MMPs EXPRESSION IN LLAMA REPRODUCTIVE TRACT

ZAMPINI, RENATO; ARGAÑARAZ, MARTÍN E; APICHELA, SILVANA A.; MICELI, DORA C.

Sevilla

Simposio; 5TH EUROPEAN SYMPOSIUM ON SOUTH AMERICAN CAMELIDS AND 1ST EUROPEAN MEETING ON FIBRE ANIMALS; 2010

MInisterio de Medio Ambiente y Medio Rural y Marino. España

The knowledge of the reproductive biology in South American camelids (SAC) is essential for developing new reproductive biotechnologies and for increasing the rate of genetic improvement. Our particular interests are the molecules involved in fertilization, pre-implantational embryo development and embryo implantation, which take place in the oviduct and uterine horns. The oviduct provides an optimal environment for crucial events that occur during the early reproductive stage, leading to pregnancy. These occurrences include transportation of gametes, fertilization, embryos early development and embryo implantation. Different proteolitic systems, which participate in the reproductive process, are synthezed and secreted by the oviductal epithelium. Among them, MMPs are enzymes that hydrolyze components of the extracellular matrix. These proteins are proposed to be involved regulating the structural changes in the uterus and ovary during the estrous cycle and pregnancy. The aim of this work was to identify the nucleotide sequences of some MMPs and analyze their expression in the reproductive tract of llama. Ovary, oviduct and uterine horns were obtained from llama females belonging to the INTA Abra Pampa (Jujuy, Argentine). Samples were homogenized with lysis buffer, stored in liquid N2, transported to the lab and then, processed for total RNA extraction. Since the SAC genome or EST (Expression sequence tag) are scarce or not available MMP1, MMP2, MMP8 and MMP9 primers were designed from mRNA known sequence of other mammals mRNA (cow, pig, horse, rat and human). RT-PCR of oviducts was performed. MMP1, MMP2 and MMP9 amplification products with the expected size, were cloned and sequenced. The amplicon sequences showed a high identity percentage with their counterparts in other species, confirming that they corresponded to MMP1, MMP2 and MMP9. MMP8 was no detected in llama oviduct. . Specific primers were designed and semiquantitative relative RT-PCR was performed in ovary, oviduct (uterino-tubal junction, isthmus and ampulla) and uterine horns. We observed the expression of MMP1, MMP2 and MMP9 all along the reproductive tract. Interestingly, the intensity of all the MMPs studied increased from ampulla region to the uterus, which could be related to higher activity in the caudal oviduct and uterus. This is the first time that llama MMPs were isolated and sequenced; moreover, MMP9 has only been reported in mammalian uterus and ovary. MMPs ESTs are a significant contribution to the knowledge of llama reproductive tract transcriptome, and will set the basis for numerous molecular studies. MMPs presence in the reproductive tract suggests that they might be involved in reproductive events. Further studies are required to establish their functions.