• Applied Microscopy
• /
• v.31 no.3
• /
• pp.257-266
• /
• 2001

To investigate the process of spermoigenesis and glycogen effect during the spermatogenesis of Rana catesbeiana, the morphological characteristics of the testes were examined by light and transmission electron microscopy. Spermiogenesis of R. catesbeiana was divided into three stages on the basis of the features of the nucleus and the cytoplasm organelles. Except for the primary spermatogonia, the phases from the spermatocytogenesis to the spermatids before spermiation phase were surrounded by spermatocyst. Especially , the glycogen particles were not observed until in the stage of spermatocytogenesis, but from the early spermatids to the maturation phase were observed in the nucleus, acrosome and cytoplasm of the spermatids. The present result suggests that the glycogen may play an important role in the sperm maturation, and as a source of the energy in the wave-movement of sperm tail.

• Journal of Animal Science and Technology
• /
• v.65 no.2
• /
• pp.401-411
• /
• 2023

Many studies have been conducted to improve technology for semen cryopreservation in pigs. However, computer-assisted analysis of sperm motility and morphology is insufficient to predict the molecular function of frozen-thawed semen. More accurate expression patterns of boar sperm proteins may be derived using the isobaric tags for relative and absolute quantification (iTRAQ) technique. In this study, the iTRAQ-labeling system was coupled with liquid chromatography tandem-mass spectrometry (LC-MS/MS) analysis to identify differentially expressed CM10-fractionated proteins between fresh and frozen-thawed boar semen. A total of 76 protein types were identified to be differentially expressed, among which 9 and 67 proteins showed higher and lower expression in frozen-thawed than in fresh sperm samples, respectively. The classified functions of these proteins included oxidative phosphorylation, mitochondrial inner membrane and matrix, and pyruvate metabolic processes, which are involved in adenosine triphosphate (ATP) synthesis; and sperm flagellum and motile cilium, which are involved in sperm tail structure. These results suggest a possible network of biomarkers associated with survival after the cryopreservation of Duroc boar semen.

• Fisheries and Aquatic Sciences
• /
• v.10 no.4
• /
• pp.196-199
• /
• 2007

The spermatozoa of Leiocassis nitidus are relatively simple cells composed of a spherical head, a short midpiece, and a tail, as in most Siluriformes. The ultrastructure is characterized by the following features: Acrosome absent, as in most teleosts; around nucleus about $1.8\;{\mu}m$ long, with a deep nuclear fossa containing the proximal and distal centrioles and mitochondria. Two centrioles approximately $180^{\circ}$ from each other; 10 or more mitochondria surrounding the axoneme (with a 9+2 microtubular pattern), arranged in two layers in the postnuclear cytoplasm and separated from the axoneme by the cytoplasmic canal. Two lateral fins on the same plane as the two central microtubules; doublets 3 and 8, which are ultrastructural characteristics of the sperma tail unlike other siluroids laking the lateral fins.

• Applied Microscopy
• /
• v.26 no.3
• /
• pp.277-291
• /
• 1996

The structures of the egg envelope in fertilized eggs of three species of characidae, head and tail light fish (Hemigrammus ocellifer), black tetra (Gymnocorymbus ternetzi), and buenos aires tetra (Hemigrammus caudovittatus) were studied using light and electron microscopes. The fertilized eggs in all species were colorless, transparent, spherical and non-floted type. The egg envelopes have a single micropyle resembling the pathway of sperm in the area of the animal pole. The micropyle was surrounded by protruded lines of the egg envelope in a radiated form. Egg envelopes of fertilized eggs in both head and tail light fish and buenos aires tetra consisted of three distinct layers; an outer layer, a middle layer and an inner layer. And that of blacktetra consisted of two layers; an outer layer and an inner layer. Also, an outer layers of both head and tail light fish and black tetra were adhesive types but, in that of buenous aires tetra was non-adhesive type. An outer surface of egg envelope in black tetra was arranged by pores regularly. In that of head and tail light fish and buenos aires tetra have a rough side. An inner layer of egg envelope in fertilized eggs consisted of lamellae alternating with interlamellae of lower electron density; an inner layer of fertilized eggs in head and tail light fish consisted of three layers, that of black tetra was four layers, and that of buenos aires tetra was five layers.

• Clinical and Experimental Reproductive Medicine
• /
• v.25 no.3
• /
• pp.331-340
• /
• 1998

We determined the incidence of activation, male pronuclear formation and apposition of pronuclei in porcine oocytes following intracy-toplasmic injection of various porcine sperm components and foreign species spermatozoa, such as mouse, human or cattle. The porcine oocytes were activated by injection of a spermatozoon or an isolated sperm head. Neither isolated sperm tail nor perinuclear material removed sperm head activated oocytes. Because injection of mouse, bovine or human spermatozoon activated porcine oocytes, the sperm born activation factors is not strict species specific. Male pronuclear formation and pronuclear apposition were observed in the porcine oocytes following injection of porcine, bovine, mouse or human spermatozoa. The electrical stimulation following sperm cell injection did not enhance the incidence of male pronuclear formation nor pronuclear apposition comparent with sperm cell injection alone (p>0.1). Mitosis and two cell division in some oocytes were observed at 20 to 24 h after injection of porcine spermatozoon. However, none of oocytes following injection of mouse, bovine or human spermatozoa developed to the mitotic metaphase or normally divided to the two cell stage. These results suggested that the oocyte activating factor(s) presented in the perinuclear material and it is not species specific for the porcine oocyte.

• Applied Microscopy
• /
• v.31 no.3
• /
• pp.245-256
• /
• 2001

The morphological characteristics of spermatogenic cells during the spermiogenesis of Paradoxornis webbiana were studied by transmission electron microscope. Spermiogenesis of P. webbiana was divided into ten phase. The chromatin granules became fibrous granules at the Golgi phase, gradually condensed at the cap phases, condensed as a stick at the acrosomal phase, and finally, a perfect nucleus was formed at the maturation phase. The formation of sperm tail began at the early Golgi phase, and completed at the late maturation phase. In particular, the dense materials existed in the sperm neck, which is wedged between the tip of segmented columns and the first mitochondria of the middle piece. The axone in the neck were surrounded by the dense materials. The axonema in spermatozoon contains a 9+2 arrangement of microtubules: 9 doublets, and 2 central single microtubules. Mitochondrial bundles of middle piece were composed of a pair of arms, which surrounded the axone of the middle piece by the $15^{\circ}$ angled-helical structure. The outer membrane of mitochondria were surrounded by microtubules in plasma membrane of the sperm. The undulating membrane had a helical structure, and the sperm plasma membrane was surrounded by undulating membrane.

• Applied Microscopy
• /
• v.22 no.2
• /
• pp.97-117
• /
• 1992

In order to study process of spermiogenesis of the Korean greater horseshoe bat, Rhinolophus ferrumequinum korai, the cycle of seminiferous epithelium was examined by the light and electron microscope and the following results were obtained based on the epithelial cell differentiation. 1. Spermiogenesis occurred from early July to mid-Octber, and spermatogenic activity was vigorous from mid-August to late September. Spermatocytes including spermatogonia were found to be degenerated in only July. It is deduced that the degeneration serves as the mechanism to regulate effective use of energy to prepare for mating and hibernating periods, and regulation of breeding cycle. 2. Spermiogenesis of the Korean greater horseshoe bat was divided according to differentiation of the cell structure, into Golgi, cap, acrosome, maturation and spermiation phases; Golgi, cap and spermiation phases were further divided into two steps of early and late phase respectively, and acrosome phase into three steps of early, mid and late phases, and maturation phase has only one step. Hence, the spermiogenesis consists of ten phases. The first research was done in this article on the changes of chromatin with nucleus, the time of appearance and disappearance of chromatin granules, in case of Korean greater horseshoe bat (Rhinolophus ferrumequinum korai). Chromatin granule began to be condensed in late Golgi and the condensation proceeded to form an irregular mass of a electron-dense chromatin in a form of circular cylinder in the center of nucleus at the phase of maturation. Finally, the chromatin condensation proceeded and perfect nucleus of sperm with homogeneous density was formed when the sperm was separated from Sertoli cell. Therefore, appearance and disappearance of chromatin granules occurred in the period of time between late Golgi and maturation phase, The tail of sperm began to develop in early cap phase, Numerous lipid droplets were obseved in the cytoplasm of spermatids during the maturation phase, which seemed to be used as energy source necessary to make mature sperm during spermiogenesis.

• Journal of Embryo Transfer
• /
• v.30 no.3
• /
• pp.213-218
• /
• 2015

In the study for a differentiation and development of spermatogonial cells, the researchers should commonly require a simple, fast and reasonable method that could evaluate the developmental stage of male germ cells without any damage and also relentlessly culture them so far as a cell stage aiming at experimental applications. For developing the efficient method to identify the stage of sperm cells, the morphological characteristics of sperm cells were investigated by staining the cells with blue fluorescent dye Hoechst 33258, and a criterion for male germ cell classification was elicited from results of the previous investigation, then the efficiency of the criterion was verified by applying it to assort the germ cells recovered from male mice in age from 6 to 35 days. As morphological characteristics, spermatogonia significantly differed from spermatocytes in size, appearance and fluorescent patches of nucleus, and spermatids could also be distinguished from spermatozoa by making a difference in the volume and shape of nucleus and the shape and fluorescence of tail. Aforesaid criterion was applicable for classifying in vitro cultured sperm cells by verifying its efficiency and propriety for assorting the stages of testicular germ cells. However, the fluorescent staining showed that germ cells in mouse testis should be dramatically differentiated and developed at 21 days and 35 days of age, which were known as times of sexual puberty and maturity in male mice, respectively. In conclusion, the results indicated that this simple criterion for sperm cell classification using fluorescence staining with Hoechst 33258 may be highly efficient and reasonable for spermatogenesis study.

• The Korean Journal of Zoology
• /
• v.24 no.2
• /
• pp.65-75
• /
• 1981

The formation of the acrosome during spermatogenesis in Gerris paludum was studied. The Golgi bodies are dispersed randomly in the cytoplasm at the early stage of the spermatocyte and get together to form several group of many bodies, and then they are equally divided into the spermatids by the meiotic divisions. The acroblast first appears in the form of a vesicle and soon an acrosomal granule is differentiated within it. The acroblast is separated from the acrosomal granule at the posterior of the nucleus and is finally sloughed off along the tail filament. The acrosome, after moving to the side of the nucleus opposite the mitochondrial derivatives, differentiates into two zones. The two basal bodies and the differentiated tip originate from the sheath. The basal bodies appear at the proximal part of the sheath simply in contact with the core on one side. During elongation and and narrowing of the acrosomes of the spermatids, they surround the one side at the base of the acrosome and finally all the other are immediately adjacent to the nucleus. The differentiated tip continues to the sheath at the anterior of the cores and is elongated prior to the two basal bodies. They appear to be contiguous twin-tubes, not a single granule in the later stage of the spermatids, and a group of the basal bodies in the sperm bundle.

• Journal of Animal Reproduction and Biotechnology
• /
• v.34 no.4
• /
• pp.272-279
• /
• 2019

The study was conducted to evaluate the seminal attributes and cryobanking of Achhami (Bos indicus) bull semen. Of two Achhami bulls, 8 ejaculates from each bull were evaluated for seminal attributes. For semen freezing and cryo-banking, 4 ejaculates (having ≥2 mL semen volume, ≥75% of sperm motility and ≥1,000 × 10⁶ cells/mL of sperm concentration) from each bull were used. Semen samples were diluted in egg-yolk-tris-citrate extender using a two-step dilution protocol, and were frozen in liquid nitrogen (LN₂) vapour in a styrofoam box. The mean semen volume, colour, sperm mass activity, motility, viability, concentration, abnormal acrosome, midpiece and tail and, abnormal head of two Achhami bulls were 4.4 ± 0.5 mL vs. 2.5 ± 0.2 mL, 2.5 ± 0.1 vs. 2.4 ± 0.1, 3.5 ± 0.1 vs. 3.5 ± 0.1, 77.0 ± 1.1% vs. 78.3 ± 1.3%, 94.4 ± 0.5% vs. 91.0 ± 0.6%, 1137.7 ± 73.7 × 10⁶ cells/mL vs. 1060.0 ± 44.3 × 10⁶ cells/mL, 10.2 ± 0.5% vs. 10.3 ± 0.5% and 6.7 ± 0.5% vs. 8.2 ± 0.3%, respectively. The post-thawed sperm motility and viability were 53.0 ± 2.0% vs. 50.0 ± 0.0% and 80.2 ± 0.4% vs. 73.2 ± 0.7%, while evaluating by computer-assisted sperm analysis (CASA) system, the percentage of the progressive motility, fast motility, slow motility, local motility and immotile sperm were 75%, 68%, 7.4%, 16.6% and 8.6%, respectively. A total number of 620 doses semen straw were cryo-banked. Due to the acceptable post-thawed sperm motility and viability recorded, cryopreservation of Achhami semen is hereby recommended so as to preserve the Achhami breed. For further validation, the fertility will be observed from the produced frozen semen.