McDaniel C D, Bramwell R K, Howarth B
Department of Poultry Science, University of Georgia, Athens 30602, USA.
J Reprod Fertil. 1997 Jan;109(1):173-9. doi: 10.1530/jrf.0.1090173.
A technique was developed to determine the total number of spermatozoa stored in the uterovaginal junction of hens. After insemination of spermatozoa treated with the nuclear fluorescent dye bisbenzimide, oviductal tissue was collected from hens and homogenized. Samples of homogenate were dried, and the number of spermatozoa mm-2 was determined with the use of a fluorescence microscope. When spermatozoa were added to excised uterovaginal junction tissue before homogenization, results indicated a 1:1 linear relationship between actual numbers of spermatozoa added to the tissue and calculated numbers of spermatozoa added to the tissue. This new technique was used to show that insemination of hens with 25, 50 or 100 x 10(6) spermatozoa resulted in a linear increase in the number of spermatozoa stored in the uterovaginal junction. Insemination of hens with 328 x 10(6) spermatozoa produced no increase in uterovaginal junction storage of spermatozoa over insemination with 100 x 10(6) spermatozoa. At the maximum sperm storage tubule filling dose of 100 x 10(6) spermatozoa, only 0.22% of the spermatozoa inseminated were found in the uterovaginal junction 24 h after insemination. Treatment of spermatozoa with bisbenzimide had no detrimental effects on fertility or penetration rates when compared with untreated (control) spermatozoa. However, when spermatozoa were treated with bisbenzimide, hatchability of fertile eggs was reduced. In conclusion, this new fluorescence technique appears to be valuable in determining the total number of spermatozoa stored in the uterovaginal junction of hens.
已开发出一种技术来确定母鸡子宫阴道连接处储存的精子总数。在用核荧光染料双苯甲酰亚胺处理精子后进行授精，从母鸡收集输卵管组织并匀浆。匀浆样品干燥后，使用荧光显微镜确定每平方毫米的精子数量。当在匀浆前将精子添加到切除的子宫阴道连接处组织时，结果表明添加到组织中的实际精子数量与计算得出的添加到组织中的精子数量之间呈1:1线性关系。这项新技术用于表明，用25、50或100×10⁶个精子对母鸡进行授精会导致子宫阴道连接处储存的精子数量呈线性增加。用328×10⁶个精子对母鸡进行授精，与用100×10⁶个精子授精相比，子宫阴道连接处的精子储存量没有增加。在最大精子储存小管充盈剂量为100×10⁶个精子时，授精后24小时在子宫阴道连接处仅发现0.22%的授精精子。与未处理（对照）的精子相比，用双苯甲酰亚胺处理精子对受精率或穿透率没有不利影响。然而，当精子用双苯甲酰亚胺处理时，受精卵的孵化率降低。总之，这种新的荧光技术在确定母鸡子宫阴道连接处储存的精子总数方面似乎很有价值。
