![]() ![]() beta-cyclodextrins initiate transmembrane signaling leading to an increase in protein tyrosine phosphorylation and capacitation. Cholesterol efflux-mediated signal transduction in mammalian sperm. Influence of serum albumin on the fertilizing ability in vitro of rat spermatozoa. Strain differences in superovulatory response, embryo development and efficiency of transgenic rat production. Optimal conditions for successful in vitro fertilization and subsequent embryonic development in Sprague–Dawley rats. Establishment of sperm cryopreservation and in vitro fertilisation protocols for rats. ![]() Nakagata, N., Mikoda, N., Nakao, S., Nakatsukasa, E. Extracellular ATP and dibutyryl cAMP enhance the freezability of rat epididymal sperm. Generation of live rats produced by in vitro fertilization using cryopreserved spermatozoa. Generation of live rat offspring by intrauterine insemination with epididymal spermatozoa cryopreserved at −196 degrees C. Nakatsukasa, E., Inomata, T., Ikeda, T., Shino, M. Changes in rat spermatozoa function after cooling, cryopreservation and centrifugation processes. Osmotic tolerance limits and effects of cryoprotectants on the motility, plasma membrane integrity and acrosomal integrity of rat sperm. Motility initiation of quiescent spermatozoa from rat caudal epididymis: effects of pH, viscosity, osmolality and inhibitors. Cryopreservation of mouse sperm for genome banking. Cryopreservation of mouse spermatozoa: double your mouse space. Archiving genetically altered animals: a review of cryopreservation and recovery methods for genome edited animals. National BioResource Project-Rat and related activities. Progress and prospects in rat genetics: a community view. ssODN-mediated knock-in with CRISPR–Cas for large genomic regions in zygotes. Simultaneous generation and germline transmission of multiple gene mutations in rat using CRISPR–Cas systems. Heritable gene targeting in the mouse and rat using a CRISPR–Cas system. The Laboratory Rat 3rd edn (Academic Press, 2020). This sperm cryopreservation and in vitro fertilization protocol for rats will provide an efficient system for the archiving and production of genetically modified rats for the transgenic community. This protocol can be easily implemented by researchers and technicians with experience in reproductive engineering technology it can also be used, albeit with some practice, by researchers and technicians who have no experience in reproductive techniques. This process takes approximately 1 month to produce live pups from cryopreserved sperm. This protocol consists of three main steps: preparation of cryopreserved sperm, in vitro fertilization using cryopreserved sperm and embryo transfer. In this optimized protocol, treatment of frozen–thawed rat sperm with a high concentration of bovine serum albumin (40 mg/ml) results in a high in vitro fertilization rate. Here we describe an improved in vitro fertilization protocol to enhance the fertilization rate of cryopreserved sperm in major strains of rats. Previously, we have reported a protocol for rat sperm cryopreservation and in vitro fertilization using frozen–thawed sperm. Sperm cryopreservation is the most efficient and robust means to archive genetic resources, and this technique reduces the number of animals required for colony management. This development has increased the demand for efficient preservation and production of rat resources. Recently, genome editing technology has facilitated the generation of genetically modified rats worldwide. Laboratory rats have been used in biomedical research for over 170 years. ![]()
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