Revealing cellular and molecular transitions in neonatal germ cell differentiation using single cell RNA sequencing

Jinyue Liao, Shuk Han Ng, Alfred Chun Luk, Hoi Ching Suen, Yan Qian, Annie Wing Tung Lee, Jiajie Tu, Jacqueline Chak Lam Fung, Nelson Leung Sang Tang, Bo Feng, Wai Yee Chan, Pierre Fouchet, Robin M. Hobbs, Tin Lap Lee

Research output: Contribution to journalArticleResearchpeer-review

3 Citations (Scopus)

Abstract

Neonatal germ cell development provides the foundation of spermatogenesis. However, a systematic understanding of this process is still limited. To resolve cellular and molecular heterogeneity in this process, we profiled single cell transcriptomes of undifferentiated germ cells from neonatal mouse testes and employed unbiased clustering and pseudotime ordering analysis to assign cells to distinct cell states in the developmental continuum. We defined the unique transcriptional programs underlying migratory capacity, resting cellular states and apoptosis regulation in transitional gonocytes. We also identified a subpopulation of primitive spermatogonia marked by CD87 (plasminogen activator, urokinase receptor), which exhibited a higher level of self-renewal gene expression and migration potential. We further revealed a differentiation-primed state within the undifferentiated compartment, in which elevated Oct4 expression correlates with lower expression of self-renewal pathway factors, higher Rarg expression, and enhanced retinoic acid responsiveness. Lastly, a knockdown experiment revealed the role of Oct4 in the regulation of gene expression related to the MAPK pathway and cell adhesion, which may contribute to stem cell differentiation. Our study thus provides novel insights into cellular and molecular regulation during early germ cell development.

Original languageEnglish
Article numberdev174953
Number of pages15
JournalDevelopment (Cambridge)
Volume146
Issue number6
DOIs
Publication statusPublished - 27 Mar 2019

Keywords

  • Differentiation
  • Gonocytes
  • Single cell
  • Spermatogonia
  • Transcriptomes

Cite this

Liao, J., Ng, S. H., Luk, A. C., Suen, H. C., Qian, Y., Lee, A. W. T., ... Lee, T. L. (2019). Revealing cellular and molecular transitions in neonatal germ cell differentiation using single cell RNA sequencing. Development (Cambridge), 146(6), [dev174953]. https://doi.org/10.1242/dev.174953
Liao, Jinyue ; Ng, Shuk Han ; Luk, Alfred Chun ; Suen, Hoi Ching ; Qian, Yan ; Lee, Annie Wing Tung ; Tu, Jiajie ; Fung, Jacqueline Chak Lam ; Tang, Nelson Leung Sang ; Feng, Bo ; Chan, Wai Yee ; Fouchet, Pierre ; Hobbs, Robin M. ; Lee, Tin Lap. / Revealing cellular and molecular transitions in neonatal germ cell differentiation using single cell RNA sequencing. In: Development (Cambridge). 2019 ; Vol. 146, No. 6.
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abstract = "Neonatal germ cell development provides the foundation of spermatogenesis. However, a systematic understanding of this process is still limited. To resolve cellular and molecular heterogeneity in this process, we profiled single cell transcriptomes of undifferentiated germ cells from neonatal mouse testes and employed unbiased clustering and pseudotime ordering analysis to assign cells to distinct cell states in the developmental continuum. We defined the unique transcriptional programs underlying migratory capacity, resting cellular states and apoptosis regulation in transitional gonocytes. We also identified a subpopulation of primitive spermatogonia marked by CD87 (plasminogen activator, urokinase receptor), which exhibited a higher level of self-renewal gene expression and migration potential. We further revealed a differentiation-primed state within the undifferentiated compartment, in which elevated Oct4 expression correlates with lower expression of self-renewal pathway factors, higher Rarg expression, and enhanced retinoic acid responsiveness. Lastly, a knockdown experiment revealed the role of Oct4 in the regulation of gene expression related to the MAPK pathway and cell adhesion, which may contribute to stem cell differentiation. Our study thus provides novel insights into cellular and molecular regulation during early germ cell development.",
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author = "Jinyue Liao and Ng, {Shuk Han} and Luk, {Alfred Chun} and Suen, {Hoi Ching} and Yan Qian and Lee, {Annie Wing Tung} and Jiajie Tu and Fung, {Jacqueline Chak Lam} and Tang, {Nelson Leung Sang} and Bo Feng and Chan, {Wai Yee} and Pierre Fouchet and Hobbs, {Robin M.} and Lee, {Tin Lap}",
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Liao, J, Ng, SH, Luk, AC, Suen, HC, Qian, Y, Lee, AWT, Tu, J, Fung, JCL, Tang, NLS, Feng, B, Chan, WY, Fouchet, P, Hobbs, RM & Lee, TL 2019, 'Revealing cellular and molecular transitions in neonatal germ cell differentiation using single cell RNA sequencing', Development (Cambridge), vol. 146, no. 6, dev174953. https://doi.org/10.1242/dev.174953

Revealing cellular and molecular transitions in neonatal germ cell differentiation using single cell RNA sequencing. / Liao, Jinyue; Ng, Shuk Han; Luk, Alfred Chun; Suen, Hoi Ching; Qian, Yan; Lee, Annie Wing Tung; Tu, Jiajie; Fung, Jacqueline Chak Lam; Tang, Nelson Leung Sang; Feng, Bo; Chan, Wai Yee; Fouchet, Pierre; Hobbs, Robin M.; Lee, Tin Lap.

In: Development (Cambridge), Vol. 146, No. 6, dev174953, 27.03.2019.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Revealing cellular and molecular transitions in neonatal germ cell differentiation using single cell RNA sequencing

AU - Liao, Jinyue

AU - Ng, Shuk Han

AU - Luk, Alfred Chun

AU - Suen, Hoi Ching

AU - Qian, Yan

AU - Lee, Annie Wing Tung

AU - Tu, Jiajie

AU - Fung, Jacqueline Chak Lam

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AU - Feng, Bo

AU - Chan, Wai Yee

AU - Fouchet, Pierre

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AU - Lee, Tin Lap

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AB - Neonatal germ cell development provides the foundation of spermatogenesis. However, a systematic understanding of this process is still limited. To resolve cellular and molecular heterogeneity in this process, we profiled single cell transcriptomes of undifferentiated germ cells from neonatal mouse testes and employed unbiased clustering and pseudotime ordering analysis to assign cells to distinct cell states in the developmental continuum. We defined the unique transcriptional programs underlying migratory capacity, resting cellular states and apoptosis regulation in transitional gonocytes. We also identified a subpopulation of primitive spermatogonia marked by CD87 (plasminogen activator, urokinase receptor), which exhibited a higher level of self-renewal gene expression and migration potential. We further revealed a differentiation-primed state within the undifferentiated compartment, in which elevated Oct4 expression correlates with lower expression of self-renewal pathway factors, higher Rarg expression, and enhanced retinoic acid responsiveness. Lastly, a knockdown experiment revealed the role of Oct4 in the regulation of gene expression related to the MAPK pathway and cell adhesion, which may contribute to stem cell differentiation. Our study thus provides novel insights into cellular and molecular regulation during early germ cell development.

KW - Differentiation

KW - Gonocytes

KW - Single cell

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KW - Transcriptomes

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JO - Development

JF - Development

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