A Noninvasive Ocular (Tear) Sampling Method for Genetic Ascertainment of Transgenic Mice and Research Ethics Innovation

Abstract

Animal models, animal welfare and research ethics are both facilitators and gatekeepers for Big Data generation in genomics and multi-omics R&D. Safeguarding animal welfare is also a research ethics issue that can benefit from technical innovations in biosample collection in particular. Animal welfare draws from the guiding principles of 3R, namely, ''Replacement'' (methods avoiding the use of animals in research), ''Reduction'' (methods using fewer animals or derive more information from the same number of animals), and ''Refinement'' (methods removing or minimizing pain or distress). We report here that noninvasive ocular (tear) sampling for genetic ascertainment of transgenic mice can serve as an innovative ethical safeguard for animal welfare, and as a veritable alternative to the surgical tail biopsies, ear puncture, or blood sampling from the weanling transgenic mice. We compared ocular versus tail biopsy sampling in regard to ascertainment, by genotyping, of apolipoprotein E-deficient (ApoE-/-) transgenic weanling mice (n = 60) by one-round polymerase chain reaction analysis. We found that ocular sampling compares to the results obtained by tail sampling with the obvious benefit of being noninvasive and improving the 3R, especially for the Refinement principle of animal welfare. To place the importance of this new biosample collection approach into further context, transgenic mice research and animal models are at the epicenter of Big Data translation to health innovation. We suggest that ocular sampling is considered and evaluated further in transgenic mice models, not to mention warrant exploration for applications in other types of animal models that require noninvasive biosample collection.

An eye on the dog as the scientist's best friend for translational research in ophthalmology: Focus on the ocular surface

“…249,257 In addition to their conventional use for aqueous tear assessment, Schirmer strips can retrieve tear fluid for bioanalytical purposes in humans, 182,216,219,220,228,231,258 dogs, 190,209,211,214,215,240,259 and small laboratory species. 244,260,261 For instance, Schirmer strips were used for indepth characterization of proteomics in human 173 and canine tears, 190 and can also successfully recover specific analytes such as cytokines, 200,258 clusterin, 182 and xenobiotics. 211,[213][214][215]244,262 Absorbent sponges exist in different material types such as cellulose, 234,236,263,264 polyvinyl acetal, 209,214,228,236,244,264-266 polyester, 239,242,266 and polyrurethane.…”

“…41 cellulose filter paper and possess specific characteristics to promote tolerance (5‐mm width × 35‐mm length, 0.22‐mm thick, 20–25 µm porosity, foldable extremity for ease of insertion) 249,257 . In addition to their conventional use for aqueous tear assessment, Schirmer strips can retrieve tear fluid for bioanalytical purposes in humans, 182,216,219,220,228,231,258 dogs, 190,209,211,214,215,240,259 and small laboratory species 244,260,261 . For instance, Schirmer strips were used for in‐depth characterization of proteomics in human 173 and canine tears, 190 and can also successfully recover specific analytes such as cytokines, 200,258 clusterin, 182 and xenobiotics 211,213–215,244,262 …”

Refinement in the European Union: A Systematic Review

“…A total of 384 references were identified by electronic search; 338 full-text studies were evaluated in accordance with the eligibility criteria and 290 were excluded. Finally, 48 studies [ 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 ] complied with all the established eligibility criteria and were included in the study ( Figure 1 ).…”

An Eye on the Dog as the Scientist’s Best Friend for Translational Research in Ophthalmology: Focus on the Ocular Surface

“…197,207 In addition to their conventional use for aqueous tear assessment, Schirmer strips can retrieve tear fluid for bioanalytical purposes in humans, 144,161,165,166,173,176,208 dogs, 143,149,156,159,160,186,209 and small laboratory species. 191,210,211 For instance, Schirmer strips were used for in-depth characterization of proteomics in human 162 and canine tears, 149 and can also successfully recover specific analytes such as cytokines, 208,212 clusterin, 144 and xenobiotics. 156,[158][159][160]191,213 Absorbent sponges exist in different material types such as cellulose, 179,181,214,215 polyvinyl acetal, 143,159,173,181,191,214,216,217 polyester, 185,188,217 and polyrurethane.…”