PMID- 23059640
OWN - NLM
STAT- MEDLINE
DCOM- 20130924
LR  - 20211021
IS  - 1348-4400 (Electronic)
IS  - 0916-8818 (Print)
IS  - 0916-8818 (Linking)
VI  - 59
IP  - 1
DP  - 2013
TI  - Sexing of dog sperm by fluorescence in situ hybridization.
PG  - 92-6
AB  - Effective preselection of sex has been accomplished in several species of 
      livestock and also in humans using the flow cytometric sperm sorting method. A 
      guaranteed high sorting accuracy is a key prerequisite for the widespread use of 
      sperm sexing. The standard validation method is flow cytometric remeasurement of 
      the DNA content of the sexed sperm. Since this method relies on the same 
      instrument that produced the original sperm separation, it is not truly 
      independent. Therefore, to be able to specifically produce either male or female 
      offspring in the dog, we developed a method of direct visualization of sex 
      chromosomes in a single sperm using fluorescence in situ hybridization (FISH) as 
      a validation method. Denaturation of canine spermatozoa by immersion in 1 M NaOH 
      for 4 min yielded consistent hybridization results with over 97% hybridization 
      efficiency and a good preservation of sperm morphology. There was no significant 
      difference between the theoretical ratio (50:50) and the observed ratio of X- and 
      Y-chromosome-bearing spermatozoa in any of the three dogs. In addition, the mean 
      purities of flow-sorted sex chromosomes in spermatozoa of the three dogs were 
      90.8% for the X chromosome fraction and 89.6% for the Y chromosome fraction. This 
      sorting was evaluated by using the dual color FISH protocol. Therefore, our 
      results demonstrated that the FISH protocol worked reliably for both unsorted and 
      sexed sperm samples.
FAU - Oi, Maya
AU  - Oi M
AD  - Research Unit for Functional Genomics, National Research Center for Protozoan 
      Diseases, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido 
      080-8555, Japan.
FAU - Yamada, Keisuke
AU  - Yamada K
FAU - Hayakawa, Hiroyuki
AU  - Hayakawa H
FAU - Suzuki, Hiroshi
AU  - Suzuki H
LA  - eng
PT  - Journal Article
DEP - 20121011
PL  - Japan
TA  - J Reprod Dev
JT  - The Journal of reproduction and development
JID - 9438792
SB  - IM
MH  - Animals
MH  - Dogs
MH  - Female
MH  - Flow Cytometry
MH  - In Situ Hybridization
MH  - *In Situ Hybridization, Fluorescence
MH  - Male
MH  - Reproducibility of Results
MH  - Sex Determination Analysis
MH  - Sex Preselection/*methods
MH  - Spermatozoa/*cytology
MH  - X Chromosome
MH  - Y Chromosome
PMC - PMC3943228
EDAT- 2012/10/13 06:00
MHDA- 2013/09/26 06:00
PMCR- 2013/02/01
CRDT- 2012/10/13 06:00
PHST- 2012/10/13 06:00 [entrez]
PHST- 2012/10/13 06:00 [pubmed]
PHST- 2013/09/26 06:00 [medline]
PHST- 2013/02/01 00:00 [pmc-release]
AID - DN/JST.JSTAGE/jrd/2012-098 [pii]
AID - 2012-098 [pii]
AID - 10.1262/jrd.2012-098 [doi]
PST - ppublish
SO  - J Reprod Dev. 2013;59(1):92-6. doi: 10.1262/jrd.2012-098. Epub 2012 Oct 11.