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Expression studies of neuronatin in prenatal and postnatal rat pituitary

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Abstract

The pituitary gland, an indispensable endocrine organ that synthesizes and secretes pituitary hormones, develops with the support of many factors. Among them, neuronatin (NNAT), which was discovered in the neonatal mouse brain as a factor involved in neural development, has subsequently been revealed to be coded by an abundantly expressing gene in the pituitary gland but its role remains elusive. We analyze the expression profile of Nnat and the localization of its product during rat pituitary development. The level of Nnat expression was high during the embryonic period but remarkably decreased after birth. Immunohistochemistry demonstrated that NNAT appeared in the SOX2-positive stem/progenitor cells in the developing pituitary primordium on rat embryonic day 11.5 (E11.5) and later in the majority of SOX2/PROP1 double-positive cells on E13.5. Thereafter, during pituitary embryonic development, Nnat expression was observed in some stem/progenitor cells, proliferating cells and terminally differentiating cells. In postnatal pituitaries, NNAT-positive cells decreased in number, with most coexpressing Sox2 or Pit1, suggesting a similar role for NNAT to that during the embryonic period. NNAT was widely localized in mitochondria, peroxisomes and lysosomes, in addition to the endoplasmic reticulum but not in the Golgi. The present study thus demonstrated the variability in expression of NNAT-positive cells in rat embryonic and postnatal pituitaries and the intracellular localization of NNAT. Further investigations to obtain functional evidence for NNAT are a prerequisite.

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Abbreviations

ACTH:

Adrenocorticotropic hormone

DAPI:

4′ 6-Diamidino-2-phenylindole

ER:

Endoplasmic reticulum

EYFP:

Enhanced yellow fluorescent protein

FITC:

Fluorescein isothiocyanate

FSHβ:

Follicle-stimulating hormone

GFP:

Green fluorescent protein

GH:

Growth hormone

LHβ:

Luteinizing hormone

MCL:

Marginal cell layer

NNAT:

Neuronatin

PCR:

Polymerase chain reaction

PDI:

Protein disulfide isomerase

PIT1:

Pituitary-specific transcription factor 1

PRL:

Prolactin

PROP1:

Prophet of PIT1

PRRX1:

Paired-related homeobox 1

PRRX2:

Paired-related homeobox 2

SERCA2:

Sarco/endoplasmic reticulum Ca2+ ATPase isoform 2

SOX2:

Sex-determining region Y-box 2

TBP:

TATA-box-binding protein

TSHβ:

Thyroid-stimulating hormone

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Acknowledgments

The authors thank Dr. A.F. Parlow and the NIDDK for antibodies against pituitary hormones, Dr. S. Tanaka at Shizuoka University for the antibody against human adrenocorticotropic hormone and GH, Dr. P. Mellon for providing us with LβT2 cells and Mr. K. Kawai for his excellent technical assistance.

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Authors and Affiliations

  1. Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa, 214-8571, Japan

    Naoko Kanno, Saishu Yoshida, Hideji Yako, Hiroki Ueharu, Naoto Nishimura & Yukio Kato

  2. Organization for the Strategic Coordination of Research and Intellectual Property, Meiji University, Kanagawa, 214-8571, Japan

    Masashi Higuchi, Mo Chen & Takako Kato

  3. Institute of Reproduction and Endocrinology, Meiji University, Kanagawa, 214-8571, Japan

    Masashi Higuchi, Mo Chen, Takako Kato & Yukio Kato

Authors
  1. Naoko Kanno
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  2. Masashi Higuchi
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  3. Saishu Yoshida
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  4. Hideji Yako
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  5. Mo Chen
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  6. Hiroki Ueharu
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  7. Naoto Nishimura
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  8. Takako Kato
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  9. Yukio Kato
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Corresponding author

Correspondence to Yukio Kato.

Additional information

This work was partially supported by JSPS KAKENHI Grants, nos. 21380184 to Y.K. and 24580435 to T.K., by the MEXT-Supported Program for the Strategic Research Foundation at Private Universities, 2014–2018 and by a research grant (A) to Y.K. from the Institute of Science and Technology, Meiji University. This study was also supported by the Meiji University International Institute for BioResource Research (MUIIR).

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Kanno, N., Higuchi, M., Yoshida, S. et al. Expression studies of neuronatin in prenatal and postnatal rat pituitary. Cell Tissue Res 364, 273–288 (2016). https://doi.org/10.1007/s00441-015-2325-2

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