RESUMEN
Colony management of gene-modified animals is time-consuming, costly and affected by random events related to Mendelian genetics, fertility and litter size. Careful planning is mandatory to ensure successful outcomes using the least number of animals, hence adhering to the 3R principles of animal welfare. Here we have developed an R package, accessible also through an interactive public website, that optimizes breeding design by providing information about the optimal number of breedings needed to obtain defined breeding outcomes, taking into account specific species, strain, or line properties and success probability. Our software also enables breeding planning for balanced male-to-female ratio or single-sex experiments. We show that, for single-sex designs, the necessary number of breedings is at least doubled compared to the use of all born animals. While the presented tool provides preset parameters for the laboratory mouse, it can be readily used for any other species.
Asunto(s)
Programas Informáticos , Embarazo , Animales , Ratones , Femenino , Masculino , Tamaño de la Camada/genéticaRESUMEN
Molecular events occurring in stepwise progression from pre-malignant lesions (pancreatic intraepithelial neoplasia; PanIN) to the development of pancreatic ductal adenocarcinoma (PDAC) are poorly understood. Thus, characterization of early PanIN lesions may reveal markers that can help in diagnosing PDAC at an early stage and allow understanding the pathology of the disease. We performed the molecular and histological assessment of patient-derived PanINs, tumor tissues and pancreas from mouse models with PDAC (KC mice that harbor K-RAS mutation in pancreatic tissue), where we noted marked upregulation of gastrokine (GKN) proteins. To further understand the role of gastrokine proteins in PDAC development, GKN-deficient KC mice were developed by intercrossing gastrokine-deficient mice with KC mice. Panc-02 (pancreatic cancer cells of mouse origin) were genetically modified to express GKN1 for further in vitro and in vivo analysis. Our results show that gastrokine proteins were absent in healthy pancreas and invasive cancer, while its expression was prominent in low-grade PanINs. We could detect these proteins in pancreatic juice and serum of KC mice. Furthermore, accelerated PanIN and tumor development were noted in gastrokine deficient KC mice. Loss of gastrokine 1 protein delayed apoptosis during carcinogenesis leading to the development of desmoplastic stroma while loss of gastrokine 2 increased the proliferation rate in precursor lesions. In summary, we identified gastrokine proteins in early pancreatic precursor lesions, where gastrokine proteins delay pancreatic carcinogenesis.