Toward using stable spermatozoal RNAs for prognostic assessment of male factor fertility.
OBJECTIVE
To establish the stability of spermatozoal RNAs as a means to validate their use as a male fertility marker.


METHODS
Semen samples were randomly selected for 1 of 3 cryopreservation treatments.


METHODS
An academic research environment.


METHODS
Men aged 19 to 55 years who had fathered a child by natural conception within the past 6 months.


METHODS
Ejaculates were collected by masturbation and total spermatozoan RNA was isolated from two semen samples of ideal quality; one sample of medium quality, having been subjected to an additional freeze-thaw cycle, and two samples of poor quality, having been subjected to a third freeze-thaw cycle.


METHODS
Labeled cDNAs were generated and then used to interrogate Atlas Nylon Human Toxicology 1.2 microarrays. The spermatozoan transcriptomes were compared using a binomial approach.


RESULTS
The analysis identified a total of 228 unique spermatozoal transcripts among all samples. The medium quality sample shared 98% and 39% of its RNAs with the ideal and poor quality samples, respectively. A set of 36 RNAs resistant to insult were observed, some of which have been implicated in regulating male fertility, when all individuals were compared.


CONCLUSIONS
These results support the view that a population of spermatozoal RNAs is rapidly degraded in response to insult, whereas another population appears protected from such damage. Because spermatozoal RNAs echo the gene expression of spermatogenesis, the latter is likely to prove useful as a clinical maker of fertility status.
Ostermeier
G Charles
GC
Department of Obstetrics and Gynecology, Wayne State University, Detroit, Michigan, USA.
Goodrich
Robert J
RJ
Diamond
Michael P
MP
Dix
David J
DJ
Krawetz
Stephen A
SA
D000328:Adult; D001483:Base Sequence / Q000235:genetics; D015415:Biomarkers / Q000032:analysis; D015925:Cryopreservation / Q000706:statistics & numerical data; D006367:HeLa Cells; D006801:Humans; D007248:Infertility, Male / Q000175:diagnosis* / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D011379:Prognosis; D012313:RNA / Q000235:genetics*; D013094:Spermatozoa / Q000502:physiology*
D015415:Biomarkers; D012313:RNA