1. Investigación

Introduction: Learning molecular genetics techniques is part of the program of the genetics course of the veterinary degree. It is essential to maintain motivation and interest in genetics practice by using a common thread that connects with the students’ interests. This work describes the design, implementation, and results of a gamification strategy developed during two practical sessions in which the student becomes a geneticist to identify which dog a faecal sample collected from the street belongs to, as some municipalities currently do to promote hygiene and public health in their streets. The aim was for students to understand basic concepts and techniques in molecular genetics during these two sessions. This included learning how to extract DNA from different types of samples, describing the amplification using Random Amplification Polymorphic DNA (RAPD), and getting a better understanding of molecular markers and the theory behind Polymerase Chain Reaction (PCR). Methodology: In the first practice, students extracted DNA from fresh dog faeces, while also discussing various DNA sources and extraction methods. Then they used the extracted DNA to create a simulated database of fictional dogs associated with students. They quantified the DNA, analysed its quality, and prepared a dilution to 10ng/μL. In the second practice, the students used RAPD to identify individuals by matching DNA from a simulated collected faeces sample to their fictional dogs DNA database. They performed amplification reactions with various primers pairs, followed by gel electrophoresis, to compare DNA band patterns and identify the dog and the fictional student associated with the uncollected dog faeces. The advantages and limitations of the RAPD technique were discussed, along with its potential applications in veterinary science and genetics. Results and discussion: The students were successful in extracting DNA with concentrations over 100 ng/μL in most cases as well as a good purity with respect to proteins. However, it was found that there was usually a low quality of DNA with respect to salts, although this did not influence the results of the second practice. They were able to generate reproducible RAPD profiles with all primer pairs. The unknown individual could be easily recognized within the database. Conclusions: It is concluded that this educational proposal is an effective option for teaching DNA extraction and the RAPD technique, as well as for many molecular genetics terms and concepts and contributes to the comprehensive training of future veterinary professionals. Additionally, the educational and social value of the practices are highlighted, as they promote interest in science, respect for the environment, and civic responsibility.

The lens proteome undergoes dramatic composition changes during development and maturation. A defective developmental process leads to congenital cataracts that account for about 30% of cases of childhood blindness. Gene mutations are associated with approximately 50% of early-onset forms of lens opacity, with the remainder being of unknown etiology. To gain a better understanding of cataractogenesis, we utilized a transgenic mouse model expressing a mutant ubiquitin protein in the lens (K6W-Ub) that recapitulates most of the early pathological changes seen in human congenital cataracts. We performed mass spectrometry-based tandem-mass-tag quantitative proteomics in E15, P1, and P30 control or K6W-Ub lenses. Our analysis identified targets that are required for early normal differentiation steps and altered in cataractous lenses, particularly metabolic pathways involving glutathione and amino acids. Computational molecular phenotyping revealed that glutathione and taurine were spatially altered in the K6W-Ub cataractous lens. High-performance liquid chromatography revealed that both taurine and the ratio of reduced glutathione to oxidized glutathione, two indicators of redox status, were differentially compromised in lens biology. In sum, our research documents that dynamic proteome changes in a mouse model of congenital cataracts impact redox biology in lens. Our findings shed light on the molecular mechanisms associated with congenital cataracts and point out that unbalanced redox status due to reduced levels of taurine and glutathione, metabolites already linked to age-related cataract, could be a major underlying mechanism behind lens opacities that appear early in life.