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Tob1 is expressed in developing and adult gonads and is associated with the P-body marker, Dcp2

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Abstract

Tob1 is a member of the BTG/TOB family of proteins with established antiproliferative function. In Danio rerio and Xenopus laevis, the Tob1 gene is expressed from the one-cell stage through to early gastrula stages, followed in later development by discrete expression in many tissues including the notochord and somites. In both mouse and human, Tob1 is expressed in many adult tissues including the testis and ovary; however, the specific cell types are unknown. We examine Tob1 gene expression in mouse in developing germ cells and in sorted male germ cells (gonocytes, spermatogonia, pachytene spermatocytes and round spermatids) by reverse transcription and droplet digital polymerase chain reaction (RT-ddPCR) and in adult ovary and testis by immunofluorescence with anti-Tob1 protein staining. By RT-ddPCR, Tob1 expression was low in developing male germ cells but was highly expressed in round spermatids. In developing female germ cells undergoing entry into meiosis, it increased 10-fold. Tob1 was also highly expressed in round spermatids and in oocytes in all stages of folliculogenesis. Notably, a marker for P-bodies, Dcp-2, was also highly expressed in round spermatids and all oocyte stages examined. The cytoplasmic presence of Tob1 protein in round spermatids and oocytes and the association of Tob1 protein with Dcp2 in both cell types suggest that Tob1 protein plays a role in post-transcriptional mechanisms.

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Abbreviations

CCR4:

Carbon catabolite repression 4

CAF1:

Ccr4p associated factor

CPEB1:

Cytoplasmic polyadenylation binding protein 1

PABP:

Poly(A)binding protein:

PAM2:

Poly(A)-binding protein interacting motif 2

NOT:

Negative on TATA-less

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Authors and Affiliations

  1. Department of Anatomy and Neuroscience, University of Melbourne, Melbourne, Australia

    Farnaz Shapouri, Shaghayegh Saeidi, Robb U. de Iongh, Franca Casagranda & Gary R. Hime

  2. Hudson Institute of Medical Research, Monash University, Melbourne, Australia

    Patrick S. Western

  3. School of Environmental and Life Sciences, University of Newcastle, Newcastle, Australia

    Eileen A. McLaughlin & Jessie M. Sutherland

  4. School of Biosciences, University of Melbourne, Parkville, 3010, VIC, Melbourne, Australia

    Mary Familari

Authors
  1. Farnaz Shapouri
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  2. Shaghayegh Saeidi
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  3. Robb U. de Iongh
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  4. Franca Casagranda
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  5. Patrick S. Western
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  6. Eileen A. McLaughlin
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  7. Jessie M. Sutherland
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  8. Gary R. Hime
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  9. Mary Familari
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Correspondence to Mary Familari.

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The authors have no conflicts of interest in presenting the material, information or techniques described in this paper.

Additional information

Gary R. Hime and Mary Familari are joint senior authors.

These studies were supported by funds provided to M.F. by the School of Biosciences, University of Melbourne.

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Figure S1

In situ hybridisation with Tob1 sense (a, c) and antisense (b, d) riboprobes in mouse ovary (a, b) and mouse testis (c, d). a In frozen sections of the ovary, hybridisation signal with the sense probe is present in the follicular granulosa cells of an antral follicle but not in the oocyte cytoplasm (asterisk).b The antisense probe shows intense labelling in follicular cells but specific staining is present in the oocyte cytoplasm (asterisk). c In paraffin sections of the testis, no specific staining is observed with the sense probe in the seminiferous tubules. d Intense specific labelling is detected with the antisense probe in the round spermatids (solid arrowheads) with a weaker signal being detected in elongating spermatids (open arrowheads). Inset in d Higher magnification of labelled round spermatids (solid arrowhead) and elongating spermatids (open arrowhead). Bar 100 μm (a–d), 50 μm (inset). (GIF 109 kb)

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Shapouri, F., Saeidi, S., de Iongh, R.U. et al. Tob1 is expressed in developing and adult gonads and is associated with the P-body marker, Dcp2. Cell Tissue Res 364, 443–451 (2016). https://doi.org/10.1007/s00441-015-2328-z

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  • DOI: https://doi.org/10.1007/s00441-015-2328-z

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