Refinement and reduction in production of genetically modified mice

Robinson, Vicky; Morton, David B.; Anderson, Diana; Carver, Jan; Francis, R. J.; Hubrecht, Robert; Jenkins, Elizabeth S.; Mathers, Kathleen; Raymond, Ruth; Rosewell, Ian; Wallace, James A.; Wells, Dominic J.

doi:10.1258/002367703766452723

Cited by 55 publications

(42 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…This indicates that the most important time for assessing these differences is during the early postoperative period and further study should focus on the time period between one and 24 h. The Joint Working Group recommendations indicated a preference for scrotal approach versus abdominal surgery, due to the comparatively smaller incision needed to access the vas deferens. 2 However, this still requires tissue tearing that could result in additional pain, and could explain the relatively similar findings with respect to changes in normal behaviour. It is possible, indeed likely, that surgeon skill rather than technique is more significant in predicting surgical outcomes.…”

Section: Discussionmentioning

confidence: 89%

A comparison of abdominal and scrotal approach methods of vasectomy and the influence of analgesic treatment in laboratory mice

et al. 2012

View full text Add to dashboard Cite

Vasectomized mice are needed in the production of genetically-modified animals. The BVAAWF/FRAME/RSPCA/UFAW Joint Working Group on Refinement recommended that vasectomy should be performed via an incision in the scrotal sac, rather than via laparotomy, arguing that the former could be less painful due to minimal tissue trauma. This study was undertaken to assess the validity of this recommendation. Mice underwent vasectomy via either abdominal or scrotal approach surgery. Mice were filmed for 15 min presurgery and at one, 24 and 48 h postsurgery. Data were obtained using automated behaviour recognition software (HomeCageScan). Meloxicam was administered either alone or combined with acetaminophen prior to surgery. A third group received only saline subcutaneously. Postsurgery behaviour changes were compared between groups at each time point. Exploratory behaviours such as rearing, walking and sniffing were most greatly reduced at one hour following surgery whereas the duration of grooming increased. By 48 h these changes had largely subsided. Results indicated mice undergoing scrotal approach surgery fared better at one hour postsurgery, but the magnitude of this was relatively insignificant compared with the overall effects of surgery. If the observed behaviour changes resulted from pain, results suggested there was no significant advantage of scrotal versus abdominal approach vasectomy. These and other recently obtained data on the effects of non-steroidal anti-inflammatory drugs (NSAIDs) in mice suggest considerably larger doses of these or more potent analgesics, more precise monitoring of surgical outcomes, or a combination of these factors are needed to determine the extent of pain experienced by mice undergoing vasectomy. Of the total number of animals used in regulated procedures in the UK, genetically-modified animals has increased from 8% (1995) to 54% in 2010. 1 Almost all of these animals were mice, of which a significant proportion being males undergoing vasectomy for inducing pseudopregnancy in females for transgenic animal production. It is presently unknown whether these mice, and those currently involved in these types of studies, experience pain. Despite virtually no supporting evidence the BVAAWF/FRAME/RSPCA/ UFAW Joint Working Group on Refinement 2 recommended vasectomy should be performed via scrotal approach surgery, where the vas deferens could be accessed via a relatively minor incision. By comparison with laparotomy, avoiding incising the abdominal muscles bearing the weight of the viscera (ca. 30% of body mass) 2 was considered likely to be less painful. However, scrotal approach vasectomy also involves dissection and tearing of tissue to access the vas deferens, so significant postoperative pain remains likely.Behavioural analysis is a key method of identifying pain postsurgery in rats and mice. In addition to suppression of normal behaviours including walking and rearing, a range of discrete identifiable acts have been characterized following vasectomy including; belly pressin...

show abstract

Section: Discussionmentioning

confidence: 89%

A comparison of abdominal and scrotal approach methods of vasectomy and the influence of analgesic treatment in laboratory mice

et al. 2012

View full text Add to dashboard Cite

show abstract

“…5 -7 Toe clipping has been widely used in the past, but is currently not recommended due to welfare and ethical grounds. 1,8,9 Recommendations and guidelines indicate toe clipping as a method only acceptable for altricial neonates ( 7 postnatal day [ pnd]), when no other methods are feasible 9,10 and several Institutional Animal Care and Use Committees (IACUC) only allow toe clipping when strong scientific justification is presented. 11 -15 Toe clipping can be applied to newborn mice as early as one day after birth, but it is easier to perform between pnd 5 and 7, when the toes are larger but ossification is not yet complete.…”

mentioning

confidence: 99%

Identification methods in newborn C57BL/6 mice: a developmental and behavioural evaluation

et al. 2010

View full text Add to dashboard Cite

The use of group-housed rodents in many fields of biomedical research imposes a need to identify individuals in a cage. Few studies have been designed to assess possible negative effects of identification methods of newborn mice on their development and wellbeing. In the present study, three different identification methods were applied to newborn C57BL/6J mice on postnatal day ( pnd) 5 (toe clipping, toe tattoo ink puncture and subcutaneous implantation of a small transponder). All identification methods used proved to be effective for long-term marking of individual animals. Newborn mice showed the least reaction to toe clipping followed by toe tattoo ink puncture and transponder implantation was the most distressful individual identification procedure in newborn mice. Importantly, clipped toe tissue proved to be enough for genotyping purposes. No overall consistent differences in somatic and neurological reflex development during the postnatal period were shown as a result of the newborn individual identification procedures used. Further, none of the methods interfered significantly with the adult animals' general normal behaviour (e.g. ability to move, grasp, climb) and sensory -motor functions as assessed with a simplified SHIRPA battery of tests, as well as Rotarod and Elevated Plus Maze tests. Postmortem thymus and adrenal gland weights gave no indication of chronic stress as a consequence of the identification method. We conclude that toe clipping might even be advisable in newborn mice at a very young age, when genotyping is needed. Toe tattoo ink puncture is also a good identification method for newborn mice and transponder implantation should only be used in older newborns or applied at weaning.

show abstract

“…The greatest advantage is that embryos at this stage permit postthaw viability to be accurately assessed by culture before transfer into recipient pseudopregnant females [20,24,29]. Therefore, in this study, we adopted freezing of 8-cell embryos.…”

Section: Discussionmentioning

confidence: 99%

“…a bank of cryopreserved embryos also assists the management of animal facilities by providing protection against loss from breeding failure, catastrophe, infectious disease outbreaks, or genetic drift [9,18,27]. Cryopreservation also avoids the logistics associated with the transport of live mice from one place to another [20]. Transport of animals require appropriate transport cages, proper environmental conditions, food, a water source, road connectivity, regulatory permission to transport the animals, and quarantine.…”

Section: Introductionmentioning

confidence: 99%

An Attempt of Cryopreservation of Mouse Embryos at the ACTREC Laboratory Animal Facility in India

Thorat

Ingle

2012

Exp. Anim.

View full text Add to dashboard Cite

Abstract:Cryopreservation is the long-term storage of viable cells/tissue in liquid nitrogen. The present study was conducted to freeze 8-cell-to morula-stage mouse embryos from the ACTREC Laboratory Animal Facility using a "slow freezing and fast revival" method. In all, 4,088 embryos were collected from 495 donor female mice of ten different strains. An average recovery of 8 embryos per donor mouse were recorded. Of the 4,088 embryos, 3,946 embryos of normal morphology were frozen in 173 straws. They were cooled down using a controlled-rate freezing assembly, and the straws were directly plunged into liquid nitrogen for long-term storage. Out of these 3,946 frozen embryos, 2,650 were found to be viable after fast revival. The highest survival rate, 81%, was recorded in B6D2F1 hybrid mice, whereas the lowest rate, 51%, was recorded in the S/RV/Cri-ba mutant strain. Out of 2,650 viable embryos, 2,359 embryos (89%) developed to the blastocyst stage after 24 h of incubation in a CO 2 incubator. The developed blastocysts were transferred surgically into 101 pseudopregnant female mice, of which 49 (48.5%) females were found to be pregnant. The highest percentage of pregnancy, 75%, was recorded in C57BL/6NCrl and NIH-III mice, whereas no pregnant recipients were recorded in Ptch, C3H/HeNCrl and NOD SCID mice. Based on the deliveries of these 49 females, an average of 4 young were delivered per female. Improvement in efficiency of freezing, thawing, and surgical transfer of embryos into pseudopregnant females is one of the challenges in such studies.

show abstract

Refinement and reduction in production of genetically modified mice

Cited by 55 publications

References 42 publications

A comparison of abdominal and scrotal approach methods of vasectomy and the influence of analgesic treatment in laboratory mice

A comparison of abdominal and scrotal approach methods of vasectomy and the influence of analgesic treatment in laboratory mice

Identification methods in newborn C57BL/6 mice: a developmental and behavioural evaluation

An Attempt of Cryopreservation of Mouse Embryos at the ACTREC Laboratory Animal Facility in India

Contact Info

Product

Resources

About