In this study, we infected Holstein calves with Ostertagia ostertagi stage 3 larvae (L3) and determined gene expression profiles of abomasal fundic and pyloric mucosa by RNA sequencing (RNA-seq) at 3-5, 7-9, 10, and 21 days post-infection (dpi), which represent late L3 and early L4 (between 3-5 dpi), mid to late L4 (between 7-9 and 10 dpi) and adult stages (21 dpi) of the parasitic stage. Bioinformatics analyses were performed to profile the transcriptomic changes over time as well as between cattle abomasal tissues. The results will help understand the gastric responses of the host, especially the immune responses, at different phases of early nematode infection. Such an undertaking is crucial for an in-depth comprehension of host responses, where immunologic reagents for cattle are highly limited. The RNA-seq datasets generated in this study provide a vital data resource, allowing for future comparative analyses to similar data resources or using more advanced analytical technologies in mechanisms of host-parasite interactions.