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Home / Equine Research Bank / PLoS One / The Lin28 Expression in Stallion Testes.
PloS one2016; 11(10); e0165011; doi: 10.1371/journal.pone.0165011

The Lin28 Expression in Stallion Testes.

Abstract: The molecular markers for specific germ cell stages can be utilized for identifying, monitoring, and separating a particular stage of germ cells. The RNA-binding protein Lin28 is expressed in gonocytes of human fetal testes. The Lin28 expression is restricted to a very small population of spermatogonial cells in human, mice, and monkey. The main objective of this study was to investigate the expression pattern of Lin28 in stallion testes at different reproductive stages. Based on the presence or absence of full spermatogenesis and lumina in seminiferous tubules, the testicular samples were categorized into two reproductive stages pre-pubertal and post-pubertal. We performed a reverse transcription polymerase chain reaction to confirm the presence of Lin28 mRNA in the testicular tissues and a western blot analysis to verify the cross-reactivity of rabbit Lin28 antibody with horse testicular tissue. For immunohistochemistry, Lin28 (rabbit anti-human), GATA4 (goat anti-human) or DAZL (goat anti-human) antibodies were used. The results of RT-PCR confirmed the expression of Lin28 mRNA in the stallion testes. The western blot analysis showed that the expression of 28 kDa Lin28 protein was localized in the cytoplasm of spermatogonia at both reproductive stages. The numbers of Lin28-positive germ cells per 1000 Sertoli cells in pre- and post-pubertal stages were 253 ± 8.66 and 29.67 ± 2.18, respectively. At both reproductive stages, all Lin28 positive cells showed no co-stained with GATA4 antibody, whereas only some of the Lin28-positive germ cells showed co-staining with DAZL antibody. The results from whole-mount staining showed that the Lin28 expression was limited to Asingle (As) and Apaired (Apr) spermatogonia. In conclusion, Lin28 might be utilized as a molecular marker for undifferentiated spermatogonial stem cells when used with DAZL antibody.
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Publication Date: 2016-10-31 PubMed ID: 27798668PubMed Central: PMC5087892DOI: 10.1371/journal.pone.0165011Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research aims to investigate the pattern of Lin28 expression in stallion testes during different reproductive stages. Findings could suggest the potential of Lin28 as a molecular marker for undifferentiated spermatogonial stem cells when used with a DAZL antibody.

Objective of the Study

  • The research’s primary aim was to investigate the expression pattern of Lin28 in stallion testes at different stages of reproduction. Lin28 is an RNA-binding protein expressed in gonocytes of human fetal testes, with its expression limited to a few spermatogonial cells in humans, mice, and monkeys. Understanding this pattern could aid in the development of specific germ cell stage markers necessary for identification, surveillance, and separation of certain germ cell stages.

Methodology of the Study

  • To achieve the objective, the researchers divided testicular samples into two reproductive stages: pre-pubertal and post-pubertal. This division was based on the presence or absence of complete spermatogenesis and lumina in the seminiferous tubules.
  • The team used reverse transcription polymerase chain reaction (RT-PCR) to confirm Lin28 mRNA’s presence in testicular tissues and western blot analysis to verify the cross-reactivity of rabbit Lin28 antibody with horse testicular tissue.
  • Three kinds of antibodies were used for immunohistochemistry: Lin28 (rabbit anti-human), GATA4 (goat anti-human), and DAZL (goat anti-human).

Findings of the Study

  • The RT-PCR results verified Lin28 mRNA expression in the stallion testes. Furthermore, western blot analysis demonstrated the expression of a 28 kDa Lin28 protein, localized in the cytoplasm of spermatogonia at both reproductive stages.
  • The researchers quantified Lin28-positive germ cells in terms of the number per 1000 Sertoli cells in pre- and post-pubertal stages. These figures were 253 ± 8.66 and 29.67 ± 2.18, respectively.
  • At both reproductive stages, all Lin28 positive cells did not show co-staining with the GATA4 antibody, while only a portion of these cells displayed co-staining with DAZL antibody. This pattern confirms a specific binding pattern of the Lin28 and DAZL antibodies, lending credibility to using these as potential molecular marker combinations.
  • Whole-mount staining results showed Lin28 expression to be confined to Asingle (As) and Apaired (Apr) spermatogonia.

Conclusion of the Study

  • Based on the above findings, the study concluded that Lin28 can potentially be utilized as a molecular marker for undifferentiated spermatogonial stem cells when used in combination with the DAZL antibody. As such, these findings contribute significant insights towards the understanding and potential manipulation of reproductive processes.

Cite This Article

APA
Lee G, Jung H, Yoon M. (2016). The Lin28 Expression in Stallion Testes. PLoS One, 11(10), e0165011. https://doi.org/10.1371/journal.pone.0165011

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 11
Issue: 10
Pages: e0165011
PII: e0165011

Researcher Affiliations

Lee, Geumhui
  • Department of Horse, Companion, and Wild Animal Science, Kyungpook National University, Sangju, Republic of Korea.
Jung, Heejun
  • Department of Animal Science and Biotechnology, Kyungpook National University, Sangju, Republic of Korea.
Yoon, Minjung
  • Department of Horse, Companion, and Wild Animal Science, Kyungpook National University, Sangju, Republic of Korea.
  • Department of Animal Science and Biotechnology, Kyungpook National University, Sangju, Republic of Korea.

MeSH Terms

  • Animals
  • Biomarkers
  • GATA4 Transcription Factor / genetics
  • GATA4 Transcription Factor / metabolism
  • Gene Expression
  • Germ Cells / metabolism
  • Horses
  • Immunohistochemistry
  • Male
  • Protein Binding
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA-Binding Proteins / genetics
  • Reproduction / genetics
  • Seminiferous Tubules / metabolism
  • Sexual Maturation / genetics
  • Spermatogonia / metabolism
  • Testis / metabolism

Conflict of Interest Statement

The authors have declared that no competing interests exist.

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Citations

This article has been cited 7 times.
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