VASA (DDX4) is a Putative Marker for Spermatogonia, Spermatocytes and Round Spermatids in Stallions

Kim, Junyoung; Jung, Hyun-Key; Yoon, Minjung

doi:10.1111/rda.12632

Cited by 39 publications

(30 citation statements)

References 17 publications

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“…The GATA4 has been used as a molecular marker for Sertoli cells [4, 11, 12]. As expected, GATA4 positive nucleus of Sertoli cells were found adjacent to the basement membrane of seminiferous tubules in the pre-pubertal testes (Fig 3E, 3F, 3I and 3J), but they were located between germ cells in the post-pubertal stages (Fig 3G, 3H, 3K and 3L).…”

Section: Resultssupporting

confidence: 53%

“…Based on the age and presence of lumina in the cross-sections of the seminiferous tubules of stallions, their reproductive stages were categorized as follows: pre-pubertal (< 1.5 yr, n = 3) and post-pubertal (2–4 yr, n = 3) [4, 11]. …”

Section: Methodsmentioning

confidence: 99%

“…Immunofluorescence was performed as previously described [11] with minor modification. Briefly, 5-μm sections of the testicular tissues were treated with xylene (Duksan) to remove the paraffin and then rehydrated in a graded series of ethanol washes.…”

Section: Methodsmentioning

confidence: 99%

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The Lin28 Expression in Stallion Testes

2016

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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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Section: Resultssupporting

confidence: 53%

Section: Methodsmentioning

confidence: 99%

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The Lin28 Expression in Stallion Testes

2016

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“…The anti-γ-H2AX was used to detect DNA double strand breaks (DSBs) in the nucleus 40 . The anti-GENA and anti-MVH antibodies were used to detect germ cells 41,42 . The anti-GFRα1 antibodies were also used to detect spermatogonial stem cells (SSCs) 43,44 .…”

Section: Methodsmentioning

confidence: 99%

High-precision microbeam radiotherapy reveals testicular tissue-sparing effects for male fertility preservation

Fukunaga

Kaminaga

Sato

et al. 2019

Sci Rep

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Microbeam radiotherapy (MRT) is based on a spatial fractionation of synchrotron X-ray microbeams at the microscale level. Although the tissue-sparing effect (TSE) in response to non-uniform radiation fields was recognized more than one century ago, the TSE of MRT in the testes and its clinical importance for preventing male fertility remain to be determined. In this study, using the combination of MRT techniques and a unique ex vivo testes organ culture, we show, for the first time, the MRT-mediated TSE for the preservation of spermatogenesis. Furthermore, our high-precision microbeam analysis revealed that the survival and potential migration steps of the non-irradiated germ stem cells in the irradiated testes tissue would be needed for the effective TSE for spermatogenesis. Our findings indicated the distribution of dose irradiated in the testes at the microscale level is of clinical importance for delivering high doses of radiation to the tumor, while still preserving male fertility.

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“…Both Leydig cell-specific-(INSL3; HSD17B3) as well as germ-cell specific genes (e.g. FATE1; DMRT1; DDX4) (22,24,25) were overexpressed in TART compared to adult adrenal tissue. The overall expression of fourteen of those 31 testis-specific genes was higher in testis compared to TART tissue.…”

Section: Tart Express Both Adrenal-and Testis-specific Genesmentioning

confidence: 99%

Transcriptional comparison of Testicular Adrenal Rest Tumors with fetal and adult tissues

Schröder

Herwaarden

et al. 2020

Preprint

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Testicular Adrenal Rest Tumors (TART) are a common complication of unknown origin in patients with Congenital Adrenal Hyperplasia. These benign tumors may derive from cells of adrenal origin or from pluripotent progenitor cells from the fetal adrenogonadal primordium. By comparing the transcriptome of TART with fetal-and adulttestis and adrenal tissues, this study aims to unravel the origin of TART. Targeted transcriptome sequencing was followed by unsupervised clustering-, differential expression-, functional enrichment-and pathway analyses. Immunohistochemistry demonstrated co-expression of adrenal-specific CYP11B1 and testis-specific HSD17B3 in TART, indicating the existence of a distinct TART cell exhibiting both adrenal-and testicular characteristics. Principal component analysis indicated that the TART transcriptome was distinct from either adrenal or testis fetal tissue, making a progenitor-like phenotype of TART unlikely. Rather, TART appears to originate from -or differentiate into-a mature cell type, with both adrenal-and testicular characteristics. The present study, the first to describe the TART transcriptome, expands knowledge about the identity and functional characteristics of TART and identifies clinically targetable pathways associated with fibrosis.

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VASA (DDX4) is a Putative Marker for Spermatogonia, Spermatocytes and Round Spermatids in Stallions

Cited by 39 publications

References 17 publications

The Lin28 Expression in Stallion Testes

The Lin28 Expression in Stallion Testes

High-precision microbeam radiotherapy reveals testicular tissue-sparing effects for male fertility preservation

Transcriptional comparison of Testicular Adrenal Rest Tumors with fetal and adult tissues

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