCL can be stem/progenitor cells that provide stem/progenitor cells to the AL-side MCL via SLUG-mediated EMT and cell migration.
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- 2021
10. Cluster of differentiation (CD) 9-positive mouse pituitary cells are adult stem/progenitor cells
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Rumi Hasegawa, Takashi Nakakura, Takehiro Tsukada, Saishu Yoshida, Shu Takigami, Shunji Ohsako, Kotaro Horiguchi, and Ken Fujiwara
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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
11. 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.
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- 2016
12. Isolation and characterization of cluster of differentiation 9-positive ependymal cells as potential adult neural stem/progenitor cells in the third ventricle of adult rats
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Rumi Hasegawa, Takako Kato, Saishu Yoshida, Kotaro Horiguchi, Yukio Kato, Shu Takigami, and Shunji Ohsako
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0301 basic medicine ,Male ,Histology ,Ependymal Cell ,Biology ,Tetraspanin 29 ,Pathology and Forensic Medicine ,03 medical and health sciences ,0302 clinical medicine ,Neural Stem Cells ,Neurosphere ,Ependyma ,medicine ,Animals ,Progenitor cell ,Rats, Wistar ,In Situ Hybridization ,Cell Proliferation ,Third Ventricle ,Third ventricle ,Cluster of differentiation ,Stem Cells ,Cell Differentiation ,Cell Biology ,Immunohistochemistry ,Oligodendrocyte ,Cell biology ,Rats ,030104 developmental biology ,medicine.anatomical_structure ,Neuron ,030217 neurology & neurosurgery ,Astrocyte - Abstract
Ependymal cells located above the ventricular zone of the lateral, third, and fourth ventricles and the spinal cord are thought to form part of the adult neurogenic niche. Many studies have focused on ependymal cells as potential adult neural stem/progenitor cells. To investigate the functions of ependymal cells, a simple method to isolate subtypes is needed. Accordingly, in this study, we evaluated the expression of cluster of differentiation (CD) 9 in ependymal cells by in situ hybridization and immunohistochemistry. Our results showed that CD9-positive ependymal cells were also immunopositive for SRY-box 2, a stem/progenitor cell marker. We then isolated CD9-positive ependymal cells from the third ventricle using the pluriBead-cascade cell isolation system based on antibody-mediated binding of cells to beads of different sizes and their isolation with sieves of different mesh sizes. As a result, we succeeded in isolating CD9-positive populations with 86% purity of ependymal cells from the third ventricle. We next assayed whether isolated CD9-positive ependymal cells had neurospherogenic potential. Neurospheres were generated from CD9-positive ependymal cells of adult rats and were immunopositve for neuron, astrocyte, and oligodendrocyte markers after cultivation. Thus, based on these findings, we suggest that the isolated CD9-positive ependymal cells from the third ventricle included tanycytes, which are special ependymal cells in the ventricular zone of the third ventricle that form part of the adult neurogenic and gliogenic niche. These current findings improve our understanding of tanycytes in the adult third ventricle in vitro.
- Published
- 2019
13. CXCL10/CXCR3 signaling mediates inhibitory action by interferon-gamma on CRF-stimulated adrenocorticotropic hormone (ACTH) release
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Takehiro Tsukada, Takashi Yashiro, Kozue Tateno, Shunji Ohsako, Kotaro Horiguchi, Masashi Higuchi, Shu Takigami, Ken Fujiwara, Rumi Hasegawa, Takako Kato, Saishu Yoshida, and Yukio Kato
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Male ,0301 basic medicine ,Agonist ,endocrine system ,medicine.medical_specialty ,Pro-Opiomelanocortin ,Receptors, CXCR3 ,Histology ,Corticotropin-Releasing Hormone ,medicine.drug_class ,medicine.medical_treatment ,S100 Calcium Binding Protein beta Subunit ,Adrenocorticotropic hormone ,CXCR3 ,Pathology and Forensic Medicine ,Interferon-gamma ,03 medical and health sciences ,Paracrine signalling ,Adrenocorticotropic Hormone ,Anterior pituitary ,Pituitary Gland, Anterior ,Internal medicine ,medicine ,Animals ,CXCL10 ,Rats, Wistar ,Cells, Cultured ,Inflammation ,Chemistry ,hemic and immune systems ,Cell Biology ,Rats ,Chemokine CXCL10 ,030104 developmental biology ,Endocrinology ,Cytokine ,medicine.anatomical_structure ,Corticotropic cell ,Rats, Transgenic ,hormones, hormone substitutes, and hormone antagonists ,Signal Transduction - Abstract
Secretion of hormones by the anterior pituitary gland can be stimulated or inhibited by paracrine factors that are produced during inflammatory reactions. The inflammation cytokine interferon-gamma (IFN-γ) is known to inhibit corticotropin-releasing factor (CRF)-stimulated adrenocorticotropin (ACTH) release but its signaling mechanism is not yet known. Using rat anterior pituitary, we previously demonstrated that the CXC chemokine ligand 10 (CXCL10), known as interferon-γ (IFN-γ) inducible protein 10 kDa, is expressed in dendritic cell-like S100β protein-positive (DC-like S100β-positive) cells and that its receptor CXCR3 is expressed in ACTH-producing cells. DC-like S100β-positive cells are a subpopulation of folliculo-stellate cells in the anterior pituitary. In the present study, we examine whether CXCL10/CXCR3 signaling between DC-like S100β-positive cells and ACTH-producing cells mediates inhibition of CRF-activated ACTH-release by IFN-γ, using a CXCR3 antagonist in the primary pituitary cell culture. We found that IFN-γ up-regulated Cxcl10 expression via JAK/STAT signaling and proopiomelanocortin (Pomc) expression, while we reconfirmed that IFN-γ inhibits CRF-stimulated ACTH-release. Next, we used a CXCR3 agonist in primary culture to analyze whether CXCL10 induces Pomc-expression and ACTH-release using a CXCR3 agonist in the primary culture. The CXCR3 agonist significantly stimulated Pomc-expression and inhibited CRF-induced ACTH-release, while ACTH-release in the absence of CRF did not change. Thus, the present study leads us to an assumption that CXCL10/CXCR3 signaling mediates inhibition of the CRF-stimulated ACTH-release by IFN-γ. Our findings bring us to an assumption that CXCL10 from DC-like S100β-positive cells acts as a local modulator of ACTH-release during inflammation.
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- 2015
14. Isolation and characterisation of CD9-positive pituitary adult stem/progenitor cells in rats
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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.
- Published
- 2017
15. Degradation of serum microRNAs during transient storage of serum samples at 4℃
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Toshiko Aiso, Hiroaki Ohnishi, Akiko Yamaki, and Shu Takigami
- Subjects
0301 basic medicine ,business.industry ,Clinical Biochemistry ,Pilot Projects ,General Medicine ,Serum samples ,Real-Time Polymerase Chain Reaction ,Specimen Handling ,Cold Temperature ,03 medical and health sciences ,Circulating MicroRNA ,MicroRNAs ,030104 developmental biology ,0302 clinical medicine ,Transient storage ,030220 oncology & carcinogenesis ,microRNA ,Immunology ,Medicine ,Humans ,business - Abstract
Background Numerous studies demonstrate the potential of circulating microRNAs as non-invasive biomarkers for several diseases. Circulating microRNAs are much more stable than mRNAs and remain largely intact even after prolonged incubation at room temperature. However, recent reports show that microRNAs in serum or plasma samples have diverse stabilities. The aim of this pilot study is to evaluate the stabilities of miR-92a, miR-122 and miR-145 in serum during transient storage at 4℃ before freezing. Methods Serum samples were stored for 24 h at 4℃, and then RNA was extracted from whole serum or extracellular vesicles in serum. Total Exosome Isolation Reagent (from serum) was used for the fractionation of extracellular vesicles. Reverse transcription and real-time PCR of microRNAs were performed using the TaqMan MicroRNA Assays for miR-92a, miR-122 and miR-145. Results MiR-122 and miR-145 were degraded rapidly in serum; the concentrations dropped to 35.9% ( P Conclusions MiR-122 and miR-145 in serum are extremely unstable and could be degraded during transient storage of serum at 4℃ prior to freezing.
- Published
- 2017
16. Morphological assessment of the luminal surface of olfactory epithelium in mice deficient in tissue plasminogen activator following bulbectomy
- Author
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Shigeo Ookawara, Nobuko Makino, Takeharu Kanazawa, Toshio Ishikawa, Masumi Ichikawa, Jun Mimuro, Osamu Matsuo, Yoichi Sakata, Keiichi Ichimura, Yasushi Ohta, Kazuo Katoh, Seiji Madoiwa, and Shu Takigami
- Subjects
Pathology ,medicine.medical_specialty ,Plasmin ,business.industry ,Regeneration (biology) ,General Medicine ,Olfaction ,Olfactory Bulb ,Tissue plasminogen activator ,Olfactory bulb ,Mice, Inbred C57BL ,Mice ,medicine.anatomical_structure ,Microscopy, Electron, Transmission ,Olfactory Mucosa ,Otorhinolaryngology ,Tissue Plasminogen Activator ,medicine ,Animals ,Regeneration ,Neuron ,business ,Receptor ,Olfactory epithelium ,medicine.drug - Abstract
Objective:This study aimed to investigate the function of tissue plasminogen activator in the olfactory epithelium of mice following neural injury.Method:Transmission electron microscopy was used to study the changes in the morphology of the olfactory epithelium 1–7 days after surgical ablation of the olfactory bulb (bulbectomy).Results:Prior to bulbectomy, a uniformly fine material was observed within some regions of the olfactory epithelium of mice deficient in tissue plasminogen activator. At 2–3 days after bulbectomy, there were degenerative changes in the olfactory epithelium. At 5–7 days after bulbectomy, we noted drastic differences in olfactory epithelium morphology between mice deficient in tissue plasminogen activator and wild-type mice (comparisons were made using findings from a previous study). The microvilli seemed to be normal and olfactory vesicles and receptor neuron dendrites were largely intact in the olfactory epithelium of mice deficient in tissue plasminogen activator.Conclusion:The tissue plasminogen activator plasmin system may inhibit the regeneration of the olfactory epithelium in the early stages following neural injury.
- Published
- 2012
17. Estrogen Suppresses Retinaldehyde Dehydrogenase 1 Expression in the Anterior Pituitary Glands of Female Rats
- Author
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Shu Takigami, Motoshi Kikuchi, Ken Fujiwara, Takashi Yashiro, and Bulgan Davaadash
- Subjects
Male ,Pituitary gland ,medicine.medical_specialty ,medicine.drug_class ,Endocrinology, Diabetes and Metabolism ,Retinoic acid ,Down-Regulation ,Tretinoin ,In situ hybridization ,Biology ,Aldehyde Dehydrogenase 1 Family ,Gene Expression Regulation, Enzymologic ,chemistry.chemical_compound ,Paracrine signalling ,Endocrinology ,Anterior pituitary ,Pituitary Gland, Anterior ,Internal medicine ,medicine ,Animals ,RNA, Messenger ,Rats, Wistar ,Estrous cycle ,Messenger RNA ,Age Factors ,Retinal Dehydrogenase ,Estrogens ,Rats ,medicine.anatomical_structure ,chemistry ,Estrogen ,Female - Abstract
Retinoic acid (RA) plays a critical role in cell growth and tissue development. RA is also a regulating factor of pituitary function. RA is synthesized from retinoids through oxidation processes. The oxidation of retinal to RA is catalyzed by the retinaldehyde dehydrogenases (RALDHs), including RALDH1, RALDH2 and RALDH3. Recently, we demonstrated that RALDH1 is expressed in the anterior pituitary glands of adult male rats. However, the expression of RALDH1 in the female pituitary gland and the regulation of RALDH1 expression have not been determined. Therefore, we examined the expression of RALDH1 mRNA in the pituitary glands of adult female rats. By in situ hybridization with digoxigenin-labeled cRNA probes and quantitative real-time PCR analysis, we found that the expression level of RALDH1 was significantly lower in estrus as compared to proestrus, metestrus and diestrus. RALDH1 mRNA levels increased after ovariectomy and decreased remarkably after a 1-week treatment with 17beta-estradiol implants. Estradiol (0.01-100 microg per rat) dose-dependently decreased the expression of RALDH1 in the pituitary glands after 24 hours of subcutaneous administration. These results clearly show that RALDH1 mRNA expression is suppressed by estrogen. We speculate that the generation of RA is regulated by estrogen and that RA plays a role in the estrus cycle through paracrine and/or autocrine mechanisms in the anterior pituitary gland of female rats.
- Published
- 2008
18. In vivo correlation between c-Fos expression and corticotroph stimulation by adrenocorticotrophic hormone secretagogues in rat anterior pituitary gland
- Author
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Shu Takigami, Motoshi Kikuchi, Takashi Yashiro, and Ken Fujiwara
- Subjects
Male ,endocrine system ,medicine.medical_specialty ,Histology ,Somatotropic cell ,Corticotropin-Releasing Hormone ,Lactotrophs ,Population ,Stimulation ,Biology ,c-Fos ,Pathology and Forensic Medicine ,Rats, Sprague-Dawley ,Prolactin cell ,Adrenocorticotropic Hormone ,Anterior pituitary ,Stress, Physiological ,Internal medicine ,medicine ,Animals ,education ,Corticotrophs ,education.field_of_study ,Cell Biology ,Prolactin ,Rats ,medicine.anatomical_structure ,Endocrinology ,Pituitary Gland ,biology.protein ,Corticotropic cell ,Pituitary Hormone-Releasing Hormones ,Proto-Oncogene Proteins c-fos ,hormones, hormone substitutes, and hormone antagonists ,Hormone - Abstract
In the anterior pituitary gland, c-Fos expression is evoked by various stimuli. However, whether c-Fos expression is directly related to the stimulation of anterior pituitary cells by hypothalamic secretagogues is unclear. To confirm whether the reception of hormone-releasing stimuli evokes c-Fos expression in anterior pituitary cells, we have examined c-Fos expression of anterior pituitary glands in rats administered with synthetic corticotrophin-releasing hormone (CRH) intravenously or subjected to restraint stress. Single intravenous administration of CRH increases the number of c-Fos-expressing cells, and this number does not change even if the dose is increased. Double-immunostaining has revealed that most of the c-Fos-expressing cells contain adrenocorticotrophic hormone (ACTH); corticotrophs that do not express c-Fos in response to CRH have also been found. However, restraint stress evokes c-Fos expression in most of the corticotrophs and in a partial population of lactotrophs. These results suggest that c-Fos expression increases in corticotrophs stimulated by ACTH secretagogues, including CRH. Furthermore, we have found restricted numbers of corticotrophs expressing c-Fos in response to CRH. Although the mechanism underlying the different responses to CRH is not apparent, c-Fos is probably a useful immunohistochemical marker for corticotrophs stimulated by ACTH secretagogues.
- Published
- 2007
19. Expression of retinaldehyde dehydrogenase 1 in the anterior pituitary glands of adult rats
- Author
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Ken Fujiwara, Shu Takigami, Takashi Yashiro, Motoshi Kikuchi, and Tom Kouki
- Subjects
Male ,Pituitary gland ,medicine.medical_specialty ,Histology ,Basophil cell ,Retinoic acid ,In situ hybridization ,Biology ,Polymerase Chain Reaction ,Aldehyde Dehydrogenase 1 Family ,Pathology and Forensic Medicine ,chemistry.chemical_compound ,Paracrine signalling ,Anterior pituitary ,Pituitary Gland, Anterior ,Internal medicine ,medicine ,Animals ,RNA, Messenger ,Rats, Wistar ,Autocrine signalling ,In Situ Hybridization ,Age Factors ,Retinal Dehydrogenase ,Cell Biology ,Embryo, Mammalian ,Rats ,medicine.anatomical_structure ,Endocrinology ,chemistry ,embryonic structures ,Female ,Endocrine gland - Abstract
Retinoic acid (RA) plays a critical role in cell growth and tissue development and is also a regulatory factor of pituitary function. However, whether RA is generated in the pituitary gland and plays a role as a paracrine and/or autocrine factor is generally unknown. RA is synthesized from retinoids through oxidation processes. Dehydrogenases that catalyze the oxidation of retinal to RA are members of the retinaldehyde dehydrogenase (RALDH) family. Recently, we demonstrated that RALDH2 and RALDH3, but not RALDH1, were expressed in the developing anterior pituitary gland of rats, but the expression of RALDHs in the adult pituitary gland was not determined. Therefore, we have now examined the expression of RALDH1, RALDH2, and RALDH3 mRNAs in the pituitary gland of adult rats. Analysis by quantitative real-time polymerase chain reaction of adult pituitary glands has revealed a high level of RALDH1 mRNA but not of RALDH2 mRNA or RALDH3 mRNA. We have also detected mRNA expression for RALDH1 in the anterior pituitary gland by in situ hybridization with digoxigenin-labeled cRNA probes. Double-staining for RALDH1 mRNA and pituitary hormones or S-100 protein, a marker of folliculo-stellate cells (FS-cells), has revealed RALDH1 mRNA expression in a portion of prolactin-producing cells, marginal layer cells, and FS-cells. Our results suggest that RA is generated in the adult anterior pituitary gland, and that it may act locally on pituitary cells.
- Published
- 2007
20. An In Vivo and In Vitro Immunohistochemical Study of Integrin Alpha Subunit Localization in the Rat Anterior Pituitary
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Shu Takigami, Megumi Yatabe, Atsushi Sakamoto, Takashi Yashiro, Kaori Sunaga, and Motoshi Kikuchi
- Subjects
medicine.medical_specialty ,Histology ,biology ,Physiology ,Cell adhesion molecule ,Protein subunit ,Integrin ,Cell Biology ,Biochemistry ,Pathology and Forensic Medicine ,Cell biology ,Cell membrane ,medicine.anatomical_structure ,Endocrinology ,Anterior pituitary ,Cytoplasm ,Laminin ,Internal medicine ,biology.protein ,medicine ,G alpha subunit - Abstract
Different types of integrin alpha subunit were investigated immunohistochemically to determine the relationship between the cell adhesion molecule and the local morphology of the rat anterior pituitary using anterior pituitary tissues and primary cultured cells. Alpha 3 subunit was shown to be the main integrin subunit in the anterior pituitary. In vitro, immunoreactions of α3 subunit appeared mainly surrounding cell clusters. The distribution roughly overlapped with immunoreaction of laminin, while it was also observed where expression of laminin was negative. When dispersed cells were primary cultured for 2, 3 or 5 days, immunoreaction of α3 subunit was clearly detected in the vicinity of cell membrane throughout the period; while that of α5 subunit was not observed on day 2, it was observed as dots in cytoplasm on day 3 and more distinctly as lines in the vicinity of the cell membrane on day 5. Alpha 5 subunit was not detectable in vivo. These results indicate the possibility that integrin α3 subunit not only binds the cells to the basement membrane but also to other extracellular matrix or directly to adjacent cells in the anterior pituitary. In addition, integrin α5 subunit may supplement the function of α3 subunit under certain conditions.
- Published
- 2005
21. Fetal development of vomeronasal system in the goat
- Author
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Shu Takigami, Toshiya Osada, Yoshihiro Wakabayashi, Masumi Ichikawa, Satoshi Ohkura, Atsushi Ikai, Hiroaki Okamura, and Shunji Ohsako
- Subjects
Male ,Pathology ,medicine.medical_specialty ,Vomeronasal organ ,Rodent ,GTP-Binding Protein alpha Subunits, Gi-Go ,Biology ,Embryonic and Fetal Development ,Developmental Neuroscience ,Pregnancy ,Opossum ,biology.animal ,medicine ,Animals ,Neural Cell Adhesion Molecules ,Neurons ,Fetus ,Goats ,Age Factors ,biology.organism_classification ,Immunohistochemistry ,Olfactory Bulb ,Axons ,Cell biology ,Olfactory bulb ,Microscopy, Electron ,medicine.anatomical_structure ,biology.protein ,Female ,Neural cell adhesion molecule ,Neuron ,Vomeronasal Organ ,Olfactory marker protein ,Developmental Biology - Abstract
Our previous study morphologically revealed that the adult goat vomeronasal (VN) system was different from the rodent and opossum one, and at least two types of VN systems exist in mammals. However, it remains unknown whether the developments in both types of VN systems are ontogenetically distinct and when the goat VN system is established. In this study, we morphologically observed the fetal development of the goat accessory olfactory bulb (AOB) and VN neuron. In the fetus, Gi2-expressing VN terminals terminated at glomeruli throughout the AOB, and no immunoreactivities for Go were detected in the nerve terminals reaching into AOB. The layer structure of AOB rapidly developed in the latter half of gestation. In the VN organ (VNO), at the middle stage of gestation, the dendritic processes of VN neuron were exposed in the VN lumen, and scattered and thin microvilli existed on the protrusion of the VN neuron. In the apical part of dendritic processes, no clear vesicle existed. However, the immunohistochemistry of an olfactory marker protein (OMP) revealed that a few VN neurons with OMP exist in VN sensory epithelium (VSE) before birth, although marked immunoreactivities were detected in adult VSE. Fetal VN neurons appeared to be underdeveloped. These results suggest that the goat VN system is ontogenetically distinct from the rodent and opossum VN systems, and is underdeveloped before birth. The goat VN system will develop and mature during the early postnatal period similar to the rodent and opossum VN systems.
- Published
- 2004
22. 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
- Subjects
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
23. Changes in E- and N-cadherin expression in developing rat adenohypophysis
- Author
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Shu Takigami, Atsushi Sakamoto, Tom Kouki, Ken Fujiwara, Megumi Yatabe, Takashi Yashiro, and Motoshi Kikuchi
- Subjects
medicine.medical_specialty ,Histology ,Time Factors ,Population ,Histogenesis ,Rats, Sprague-Dawley ,Anterior pituitary ,Pituitary Gland, Anterior ,Internal medicine ,medicine ,Animals ,education ,Fluorescent Antibody Technique, Indirect ,Ecology, Evolution, Behavior and Systematics ,education.field_of_study ,biology ,Cadherin ,Cadherins ,Embryo, Mammalian ,Primary and secondary antibodies ,Embryonic stem cell ,Cell biology ,Rats ,Endocrinology ,medicine.anatomical_structure ,Animals, Newborn ,biology.protein ,Immunohistochemistry ,Pars tuberalis ,Anatomy ,Biotechnology - Abstract
Recently, we showed that hormone-producing cells express N-cadherin, while folliculo-stellate cells and marginal layer cells express E-cadherin in the adult rat anterior pituitary gland. These cells are believed to originate from a single cell population of the adenohypophyseal placode. In the present study, we immunohistochemically examined the divergence of cadherin types during pituitary histogenesis. Pituitary glands were excised from rats of embryonic day 11 (E11) through postnatal day 60 (P60) and paraffin sections were prepared. E- and N-cadherins were immunostained sequentially using monoclonal and polyclonal primary antibodies and fluorescent secondary antibodies. At E11, E-cadherin was expressed over oral epithelium, while N-cadherin expression was limited to the primordium of Rathke's pouch. When Rathke's pouch was formed at E13, E- and N-cadherin were broadly expressed in the entire cell population. N-cadherin was expressed particularly intensely in the layer of cells that faced the lumen. From E14 through E16, the majority of cells expressed both types of cadherins; however, the cell population to become the pars tuberalis expressed N-cadherin but not E-cadherin. From E18 through E20, when many hormone-producing cells appear, the number of cells that expressed N-cadherin alone increased. However, some cell populations in the pars distalis and multilayered marginal cells still expressed both cadherins. After birth, most of the cells came to express only one of the cadherin types. These results may suggest that undetermined adenohypophyseal cells express both E- and N-cadherin, but come to express either E- or N-cadherin during cytogenesis.
- Published
- 2007
24. Distinctive localization of N- and E-cadherins in rat anterior pituitary gland
- Author
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Megumi Yatabe, Shu Takigami, Motoshi Kikuchi, Ken Fujiwara, Tsuyoshi Soji, Takashi Yashiro, and Atsushi Sakamoto
- Subjects
Male ,medicine.medical_specialty ,Cell ,Heterologous ,Biology ,Rats, Sprague-Dawley ,Anterior pituitary ,Pituitary Gland, Anterior ,Internal medicine ,medicine ,Homologous chromosome ,Animals ,Cells, Cultured ,Cell adhesion molecule ,Cadherin ,Cell Membrane ,Cadherins ,Agricultural and Biological Sciences (miscellaneous) ,Immunohistochemistry ,Cell biology ,Rats ,Endocrinology ,medicine.anatomical_structure ,Anatomy ,Lumen (unit) - Abstract
In the rat anterior pituitary gland, folliculo-stellate cells aggregate preferably to form pseudofollicles, and each type of hormone-producing cell shows adhesive affinity with particular types of heterologous hormone-producing cells. Distribution of cadherin types in the rat anterior pituitary was examined immunohistochemically to clarify the unique cell arrangements caused by homologous and heterologous affinities among cells. N- and E-cadherins were detected continuously along cell membranes, while P-cadherin was not. N- and E-cadherins showed distinct isolation in localization, with N-cadherins localized in hormone-producing cells of distal and intermediate lobes in various amounts, and E-cadherins limited to folliculo-stellate cells and marginal layer cells facing the residual lumen of Rathke's pouch. A similar distribution of cadherins was observed in cell clusters of primary cultured anterior pituitary cells. These findings suggest that differential expression of cell adhesion molecules may be partially responsible for localization of hormone-producing cells and folliculo-stellate cells.
- Published
- 2006
25. Expression of retinaldehyde dehydrogenase (RALDH)2 and RALDH3 but not RALDH1 in the developing anterior pituitary glands of rats
- Author
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Shu Takigami, Ken Fujiwara, Takashi Yashiro, Motoshi Kikuchi, Toshihiko Yada, and Fumihiko Maekawa
- Subjects
Pituitary gland ,medicine.medical_specialty ,Histology ,Retinoic acid ,In situ hybridization ,Biology ,Aldehyde Dehydrogenase 1 Family ,Gene Expression Regulation, Enzymologic ,Pathology and Forensic Medicine ,Paracrine signalling ,chemistry.chemical_compound ,Anterior pituitary ,Pituitary Gland, Anterior ,Internal medicine ,medicine ,Animals ,RNA, Messenger ,Rats, Wistar ,Autocrine signalling ,Messenger RNA ,Gene Expression Regulation, Developmental ,Retinal Dehydrogenase ,Retinal ,Cell Biology ,Aldehyde Oxidoreductases ,Rats ,medicine.anatomical_structure ,Endocrinology ,chemistry ,embryonic structures - Abstract
Retinoic acid (RA) plays an important role in cell growth and tissue development and is also a regulating factor of pituitary function. However, whether RA is generated in the pituitary gland and plays a role as a paracrine and/or autocrine hormone is generally unknown. RA is synthesized from retinoids through oxidation processes. Dehydrogenases catalyzing the oxidation of retinal to RA are members of the retinaldehyde dehydrogenase (RALDH) family. In this study, we examined the expression of RALDH1, RALDH2, and RALDH3 mRNA in the rat embryonic pituitary gland. By in situ hybridization with digoxigenin-labeled cRNA probes, we detected mRNA expression for RALDH2 and RALDH3, but not RALDH1. The expression of RALDH2 and RALDH3 was located in Rathke’s pouch at embryonic day 12.5 (E12.5) and subsequently in the developing anterior pituitary gland. We also used quantitative real-time polymerase chain reaction to analyze RALDH2 and RALDH3 mRNA expression levels during the development of the pituitary gland. We found that pituitary RALDH2 and RALDH3 mRNA levels were high at E17.5 and decreased markedly after birth. Our study is the first to show that RALDH2 and RALDH3, but not RALDH1, are expressed in the embryonic anterior pituitary gland of the rat.
- Published
- 2006
26. Microdissected region-specific gene expression analysis with methacarn-fixed, paraffin-embedded tissues by real-time RT-PCR
- Author
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Haruka Fujita, Makoto Shibutani, Shu Takigami, Natsumi Kato, Masao Hirose, Kyoung-Youl Lee, Hironori Takagi, and Kunitoshi Mitsumori
- Subjects
0301 basic medicine ,Pathology ,medicine.medical_specialty ,Histology ,H&E stain ,Cell Count ,Biology ,Rats, Sprague-Dawley ,03 medical and health sciences ,Fixatives ,Gene expression ,medicine ,Animals ,Frozen Sections ,RNA, Messenger ,Coloring Agents ,Hematoxylin ,Gene ,Fixative ,Microdissection ,Acetic Acid ,030102 biochemistry & molecular biology ,Tissue Embedding ,Reverse Transcriptase Polymerase Chain Reaction ,Gene Expression Profiling ,Methanol ,RNA ,Molecular biology ,Staining ,Rats ,030104 developmental biology ,Real-time polymerase chain reaction ,Animals, Newborn ,Paraffin ,Chloroform ,Anatomy - Abstract
We have previously shown methacarn to be a versatile fixative for analysis of proteins, DNA, and RNA in paraffin-embedded tissues (PETs). In this study we analyzed its suitability for quantitative mRNA expression analysis of microdissected PET specimens using a real-time RT-PCR technique. Fidelity of expression in the methacarn-fixed PET sections, with reference to dose-dependent induction of cytochrome P450 2B1 in the phenobarbital-treated rat liver, was high in comparison with the unfixed frozen tissue case, even after hematoxylin staining. RNA yield from methacarn-fixed PET sections was equivalent to that in unfixed cryosections and was also not significantly affected by hematoxylin staining. Correlations between the expression levels of target genes and input amounts of extracted RNA in the range of 1–1000 pg were very high (correlation coefficients >0.98), the regression curves being similar to those with unfixed cryosections. Although cell numbers should be optimized for each target gene/tissue, ≥200 cells were necessary for accurate measurement in 10-μm-thick rat liver sections judging from the variation of measured value in small microdissected areas. These results indicate high performance with methacarn, close to that of unfixed tissues, regarding quantitative expression analysis of mRNAs in microdissected PET-specimens. (J Histochem Cytochem 52:903–913, 2004)
- Published
- 2004
27. Morphological evidence for two types of Mammalian vomeronasal system
- Author
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Yuji Mori, Yoshikuni Tanioka, Shu Takigami, and Masumi Ichikawa
- Subjects
Olfactory system ,Male ,Vomeronasal organ ,Physiology ,Central nervous system ,Guinea Pigs ,Sensory system ,Biology ,Cross Reactions ,GTP-Binding Protein alpha Subunits, Gi-Go ,Behavioral Neuroscience ,Dogs ,Physiology (medical) ,biology.animal ,Proto-Oncogene Proteins ,medicine ,Animals ,Horses ,Shrews ,Marmoset ,Callithrix ,Accessory Olfactory Bulb ,Immunohistochemistry ,Olfactory Bulb ,Sensory Systems ,Olfactory bulb ,medicine.anatomical_structure ,Sex pheromone ,Female ,GTP-Binding Protein alpha Subunit, Gi2 ,Vomeronasal Organ ,Neuroscience - Abstract
The vomeronasal (VN) systems of rodents and opossums are of the segregated type, i.e alpha-subtype G protein Gi2- or Go-expressing VN neurons, which are sensory cells, project discretely to the rostral or caudal region of the accessory olfactory bulb (AOB). Although this zone-specific projection is believed to be a common feature for processing pheromones in mammals, we previously found a uniform-type VN system in goat in which only Gi2-expressing VN axons terminate at the AOB. In most mammals, it remains unclear whether their VN systems are of the segregated or uniform type. Therefore, we investigated morphologically the VN systems of different mammalian species (dog, horse, musk shrew and common marmoset). Consequently, all VN axons of the examined animals were positively stained with immunohistochemistry for Gi2 in the same way as that in the goat. On the other hand, we observed immunoreactivities against Go in the olfactory axons, but not in the VN axons. These results suggest that many mammals have uniform-type VN systems, and at least two types of VN systems exist in terrestrial mammals. This morphological evidence will help us determine the processing function of VN systems.
- Published
- 2004
28. Diverse developmental toxicity of di-n-butyl phthalate in both sexes of rat offspring after maternal exposure during the period from late gestation through lactation
- Author
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Makoto Shibutani, Shu Takigami, Chikako Uneyama, Hironori Takagi, Masao Hirose, Natsumi Kato, and Kyoung-Youl Lee
- Subjects
Male ,endocrine system ,medicine.medical_specialty ,Pituitary gland ,Sex Differentiation ,Offspring ,Developmental toxicity ,Estrous Cycle ,Gestational Age ,Growth ,Biology ,Toxicology ,Mammary Glands, Animal ,Pregnancy ,Internal medicine ,Lactation ,Endocrine Glands ,medicine ,Animals ,Genitalia ,Sexual Maturation ,Gonadal Steroid Hormones ,Estrous cycle ,Sex Characteristics ,Anogenital distance ,Body Weight ,Organ Size ,Immunohistochemistry ,Prolactin ,Dibutyl Phthalate ,Diet ,Rats ,Pituitary Hormones ,medicine.anatomical_structure ,Endocrinology ,Prenatal Exposure Delayed Effects ,Female ,Luteinizing hormone - Abstract
To evaluate developmental toxicity of di- n -butyl phthalate (DBP) with exposure during the period from late gestation to following lactation, maternal rats were given DBP at dietary concentrations of 0, 20, 200, 2000 and 10,000 ppm from gestational day 15 to postnatal day (PND) 21. At 10,000 ppm, male offspring showed a decreased neonatal anogenital distance and retention of nipples (PND 14), while females showed a slight non-significant delay in the onset of puberty. At PND 21, reduction of testicular spermatocyte development was evident from 20 ppm, as well as mammary gland changes at low incidence in both sexes. At this time point, population changes of pituitary hormone-immunoreactive cells were observed at 10,000 ppm with a similar pattern of increase in the percentages of luteinizing hormone (LH)-positive and decrease in follicle-stimulating hormone (FSH) and prolactin producing cells in both sexes, effects also being evident on FSH from 200 ppm and LH from 2000 ppm in females. During postnatal week (PNW) 8–11, marginal increase of the number of cases with extended diestrus was found at 10,000 ppm. At adult stage necropsy, testicular lesions appeared to be very faint in most cases, but degeneration and atrophy of mammary gland alveoli were observed in males from 20 ppm. Although without clear monotonic dose-dependence, relative pituitary weights were increased with the intermediate doses in males at PNW 11. In females, relative pituitary weights were decreased after 10,000 ppm at PNW 11, and from 200 ppm at PNW 20. The proportion of FSH-positive cells in the pituitaries at PNW 11 was increased in both sexes at 10,000 ppm. Thus, developmental exposure to DBP affected female sexual development involving pituitary function, while in males testicular toxicity was mostly reversible but mammary gland toxicity was persistent at a dose level as low as 20 ppm.
- Published
- 2004
29. Lack of modifying effects of genistein on disruption of the reproductive system by perinatal dietary exposure to ethinylestradiol in rats
- Author
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Kunitoshi Mitsumori, Makoto Shibutani, Kyoung-Youl Lee, Shu Takigami, Hironori Takagi, Chikako Uneyama, Hwi Cheul Lee, Masugi Nishihara, and Masao Hirose
- Subjects
Male ,medicine.medical_specialty ,Sex Differentiation ,Litter Size ,medicine.drug_class ,media_common.quotation_subject ,Genistein ,Administration, Oral ,Estrous Cycle ,Phytoestrogens ,Biology ,Toxicology ,Ethinyl Estradiol ,chemistry.chemical_compound ,Oral administration ,Pregnancy ,Internal medicine ,Ethinylestradiol ,Adrenal Glands ,Testis ,medicine ,Animals ,Testosterone ,Reproductive system ,media_common ,Estrous cycle ,Epididymis ,Reproduction ,Body Weight ,Ovary ,Uterus ,Estrogens ,Rats, Inbred Strains ,Rats ,Endocrinology ,chemistry ,Estrogen ,Pituitary Gland ,Prenatal Exposure Delayed Effects ,Gestation ,Female ,medicine.drug - Abstract
We previously found that effects of perinatal dietary exposure to ethinylestradiol (EE) on the rat reproductive system differ depending on the diet used, with a more pronounced estrogenic impact with a regular diet that includes soy-derived proteins than with a soy-free (SF) diet. The present study was performed to examine whether genistein (GEN), a soy-derived major phytoestrogen, acts synergistically with EE. Maternal rats were fed SF diet without chemical (control) or containing 0.5-ppm EE, 0.5-ppm EE + 100-ppm GEN, 0.5-ppm EE + 1250-ppm GEN, or 1250-ppm GEN, from gestational day 15 to postnatal day (PND) 11. EE reduced serum testosterone in males at PND 3, and affected the onset of puberty of both sexes and estrous cyclicity and reproductive system in females, irrespective of co-administration of GEN. GEN alone also affected estrous cyclicity and the reproductive system in females. However, no combination effects of GEN with EE were evident, suggesting no synergism between the two.
- Published
- 2003
30. Subchronic toxicity study of dietary N-acetylglucosamine in F344 rats
- Author
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Masao Hirose, Kyoung-Youl Lee, Hironori Takagi, Takuro Arimura, Natsumi Kato, Chikako Uneyama, Makoto Shibutani, and Shu Takigami
- Subjects
Male ,medicine.medical_specialty ,No-observed-adverse-effect level ,F344 rats ,Administration, Oral ,Toxicology ,Acetylglucosamine ,chemistry.chemical_compound ,Oral administration ,Internal medicine ,medicine ,N-Acetylglucosamine ,Animals ,No-Observed-Adverse-Effect Level ,Hematology ,Dose-Response Relationship, Drug ,business.industry ,Body Weight ,General Medicine ,Organ Size ,Rats, Inbred F344 ,Diet ,Rats ,Dose–response relationship ,Endocrinology ,chemistry ,Aminosugar ,Toxicity ,Female ,business ,Food Science - Abstract
A subchronic toxicity study of N-acetylglucosamine (GlcNAc), a monomeric form of chitin, was conducted in groups of 10 male and 10 female F344 rats fed pelleted diets containing 0, 0.625, 1.25, 2.5 or 5% concentrations for 13 weeks. All animals survived until the end of the experiment. Slight, non-significant increase in body weights was observed in males receiving 0.625, 1.25 or 2.5% from week 4 until the end of the experiment, when significant elevation was found for the males receiving 0.625, 1.25 or 2.5% at the terminal sacrifice to result in decreased relative weights of many organs in these cases. However, there were no obvious indications of toxicity in any group receiving GlcNAc in terms of clinical signs, food intake, hematology, serum biochemistry, and histopathological findings. Thus, it was concluded that orally administered GlcNAc exerts no obvious toxicity to F344 rats at concentrations up to 5% in the diet for 13 weeks. Based on the present toxicity data, > or =5% was determined to be a no-observed-adverse-effect level, translated into 2476 and 2834 mg/kg/day for male and female rats, respectively.
- Published
- 2003
31. Alarm pheromone enhances stress-induced hyperthermia in rats
- Author
-
Takefumi Kikusui, Yukari Takeuchi, Shu Takigami, and Yuji Mori
- Subjects
Olfactory system ,Hyperthermia ,Male ,medicine.medical_specialty ,Fever ,Central nervous system ,Experimental and Cognitive Psychology ,Biology ,Pheromones ,Body Temperature ,Behavioral Neuroscience ,ALARM ,Sniffing ,Heart Rate ,Internal medicine ,Heart rate ,medicine ,Animals ,Rats, Wistar ,Behavior, Animal ,medicine.disease ,Immunohistochemistry ,Olfactory Bulb ,Olfactory bulb ,Rats ,Endocrinology ,medicine.anatomical_structure ,Pheromone ,Proto-Oncogene Proteins c-fos ,Stress, Psychological - Abstract
Behavioral and physiological effects of alarm pheromones emanating from stressed conspecific animals were investigated. Experimentally naive male Wistar rats were exposed to the test chambers containing alarm pheromones, which had been released by other rats receiving foot shocks in the same chamber beforehand. Along with behavioral analysis, the heart rate (HR) and core body temperature (cBT) were measured simultaneously using a biotelemetory system. Exposure to the alarm pheromones increased freezing, sniffing and walking and decreased resting as compared with rats exposed to control odors. In addition, these pheromone-exposed animals showed consistent increases in body temperature, i.e., stress-induced hyperthermia. After exposure to the alarm substances, immunoreactivity to nuclear Fos protein in the mitral cell layer in the accessory olfactory bulb (AOB) also increased compared with the reaction to control odors. These results suggest that an alarm pheromone enhances stress responses of conspecific animals both behaviorally and physiologically, and that these effects are mediated via activation of the AOB.
- Published
- 2001
32. Projection pattern of vomeronasal neurons to the accessory olfactory bulb in goats
- Author
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Masumi Ichikawa, Shu Takigami, and Yuji Mori
- Subjects
Olfactory system ,Male ,Rodent ,Vomeronasal organ ,Physiology ,Central nervous system ,Blotting, Western ,Guinea pig ,Behavioral Neuroscience ,Opossum ,Physiology (medical) ,biology.animal ,medicine ,Animals ,Neurons ,biology ,Goats ,Anatomy ,biology.organism_classification ,Immunohistochemistry ,Olfactory Bulb ,Sensory Systems ,Olfactory bulb ,Rats ,medicine.anatomical_structure ,Female ,sense organs ,Vomeronasal Organ - Abstract
Goats have a well-developed vomeronasal (VN) system and exhibit pheromone-induced reproductive facilitation, but there are no reports on the projection pattern of VN neurons in this species. Rodent, guinea pig and opossum accessory olfactory bulbs (AOBs) have been shown to have a segregated pattern of projection of the VN neurons, which express the two alpha-subtypes of the G-protein, namely Gi2 and Go, to the rostral and caudal regions of the AOB, respectively. In this study we investigated the projection pattern of VN nerve terminals by immunocytochemical staining of the goat vomeronasal organ (VNO) and the AOB with antibodies to Gi2 and Go. Gi2-immunoreactivity was found on the luminal surface of the sensory epithelium of the VNO, and in the VN nerve and glomerular layer throughout the AOB. On the other hand, Go-immunoreactivity was not identified in either the VNO or the VN nerve layer of the AOB. These results indicate that the projection pattern of VN neurons from the VNO to the AOB in the goat is considerably different from that in rodents which show a distinct segregated pattern.
- Published
- 2000
33. Morphological alterations of neuronal dendrites in developmental disorder model
- Author
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Takako Takemiya, Takashi Yamauchi, Kanato Yamagata, Hiroko Sugiura, Okio Hino, Yoshiyuki Yoshimura, Shu Takigami, and Shin Yasuda
- Subjects
Developmental disorder ,General Neuroscience ,medicine ,General Medicine ,Biology ,medicine.disease ,Neuroscience - Published
- 2009
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