Fertilization of C57BL/6 mouse sperm collected from cauda epididymides after preservation or transportation at 4 degrees C using laser-microdissected oocytes.
The C57BL/6 mouse is commonly used to produce transgenic and knockout strains for biomedical research. However, the motility and fertility of its sperm decrease markedly with freezing. Short-term preservation of sperm without freezing can avoid this. Furthermore, such samples can be transported safety without the special skills or equipment needed for the transportation of live animals or frozen products. We evaluated the motility and fertility of sperm collected from cauda epididymides after preservation or transportation at 4 degrees C. Oocytes with the zona pellucida subjected to laser-microdissection were used to assist fertilization in vitro. Although the motility of sperm gradually decreased with storage (P<0.05), no disruption of the sperm plasma membrane was seen. The proportion of zona-intact oocytes fertilized with sperm preserved for 0, 24, 48 and 72h were 70, 14, 5 and 1%, respectively. On the other hand, 45, 20 and 14% of laser-microdissected oocytes were fertilized by sperm preserved for 24, 48 and 72h, respectively (P<0.05). The fertility of sperm collected from cauda epididymides of two transgenic strains after transportation at 4 degrees C were also significantly increased using laser-microdissected oocytes rather than zona-intact oocytes (57 and 68% vs. 5%, P<0.05). Efficient production of offspring from sperm preserved or transported at 4 degrees C was achieved using laser-microdissected oocytes. Thus the fertility of sperm preserved or transported at 4 degrees C could be maintained, although motility gradually decreased with storage. Laser-microdissected oocytes will contribute to the efficient production of embryos and offspring using such preserved sperm samples.
Kaneko
Takehito
T
Division of Reproductive Engineering, Center for Animal Resources and Development (CARD), Kumamoto University, Honjo, Kumamoto, Japan. take@kumamoto-u.ac.jp
Fukumoto
Kiyoko
K
Haruguchi
Yukie
Y
Kondo
Tomoko
T
Machida
Hiromi
H
Koga
Mika
M
Nakagawa
Yoshiko
Y
Tsuchiyama
Shuuji
S
Saiki
Kiyora
K
Noshiba
Shiho
S
Nakagata
Naomi
N
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D015925:Cryopreservation / Q000379:methods*; D004822:Epididymis / Q000473:pathology*; D005260:Female; D005306:Fertilization; D005307:Fertilization in Vitro / Q000379:methods*; D005615:Freezing; D007834:Lasers; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008954:Models, Biological; D009865:Oocytes / Q000378:metabolism; D013094:Spermatozoa / Q000378:metabolism*