An In Vivo and In Vitro Immunohistochemical Study of Integrin Alpha Subunit Localization in the Rat Anterior Pituitary

Kikuchi, Motoshi; Sunaga, Kaori; Yatabe, Megumi; Takigami, Shu; Sakamoto, Atsushi; Yashiro, Takashi

doi:10.1267/ahc.38.381

Cited by 3 publications

(3 citation statements)

References 25 publications

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“…We are the first to note that FS cells expressed integrin-a1, -a3, -a6, and -b1 (Fig. 7A); it was previously reported that integrin a5 was expressed in anterior pituitary cells in vivo and in vitro (Horacek et al 1994, Kikuchi et al 2005. Integrins comprise an a-and a b-subunit, which form a heterodimer.…”

Section: Discussionmentioning

confidence: 50%

Living-cell imaging of transgenic rat anterior pituitary cells in primary culture reveals novel characteristics of folliculo-stellate cells

Horiguchi¹,

Kikuchi²,

Kusumoto³

et al. 2009

Journal of Endocrinology

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Folliculo-stellate (FS) cells in the anterior pituitary gland appear to possess multifunctional properties. Recently, the development of transgenic rats (S100b-green fluorescent protein (GFP) rats) that express GFP specifically in FS cells in the anterior pituitary gland has allowed us to distinguish and observe living FS cells in other kinds of pituitary cells. We used S100b-GFP rats to investigate the topographic affinity of FS cells for other pituitary cells. We observed living FS cells in enzymatically dispersed anterior pituitary cells of S100b-GFP rats under a fluorescent microscope, and noted that FS cells markedly extended and contracted cytoplasmic processes and formed interconnections with neighboring FS cells. In addition, FS cells adhered to small clusters of GFPnegative cells, which were primarily hormone-producing cells, and these clusters further aggregated during the course of cytoplasmic contraction. In the presence of laminin, fibronectin, and varying types of collagen, FS cells showed marked changes in shape and specific proliferative activity; however, GFP-negative cells did not. On reverse transcription-PCR analysis and immunohistochemistry, FS cells were shown to express integrin subunits, which are the cell surface receptors for extracellular matrix (ECM). In the anterior pituitary gland, FS cells and the various types of hormoneproducing cells generate a unique topography in the presence of basement membrane components and interstitial collagens. The novel characteristics of FS cells observed in the present study suggest that in the anterior pituitary gland, FS cells play important roles in determining and/or maintaining local cellular arrangement in the presence of ECM components.

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Section: Discussionmentioning

confidence: 50%

Living-cell imaging of transgenic rat anterior pituitary cells in primary culture reveals novel characteristics of folliculo-stellate cells

Horiguchi¹,

Kikuchi²,

Kusumoto³

et al. 2009

Journal of Endocrinology

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“…In contrast to other tissues, the specific laminin isoforms expressed in the anterior pituitary have not been identified, although several studies have confirmed the presence of laminin protein in this gland [ 10 , 11 , 13 , 19 ]. In the present study, we used RT-PCR to identify laminin chains expressed in the anterior pituitary.…”

Section: Introductionmentioning

confidence: 99%

Laminin Isoforms and Laminin-Producing Cells in Rat Anterior Pituitary

Ramadhani

Tsukada

Fujiwara

et al. 2012

Acta Histochem. Cytochem

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Laminin is a key component of the basement membrane and is involved in the structural scaffold and in cell proliferation and differentiation. Research has identified 19 laminin isoforms, which are assemblies of α, β, and γ chains (eg, the α1, β1, and γ1 chains form the laminin 111 isoform). Although laminin is known to be present in the anterior pituitary, the specific laminin isoforms have not been identified. This study used molecular biological and histochemical techniques—namely, RT-PCR, immunohistochemistry, and in situ hybridization—to identify the laminin isoforms and laminin-producing cells in rat anterior pituitary. RT-PCR showed that laminin α1, α3, and α4 genes were expressed in anterior pituitary. Immunohistochemistry revealed laminin α1 in gonadotrophs and laminin α4 in almost all vascular endothelial cells. Laminin α3 was seen in a subset of vascular endothelial cells. We then performed in situ hybridization to localize β and γ chains in these cells and found that laminin β1, β2, and γ1 were expressed in gonadotrophs and that laminin β1 and γ1 were expressed in endothelial cells. In conclusion, we identified gonadotroph-type (laminin 111 and 121) and vascular-type (laminin 411 and 311) laminin isoforms in rat anterior pituitary.

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“…Cultured cells were fixed with 4% paraformaldehyde in 25 mM phosphate buffer (pH 7.4) for 30 min at room temperature. The procedure for immunostaining was essentially the same as in Kikuchi et al , 2005 [ 8 ]. Primary antibodies used in experiments were rabbit anti-rat growth hormone (GH; Dr. K. Wakabayashi, Gunma University, Gunma, Japan, dilution of 1:12,800), anti-ovine luteinizing hormone (LH) β subunit (Dr. K. Wakabayashi, dilution of 1:12,800), anti-rat prolactin (NIH, Bethesda, MD, USA, dilution of 1:12,800), anti-rat thyroid-stimulating hormone (TSH) β subunit (NIH, dilution of 1:3,200), anti-porcine adrenocorticotrophic hormone 1–39 (ACTH; Dr. Nakamura, Hokkaido University, Hokkaido, Japan, dilution of 1:3,200), and anti-S100B protein (DAKO, Glostrup, Denmark, dilution of 1:500).…”

Section: Methodsmentioning

confidence: 99%

Live Staining and Isolation of Specific Hormone-Producing Cells from Rat Anterior Pituitary by Cytochemistry with Lectins and Cholera Toxin B Subunit

Kikuchi

Kusumoto

Fujiwara

et al. 2011

Acta Histochem. Cytochem

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Anterior pituitary glands contain five types of hormone-producing cells. Distinguishing and isolating specific types of living cells are essential for studying their function. Although many such attempts have been made, the results have been disappointing. In the present study, we labeled specific types of living hormone-producing cells by using potential differences in sugar chains on the cell surfaces. Cytochemical analysis with lectins and cholera toxin B subunit revealed that PNA, S-WGA, and cholera toxin B subunit recognized sugar chains specific to prolactin cells, ACTH cells, and GH cells, respectively, and that UEA-I recognized most of prolactin cells and GH cells. Next, fluorescence-activated cell sorting was used to isolate GH cells labeled by fluoresceinated cholera toxin B. The purity of the GH cell fraction estimated by immunocytochemistry and quantitative real-time PCR for cell type-specific genes was more than 98%, which was higher than that reported in earlier studies, including those using transgenic animals. We conclude that cytochemistry with lectins and cholera toxin B subunit is a straightforward, acceptable method of isolating specific types of anterior pituitary cells and that the cells isolated by this method can serve as useful materials in the study of anterior pituitary cells.

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An In Vivo and In Vitro Immunohistochemical Study of Integrin Alpha Subunit Localization in the Rat Anterior Pituitary

Cited by 3 publications

References 25 publications

Living-cell imaging of transgenic rat anterior pituitary cells in primary culture reveals novel characteristics of folliculo-stellate cells

Living-cell imaging of transgenic rat anterior pituitary cells in primary culture reveals novel characteristics of folliculo-stellate cells

Laminin Isoforms and Laminin-Producing Cells in Rat Anterior Pituitary

Live Staining and Isolation of Specific Hormone-Producing Cells from Rat Anterior Pituitary by Cytochemistry with Lectins and Cholera Toxin B Subunit

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