Abstract
Histamine synthesis in male reproductive tissues remains largely unknown. The interaction between stem cell factor and its receptor, c-Kit, has been found to be essential for the maturation of male germ cells and peripheral mast cells. Based on this analogy, we investigated the expression of histidine decarboxylase (HDC), the rate-limiting enzyme of histamine synthesis, in mouse male germ cells. Immunohistochemical analyses revealed that HDC is localized in the acrosomes of spermatids and spermatozoa. In the testis, epididymis, and spermatozoa, a significant amount of histamine and HDC activity were detected. W/W(V) mice, known to lack most of their germ cells in the seminiferous tubules, were found to lack HDC protein expression as well as HDC activity in the testis. An in vitro acrosome reaction induced by a calcium ionophore, caused the release of histamine from epididymal spermatozoa. Our observations indicate that histamine is produced in and released from the acrosomes.
Highlights
During spermatogenesis [8, 9]
The abundant expression of histidine decarboxylase (HDC) mRNA was demonstrated in the mouse testis by Northern blot analysis
HDC mRNA and protein were detected in the cells inside the seminiferous tubules
Summary
Animals—Adult male ICR mice (9 –12 weeks of age), WBB6F1-W/WV (W/WV), and WBB6F1-W/Wϩ (W/Wϩ) mice (8 weeks of age) were obtained from the Shizuoka Agricultural Cooperative Association for Laboratory Animals (Hamamatsu, Japan). RT-PCR—Various male reproductive tissues and stomach were collected from ICR mice, immediately frozen in liquid nitrogen and stored at Ϫ80 °C until use. Hybridization was performed with a 32P-labeled cDNA fragment specific for mouse HDC (PvuII-digested fragment) at 65 °C in 6ϫ SSC (1ϫ SSC is composed of 0.15 M NaCl and 0.015 M sodium citrate), 0.5% SDS, and 5ϫ Denhardt’s solution. Hybridization was carried out in a buffer containing 50% formamide, 2ϫ SSC, 10 mM Tris-HCl, pH 7.5, 1ϫ Denhardt’s solution, 10% dextran sulfate, 0.2% SDS, 100 mM dithio-.
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