Abstract
This experiment was designed to investigate leptin receptors (Ob-R and Ob-Rb) mRNA in ram epididymal spermatozoa by RT-PCR. Ten testes were obtained from abattoir and epididymal spermatozoa recovery was performed. To purify spermatozoa, motile sperm were isolated by the swim-up procedure. Total RNA was isolated from epididymal spermatozoa and placental cotyledon, as a positive control, and then they were purified. Specific bands (98 and 308 bp) for Ob-R and Ob-Rb were detected after RT-PCR in both epididymal spermatozoa and placental cotyledon. We may conclude that Ob-R and Ob-Rb mRNA are present in ram epididymal spermatozoa and leptin perhaps exerts physiological effects, as already demonstrated in human.
Highlights
Leptin is a 16 kDa protein and mainly secreted by white adipose tissue [1]
We may conclude that Ob-R and Ob-Rb mRNA are present in ram epididymal spermatozoa and leptin perhaps exerts physiological effects, as already demonstrated in human
Since the primers used for the amplification of Ob-R and Ob-Rb were designed from nucleotide sequences within one specific exon of gene, the isolated RNA samples were directly used in RT-PCR after DNaseI treatment to demonstrate the absence of genomic DNA contamination in the RNA probes (Figure 1, lanes 3 and 4)
Summary
Leptin is a 16 kDa protein and mainly secreted by white adipose tissue [1]. Leptin modulates different systems via neuroendocrine regulation, including adrenal, thyroid and gonadal axes. Other functions of leptin have been suggested recently in peripheral organs that leptin independently interacts with leptin receptor in peripheral tissue, as well as reproductive organs [2]. Recent observations in female suggest that leptin is involved in relaying energy status to reproduction [3,4,5,6,7]. Various studies have pointed out a direct role of leptin in the control of testicular function, but its actual role in the regulatory network controlling male reproduction is still a matter of debate [3,6,8]. At least five Ob-R isoforms (Ob-Ra-e) have been cloned from rodents and exhibit widespread distribution both in central and peripheral tissues [6,9]. Ob-Rb was reported in most of the farm animal tissues [10,13]
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