PMID- 16104034
OWN - NLM
STAT- MEDLINE
DCOM- 20060120
LR  - 20061115
IS  - 0275-2565 (Print)
IS  - 0275-2565 (Linking)
VI  - 66
IP  - 4
DP  - 2005 Aug
TI  - Flow cytometric sorting of fresh and frozen-thawed spermatozoa in the western 
      lowland gorilla (Gorilla gorilla gorilla).
PG  - 297-315
AB  - We adapted flow cytometry technology for high-purity sorting of X 
      chromosome-bearing spermatozoa in the western lowland gorilla (Gorilla gorilla 
      gorilla). Our objectives were to develop methodologies for liquid storage of 
      semen prior to sorting, sorting of liquid-stored and frozen-thawed spermatozoa, 
      and assessment of sorting accuracy. In study 1, the in vitro sperm 
      characteristics of gorilla ejaculates from one male were unchanged (P > 0.05) 
      after 8 hr of liquid storage at 15 degrees C in a non-egg yolk diluent 
      (HEPES-buffered modified Tyrode's medium). In study 2, we examined the efficacy 
      of sorting fresh and frozen-thawed spermatozoa using human spermatozoa as a model 
      for gorilla spermatozoa. Ejaculates from one male were split into fresh and 
      frozen aliquots. X-enriched samples derived from both fresh and frozen-thawed 
      human semen were of high purity, as determined by fluorescence in situ 
      hybridization (FISH; 90.7%+/-2.3%, overall), and contained a high proportion of 
      morphologically normal spermatozoa (86.0%+/-1.0%, overall). In study 3, we 
      processed liquid-stored semen from two gorillas for sorting using a modification 
      of methods for human spermatozoa. The sort rate for enrichment of X-bearing 
      spermatozoa was 7.3+/-2.5 spermatozoa per second. The X-enriched samples were of 
      high purity (single-sperm PCR: 83.7%) and normal morphology (79.0%+/-3.9%). In 
      study 4 we examined frozen-thawed gorilla semen, and the sort rate (8.3+/-2.9 
      X-bearing sperm/sec), purity (89.7%), and normal morphology (81.4%+/-3.4%) were 
      comparable to those of liquid-stored semen. Depending on the male and the type of 
      sample used (fresh or frozen-thawed), 0.8-2.2% of gorilla spermatozoa in the 
      processed ejaculate were present in the X-enriched sample. These results 
      demonstrate that fresh or frozen-thawed gorilla spermatozoa can be flow 
      cytometrically sorted into samples enriched for X-bearing spermatozoa.
CI  - Copyright 2005 Wiley-Liss, Inc.
FAU - O'Brien, J K
AU  - O'Brien JK
AD  - Centre for Advanced Technologies in Animal Genetics and Reproduction, Faculty of 
      Veterinary Science, University of Sydney, Sydney, Australia. 
      justineo@vetsci.usyd.edu.au
FAU - Stojanov, T
AU  - Stojanov T
FAU - Crichton, E G
AU  - Crichton EG
FAU - Evans, K M
AU  - Evans KM
FAU - Leigh, D
AU  - Leigh D
FAU - Maxwell, W M C
AU  - Maxwell WM
FAU - Evans, G
AU  - Evans G
FAU - Loskutoff, N M
AU  - Loskutoff NM
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Am J Primatol
JT  - American journal of primatology
JID - 8108949
SB  - IM
MH  - Analysis of Variance
MH  - Animals
MH  - *Animals, Zoo
MH  - *Cryopreservation
MH  - Flow Cytometry/*methods
MH  - *Gorilla gorilla
MH  - Male
MH  - Spermatozoa/*cytology
MH  - *X Chromosome
EDAT- 2005/08/17 09:00
MHDA- 2006/01/21 09:00
CRDT- 2005/08/17 09:00
PHST- 2005/08/17 09:00 [pubmed]
PHST- 2006/01/21 09:00 [medline]
PHST- 2005/08/17 09:00 [entrez]
AID - 10.1002/ajp.20158 [doi]
PST - ppublish
SO  - Am J Primatol. 2005 Aug;66(4):297-315. doi: 10.1002/ajp.20158.