Role of leptin in reproductive function of male dog

Studies investigating leptins role in canine male reproductive function has not previously been done. For that particular reason, this evaluation of a previous pilot study was designed to investigate the presence of leptin and leptin receptors in canine testis, epididymis and spermatozoa. Obesity is currently the most frequent nutritional disease existing in companion animals, however this increasingly issue is not given proper attention. In obese animals an increased amount of adipose tissue is found. With an excess of adipocytes, an increased level of leptin will be produced. An interesting question is therefore raised, and it could possibly be of great value if we were capable of finding out whether leptin demonstrates an impact towards the male reproductive function. However, this issue should be investigated by first examining the role of leptin in the function of the male genital tract of healthy dogs in normal condition. In this research we obtained samples from six healthy and mature dogs. The samples were collected through routinely neutering and manual semen collection. In this thesis, we performed the analysis of results from a previously performed sample collection and immunohistochemical examination. I compared these results to the data in literature in other species. Testis and epididymis were fixed in formalin and finally embedded in paraffin prior to immunohistochemistry identification. We used an indirect immunoperoxidase method to perform the immunohistochemistry. Rabbit polyclonal antibodies were used for detection of leptin and goat polyclonal antibodies for leptin receptors. Murine ovaries with CL and canine CL were used as positive controls, and a chromogenic agent was applied to make it possible to evaluate the result. The slides were assessed under 100x magnification. A full sperm-rich portion of semen was collected by manual stimulation. Both epididymal and ejaculated spermatozoa were further purified using the swim-up technique prior to immunocytochemistry identification. The samples were collected, purified and processed and 4 the immunocytochemical expression of the antibodies was obtained through the same indirect immunoperoxidase method as used in immunohistochemistry. The Objective of this study is to investigate if it is possible to detect leptin and its receptors in the male reproductive tract of canines with the use of immunohistochemistry and immunocytochemistry