ABSTRACT
In equine parturition, the role of progestins along with the nuclear progesterone receptor (nPR) signaling pathway in the placenta is not completely clarified. The progestins play an integral role in maintaining myometrial quiescence during the late stage of pregnancy via acting on nPR isoforms (PRA and PRB; PRB is more active than PRA). The current study aimed to determine the PRA and PRB expressions in the term equine placenta at the gene and protein levels. Six term equine placentas were used in this study. Reverse transcription polymerase chain reaction (RT-PCR) was used to quantify the mRNA expression for PRA and PRB. The protein expression was detected using the Western Blot technique. The results revealed that the mRNA and protein expressions for PRA were significantly higher (P < 0.0001) in the term equine placental tissue compared to the mRNA and protein expressions of PRB. These results demonstrated that nPRs are detectable in the term placenta of mares and PRA is the dominant isoform expressed. The present findings raised the possibility that the PRA plays an important role in the parturition process and expulsion of the placenta in mares.
ABSTRACT
Pigs with severe combined immunodeficiency (SCID) are an emerging biomedical animal model. Swine are anatomically and physiologically more similar to humans than mice, making them an invaluable tool for preclinical regenerative medicine and cancer research. One essential step in further developing this model is the immunological humanization of SCID pigs. In this work we have generated T- B- NK- SCID pigs through site directed CRISPR/Cas9 mutagenesis of IL2RG within a naturally occurring DCLRE1C (ARTEMIS)-/- genetic background. We confirmed ART-/-IL2RG-/Y pigs lacked T, B, and NK cells in both peripheral blood and lymphoid tissues. Additionally, we successfully performed a bone marrow transplant on one ART-/-IL2RG-/Y male SCID pig with bone marrow from a complete swine leukocyte antigen (SLA) matched donor without conditioning to reconstitute porcine T and NK cells. Next, we performed in utero injections of cultured human CD34+ selected cord blood cells into the fetal ART-/-IL2RG-/Y SCID pigs. At birth, human CD45+ CD3ε+ cells were detected in cord and peripheral blood of in utero injected SCID piglets. Human leukocytes were also detected within the bone marrow, spleen, liver, thymus, and mesenteric lymph nodes of these animals. Taken together, we describe critical steps forwards the development of an immunologically humanized SCID pig model.
Subject(s)
Bone Marrow Transplantation , Interleukin Receptor Common gamma Subunit/genetics , Severe Combined Immunodeficiency/genetics , Animals , Animals, Genetically Modified , Antigens, CD34 , CRISPR-Cas Systems , Cell Differentiation , Chimera , DNA-Binding Proteins/deficiency , Disease Models, Animal , Gene Targeting , Genetic Engineering , Graft Survival , Host vs Graft Reaction , Humans , Killer Cells, Natural , Models, Animal , Swine , T-Lymphocytes/metabolism , Transplantation, HeterologousABSTRACT
Laparoscopic artificial insemination (LAI) is an intrauterine method of insemination, especially utilized in the small ruminant species to bypass their unique anatomically tortuous cervix. There are several advantages of LAI that include efficient use of processed semen leading to higher pregnancy rates. Success of LAI programs depends on proper implementation of estrus synchronization programs, patient selection and thorough knowledge of the reproductive physiology. In addition, proper equipment and surgical expertise help in reducing patient morbidity and mortality rates. LAI can be associated with several complications as a result of inadequate patient preparation, poor technique or equipment failure. Hence, a thorough planning is essential to carry out the procedure safely and with consistent success rates. Addition of LAI to a small ruminant/food animal practice can be quite profitable and professionally fulfilling, as long as an appropriate investment in equipment and adequate training of veterinarians and technical staff is implemented. Though the technique for performing LAI has been mentioned briefly through various research articles, this article serves as the first comprehensive review of the technique, equipment used, associated complications and useful practical tips that could serve as a guideline for clinicians interested in adding this service to their practice. The article also contains some novel research supported ideas to provide laparoscopic insufflation that have been recently developed.
