Evaluation of tail biopsy collection in laboratory mice (Mus musculus): vertebral ossification, DNA quantity, and acute behavioral responses.
A preferred method to genotype genetically engineered mice is through collection of distal tail tissue (tail biopsy) followed by DNA isolation. Currently, general or local anesthesia (or both) is recommended for biopsy after 3 wk of age, the time after which tail vertebrae are considered to be ossified. Our objective was to rigorously evaluate vertebral development, DNA content, and acute behavioral responses at different ages by harvesting tail biopsies of different lengths. We evaluated laboratory mice from 5 inbred strains and 1 outbred stock at each of 12 ages (3 to 42 d of age). Biopsies of 5-, 10-, and 15-mm lengths were obtained. Vertebrae were graded according to level of ossification by using complementary modalities of high-resolution microradiography, microcomputed tomography, and histology. Vertebral development progressed at different rates among the strains, with mature tail vertebrae containing endplates detectable in the tail of some strains by 10 d of age. Within the distal 2 mm of tail, end plates were not identified before 21 d of age. DNA yield (DNA weight/tissue weight) was greatest from the 5-mm biopsy harvest. Acute behavioral responses to biopsy varied by age and strain, and these differences were associated with vertebral maturation. Vertebral development progressed most rapidly in C57BL/6 mice, which also demonstrated the highest response rate to biopsy, whereas BALB/c mice had slower vertebral development and were less responsive. These findings support the collection of minimal lengths of tail tissue from mice at ages younger than 17 d, unless anesthesia or analgesia is provided.
Hankenson
F Claire
FC
University Laboratory Animal Resources and Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA. fclaire@upenn.edu
Garzel
Laura M
LM
Fischer
David D
DD
Nolan
Bonnie
B
Hankenson
Kurt D
KD
D000375:Aging; D000698:Analgesia / Q000662:veterinary; D000758:Anesthesia / Q000662:veterinary; D000818:Animals; D001522:Behavior, Animal; D001706:Biopsy / Q000009:adverse effects / Q000379:methods / Q000662:veterinary*; D004247:DNA / Q000032:analysis*; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D010012:Osteogenesis; D013131:Spine / Q000254:growth & development / Q000473:pathology; D013623:Tail / Q000737:chemistry / Q000473:pathology*
D004247:DNA