Effects of indented zona pellucida on oocyte growth and development explored from changes of gene expression in cumulus cells.

Abstract

Abnormal Zona Pellucida (ZP) of human oocytes is an extracellular oocyte abnormality leading to subfertility or infertility, among which indented ZP (iZP) is a common clinical case, and there is currently no effective clinical solution. The study aimed to find out the influence of this abnormal ZP on the growth and development of GC and further explore its influence on the growth and development of oocytes, hoping to provide new ideas for the etiology and treatment of such patients. In this study, we collected granulosa cells GC from oocytes with iZP(four cases) and GC from oocytes with a normal appearance of the ZP(eight cases) during ICSI treatment cycles, and submitted them to transcriptomic analysis using next-generation RNA sequencing (RNAseq). 177 Differentially Expressed Genes (DEG) were identified by RNAseq analysis of Granulosa Cells (GC) from oocytes with a normal ZP morphological appearance and those with iZP. Correlation analysis of these DEGs showed that the expression levels of the immune factor CD274 and the inflammatory factors IL4R and IL-7R, which are positively associated with ovulation, were significantly down-regulated in the GC of oocytes with iZP. Hippo, PI3K-AKT, Ras and calcium signaling pathways related to oocyte growth and development, NTRK2 and its ligands (BDNF and NT5E) from the neurotrophin family that are trophic to the oocyte were also significantly down-regulated in the GC of oocytes with iZP. In addition, the expression of cadherin family members CDH6, CDH12 and CDH19 were significantly down-regulated in DEGs, and the down-regulation of these proteins may affect the gap junction between Granulosa cells and oocytes. IZP might cause obstacles to dialogue and material exchange between GC and oocytes and further affect the growth and development of oocytes.