CONICET-Universidad de Buenos Aires, Instituto de Investigaciones en Producción Animal (INPA), Buenos Aires, Argentina.
Fertilidad San Isidro, Buenos Aires, Argentina.
Open Vet J. 2024 May;14(5):1191-1198. doi: 10.5455/OVJ.2024.v14.i5.14. Epub 2024 May 31.
The intracytoplasmic sperm injection (ICSI) technique has low efficiency in cattle. This has mainly been attributed to the oocyte activation failure due to oocyte and/or sperm factors.
Our aim was to evaluate the effect of conventional ICSI and Piezo-ICSI with bull or human sperm on bovine oocyte activation and embryo development and to assess its relationship with the phospholipase C zeta (PLCɀ) activity of both species.
matured bovine oocytes were randomly divided into five groups and were fertilized as follows: conventional ICSI using bovine sperm with chemical activation (control), conventional ICSI using bovine sperm, Piezo-ICSI using bovine sperm, conventional ICSI using human sperm, and Piezo-ICSI using human sperm. PLCɀ activity was determined in bull and human sperm samples.
Within the groups using bull sperm, the oocytes fertilized by conventional ICSI had the lowest values of 2 pronuclei (PN) formation and cleavage, Piezo-ICSI increased both percentages and ICSI + chemical activation presented the highest 2 PN, cleavage, and blastocyst rates ( < 0.05). Within the groups using human sperm, the oocytes fertilized by Piezo-ICSI presented higher 2 PN and cleavage rates than those activated by conventional ICSI ( < 0.05). Piezo-ICSI with human sperm increased bovine oocyte activation as much as conventional ICSI + chemical activation with bovine sperm ( < 0.05). Higher values of PLCɀ activity were found in human sperm compared with bovine sperm ( < 0.05).
Our results suggest that the higher stability of the bovine sperm in combination with its relatively low content of PLCɀ impairs bovine oocyte activation after ICSI.
胞质内精子注射(ICSI)技术在牛中的效率较低。这主要归因于卵母细胞和/或精子因素导致的卵母细胞激活失败。
我们的目的是评估常规 ICSI 和 Piezo-ICSI 联合公牛或人精子对牛卵母细胞激活和胚胎发育的影响,并评估其与两种精子的磷脂酶 C zeta(PLCɀ)活性的关系。
成熟的牛卵母细胞被随机分为五组,并进行如下受精:使用化学激活的牛精子进行常规 ICSI(对照)、使用牛精子进行常规 ICSI、使用牛精子进行 Piezo-ICSI、使用人精子进行常规 ICSI 和使用人精子进行 Piezo-ICSI。测定了公牛和人精子样本中的 PLCɀ 活性。
在使用公牛精子的组内,常规 ICSI 受精的卵母细胞的 2 原核(PN)形成和分裂率最低,Piezo-ICSI 提高了这两个百分比,ICSI+化学激活呈现出最高的 2 PN、分裂和囊胚率(<0.05)。在使用人精子的组内,Piezo-ICSI 受精的卵母细胞的 2 PN 和分裂率高于常规 ICSI 受精的卵母细胞(<0.05)。Piezo-ICSI 联合人精子增加了牛卵母细胞的激活,与人精子的常规 ICSI+化学激活效果相当(<0.05)。人精子中的 PLCɀ 活性值高于牛精子(<0.05)。
我们的结果表明,与相对较低的 PLCɀ 含量相结合,牛精子的更高稳定性会损害 ICSI 后牛卵母细胞的激活。