Dynamic Hedgehog signalling pathway activity in germline stem cells

Z. Sahin, A. Szczepny, E. A. Mclaughlin, M. L. Meistrich, W. Zhou, I. Ustunel, K. L. Loveland

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Summary: Although the contribution of Hedgehog (Hh) signalling to stem cell development and oncogenesis is well recognised, its importance for spermatogonial stem cells (SSCs) has not been established. Here we interrogate adult rat SSCs using an established model in which only undifferentiated spermatogonial cells remain in the testis at 15 weeks following irradiation, and spermatogonial differentiation is induced within 4 weeks by gonadotrophin-releasing hormone antagonist (GnRH-ant) administration. Synthesis of Hh pathway components in untreated adult rat testes was compared with that in irradiated testes prior to and after GnRH-ant exposure using in situ hybridization. In adult testes with complete spermatogenesis, the Desert Hedgehog ligand transcript, Dhh, was detected in Sertoli cells, some spermatogonia and in spermatocytes by in situ hybridization. Spermatogenic cells were identified as sites of Hh signalling through detection of transcripts encoding the Hh receptor, Ptc2 transcripts and proteins for the key downstream target of Hh signalling, Gli1 and the Hh transcriptional activator, Gli2. Remarkably, the undifferentiated spermatogonia present in irradiated adult rat testes contained Dhh in addition to Ptc2, Gli1 and Gli2, revealing the potential for an autocrine Hh signalling loop to sustain undifferentiated spermatogonial cells. These transcripts became undetectable by in situ hybridization following GnRH-ant induction of spermatogonial differentiation, however, detection of Gli1 protein in spermatogonia in all groups indicates that Hh signalling is sustained. This is the first evidence of active Hh signalling in mammalian male germline stem cells, as has been documented for some cancer stem cells.

Original languageEnglish
Pages (from-to)267-274
Number of pages8
JournalAndrology
Volume2
Issue number2
DOIs
Publication statusPublished - Mar 2014
Externally publishedYes

Fingerprint

Hedgehogs
Stem Cells
Testis
Hormone Antagonists
Spermatogonia
Gonadotropin-Releasing Hormone
In Situ Hybridization
Autocrine Communication
Spermatocytes
Neoplastic Stem Cells
Sertoli Cells
Spermatogenesis
Carcinogenesis
Ligands

Cite this

Sahin, Z., Szczepny, A., Mclaughlin, E. A., Meistrich, M. L., Zhou, W., Ustunel, I., & Loveland, K. L. (2014). Dynamic Hedgehog signalling pathway activity in germline stem cells. Andrology, 2(2), 267-274. https://doi.org/10.1111/j.2047-2927.2014.00187.x
Sahin, Z. ; Szczepny, A. ; Mclaughlin, E. A. ; Meistrich, M. L. ; Zhou, W. ; Ustunel, I. ; Loveland, K. L. / Dynamic Hedgehog signalling pathway activity in germline stem cells. In: Andrology. 2014 ; Vol. 2, No. 2. pp. 267-274.
@article{ced7db2a1d5d4447808121218d73ba60,
title = "Dynamic Hedgehog signalling pathway activity in germline stem cells",
abstract = "Summary: Although the contribution of Hedgehog (Hh) signalling to stem cell development and oncogenesis is well recognised, its importance for spermatogonial stem cells (SSCs) has not been established. Here we interrogate adult rat SSCs using an established model in which only undifferentiated spermatogonial cells remain in the testis at 15 weeks following irradiation, and spermatogonial differentiation is induced within 4 weeks by gonadotrophin-releasing hormone antagonist (GnRH-ant) administration. Synthesis of Hh pathway components in untreated adult rat testes was compared with that in irradiated testes prior to and after GnRH-ant exposure using in situ hybridization. In adult testes with complete spermatogenesis, the Desert Hedgehog ligand transcript, Dhh, was detected in Sertoli cells, some spermatogonia and in spermatocytes by in situ hybridization. Spermatogenic cells were identified as sites of Hh signalling through detection of transcripts encoding the Hh receptor, Ptc2 transcripts and proteins for the key downstream target of Hh signalling, Gli1 and the Hh transcriptional activator, Gli2. Remarkably, the undifferentiated spermatogonia present in irradiated adult rat testes contained Dhh in addition to Ptc2, Gli1 and Gli2, revealing the potential for an autocrine Hh signalling loop to sustain undifferentiated spermatogonial cells. These transcripts became undetectable by in situ hybridization following GnRH-ant induction of spermatogonial differentiation, however, detection of Gli1 protein in spermatogonia in all groups indicates that Hh signalling is sustained. This is the first evidence of active Hh signalling in mammalian male germline stem cells, as has been documented for some cancer stem cells.",
keywords = "Adult stem cells, Spermatogonial stem cells, Testicular cancer, Zinc Finger Protein Gli2, Male, Adult Stem Cells/metabolism, Hedgehog Proteins/biosynthesis, Receptors, Cell Surface/genetics, Kruppel-Like Transcription Factors/genetics, Patched Receptors, Spermatocytes/metabolism, Cell Differentiation/drug effects, Hormone Antagonists/pharmacology, Signal Transduction, Spermatogenesis/physiology, Testis/cytology, Rats, Gonadotropin-Releasing Hormone/analogs & derivatives, Rats, Sprague-Dawley, Promyelocytic Leukemia Zinc Finger Protein, Sertoli Cells/metabolism, Animals, DNA-Binding Proteins/genetics, Spermatogonia/metabolism, Zinc Finger Protein GLI1",
author = "Z. Sahin and A. Szczepny and Mclaughlin, {E. A.} and Meistrich, {M. L.} and W. Zhou and I. Ustunel and Loveland, {K. L.}",
year = "2014",
month = "3",
doi = "10.1111/j.2047-2927.2014.00187.x",
language = "English",
volume = "2",
pages = "267--274",
journal = "Andrology",
issn = "2047-2919",
publisher = "John Wiley & Sons",
number = "2",

}

Sahin, Z, Szczepny, A, Mclaughlin, EA, Meistrich, ML, Zhou, W, Ustunel, I & Loveland, KL 2014, 'Dynamic Hedgehog signalling pathway activity in germline stem cells', Andrology, vol. 2, no. 2, pp. 267-274. https://doi.org/10.1111/j.2047-2927.2014.00187.x

Dynamic Hedgehog signalling pathway activity in germline stem cells. / Sahin, Z.; Szczepny, A.; Mclaughlin, E. A.; Meistrich, M. L.; Zhou, W.; Ustunel, I.; Loveland, K. L.

In: Andrology, Vol. 2, No. 2, 03.2014, p. 267-274.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Dynamic Hedgehog signalling pathway activity in germline stem cells

AU - Sahin, Z.

AU - Szczepny, A.

AU - Mclaughlin, E. A.

AU - Meistrich, M. L.

AU - Zhou, W.

AU - Ustunel, I.

AU - Loveland, K. L.

PY - 2014/3

Y1 - 2014/3

N2 - Summary: Although the contribution of Hedgehog (Hh) signalling to stem cell development and oncogenesis is well recognised, its importance for spermatogonial stem cells (SSCs) has not been established. Here we interrogate adult rat SSCs using an established model in which only undifferentiated spermatogonial cells remain in the testis at 15 weeks following irradiation, and spermatogonial differentiation is induced within 4 weeks by gonadotrophin-releasing hormone antagonist (GnRH-ant) administration. Synthesis of Hh pathway components in untreated adult rat testes was compared with that in irradiated testes prior to and after GnRH-ant exposure using in situ hybridization. In adult testes with complete spermatogenesis, the Desert Hedgehog ligand transcript, Dhh, was detected in Sertoli cells, some spermatogonia and in spermatocytes by in situ hybridization. Spermatogenic cells were identified as sites of Hh signalling through detection of transcripts encoding the Hh receptor, Ptc2 transcripts and proteins for the key downstream target of Hh signalling, Gli1 and the Hh transcriptional activator, Gli2. Remarkably, the undifferentiated spermatogonia present in irradiated adult rat testes contained Dhh in addition to Ptc2, Gli1 and Gli2, revealing the potential for an autocrine Hh signalling loop to sustain undifferentiated spermatogonial cells. These transcripts became undetectable by in situ hybridization following GnRH-ant induction of spermatogonial differentiation, however, detection of Gli1 protein in spermatogonia in all groups indicates that Hh signalling is sustained. This is the first evidence of active Hh signalling in mammalian male germline stem cells, as has been documented for some cancer stem cells.

AB - Summary: Although the contribution of Hedgehog (Hh) signalling to stem cell development and oncogenesis is well recognised, its importance for spermatogonial stem cells (SSCs) has not been established. Here we interrogate adult rat SSCs using an established model in which only undifferentiated spermatogonial cells remain in the testis at 15 weeks following irradiation, and spermatogonial differentiation is induced within 4 weeks by gonadotrophin-releasing hormone antagonist (GnRH-ant) administration. Synthesis of Hh pathway components in untreated adult rat testes was compared with that in irradiated testes prior to and after GnRH-ant exposure using in situ hybridization. In adult testes with complete spermatogenesis, the Desert Hedgehog ligand transcript, Dhh, was detected in Sertoli cells, some spermatogonia and in spermatocytes by in situ hybridization. Spermatogenic cells were identified as sites of Hh signalling through detection of transcripts encoding the Hh receptor, Ptc2 transcripts and proteins for the key downstream target of Hh signalling, Gli1 and the Hh transcriptional activator, Gli2. Remarkably, the undifferentiated spermatogonia present in irradiated adult rat testes contained Dhh in addition to Ptc2, Gli1 and Gli2, revealing the potential for an autocrine Hh signalling loop to sustain undifferentiated spermatogonial cells. These transcripts became undetectable by in situ hybridization following GnRH-ant induction of spermatogonial differentiation, however, detection of Gli1 protein in spermatogonia in all groups indicates that Hh signalling is sustained. This is the first evidence of active Hh signalling in mammalian male germline stem cells, as has been documented for some cancer stem cells.

KW - Adult stem cells

KW - Spermatogonial stem cells

KW - Testicular cancer

KW - Zinc Finger Protein Gli2

KW - Male

KW - Adult Stem Cells/metabolism

KW - Hedgehog Proteins/biosynthesis

KW - Receptors, Cell Surface/genetics

KW - Kruppel-Like Transcription Factors/genetics

KW - Patched Receptors

KW - Spermatocytes/metabolism

KW - Cell Differentiation/drug effects

KW - Hormone Antagonists/pharmacology

KW - Signal Transduction

KW - Spermatogenesis/physiology

KW - Testis/cytology

KW - Rats

KW - Gonadotropin-Releasing Hormone/analogs & derivatives

KW - Rats, Sprague-Dawley

KW - Promyelocytic Leukemia Zinc Finger Protein

KW - Sertoli Cells/metabolism

KW - Animals

KW - DNA-Binding Proteins/genetics

KW - Spermatogonia/metabolism

KW - Zinc Finger Protein GLI1

UR - http://www.scopus.com/inward/record.url?scp=84894334804&partnerID=8YFLogxK

UR - http://www.mendeley.com/research/dynamic-hedgehog-signalling-pathway-activity-germline-stem-cells-1

U2 - 10.1111/j.2047-2927.2014.00187.x

DO - 10.1111/j.2047-2927.2014.00187.x

M3 - Article

VL - 2

SP - 267

EP - 274

JO - Andrology

JF - Andrology

SN - 2047-2919

IS - 2

ER -

Sahin Z, Szczepny A, Mclaughlin EA, Meistrich ML, Zhou W, Ustunel I et al. Dynamic Hedgehog signalling pathway activity in germline stem cells. Andrology. 2014 Mar;2(2):267-274. https://doi.org/10.1111/j.2047-2927.2014.00187.x