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Abstract
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VASA, also known as DDX4, is a member of the DEAD‑box proteins and an RNA binding protein
with an ATP‑dependent RNA helicase. The VASA gene expression, which is required for human
germ cell development, may lead to infertility. Immunocytochemistry and immunohistochemistry
were used to examine the expression of VASA protein in the human testis sections of azoospermic
patients, in‑vitro and in‑silico models. Some studies of fertile humans showed VASA expression in
the basal and adluminal compartments of seminiferous tubules. Our Immunocytochemistry and
immunohistochemistry in infertile humans showed expression of VASA in the luminal compartments
of the seminiferous tubule. The immunohistochemical analysis of three human cases with different
levels of non‑obstructive azoospermia revealed a higher expression of VASA‑positive cells. For this
purpose, Enrichr and Shiny Gene Ontology databases were used for pathway enrichment analysis and
gene ontology. STRING and Cytoscape online evaluation were applied to predict proteins’ functional
and molecular interactions and performed to recognize the master genes, respectively. According
to the obtained results, the main molecular functions of the up‑regulated and downregulated genes
include the meiotic cell cycle, RNA binding, and differentiation. STRING and Cytoscape analyses
presented seven genes, i.e., DDX5, TNP2, DDX3Y, TDRD6, SOHL2, DDX31, and SYCP3, as the hub
genes involved in infertility with VASA co‑function and protein–protein interaction. Our findings
suggest that VASA and its interacting hub proteins could help determine the pathophysiology of germ
cell abnormalities and infertility.
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