Effects of canine oviduct epithelial cells on movement and capacitation of homologous spermatozoa in vitro.
In this study, the interaction between canine sperms and oviduct epithelial cells (OECs) was examined in vitro. The oviducts of eight bitches in the follicular (F-) phase and six bitches in the luteal (L-) phase were removed under halothane inhalation anesthesia. The entire oviduct was opened longitudinally, and the oviductal epithelium of bitches in the F- and L-phases was scraped with a scalpel into tissue culture medium (Eagle's MEM) containing 10% estrous bitch serum and 10% diestrous bitch serum, respectively. The OEC collected were preincubated for 24h and then coincubated with ejaculated canine sperms at 38 degrees C under an atmosphere of 5% CO2 in air. The percentages of sperms exhibiting active tail movement (% TM), hyperactivated sperms (% HA), and acrosome-reacted sperms (% AR) were investigated until 72 h after the start of coincubation. The percentage of sperms labeled with fluoresceinated Ca indicator (% Ca) was evaluated to assess the influx of Ca into sperms cytoplasm during capacitation. Canine sperms attached to both ciliated OEC and non-ciliated OEC. All of the mean % TM of the OEC-binding sperms in the F-OEC and L-OEC media after 24, 48, and 72 h of coincubation were significantly higher than the values of the freely swimming sperms (P< or =0.01). Conversely, the mean % AR and % Ca of the OEC-binding sperms were significantly lower (P<0.01). All of the mean % HA and % AR of the freely swimming sperms in the F-OEC medium after 24, 48, and 72 h of coincubation were significantly higher than the values of the sperms in the L-OEC medium (P< or =0.01). These results indicate that attachment of canine sperms to the OEC prolongs their viability and motility arid inhibits Ca influx into the sperms and sperm capacitation. These phenomena may be responsible for maintaining the active movement and the fertile life of canine sperms in homologous oviducts.
Kawakami
E
E
Department of Reproduction, Nippon Veterinary and Animal Science University, 1-7-1 Kyonan-cho, Musashino-shi, 180-8602, Tokyo, Japan. reprod@annie.ne.jp
Kashiwagi
C
C
Hori
T
T
Tsutsui
T
T
D000177:Acrosome; D000818:Animals; D001826:Body Fluids / Q000502:physiology*; D002118:Calcium / Q000378:metabolism; D004285:Dogs / Q000502:physiology*; D004847:Epithelial Cells / Q000502:physiology; D005260:Female; D005307:Fertilization in Vitro; D005498:Follicular Phase / Q000502:physiology*; D008183:Luteal Phase / Q000502:physiology*; D008297:Male; D010057:Oviducts / Q000166:cytology / Q000502:physiology*; D013075:Sperm Capacitation; D013081:Sperm Motility; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa / Q000502:physiology*; D013997:Time Factors
D002118:Calcium