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Transcriptome profiling of mice testes following low dose irradiation.

Belling KC, Tanaka M, Dalgaard MD, Nielsen JE, Nielsen HB, Brunak S, Almstrup K, Leffers H - Reprod. Biol. Endocrinol. (2013)

Bottom Line: We separated the transcripts into five unique clusters that we associated with spermatogonia, spermatocytes, early spermatids, late spermatids and somatic cells, respectively.We observed large expression changes in the somatic cell profile, which mainly could be attributed to changes in cellularity, but hyperplasia of Leydig cells may also play a role.We speculate that the possible hyperplasia may be caused by lower testosterone production and inadequate inhibin signalling due to missing germ cells.

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ABSTRACT

Background: Radiotherapy is used routinely to treat testicular cancer. Testicular cells vary in radio-sensitivity and the aim of this study was to investigate cellular and molecular changes caused by low dose irradiation of mice testis and to identify transcripts from different cell types in the adult testis.

Methods: Transcriptome profiling was performed on total RNA from testes sampled at various time points (n = 17) after 1 Gy of irradiation. Transcripts displaying large overall expression changes during the time series, but small expression changes between neighbouring time points were selected for further analysis. These transcripts were separated into clusters and their cellular origin was determined. Immunohistochemistry and in silico quantification was further used to study cellular changes post-irradiation (pi).

Results: We identified a subset of transcripts (n = 988) where changes in expression pi can be explained by changes in cellularity. We separated the transcripts into five unique clusters that we associated with spermatogonia, spermatocytes, early spermatids, late spermatids and somatic cells, respectively. Transcripts in the somatic cell cluster showed large changes in expression pi, mainly caused by changes in cellularity. Further investigations revealed that the low dose irradiation seemed to cause Leydig cell hyperplasia, which contributed to the detected expression changes in the somatic cell cluster.

Conclusions: The five clusters represent gene expression in distinct cell types of the adult testis. We observed large expression changes in the somatic cell profile, which mainly could be attributed to changes in cellularity, but hyperplasia of Leydig cells may also play a role. We speculate that the possible hyperplasia may be caused by lower testosterone production and inadequate inhibin signalling due to missing germ cells.

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IHC staining of the testis tissue pi with Leydig and Sertoli cell markers. Tgfbr3 and Hsd3b were used as Leydig cell markers and Vimentin (Vim) was used as a Sertoli cell marker. Indications of Leydig cell hyperplasia are observed at pi day 21–31, whereas the Sertoli cells do not seem affected. A detailed description of the pictures is found in the text.
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Figure 3: IHC staining of the testis tissue pi with Leydig and Sertoli cell markers. Tgfbr3 and Hsd3b were used as Leydig cell markers and Vimentin (Vim) was used as a Sertoli cell marker. Indications of Leydig cell hyperplasia are observed at pi day 21–31, whereas the Sertoli cells do not seem affected. A detailed description of the pictures is found in the text.

Mentions: Large expression changes were observed in the somatic cell cluster. We expected this to be caused mainly by changes in cellularity, but we decided to use IHC to investigate whether histological changes contributed to the observed changes in expression. We stained sections of the irradiated testis tissue with the following somatic cell-specific markers: two Leydig cell markers: Tgfbr3 and Hsd3b; a PTM cell marker: SMA; and a Sertoli cell marker: Vimentin (Figure 3).


Transcriptome profiling of mice testes following low dose irradiation.

Belling KC, Tanaka M, Dalgaard MD, Nielsen JE, Nielsen HB, Brunak S, Almstrup K, Leffers H - Reprod. Biol. Endocrinol. (2013)

IHC staining of the testis tissue pi with Leydig and Sertoli cell markers. Tgfbr3 and Hsd3b were used as Leydig cell markers and Vimentin (Vim) was used as a Sertoli cell marker. Indications of Leydig cell hyperplasia are observed at pi day 21–31, whereas the Sertoli cells do not seem affected. A detailed description of the pictures is found in the text.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3672050&req=5

Figure 3: IHC staining of the testis tissue pi with Leydig and Sertoli cell markers. Tgfbr3 and Hsd3b were used as Leydig cell markers and Vimentin (Vim) was used as a Sertoli cell marker. Indications of Leydig cell hyperplasia are observed at pi day 21–31, whereas the Sertoli cells do not seem affected. A detailed description of the pictures is found in the text.
Mentions: Large expression changes were observed in the somatic cell cluster. We expected this to be caused mainly by changes in cellularity, but we decided to use IHC to investigate whether histological changes contributed to the observed changes in expression. We stained sections of the irradiated testis tissue with the following somatic cell-specific markers: two Leydig cell markers: Tgfbr3 and Hsd3b; a PTM cell marker: SMA; and a Sertoli cell marker: Vimentin (Figure 3).

Bottom Line: We separated the transcripts into five unique clusters that we associated with spermatogonia, spermatocytes, early spermatids, late spermatids and somatic cells, respectively.We observed large expression changes in the somatic cell profile, which mainly could be attributed to changes in cellularity, but hyperplasia of Leydig cells may also play a role.We speculate that the possible hyperplasia may be caused by lower testosterone production and inadequate inhibin signalling due to missing germ cells.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Background: Radiotherapy is used routinely to treat testicular cancer. Testicular cells vary in radio-sensitivity and the aim of this study was to investigate cellular and molecular changes caused by low dose irradiation of mice testis and to identify transcripts from different cell types in the adult testis.

Methods: Transcriptome profiling was performed on total RNA from testes sampled at various time points (n = 17) after 1 Gy of irradiation. Transcripts displaying large overall expression changes during the time series, but small expression changes between neighbouring time points were selected for further analysis. These transcripts were separated into clusters and their cellular origin was determined. Immunohistochemistry and in silico quantification was further used to study cellular changes post-irradiation (pi).

Results: We identified a subset of transcripts (n = 988) where changes in expression pi can be explained by changes in cellularity. We separated the transcripts into five unique clusters that we associated with spermatogonia, spermatocytes, early spermatids, late spermatids and somatic cells, respectively. Transcripts in the somatic cell cluster showed large changes in expression pi, mainly caused by changes in cellularity. Further investigations revealed that the low dose irradiation seemed to cause Leydig cell hyperplasia, which contributed to the detected expression changes in the somatic cell cluster.

Conclusions: The five clusters represent gene expression in distinct cell types of the adult testis. We observed large expression changes in the somatic cell profile, which mainly could be attributed to changes in cellularity, but hyperplasia of Leydig cells may also play a role. We speculate that the possible hyperplasia may be caused by lower testosterone production and inadequate inhibin signalling due to missing germ cells.

Show MeSH
Related in: MedlinePlus