Establishment Of Assisted Reproductive Technique And Determination Of Some Biological Indicators Of Marmosets

Laboratory animals,especially genetically modified animals,are of great significance in biological research.Since non-human primates are closely related to human evolution,it is necessary to develop corresponding genetic engineering techniques and assisted reproductive technique(ART)becomes the base of creating transgenic animal.At present,researches on the ART of marmosets are still few in number and lack of standardization.Therefore,this paper aimed at studying ART and determining some biological indicators of common marmoset.In this paper,ART of marmosets was established.In respect of semen collection,using parallel electrodes and stimulating with voltage of 1?4 V,we determined the optimal rectal probe depth,anesthetic dose and the weight range;in respect of menstrual cycle determination,we measured sex hormone level of female marmosets with Enzyme Linked Immunosorbent Assay(ELISA)and the average days of menstrual cycle were 31.75;besides,in respect of superovulation and oocytes collection,we studied on synchronization and superovulation with combination of CLO,rhFSH and hCG hormone and collected oocytes through follicle puncturing technique,for an average of 11.3 oocytes and 76.8%on retrieve rate;in respect of external fertilization,embryo culture and cryopreservation,we obtained zygotes with IVF and the average fertilization rate was 87.5%,the 4-cells rate was 85.7%,and the 8-cells rate was 14.3%.Through the above research,we formulated 5 technical specifications.To optimize and screen microsatellite loci of the marmoset population,we optimized primer amplification conditions and obtained 20 out of 41 marker loci,then we detected them with 30 marmosets,with the result that the number of alleles was 147 totally and 7.35 averagely,the number of effective alleles was 4.04 on average;the mean observed and expectation heterozygosity was 0.15 and 0.25,respectively.Besides the number of PIC was 0.7053 on average.We determined hematological and biochemical indicators of 30 common marmosets and compared the result with date from wild marmosets and macaques breeding.There was significant difference between the female and male group in serum biochemical indexes including HDL-C,LDL-C(P<0.05).Compared with the wild marmosets,the data of HGB,MCHC,NEUT,ALT,AST and GLOB from marmosets fed by our institution were obviously increased,and the data of HCT?ACT?GGT?CREA were obviously different,compared with the macaques.In summary,we established ART system of marmosets,which provided basic work and technique storage for studying genetic modification marmosets.Besides,we screened microsatellite marker loci for the genetic quality detection and provided data for the hematological and biochemical indicators of marmosets.