Linked Life Data

12390885

Source:http://linkedlifedata.com/resource/pubmed/id/12390885

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
5
pubmed:dateCreated
2002-10-22
pubmed:abstractText
Treatment of spermatozoa with either the nonionic detergent Triton X-100 (TX) or dithiothreitol (DTT) has been suggested to confer enhanced success on intracytoplasmic sperm injection (ICSI) in mice and humans. Here, we attempted to use both reagents together, to our knowledge for the first time, and found that this caused severe chromosomal breaks in paternal pronuclei. We documented this effect further by treating mouse spermatozoa with several combinations of DTT with and without detergent. Spermatozoa were treated with vigorous pipetting to induce membrane disruption or with TX or the ionic detergent mixed alkyltrimethylammonium bromide (ATAB). Swim-up spermatozoa were used as controls. In each treatment, two samples were tested, with or without the addition of DTT during the treatment procedure. In all samples with DTT, protamine reduction was confirmed by the decondensation assay. Sperm nuclei obtained after different treatments were injected into oocytes for cytogenetic analysis, and paternal and maternal chromosomes of the zygote were visualized and examined. We found that the numbers of normal paternal karyoplates resulting from ICSI with spermatozoa treated with either DTT (87%, 153/176), TX (79%, 112/142), or ATAB (85%, 99/116) alone were similar to swim-up controls (92%, 103/112). However, only 22% (23/103) and 40% (59/149) of examined metaphases were scored as normal in TX + DTT or ATAB + DTT treatments, respectively. Spermatozoa in which the membranes were disrupted by vigorous pipetting in the presence of DTT had a slightly reduced frequency of normal chromosomes (61%, 64/104), whereas those without DTT were normal (79%, 125/159). However, this difference was not statistically significant. When spermatozoa were treated with TX + DTT in the presence of EGTA or a mixture of EGTA and EDTA, the frequency of normal chromosomes was 39% (45/114) and 47% (38/81), respectively, suggesting that endogenous sperm nucleases may play a role in chromosomal damage. Our results indicate that simultaneous treatment of spermatozoa with detergent and DTT induces extensive chromosomal breakage and, therefore, should not be attempted in ICSI.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
0006-3363
pubmed:author
pubmed:issnType
Print
pubmed:volume
67
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1532-7
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:12390885-Animals, pubmed-meshheading:12390885-Cell Nucleus, pubmed-meshheading:12390885-Chromosome Aberrations, pubmed-meshheading:12390885-DNA Damage, pubmed-meshheading:12390885-Detergents, pubmed-meshheading:12390885-Dithiothreitol, pubmed-meshheading:12390885-Drug Synergism, pubmed-meshheading:12390885-Edetic Acid, pubmed-meshheading:12390885-Egtazic Acid, pubmed-meshheading:12390885-Female, pubmed-meshheading:12390885-Karyotyping, pubmed-meshheading:12390885-Male, pubmed-meshheading:12390885-Mice, pubmed-meshheading:12390885-Mice, Inbred Strains, pubmed-meshheading:12390885-Octoxynol, pubmed-meshheading:12390885-Protamines, pubmed-meshheading:12390885-Quaternary Ammonium Compounds, pubmed-meshheading:12390885-Reducing Agents, pubmed-meshheading:12390885-Sperm Injections, Intracytoplasmic, pubmed-meshheading:12390885-Spermatozoa, pubmed-meshheading:12390885-Zygote
pubmed:year
2002
pubmed:articleTitle
Combination of dithiothreitol and detergent treatment of spermatozoa causes paternal chromosomal damage.
pubmed:affiliation
Institute for Biogenesis Research, University of Hawaii Medical School, Honolulu, Hawaii 96822, USA. szczygie@hawaii.edu
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't