The leptin receptor overlapping transcript (LepROT) has been suggested to play several roles in immunomodulatory mechanisms; however, the understanding of its role in Rana amurensis immunity is still very limited. Here, we performed hematoxylin-eosin staining, quantitative reverse-transcription polymerase chain reaction (qRT-PCR), immunofluorescence and western blotting to investigate the roles of LepROT in the immunomodulatory mechanism and the influence of its expression on the nuclear factor-kappa B (NF-κB) signaling pathway, such as the activation of IκB kinase and NF-кB, in amphibian resistance to infection with Aeromonas hydrophila (Ah). After Ah infection, the liver, lung, kidney, skin, muscle, and stomach of R. amurensis showed cell structure disturbance, bleeding, and texture abnormalities. In addition, the relative expression levels of LepROT, NF-кB, IKKα, and IKKβ were all upregulated after Ah infection; however, they showed time-dependent differential expression. The NF-кB signaling pathway exhibited robust expression levels, which might be explained by the positive feedback regulation function of LepROT. Overall, this study provides a basis for further assessment of the biological functions of LepROT and highlights its role in the regulation of immune mechanisms.

Amphibians, as a group greatly disturbed by human activities, are at increased risk of extinction. Rana dybowskii is an anuran species with both ecological and economic significance. Due to environmental changes and human overexploitation, it has been classified as Near-Threatened. This study integrates morphological and molecular immunological approaches to identify R. dybowskii populations with greater survival and disease resistance, based on 32 morphological traits and MHC class I and II polymorphism. Morphological results showed that compared with other populations, Yichun (YC) population had the highest fatness, the lowest IOD/HW, and the largest HW/SVL, HL/SVL, HW/HL, SL/TL. It indicates that YC population shows larger body size, wider vision and stronger jumping ability. The polymorphism of MHC I gene was the highest in Shangzhi (SZ) population, and the polymorphism of MHC II gene was the highest in YC population. Moreover, duplication, selection, and recombination occurred during evolution of MHC class I and II genes. Since both SZ and YC populations scored higher in this category (the variant sites, nucleotide polymorphism, amino-acid divergence/nucleotide divergence, dN/dS, Tajima’ D, etc.), they were more resistant to disease. All in all, these results indicated that YC population of the Lesser Khingan Mountains had good morphology and immune results, and R. dybowskii in the Lesser Khingan Mountains might be more suitable to be the original population of artificial breeding, which provided a theoretical basis for the realization of artificial breeding in the next step.