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Rab proteins in sperm life
Vojtová, Kristýna ; Páleníková, Veronika (advisor) ; Zelenková, Natálie (referee)
Rab proteins are the largest group of small GTPases. They work as key regulators in the intracellular vesicular transport of cells. This bachelor's thesis contains a literature review of current information on the function of Rab proteins in mammalian spermatozoa. The first part of the work is devoted to their structure, function, cycle, and diseases related to the dysfunction of Rab proteins. The second part is focused on sperm development and maturation linked with specific Rab proteins involved in individual steps during the life of the sperm. Rab proteins are important in cytoskeletal organization and cytokinesis of sperm during spermatogenesis. They participate in the acrosome biogenesis mediated by the Golgi apparatus and the morphological changes of sperm during spermiogenesis. They also play a key role in the acrosome reaction and interaction with the zona pellucida. Due to their necessity in the sperm development and maturation, Rab proteins are proposed as a marker of male fertility, which is summarized in the last part of the thesis. Keywords Rab proteins, GTPases, sperm, spermatogenesis, spermiogenesis, fertilization, infertility
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Rab-2A protein: its localization and function in human spermatozoa and fertilization
Sadílková, Lucie ; Postlerová, Pavla (advisor) ; Lánská, Eva (referee)
The Rab-2A protein belongs to the Rab family of monomeric G-proteins involved in the regulation of intracellular vesicular transport. Although several studies describing the role of Rab-2A protein in mammalian sperm have been published, the exact localization and function of this protein in male gametes have not been fully understood yet. In the diploma thesis, we were the first to describe the presence of Rab-2A in human sperm using 5C5, produced by the Laboratory of Reproductive Biology. Using the 5C5 antibody, we were able to identify several Rab-2A isoforms with molecular weights of 26, 24, 22, and 18 kDa in the human sperm lysate, confirmed by mass spectrometry analysis. Using the indirect immunofluorescence method, we localized Rab-2A in the acrosome area of both ejaculated and capacitated sperm. We noticed that Rab-2A leaves the sperm during the acrosomal exocytosis. We also found a reduced Rab-2A protein level in sperm of patients with various pathologies of ejaculate. Based on studies describing the role of Rab-2A in acrosome biogenesis and the presence of Rab-2A in bovine sperm perinuclear theca, we tried to detect Rab-2A in this protein layer in human sperm. However, the chosen method is apparently not applicable to human sperm and isolation has failed. Additionally, we demonstrated the...
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