Novel approach for the detection of the vestiges of testicular mRNA splicing errors in mature spermatozoa of Japanese Black bulls.

There is a serious problem with the reduction of male reproductive performance of the livestock in the world. We have a hypothesis that the splicing error-caused derivation of aberrant sperm motility-related proteins may be one of its causal factors. It is thought that fresh testicular tissues are n...

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Autores principales:	Taichi Noda, Mitsuhiro Sakase, Moriyuki Fukushima, Hiroshi Harayama
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b73804de6da240a09fe9feced65ce278
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spelling	oai:doaj.org-article:b73804de6da240a09fe9feced65ce2782021-11-18T07:55:57ZNovel approach for the detection of the vestiges of testicular mRNA splicing errors in mature spermatozoa of Japanese Black bulls.1932-620310.1371/journal.pone.0057296https://doaj.org/article/b73804de6da240a09fe9feced65ce2782013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23468960/?tool=EBIhttps://doaj.org/toc/1932-6203There is a serious problem with the reduction of male reproductive performance of the livestock in the world. We have a hypothesis that the splicing error-caused derivation of aberrant sperm motility-related proteins may be one of its causal factors. It is thought that fresh testicular tissues are necessary for the detection of splicing errors of the mRNA. However, it is difficult to obtain testicular tissues from a number of agriculturally important bulls by surgical methods, because such procedures may have deleterious effects on bulls' reproductive performance. The aim of this study was to examine the usefulness of mRNA fragments collected from ejaculated spermatozoa as alternative analytical samples for detection of the splicing errors. In the first experiment, we characterized the alternative splicing and splicing error of bull testicular ADCY10 mRNA which coded the synthase of the regulatory molecule for sperm motility "cAMP". In testes, the exon 11-lacking variant coding the truncated ADCY10 was derived by alternative splicing. However, splicing errors, which accompanied the frame shift in the second cyclase domain, were occasionally observed in the exon 11-lacking variant. This aberrant variant retained intronic nucleotides (4 bases, CCAG) connecting the initial part of exon 10 due to splicing errors and consequently yielded the cleavage site for a restriction enzyme (Cac8I) which recognized the nucleotide sequences (GCNNGC). In the second experiment, we recovered residual testicular mRNA fragments from ejaculated spermatozoa and observed the splicing error-caused derivation of the aberrant variant of ADCY 10. Ejaculated spermatozoa conserved mRNA fragments of the exon 11-lacking variant coding exons 9, 10, 12 and 13. Moreover, the above-mentioned aberrant variant of ADCY10 mRNA fragment was detectable by Cac8I digestion treatment using the sperm mRNAs. These results indicate the utility of sperm mRNA fragments for the detection of splicing errors in bull testicular mRNAs.Taichi NodaMitsuhiro SakaseMoriyuki FukushimaHiroshi HarayamaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 2, p e57296 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Taichi Noda Mitsuhiro Sakase Moriyuki Fukushima Hiroshi Harayama Novel approach for the detection of the vestiges of testicular mRNA splicing errors in mature spermatozoa of Japanese Black bulls.
description	There is a serious problem with the reduction of male reproductive performance of the livestock in the world. We have a hypothesis that the splicing error-caused derivation of aberrant sperm motility-related proteins may be one of its causal factors. It is thought that fresh testicular tissues are necessary for the detection of splicing errors of the mRNA. However, it is difficult to obtain testicular tissues from a number of agriculturally important bulls by surgical methods, because such procedures may have deleterious effects on bulls' reproductive performance. The aim of this study was to examine the usefulness of mRNA fragments collected from ejaculated spermatozoa as alternative analytical samples for detection of the splicing errors. In the first experiment, we characterized the alternative splicing and splicing error of bull testicular ADCY10 mRNA which coded the synthase of the regulatory molecule for sperm motility "cAMP". In testes, the exon 11-lacking variant coding the truncated ADCY10 was derived by alternative splicing. However, splicing errors, which accompanied the frame shift in the second cyclase domain, were occasionally observed in the exon 11-lacking variant. This aberrant variant retained intronic nucleotides (4 bases, CCAG) connecting the initial part of exon 10 due to splicing errors and consequently yielded the cleavage site for a restriction enzyme (Cac8I) which recognized the nucleotide sequences (GCNNGC). In the second experiment, we recovered residual testicular mRNA fragments from ejaculated spermatozoa and observed the splicing error-caused derivation of the aberrant variant of ADCY 10. Ejaculated spermatozoa conserved mRNA fragments of the exon 11-lacking variant coding exons 9, 10, 12 and 13. Moreover, the above-mentioned aberrant variant of ADCY10 mRNA fragment was detectable by Cac8I digestion treatment using the sperm mRNAs. These results indicate the utility of sperm mRNA fragments for the detection of splicing errors in bull testicular mRNAs.
format	article
author	Taichi Noda Mitsuhiro Sakase Moriyuki Fukushima Hiroshi Harayama
author_facet	Taichi Noda Mitsuhiro Sakase Moriyuki Fukushima Hiroshi Harayama
author_sort	Taichi Noda
title	Novel approach for the detection of the vestiges of testicular mRNA splicing errors in mature spermatozoa of Japanese Black bulls.
title_short	Novel approach for the detection of the vestiges of testicular mRNA splicing errors in mature spermatozoa of Japanese Black bulls.
title_full	Novel approach for the detection of the vestiges of testicular mRNA splicing errors in mature spermatozoa of Japanese Black bulls.
title_fullStr	Novel approach for the detection of the vestiges of testicular mRNA splicing errors in mature spermatozoa of Japanese Black bulls.
title_full_unstemmed	Novel approach for the detection of the vestiges of testicular mRNA splicing errors in mature spermatozoa of Japanese Black bulls.
title_sort	novel approach for the detection of the vestiges of testicular mrna splicing errors in mature spermatozoa of japanese black bulls.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/b73804de6da240a09fe9feced65ce278
work_keys_str_mv	AT taichinoda novelapproachforthedetectionofthevestigesoftesticularmrnasplicingerrorsinmaturespermatozoaofjapaneseblackbulls AT mitsuhirosakase novelapproachforthedetectionofthevestigesoftesticularmrnasplicingerrorsinmaturespermatozoaofjapaneseblackbulls AT moriyukifukushima novelapproachforthedetectionofthevestigesoftesticularmrnasplicingerrorsinmaturespermatozoaofjapaneseblackbulls AT hiroshiharayama novelapproachforthedetectionofthevestigesoftesticularmrnasplicingerrorsinmaturespermatozoaofjapaneseblackbulls
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Novel approach for the detection of the vestiges of testicular mRNA splicing errors in mature spermatozoa of Japanese Black bulls.

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