Expression of a truncated form of KIT tyrosine kinase in human spermatozoa correlates with sperm DNA integrity

• Published in: Human reproduction (Oxford) 2010-09, Vol.25 (9), p.2188-2202
• Main Authors: Muciaccia, Barbara, Sette, Claudio, Paronetto, Maria Paola, Barchi, Marco, Pensini, Simona, D'Agostino, Angela, Gandini, Loredana, Geremia, Raffaele, Stefanini, Mario, Rossi, Pellegrino
• Format: Article
• Language: English
• Subjects: Acrosome Reaction; Adult; Aged; Biological and medical sciences; Biomarkers - metabolism; Cell Shape; clusterin; Clusterin - metabolism; DNA damage; DNA Fragmentation; equatorial segment; Gene Expression; Gynecology. Andrology. Obstetrics; human spermatozoa; Humans; Male; Medical sciences; Middle Aged; Organ Specificity; Protein Isoforms - metabolism; Protein Transport; Proto-Oncogene Proteins c-kit - genetics; Proto-Oncogene Proteins c-kit - metabolism; RNA, Messenger; Semen Analysis; Sperm Head - metabolism; Sperm Head - pathology; Spermatozoa - chemistry; Spermatozoa - metabolism; Spermatozoa - pathology; Testis - cytology; Testis - metabolism; TR-KIT; Young Adult
• ISSN: 0268-1161; 1460-2350
• DOI: 10.1093/humrep/deq168

BACKGROUND TR-KIT, a truncated form of KIT (the KITL receptor), corresponding to the c-terminal half of the intracellular split tyrosine kinase domain, is expressed during the haploid stages of mouse spermatogenesis, and is one of the candidate sperm factors possibly involved in egg activation at fertilization. METHODS Immunocytochemistry of adult human testis, and studies of human semen samples from volunteer donors through immunofluorescence, confocal microscopy, flow cytometry, western blot and RT–PCR analyses were performed. RESULTS We show that the TR-KIT is expressed during spermiogenesis in the human testis, and that it is maintained in human ejaculated spermatozoa. TR-KIT is localized both in the equatorial segment and in the sub-acrosomal region of the human sperm head. The equatorial localization of the TR-KIT persists after the spontaneous acrosome reaction. Cytometric analysis of several sperm samples from volunteer donors, showed variable degrees of the TR-KIT-specific immunolabeling, and a significant inverse correlation (Pearson's coefficient, r = −0.76, P < 0.0001, n = 23) of the TR-KIT positivity with markers of sperm damage, i.e. DNA fragmentation, as revealed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labeling (TUNEL) analysis and the intense clusterin positivity. We also found less significant inverse correlation with altered head morphology (r = −0.47, P < 0.05, n = 23) and direct correlation with sperm forward motility parameters (r = 0.59, P < 0.01, n = 23). CONCLUSIONS The TR-KIT is present in the equatorial region of human spermatozoa, which is the first sperm component entering into the oocyte cytoplasm after fusion with the egg. This localization is consistent with the function previously proposed for this protein in mice. In addition, the TR-KIT represents a potential predictive parameter of human sperm quality.