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Cell type-specific localization of Ephs pairing with ephrin-B2 in the rat postnatal pituitary gland

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Abstract

Sox2-expressing stem/progenitor cells in the anterior lobe of the pituitary gland form two types of micro-environments (niches): the marginal cell layer and dense cell clusters in the parenchyma. In relation to the mechanism of regulation of niches, juxtacrine signaling via ephrin and its receptor Eph is known to play important roles in various niches. The ephrin and Eph families are divided into two subclasses to create ephrin/Eph signaling in co-operation with confined partners. Recently, we reported that ephrin-B2 localizes specifically to both pituitary niches. However, the Ephs interacting with ephrin-B2 in these pituitary niches have not yet been identified. Therefore, the present study aims to identify the Ephs interacting with ephrin-B2 and the cells that produce them in the rat pituitary gland. In situ hybridization and immunohistochemistry demonstrated cell type-specific localization of candidate interacting partners for ephrin-B2, including EphA4 in cells located in the posterior lobe, EphB1 in gonadotropes, EphB2 in corticotropes, EphB3 in stem/progenitor cells and EphB4 in endothelial cells in the adult pituitary gland. In particular, double-immunohistochemistry showed cis-interactions between EphB3 and ephrin-B2 in the apical cell membranes of stem/progenitor cell niches throughout life and trans-interactions between EphB2 produced by corticotropes and ephrin-B2 located in the basolateral cell membranes of stem/progenitor cells in the early postnatal pituitary gland. These data indicate that ephrin-B2 plays a role in pituitary stem/progenitor cell niches by selective interaction with EphB3 in cis and EphB2 in trans.

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Change history

  • 16 November 2017

    The published online version contains mistakes in Table 1, Table 2 and Fig. 2. See below for the corrected Tables and Figure.

Abbreviations

CAR:

Coxsackievirus and adenovirus receptor

DAPI:

4,6-Diamidino-2-phenylindole

Eph:

Erythropoietin-producing hepatocellular carcinoma receptor

Ephrin:

Eph family receptor interacting proteins

FITC:

Fluorescein isothiocyanate

MCL:

Marginal cell layer

PCR:

Polymerase chain reaction

PDZ:

PSD-95, Dlg, ZO-1

PFA:

Paraformaldehyde

SOX2:

Sex-determining region Y-box 2

SVZ:

Subventricular zone

TBP:

TATA-box-binding protein

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Acknowledgements

The authors wish to thank Dr. A.F. Parlow and the NIDDK for antibodies against pituitary hormones, Dr. S. Tanaka at Shizuoka University for antibodies against human ACTH and GH and K. Kawai for his excellent technical assistance. This work was partially supported by JSPS KAKENHI (grant numbers 21380184 to YK, 24580435 to TK and 16 K18818 to SY); the MEXT-Supported Program for the Strategic Research Foundation at Private Universities; a research grant (A) to YK from the Institute of Science and Technology, Meiji University; and the Meiji University International Institute for BioResource Research (MUIIR).

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Correspondence to Yukio Kato.

Additional information

A correction to this article is available online at https://doi.org/10.1007/s00441-017-2721-x.

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Yoshida, S., Kato, T., Kanno, N. et al. Cell type-specific localization of Ephs pairing with ephrin-B2 in the rat postnatal pituitary gland. Cell Tissue Res 370, 99–112 (2017). https://doi.org/10.1007/s00441-017-2646-4

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Keywords

  • Rat pituitary
  • Stem/progenitor cells
  • Niche
  • Ephrin-B2
  • EphB3