The spatial and temporal expression of Tekt1, a mouse tektin C homologue, during spermatogenesis suggest that it is involved in the development of the sperm tail basal body and axoneme

Tektins comprise a family of filament-forming proteins that are known to be coassembled with tubulins to form ciliary and flagellar microtubules. Recently we described the sequence of the first mammalian tektin protein, Tekt1 (from mouse testis), which is most homologous with sea urchin tektin C. We have now investigated the temporal and spatial expression of Tekt1 during mouse male germ cell development. By in situ hybridization analysis TEKT1 RNA expression is detected in spermatocytes and in round spermatids in the mouse testis. Immunofluorescence microscopy analysis with anti-Tekt1 antibodies showed no distinct labeling of any subcellular structure in spermatocytes, whereas in round spermatids anti-Tekt1 antibodies co-localize with anti-ANA antibodies to the centrosome. At a later stage, elongating spermatids display a larger area of anti-Tektl staining at their caudal ends; as spermiogenesis proceeds, the anti-Tekt1 staining disappears. Together with other evidence, these results provide the first intraspecies evidence that Tekt1 is transiently associated with the centrosome, and indicates that Tekt1 is one of several tektins to participate in the nucleation of the flagellar axoneme of mature spermatozoa, perhaps being required to assemble the basal body.     

七星瓢虫精子鞭毛的超微结构(英文)

应用细胞整装制备和超薄切片技术,在透射电子显微镜下检查了七星瓢虫成熟精子鞭毛的超微结构。精子鞭毛是由一个典型的9＋9＋2轴丝,两个同形结晶的线粒体衍生物,两个附体（每个附体具有两部分,一个嗜锇致密月牙体和一个海绵月牙体）和一个非结晶体组成,在鞭毛终端部,仅存的轴丝失去了两个中央微管保留了9个具有动力蛋白臂的双微管和9个附微管。     

ULTRASTRUCTURE OF THE FLAGELLUM IN SPERM OF CKCINELLA SEPTEMPUNCTATA

Abstract  Using cell whole mount preparation and ultrathin section technique, the ultrastructure of the flagellum in the sperm of Coccinella septempunctata L. was examined with transmission electron microscope. The flagellum is made up of a classic 9+9+2 axoneme containing two similar crystallized mitochondria1 derivatives, two accessory bodies, which are divided in to two portions, an osmiophilic dense crescent and a spongy one, and a non-crystalline body. At the end of the flagellum, only the axoneme is present, it loses the two central microtubules but retains the nine doublets with dynein arms and the nine accessory microtubules.     

Inside and outside of the trypanosome flagellum:a multifunctional organelle

Amongst the earliest eukaryotes, trypanosomes have developed conventional organelles but sometimes with extreme features rarely seen in other organisms. This is the case of the flagellum, containing conventional and unique structures whose role in infectivity is still enigmatic.     

Use of negative staining technique and electron microscopy for the study of structural anomalies of outer dense fibres of human flagellum

Motility disorders due to tail defects are often seen in clinical andrology. Sperm motility should be assessed with regard to the morphology of the flagellum. Since suitable longitudinal sections are rarely obtained by routine transmission electron microscopy (TEM) and in view of the importance of dense fibres in modulating sperm motility and providing tensile strength, a detailed, study of human sperm flagellum by negative staining andTEM was attempted. The study was undertaken in two groups of men (I) fertile and (II) asthenozoospermic. The study revealed that outer dense fibres extend to 50–60% of the principal piece. Normal dense fibres were seen in 83% sperms and 23% sperms in groupsI andII respectively. The characteristics seen were variation in diameter, breakage or degradation with lacking or extended endpiece. The negative staining method provides an easy and useful analytical tool for identifying the defects of dense fibres and quantifying them.     

Purification,partial characterization,and localization of Sak57, an acidic intermediate filament keratin present in rat spermatocytes,spermatids, and sperm

We have purified a 57 kDa protein (designated Sak57, for spermatogenic cell/sperm-associated keratin) from sodium dodecyl sulfate-β-mercaptoethanol(SDS-βME)-dissociated outer dense fibers isolated from rat sperm tails. Internal protein sequence analysis of Sak57 yielded two 15-mer and 10-mer fragments with 70–100% homology to human, rat, and mouse keratins and corresponding to the 1A and 2A regions of the α-helical rod domain of keratins. A multiple antigenic peptide (MAP) was constructed using the 10-mer amino acid sequence KAQYEDIAQK (corresponding to the 2A region) and used as antigen for the production of polyclonal antibodies in rabbit. Anti-MAP sera were used for further analysis of the biochemical characteristics of Sak57 in testis and sperm tails using chromatofocusing, immunobloting, and [³²P]orthophosphate-labeling. We have found that rat testis displays two immunoreactive proteins: a soluble 83 kDa protein with pl range 5.9–6.3, regarded as a precursor, and both detergent-insoluble and soluble 57 kDa protein with pl range 5.0–5.9, corresponding to the mature form Sak57. The testicular soluble form was phosphorylated. Rat sperm tail samples displayed only the Sak57 detergent-insoluble form and its pl was more acidic (4.7–4.8). Whole-mount electron microscopy of negatively stained preparations of sperm-derived Sak57 resuspended in SDS-βME revealed a rod-shaped pattern. A decrease in the concentration of SDS-βME resulted in the side-by-side aggregation of rod-shaped Sak57 forming thick bundles. Indirect immunofluorescence was used to determine the localization of Sak57 in isolated outer dense fibers, epididymal sperm, spermatids, and pachytene spermatocytes. Confocal laser scanning microscopy was used to analyze the three-dimensional arrangement of Sak57 in pachytene spermatocytes. Isolated outer dense fiber and sperm tails displayed an immunoreactive product in the form of linear clusters. In elongating spermatids (steps 10–11), Sak57 immunoreactivity was predominant in the head region whereas pachytene spermatocytes displayed a cortical cytoplasmic distribution. Results of this study demonstrate that Sak57 has the characteristics of a keratin intermediate filament and is present during meiotic and postmeiotic stages of spermatogenesis. © 1996 Wiley-Liss, Inc.