Uteshev V K, Shishova N V, Kaurova S A, Browne R K, Gakhova E N
Institute of Cell Biophysics, Russian Academy of Sciences, Pushchino, Moscow Region 142290, Russia.
Reprod Fertil Dev. 2012;24(4):599-607. doi: 10.1071/RD10324.
The use of hormonally induced spermatozoa expressed in urine (HISu) is a valuable component of reproduction technologies for amphibians. Five protocols for sampling HISu from the European common frog (Rana temporaria) were compared: (1) pituitary extracts, (2) 0.12 µg g⁻¹ luteinising hormone-releasing hormone analogue (LHRHa), (3) 1.20 µg g⁻¹ LHRHa, (4) 11.7 IU g⁻¹ human chorionic gonadotrophin (hCG) and (5) 23.4 IU g⁻¹ hCG (g⁻¹ = per gram bodyweight). From 1 to 24h after administration we assessed the number and concentration of spermatozoa in spermic urine and in holding water, and in urine the percentage of motile spermatozoa and their progressive motility. The protocol using 1.20 µg g⁻¹ LHRHa gave the highest total sperm numbers (650 × 10⁶) and the highest percentage (40%) of samples with sperm concentrations above 200 × 10⁶ mL⁻¹. The percentage motility and progressive motility was similar from all protocols. Considerable amounts of spermatozoa were expressed by R. temporaria into their holding water. We tested hormonal priming and spermiation in the common toad (Bufo bufo) using 0.13 µg g⁻¹ LHRHa administered 24h before a final spermiating dose of 12.8 IU g⁻¹ hCG. No spermatozoa were expressed in holding water. Priming resulted in 35% more spermatozoa than without; however, there were no differences in sperm concentrations. Primed B. bufo produced spermatozoa with significantly higher percentage motility, but not progressive motility, membrane integrity, or abnormal spermatozoa than unprimed males.
利用尿液中激素诱导产生的精子（HISu）是两栖动物繁殖技术的一个重要组成部分。比较了从欧洲普通青蛙（林蛙）采集HISu的五种方案：（1）垂体提取物，（2）0.12 µg g⁻¹促黄体生成素释放激素类似物（LHRHa），（3）1.20 µg g⁻¹ LHRHa，（4）11.7 IU g⁻¹人绒毛膜促性腺激素（hCG）和（5）23.4 IU g⁻¹ hCG（g⁻¹ =每克体重）。给药后1至24小时，我们评估了精液尿液和饲养水中精子的数量和浓度，以及尿液中活动精子的百分比及其前进运动能力。使用1.20 µg g⁻¹ LHRHa的方案产生的精子总数最高（650×10⁶），精子浓度高于200×10⁶ mL⁻¹的样本百分比最高（40%）。所有方案的活动精子百分比和前进运动能力相似。林蛙会向饲养水中排出大量精子。我们使用0.13 µg g⁻¹ LHRHa在最终排精剂量12.8 IU g⁻¹ hCG前24小时对普通蟾蜍（中华大蟾蜍）进行激素预处理和排精测试。饲养水中未排出精子。预处理后产生的精子比未预处理的多35%；然而，精子浓度没有差异。预处理的中华大蟾蜍产生的精子活动精子百分比显著更高，但前进运动能力、膜完整性或异常精子与未预处理的雄性相比没有差异。
