• Applied Microscopy
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• v.28 no.1
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• pp.63-71
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• 1998

The spermatozoon of Squalidus gracilis majimae is approximately $36.6{\mu}m$ in length and is characterized by a spherical nucleils with the clear chromatin, a short midpiece containing the mitochondria, and a flagellum positioned tangentially to the nucleus. An acrosome is absent as in all teleost fishes. The nucleus is about $1.9{\mu}m$ in diameter and in its periphery contains the electron-lucent chromatin distinguished from the electron-dense chromatin occupying most of the nucleus. The shallow nuclear fossa contains the proximal centriole, instead of two centrioles in deep nuclear fossa in siluroids. The proximal and distal centrioles are oriented approximately $140^{\circ}$ to each other. The mitochondria of 10 or more in number are arranged in three layers and do not surround the axoneme. The asymmetrical distribution of the mitochondria and the eccentrical position of the nucleus with regard to the tail are the general pattern of the cyprinid spermatozoa. S. gracilis majimae spermatozoa have the most mitochondria and the deepest cytoplasmic canal among cyprinid species. The flagellum lacks the lateral fins.

• Animal cells and systems
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• v.16 no.2
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• pp.104-113
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• 2012

The nucleolus is a distinct nuclear territory involved in the compartmentalization of nuclear functions. There is some evidence of a relationship between nuclear fragmentation during spermatogenesis and chromatoid body (CB) formation. The CB is a typical cytoplasmic organelle of haploid germ cells, and is involved in RNA and protein accumulation for later germ-cell differentiation. The goal of this study was to qualitatively and quantitatively describe the nucleolar cycle during the spermatogenesis of $Phrynops$ $geoffroanus$ (Reptilia Testudines), and compare this nucleolar fragmentation with CB formation in this species through the use of cytochemical and ultrastructural analysis. Qualitative analysis showed a fragmentation of the nuclear material after pachytene of the first meiotic division in the primary spermatocytes. Quantitative analysis of the nucleolar cycle revealed a significant difference in the number of nucleoli and in the size of the nucleolus between spermatogonia and early spermatids. Using ultrastructural analysis, we recorded the beginning of the CB formation process in the cytoplasm of primary spermatocytes at the same time as when nuclear fragmentation occurs. In the cytoplasm of primary spermatocytes, the CB was observed in association with mitochondrial aggregates and the Golgi complex. In the cytoplasm of early spermatids, the CB was observed in association with lipid droplets. In conclusion, our data show that the nucleolus plays a role in the CB formation process. During spermatogenesis of $P.$ $geoffroanus$, the CB is involved in some important biological processes, including acrosome formation and mitochondrial migration to the spermatozoon tail and middle piece region.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.36 no.6
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• pp.619-624
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• 2003

Genital organs and gonad development in kishi velvet shrimp (Metapenaeopsis dalei) were studied based on morphological and histological observations. The samples were collected monthly around the Ochongdo area on the western coast of Korea, from April 1997 to December 1998. Thelycum appear structurally complete after females reach a length of about 12.0 mm CL. Petasma rarely Join after males reach a length of 10.6 mm CL and a fully developed petasma shows up to a length of 12.0 mm CL. The ovaries lie on the dorsal side of the female shrimps thorax. The ovary cavity is located below the heart and above the hepatopancreas. The anterior part of the ovary is round and fused, and the posterior part shows a pair of tubule-like structures. Testes are bilaterally symmetrical, the anterior part shows a convoluted tubule, and the posterior part consists of a pair of tubules. In histological observations, ripe eggs and spermatozoon show when mature females and males reach a length of about 12.0 mm CL.

• Clinical and Experimental Reproductive Medicine
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• v.22 no.2
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• pp.149-153
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• 1995

In IVF-ET program, intracytoplasmic sperm injection(ICSI) has been performed with testicular sperm extraction(TESE) in case of no normal spermatozoon could be retrieved from the epididymis. We wished to see whether the quality of testicular sperm affect the fertilization and pregnancy rate in TESE-ICSI cycles(n=40). These cycles were classified into three groups by the total number of normal motile spermatozoa(TNMS) in the TESE sample: i) good sperm(GS) group(n=12), TNMS > 10,000; ii) moderate sperm(MS) group(n=19), 1,000 < TNMS < 10,000; iii) poor sperm(PS) group(n=9), TNMS < 1,000. Among 423 injected oocytes, 307(72.6%) oocytes were normally fertilized and 43 zygotes were cryopreserved. The fertilization rates of GS group(79.3%) and MS group(75.9%) were significantly(p<0.005) higher than PS group(60.2%). After the embryo transfer(n=40), clinical pregnancy was obtained in 14 cycles(35.0%) and on-going pregnacy in 13 cycles(32.5%). The clinical and on-going pregnancy rates were similar in each group. From these results it can be concluded that testicular spermatozoa are successfully used with ICSI in IVF-ET program in spite of very poor quality of TESE sample.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.3
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• pp.247-252
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• 1994

Intracytoplasmic sperm injection(ICSI) was known as effective method in treatments of couples who unable to be helped by conventional in vitro fertilization. In 78 treatment cycles of 78 infertile couples using ICSI performed at our infertility clinic between May and August 1994 were analyzed. These patients were classified two groups, andrological factor(AF) and non-andrological factor(non-AF) group. The AF group, which had abnormal sperm physiology, included oligozoospermia, asthenozoospermia, oligoasthenoteratozoospermia(OATS) and microsurgical epididymal sperm aspiration(MESA) patients. The non-AF group, which had abnormal oocyte physiology, included abnormal zona pellucida, poor quality of oocyte and immune factor infertile patients. A single spermatozoon was injected into the ooplasm of 776 metaphase II oocytes. The fertilization rate was 44.6%(346/776) and 319 embryos were transferred. After 73 embryo transfers(93.6% of treatment cycles) 23 pregnancies were estabilshed, i. e. pregnancy rate of 29.4% per started cycle and 31.5% per embryo transfer. Fertilization rate of AF and non-AF group was 46.2% and 35.8%, pregnancy rate was 34.5%(20/58) and 20.0%(3/15), respectively. In order to increase the pregnancy rate, assisted hatching(AHA) has done after lCSl in 47 treatment cycles. Pregnancy rate of ICSI with AHA and without AHA group was 34. 0% (16/47) and 26.9%(7/26), respectively. ICSI was more effective in andrological factor infertility and the pregnancy rate was increased by ICSI with AHA procedure.

• The Korean Journal of Malacology
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• v.28 no.1
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• pp.55-64
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• 2012

Ultrastructural studies of germ cell development during spermatogenesis and taxonomic values in mature sperm morphology of Argopecten irradians irradians were investigated by transmission electron microscopic observations. In the early stage of spermatid during spermiogenesis, a few granules and proacrosomal granules are formed by the Golgi complex. In the late stage of spermatid during spermiogenesis, a proacrosomal vesicle becomes an acrosomal vesicle in the acrosome through spermiogenesis. The sperm is approximately $ 45-48{\mu}m$ in length including a jar-shaped sperm nucleus (about $1.45{\mu}m$ long), an acrosome (about $0.34{\mu}m$ long) and tail flagellum. The axoneme of the sperm tail shows a 9+2 structure. As one of common characteristics of mature sperm morphologies in Pectinidae species in subclass Pteriomorphia, mature spermatozoon consists of the cone-shaped acrosomal vesicle and subacrosomal material on the invaginated jar-shaped nucleus. The acrosomal vesicle of this species is composed of electron high dense opaque part (material) from the base to the tip, as have seen in the species in the subclass Pteriomorphia. Exceptionally, five mitochondria are found in the sperm midpiece of this species, unlike four in most species of Pectinidae in subclass Pteriomorphia. However, the acrosomal vesicle of spermatozoa of A. irradians irradians resemble to those of other investigated Pectinidae species in subclass Pteriomorphia. Therefore, we can use sperm morphology as a tool in the resolution of taxonomic relationships within the Pectinidae species. These morphological charateristics of acrosomal vesicle belong to the family Pectinidae in the subclass Pteriomorphia.

• Applied Microscopy
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• v.28 no.2
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• pp.193-205
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• 1998

The Urechis unicinctus sperm and spermatogenic cells prepared from the testis are investigated to identify $\alpha-tubulin$ of axoneme microtubules using mouse monoclonal $anti-\alpha-tubulin$ as the first Ab and Gold(10nm) conjugated goat anti-mouse IgG as the Ab marker. The Ag-Ab reaction analyzed excellently the localization of $\alpha-tubulin$ and the gold particles incorporated with the proximal and distal centrioles, manchette microtubules, and flagellum. The gold particles can be also observed in the spermatogenic cells while the cells are still in sperm ball which is composed of a somatic cell and spermatogenic cells. The sperm ball is the functional unit of sperm production in U unicinctus testis. The spermatids are developed from the spermatogenic cells in the sperm ball and released into the testis cavity through a cortical cytoplasmic opening. The spermatid architectures are similar with the mature sperm of the testis cavity in aspects of shape of discoid acrosome, degree of nuclear condensation and ring type of mitochondrion. However, the distal centriole connecting with the flagella can be observed from the mature sperm while the both proximal and distal centrioles reveal only in the spermatids. The proximal centriole is directly connected with nuclear outer membrane during the stage of nuclear condensation and oriented perpendicularly to the distal centriole whose axis coinciding with the longitudinal axis of the spermatozoon. There are indications that the distal centriole is intimately associated with the polymerization of the flagellum. The manchette microtubules appear during spermatid development but the mature sperm have round head and no conspicuous middle piece.

• Clinical and Experimental Reproductive Medicine
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• v.46 no.1
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• pp.36-40
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• 2019

A viable spermatozoon is a prerequisite for fertilization in intracytoplasmic sperm injection (ICSI). Thus, it is crucial to select viable but immotile spermatozoa on the day of ICSI. We report conflicting results in the identification of viable but immotile spermatozoa between the eosin-nigrosin staining and the laser test, which resulted in confusion for embryologists during assisted reproductive technology (ART). Three patients' semen samples that showed no motile spermatozoa are described in this report. To identify viable spermatozoa, we used both the eosin-nigrosin test and the laser test for each sample, and repeated the semen analysis twice in each patient. Viable but immotile spermatozoa selected by the laser test were used for ICSI. Viable spermatozoa were detected by both the eosin-nigrosin and laser tests in two patients (case 1, 95.00% vs. 24.21% and 92.68% vs. 22.22%; case 2, 41.18% vs. 23.48% and 39.81% vs. 22.52%), indicating consistent results between the two methods. In the third patient, the eosin-nigrosin test yielded viability rates of 20.75% and 19.14%, while the result of the laser test was 0%. Thus, testicular aspiration was performed to collect viable sperm from this patient. Normal fertilization was achieved after the injection of viable but immotile spermatozoa selected from these patients by the laser test, resulting in the birth of two healthy babies. Our study documents a case where the eosin-nigrosin test showed a limitation in identifying viable but immotile spermatozoa for ART, while the laser test may overcome this limitation. Larger samples may be required to corroborate the clinical value of the laser test.

• Development and Reproduction
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• v.13 no.2
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• pp.115-122
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• 2009

Spermatogenesis and ultrastructural characteristics of sperm of brackish water diploid Corbicula japonica were investigated by electron microscope observations. Based on the cytological studies, the spermatozoon of this species (brackish water diploid) C japonica is approximately 55 ${\mu}m$ in length. The sperm head (about 12 ${\mu}m$ long) is elongated and tapers with a slight curve. Sperm nucleus is about 7.90 ${\mu}m$ long, and the acrosome is about 2.70 ${\mu}m$ long: The morphologies of the sperm nucleus type and the acrosome shape of this species are a long arrow-like type and long cone-like shape, respectively. The sperm head of this species (external fertilization, dioecious and oviparous species) is partially modified from that of the primitive type, as seen in triploid Corbicula species (internal fertilization, hermaphrodite and ovoviparous species), reported by some authors. However, this species produces uniflagellate spermatozoa, unlike freshwater triploid hermaphroditic clams being possessed of partially modified biflagellate spermatozoa. Diploid C japonica is similar to those of other bivalves being possessed of a short midpiece containing four mitochondria surrounding the centrioles. The axoneme of the sperm tail flagellum consists of nine pairs of microtubules at the periphery and a pair at the center. The axoneme of the sperm tail shows a 9+2 structure, and from uniflagellate sperm cross sectioned, in particular, wing-like axonernal lateral fins are observed, as seen in external fertilization fishes.

• Development and Reproduction
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• v.2 no.1
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• pp.75-80
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• 1998

메기 정자는 그 길이가 약 62.5 \mu m이며 구형의 핵, 짧은 중편 및 꼬리를 ㄱ진 일반적인 메기류 정자의 미세구조적 특징을 나타내었다. 정자는 대부분의 경골어류의 정자에서와 같이 첨체를 가지고 있지 않았으며 염색질은 완전치 농축되어 있었다. 핵와(nuclear fossa)는 약 0.6 \mu m 함입되어 있었고 그 속에 기부 중심립과 말단 중심립의 일부가 포함되어 있었다. 두 중심립은 140 \circ C의 각도로 배열되어 있었으며 말단 중심립에서 9개의 부수체가 언형질막을 향하여 배열되어 있었다. 미토콘드리아는 중편 세포질에서 2층 또는 3층으로 배열되어 있었으며 핵의 후반부와 꼬리의 기부를 둘러싸고 있었다. 꼬리는 축사만으로 구성되어 있었으며 lateral fins는 관찰되지 않았다. 메기 정자의 가장 큰 구조적 특징은 중편 세포질에 구성되어 있는 관구조(tubular structure)이었다. 대부분의 경골어류의 정자는 중편 세포질에 미토콘드리아만을 포함하고 있으나, 메기 정자에서는 중편 세포질의 전반부에 미토콘드리아가 포함되어 있고, 후반부에는 소관이 모여 망상구조를 형성하는 관구조가 잘 발달되어 있었다. 이와 같은 관구조는 현재까지 Characiformes의 정자 이외의 다른 경골어류에서는 보고된 바 없으며 이러한 구조는Characiformes과 메기류의 계통학적 관계를 연구하는데 매우 중요한 형질로 여겨진다. ^u The spermatozoa of Silurus asotus are appoximately 62.5 \mu m in length and relatively simple cells composed of spherical head, a short midpiece and a tail as in most Siluriformes. The ultrastructure of the spermatozoa of S. asotus is characterized by the following features. The nucleus measuring about 1.5 \mu m in length is depressed with a deep nuclear fossa of about 0.6 \mu m in length, two fifth of the nuclear diameter. The fossa contains the proximal centriole and the half of the distal centriole. Two centrioles form an angle of approximately 140 \circ to each other. the nine satellite rays radiate from the outer surface of the distal centriole. the mitochondrea surround the basal nucleus and the axoneme, and are arranged in two or three layers in the postnuclear cytoplasm. The lateral fins are lost in the sperm tail. The most significant feature is manifested in the midpiece. The midpece comprises two parts, the mitochondria and the tubular structure unlike other teleost fishes containing only the mitochondria. The tubular structure was reported only in the spermatozoa of Citharinus belonging to the characiformes of teleost fishes. Thus it is considered to be a good characteristics for the study of phylogenetic link between Siluriformes and Characiformes.