PMID- 10735111
OWN - NLM
STAT- MEDLINE
DCOM- 20000419
LR  - 20131121
IS  - 0093-691X (Print)
IS  - 0093-691X (Linking)
VI  - 52
IP  - 6
DP  - 1999 Oct 15
TI  - Fluorescence in situ hybridization with Y chromosome-specific probe in 
      decondensed bovine spermatozoa.
PG  - 1043-54
AB  - This study was carried out to demonstrate bovine Y chromosome-bearing spermatozoa 
      by rapid fluorescence in situ hybridization (FISH), using a digoxigenin 
      (Dig)-labeled DNA probe specific to bovine Y chromosome. Before the FISH 
      procedure, sperm heads were treated for decondensation with dithiothreitol (DTT) 
      and glutathione (GSH) with or without heparin supplementation. Concentrations of 
      either above 2 mM DTT or above 100 mM GSH induced swelling of the sperm head, 
      which resulted in sufficient detection of the Y chromosome signal in sperm nuclei 
      by rapid FISH (49.8 to 53.4%). When FISH was used with 2 mM DTT or 100 mM GSH on 
      specimens from 7 sires, the rate of detection of the Y chromosome signal varied 
      among sires (5.4 to 49.6%), especially that of the GSH treatment. Supplementation 
      of GSH with heparin (100 U/mL), however, could induce reliable, repeatable 
      detection of the Y chromosome signal in sperm nuclei of all the 7 sires (48.4 to 
      50.3%). These results show that in bovine spermatozoa decondensed with GSH and 
      heparin, rapid FISH can detect Y chromosome-bearing spermatozoa.
FAU - Kobayashi, J
AU  - Kobayashi J
AD  - Miyagi Agricultural College, Sendai, Japan.
FAU - Kohsaka, T
AU  - Kohsaka T
FAU - Sasada, H
AU  - Sasada H
FAU - Umezu, M
AU  - Umezu M
FAU - Sato, E
AU  - Sato E
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Theriogenology
JT  - Theriogenology
JID - 0421510
RN  - 0 (DNA Probes)
RN  - 9005-49-6 (Heparin)
RN  - GAN16C9B8O (Glutathione)
RN  - NQ1SX9LNAU (Digoxigenin)
RN  - T8ID5YZU6Y (Dithiothreitol)
SB  - IM
MH  - Animals
MH  - Cattle
MH  - DNA Probes
MH  - Digoxigenin
MH  - Dithiothreitol/pharmacology
MH  - Glutathione/pharmacology
MH  - Heparin/pharmacology
MH  - In Situ Hybridization, Fluorescence
MH  - Male
MH  - Sex Determination Analysis/methods/*veterinary
MH  - Sperm Head/drug effects/*ultrastructure
MH  - Spermatozoa/*cytology
MH  - *Y Chromosome
EDAT- 2000/03/29 09:00
MHDA- 2000/04/25 09:00
CRDT- 2000/03/29 09:00
PHST- 2000/03/29 09:00 [pubmed]
PHST- 2000/04/25 09:00 [medline]
PHST- 2000/03/29 09:00 [entrez]
AID - S0093-691X(99)00193-4 [pii]
AID - 10.1016/S0093-691X(99)00193-4 [doi]
PST - ppublish
SO  - Theriogenology. 1999 Oct 15;52(6):1043-54. doi: 10.1016/S0093-691X(99)00193-4.