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Transcriptome Analysis of Bull Semen with Extreme Nonreturn Rate: Use of Suppression-Subtractive Hybridization to Identify Functional Markers for Fertility¹

Dairy and Swine Research and Development Center,³ Agriculture and Agri-Food Canada, Sherbrooke, Quebec, Canada J1M 1Z3 The Semex Alliance, Inc.,⁴ Saint-Hyacinthe, Quebec, Canada J2S 7B8

ABSTRACT

Spermatozoa are terminally differentiated cells produced during the complex process of spermatogenesis. Although the role of their residual RNA content is still being debated, this transcriptome may represent a fingerprint of spermatogenesis quality. In the present study, we undertook differential transcript profiling of spermatozoa from fertile bulls with extreme nonreturn rates (NRRs): a low-fertile group, and a high-fertile group. Using the suppression-subtractive hybridization technique in combination with macroarray analysis, we also identified novel genes. Both extreme NRR index groups retained redundant identity, such as ribosomal and mitochondrial sequences, at a statistically significant level. An elevated number of 12S, 18S, and Large Chain R rRNA gene copies were found in low-fertile bulls and validated in spermatozoa by quantitative RT-PCR for a small cohort of bulls with known fertility index. Whereas the high-NRR library exhibited a large proportion (29%) of transcripts associated with known functions (e.g., metabolism, signal transduction, translation, glycosylation, and protein degradation), only 10% of the low-NRR sequences did. This difference is also conveyed by two other categories: 17% Bovine Genome and 48% unknown in the high-NRR library, compared with 3% and 80%, respectively, in the low-NRR library. Some of the unknown transcripts are similar to expressed sequence tags detected in the male reproductive organ of certain plants and retain homology to a putative human protein. Whereas the individual transcriptome profiles may be useful in fertility assessment, these findings also suggest cross-species conservation, could contribute to a better understanding of spermatogenesis, and provide new insights regarding idiopathic infertility.

gamete biology, sperm, spermatogenesis

¹Supported by the Agriculture and Agri-Food Canada and by a grant from the Dairy Cattle Genetics Research and Development (DairyGen) Council of the Canadian Dairy Network.

Correspondence: ²N. Bissonnette, Dairy and Swine Research and Development Centre, Agriculture and Agri-Food Canada, 2000 College Road, Sherbrooke, Quebec, Canada J1M 1Z3. FAX: 819 564 5507; e-mail: bissonnettenath{at}agr.gc.ca

This article has been cited by other articles:

				N. Bissonnette, J.-P. Levesque-Sergerie, C. Thibault, and G. Boissonneault Spermatozoal transcriptome profiling for bull sperm motility: a potential tool to evaluate semen quality Reproduction, July 1, 2009; 138(1): 65 - 80. [Abstract] [Full Text] [PDF]