A simple DNA disc chip in a microarray design based on modified comparative genomic hybridization for sperm DNA analysis.
OBJECTIVE
A DNA disc chip assay, based on comparative genomic hybridization, was designed to measure changes in sperm DNA intensities. The objective was to analyze the DNA integrity of hyperactive sperm cells after mild heat treatment.


METHODS
The assay based on a multiple cell comet assay was used to analyze changes in genomic DNA. Washed sperm DNA were tested on the assay and images stored in a microarray design.


METHODS
Clinical and academic research environment.


METHODS
Frozen-thawed washed sperm from different donors (n = 7).


METHODS
Discarded sperm leftover from trial washes carried out at 37 degrees and 40 degrees C were frozen and processed for the DNA disc chip assay.


METHODS
Fluorescent intensities of DNA disc chips and sperm variables.


RESULTS
Heat treatment resulted in more than eightfold increase in sperm hyperactive motility with little degradation in DNA integrity. Sperm with low hyperactivation was associated with alterations in DNA after heat treatment.


CONCLUSIONS
The DNA disc chip assay was simple, inexpensive, and permitted assisted reproduction technologies laboratories to use comparative genomic hybridization for cytogenotoxicity testing. However, the assay required manual processing, a fluorescent microscope, and computer. The data showed an association between sperm hyperactivation and DNA integrity suggesting that the hyperactivation marker may be used for selecting quality sperm for intracytoplasmic sperm injection. More studies are needed to examine temperature effects on ejaculated human sperm.
Chan
Philip J
PJ
Department of Gynecology, Center for Fertility and In Vitro Fertilization, Loma Linda University School of Medicine, Loma Linda, California 92354, USA. pchann@yahoo.com
Mann
Sylvia L
SL
Corselli
Johannah U
JU
Patton
William C
WC
King
Alan
A
Jacobson
John D
JD
D004247:DNA / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D006358:Hot Temperature; D006801:Humans; D006861:Hydrogen Peroxide / Q000008:administration & dosage / Q000494:pharmacology; D008297:Male; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D016877:Oxidants / Q000494:pharmacology; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects / Q000378:metabolism*
D016877:Oxidants; D004247:DNA; D006861:Hydrogen Peroxide