背景:附睾对精子成熟很重要,没有适当的发育,会导致男性不育。组织/器官的生物力学特性在其形态发生中起着关键作用,包括沃尔夫导管.假设囊和围绕导管的间充质/细胞外基质的结构/体积刚度是调节沃尔夫导管形态发生的主要生物力学特性。这些数据将为调节这一重要器官发育的机制提供关键信息。
目的:测量在发育过程中围绕沃尔夫导管的囊和囊和间充质的结构/体积刚度,以帕斯卡(力/面积)为单位。目的研究间充质细胞在Wolffian导管发育过程中的相对膜张力。由于Ptk7以前被证明可以调节ECM完整性和Wolffian管道伸长和卷绕,测试了Ptk7调节结构/体积刚度和间充质细胞膜张力的假设。
方法:使用原子力显微镜和灭菌器压缩装置测量结构刚度。使用膜张力荧光探针检查了胶囊和间充质内细胞膜内的生物力学特性。
结论:在发育过程中,囊和下层间充质的结构刚度(帕斯卡)相对恒定,后期胶囊显着增加。然而,这种增加可能反映了ECM和相关的间质靠近胶囊,因为管道的卷绕将它们推入或压缩到该空间中。将胶囊和间充质/ECM保持在恒定的刚度将确保导管在整个开发过程中在类似的生物力学力下继续盘绕。在发育过程中,不同Wolffian导管区域的囊内和间充质内的细胞具有不同程度的膜脂张力。假设动态变化可确保管道保持恒定的刚度,而与任何外力无关。Ptk7的损失导致在E18.5处的刚度增加,这是可推测的,这是由于间充质内ECM的完整性损失。
结论:包膜和Wolffian导管周围的间充质/细胞外基质的生物力学特性对Wolffian导管形态发生具有重要作用,从而允许附睾的适当发育和随后的男性生育能力。
BACKGROUND: The epididymis is important for sperm maturation and without its proper development, male infertility will result. Biomechanical properties of tissues/organs play key roles during their morphogenesis, including the Wolffian duct. It is hypothesized that structural/bulk stiffness of the capsule and mesenchyme/extracellular matrix that surround the duct is a major biomechanical property that regulates Wolffian duct morphogenesis. These data will provide key information as to the mechanisms that regulate the development of this important organ.
OBJECTIVE: To measure the structural/bulk stiffness in Pascals (force/area) of the capsule and the capsule and mesenchyme together that surrounds the Wolffian duct during the development. To examine the relative membrane tension of mesenchymal cells during the Wolffian duct development. Since Ptk7 was previously shown to regulate ECM integrity and Wolffian duct elongation and coiling, the hypothesis that Ptk7 regulates structural/bulk stiffness and mesenchymal cell membrane tension was tested.
METHODS: Atomic force microscopy and a microsquisher compression apparatus were used to measure the structural stiffness. Biomechanical properties within the membranes of cells within the capsule and mesenchyme were examined using a membrane-tension fluorescent probe.
CONCLUSIONS: The structural stiffness (Pascals) of the capsule and underlying mesenchyme was relatively constant during development, with a significant increase in the capsule at the later stages. However, this increase may reflect the ECM and associated mesenchyme being close to the capsule because the coiling of the duct pushed or compressed them into that space. Keeping the capsule and mesenchyme/ECM at constant stiffness would ensure that the duct will continue to coil under similar biomechanical forces throughout the development. Cells within the capsule and mesenchyme at different Wolffian duct regions during the development had varying degrees of membrane lipid tension. It is hypothesized that the dynamic changes ensure the duct is kept at a constant stiffness regardless of any external forces. Loss of Ptk7 resulted in an increase in stiffness at E18.5, which was presumable due to the loss of integrity of the ECM within the mesenchyme.
CONCLUSIONS: Biomechanical properties of the capsule and the mesenchyme/extracellular matrix that surround the Wolffian duct play an important role toward Wolffian duct morphogenesis, thereby allowing for the proper development of the epididymis and subsequent male fertility.