Sex determining of cat embryo and some feline species

Abstract

SummarySex identification in mammalian preimplantation embryos is a technique that is used currently for development of the embryo transfer industry for zootechnical animals and is, therefore, a resource for biodiversity preservation. The aim of the present study was to establish a rapid and reliable method for the sexing of preimplantation embryos in domestic cats. Here we describe the use of nested PCR identify Y chromosome-linked markers when starting from small amounts of DNA and test the method for the purpose of sexing different species of wild felids. To evaluate the efficiency of the primers, PCR analysis were performed first in blood samples of sex-known domestic cats. Cat embryos were produced both in vitro and in vivo and the blastocysts were biopsied. A Magnetic Resin System was used to capture a consistent amount of DNA from embryo biopsy and wild felid hairs. The results from nested PCR applied on cat blood that corresponded to the phenotypical sex. Nested PCR was also applied to 37 embryo biopsies and the final result was: 21 males and 16 females. Furthermore, β-actin was amplified in each sample, as a positive control for DNA presence. Subsequently, nested PCR was performed on blood and hair samples from some wild felines and again the genotyping results and phenotype sex corresponded. The data show that this method is a rapid and repeatable option for sex determination in domestic cat embryos and some wild felids and that a small amount of cells is sufficient to obtain a reliable result. This technique, therefore, affords investigators a new approach that they can insert in the safeguard programmes of felida biodiversity.

Influence of ROS on Ovarian Functions

“…They can be used in the veterinary field also [90]. ART, in fact, were recently accepted into the programs for the safeguard of endangered species from extinction [90][91][92][93]. In a feasible program it is necessary proceed in the following five steps: 1) Technique development in a domestic animal counterpart, if available; 2) characterization of species-specific reproductive biology in a targeted nondomestic animal; 3) assessment of technique feasibility for producing offspring; 4) demonstration of adequate efficiency for applied usage; 5) application of new tool for population management [90] Figs 4, 5, and 6 show cumulus oocyte complexes (COCs) from mare explanted ovaries: these tools are employed in ART to have genetic improvement in horses,…”

37,X/38,XY Mosaicism in a Cryptorchid Bengal Cat with Müllerian Duct Remnants

“…High molecular weight genomic DNA was isolated using a proteinase K/phenol extraction method. PCR amplification of the SRY gene from genomic DNA was performed using a primer pair described in Ciani et al [2008], which results in a 164-bp amplification product. Chromosomes were prepared according to standard protocols from short-term lymphocyte cultures.…”

A Case of SRY-Positive 38,XY True Hermaphroditism (XY Sex Reversal) in a Cat

“…The entire feline SRY coding region was amplified by polymerase chain reaction (PCR) from genomic DNA extracted from cultured peripheral blood lymphocytes of the affected cat and from peripheral blood of a normal male control cat. To amplify the SRY HMG box region, primers (SRYA1 and SRYA2) and reaction conditions were used as described in Ciani et al 3 The remainder was amplified with PCR primers flanking the coding region of feline SRY, designed in Primer3Plus (http://www.bioinformatics.nl/cgi-bin/ primer3plus/primer3plus.cgi). These reactions (25 ml) contained 1.25mM dNTPs, 2.0mM MgCl 2 , 1Â PCR buffer II (Applied Biosystems, Foster City, CA), 0.375 U of AmpliTaq polymerase (Applied Biosystems), 25 ng of genomic DNA, and 1.0 uM of each primer (forward, 5 0 -ACAACCTTCAACGGTTAGATAA-TTC-3 0 ; reverse, 5 0 -CCAGGCTGGATCAGGTAGAC-3 0 ).…”