• 한국해양생명과학회지
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• 제6권1호
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• pp.23-30
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• 2021

북방전복 Haliotis discus hannai 웅성생식세포의 분화과정과 정자의 형태를 미세구조적으로 기재하였다. 정자의 분화과정은 정원세포기, 정모세포기, 정세포기 및 정자기의 4단계로 구분하였다. 정원세포기에서 정모세포기로의 분화과정은 형태학적 변화가 크지 않았다. 그러나 정자변태과정 동안 염색질 응축, 핵의 형태 변화, 첨체와 중편 및 편모 형성 등의 급격한 형태학적 변화를 나타냈다. 북방전복의 정자는 두부, 중편 및 미부로 구성되며, 두부의 길이는 약 5.3 ㎛로 전자밀도가 높은 핵과 총알형의 첨체로 이루어져 있었다. 중편은 기저체와 미토콘드리아로 구성되어 있었으며, 기저체를 중심으로 5개의 미토콘드리아가 한 층으로 배열되어 있었다. 미부의 횡단면은 "9+2"의 미세소관 구조를 보였다. 이러한 형태 및 구조적 특징은 북방전복의 정자는 원시형(primitive type) 정자임을 보여주는 결과이다.

• 한국가금학회지
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• 제42권2호
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• pp.109-116
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• 2015

본 연구에서는 육종이 필요한 종계장에서 슬라이드 위에 도말된 닭 정액시료를 간편하게 염색하여 정자의 염색질 이상성과 운동성을 유추할 수 있는 기법을 확립하고자 실시하였다. 오계 정자 도말 슬라이드를 Diff-Quik 시약으로 염색하여 관찰하면 신선정액에서는 정자 생존성이 93.53%, 82.42% 및 90.63%일 때, Diff-Quik 염색질의 정상도는 87.96%, 74.25% 및 85.10%로 관찰되었다. 동일한 시료를 동결하고 융해후 정액의 생존성은 조사하면 69.58%, 61.98% 및 72.20%로 관찰되었으며, 염색질의 정상도는 58.91%, 48.49% 및 63.34%로 관찰되었다. 융해된 동결정액에서 활력이 우수한 정자를 쉽게 관찰하기 위하여 정자를 HS-1 희석액으로 재 희석하고, $37^{\circ}C$에서 가온하여 도말하면 염색된 정자의 두부에서 활력이 우수한 정자의 비율을 간단히 유추할 수 있음을 보여주었다. 특히, 신선 정자에서 정상 염색질을 가진 정자의 비율과 생존성이 상관관계는 매우 높은 것으로 판단되며, 동결정자에서도 상관관계가 높다고 추정되었다. 이러한 결과는 Diff-Quik 염색방법으로 닭 정액의 품질을 정자의 염색질의 이상성 유무로 판단할 수 있음을 보여주고 있다. 특히 본 연구에서 제시된 방법은 닭 정자의 우수성을 판단하는데 유용할 것으로 판단되며, 닭 정액 시료를 준비할 때 준비한 도말슬라이드를 현미경적 관찰에 의하여 활성화된 정자의 비율과 정상 염색질을 가진 정자의 비율을 추정하는 방법을 제시하였으며, 이는 인공 수정에 필요한 현장에서 수컷 개체의 종축 이용성을 쉽게 판단할 수 있는 근거를 마련할 수 있음을 의미한다.

• Applied Microscopy
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• 제44권1호
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• pp.1-7
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• 2014

The spermatogenesis of Siamese fighting fish, Betta splendens, belongs to Osphronemidae was investigated by light and electron microscopic observations. In primary spermatocyte stage, the nucleus was comparatively large ellipsoidal, and mitochondria showed a marked development in cytoplasm. In secondary spermatocyte stage, the germ cells were smaller than that of primary spermatocytes. The nucleus was a spherical shape and intercellular space was formed between germ cells. In spermatid stage, the early spermatids were not much different from a secondary spermatocyte. But, the chromatin condensation was occurred from the outside to the inside. The nucleus was more condensed. Intracellular space was larger than early spermatid. The mitochondria were rearranged in a middle piece, and occupied about half of the head part in early sperm. In sperm stage, the head of mature sperm was a spherical shape and had no acrosome. The flagellum was showed the typical 9+2 array of microtubules. Also, the tail of sperm had no lateral fins and outer coarse fibers. These ultrastructural characteristics can be used in classification of species.

• 한국수산과학회지
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• 제37권4호
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• pp.281-286
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• 2004

Spermatogenesis and sperm ultrastructure of the marsh clam (Corbicula japonica) were investigated by electron microscopic observations. Testis of the marsh clam consists of numerous spermatogenic follicle containing germ cells in the different developmental stages. Spermatogonia are located nearest the outer wall of the follicle, while spermtocytes and spermatids are positioned nearer to the lumen. Spematogonia are oval-shaped and about $5{\mu}m$ In diameter. Spermatogonia develop into spermatocyte, spermatid and spermatozoon. In the spermatid to about $2{\mu}m$ in diameter, cytoplasm decreases and mitochondria move to the base of the nucleus and fuse into several spheres, the centrioles become orthogonally oriented, a flagellum appears, and an acrosomal vesicle forms. The mature sperm has primitive type, consisting of a head, a midpiece and a tail. The sperm was arrow-shaped, and its head is about $8{\mu}m$ long and comprised of a long nucleus and an acrosome. The four mitochondria encircled the centrosome in midpiece. The flagellum had the classical 9+2 axoneme structure, and axonemal lateral fins in the tail were observed.

• 대한수의학회지
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• 제61권3호
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• pp.25.1-25.7
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• 2021

The axolotl has extraordinary regeneration capacity compared to other vertebrates. This remarkable potential has been attributed to its life-long neoteny, characterized by the exhibition of embryonic characteristics at the adult stage. A recent study provided a detailed morphological analysis of the sperm morphology of the Ambystoma mexicanum using routine and detailed histological techniques. The primary purpose of the present study is to describe a simple and inexpensive method for evaluating the morphology of axolotl sperm. In this study, spermatophore structures were collected and spread on slides and air-dried. The slides were stained with periodic acid Schiff, toluidine blue, Masson's trichrome, Giemsa, Spermac, and Diff-Quik dye for a morphological examination. The slides were coated with gold/palladium for a scanning electron microscopy examination. The sperm of the axolotl consisted of an elongated head, a neck, and a flagellum covered with an undulating membrane. The lengths of the midpiece, tail, and head were 8.575 ㎛, 356.544 ㎛, and 103.661 ㎛, respectively. In the flagellum part, the wavy membrane structure, whose function has not been explained, surrounds the tail. The data obtained from this study will constitute an important step in designing future research on the reproductive and regeneration capacity of the axolotl.

• Applied Microscopy
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• 제41권2호
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• pp.99-107
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• 2011

한국산 다람쥐 정자의 형태적 특징을 알아보기 위하여 광학현미경과 주사 및 투과 전자현미경으로 관찰하였다. 다람쥐 정자머리의 모양은 주걱 모양의 형태를 취하고 있었다. 정자의 전체 길이는 67.8 ${\mu}m$이었으며, 머리의 길이는 7.8 ${\mu}m$, 꼬리는 60.0 ${\mu}m$이었다. 꼬리는 경부(1.0 ${\mu}m$), 중편부(8.0 ${\mu}m$), 주편부(48.5 ${\mu}m$) 그리고 미부(2.5 ${\mu}m$)로 각각 구성되어져 있었다. 특히, 본 연구에서 정자의 중편부도 짧지만 미부의 길이는 다른 설치류 종보다도 매우 짧은 것이 특징적이다. 첨체 후 덮개는 핵의 약 1/5을 점유하고 있으며, 적도절은 핵 상단의 첨체 후 덮개와 첨체 덮개사이에 위치하고 있었다. 9개의 분절주들은 미토콘드리아에 의해 둘러싸여져 있었다. 중편부의 미토콘드리아의 회전수는 26개 이었다. 한 개의 분절주는 10~12개의 마디로 구성되어져 있었으며, 이들 경부의 각각의 분절주는 중편부의 9개의 외측치밀 섬유와 제각기 결합되어 있었다. 특히 경부의 분절주와 분절주 사이에는 수많은 satellite 섬유와 흡사한 물질들이 존재하고 있었다. 중편부의 9개의 외측 치밀 섬유는 Nos. 1, 5, 6의 섬유가 다른 섬유들(2, 3, 4, 7, 8, 9) 보다도 크다. 섬유초와 종 지주는 주편부에는 존재하고 있었으며, 미부에는 섬유초와 종 지주는 존재하지 않았다. 이상의 결과에서 볼 때, 정자두부와 꼬리의 구조적인 특징들은 정자의 진화와 더불어 종을 분류하는데 있어서 매우 중요한 정보를 제공하리라 여겨진다.

• Applied Microscopy
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• 제28권2호
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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.

• Applied Microscopy
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• 제39권2호
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• pp.89-99
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• 2009

Cricetinae 아과 Clethrionomys 아속에 속하는 한국산 대륙밭쥐(Clethrionomys rufocanus)의 정자의 형태적 특징을 알아보기 위하여 주사 및 투과형전자현미경으로 관찰하였다. C. rufocanus정자머리의 모양은 날카롭게 굽은 하나의 dorsal hook를 가지며, 전체적인 머리모양은 마치 도끼모양의 형태를 취하고 있었다. C. rufocanus정자의 전체길이는 95.8 ${\mu}m$이었으며, 정자머리의 길이는 7.8 ${\mu}m$, 꼬리는 88.0 ${\mu}m$이었다. 꼬리는 경부 (1.0 ${\mu}m$), 중편부(22.0 ${\mu}m$), 주편부와 미부 (65.0 ${\mu}m$)로 각각 구성되어져 있었다. 분절주 (segmented columns)는 약 10~12개의 마디로 이루어져 있었으며, 미토콘드리아의 수는 약 170~178개 정도이었다. 첨체 후 덮개는 핵의 1/2을 점유하고 있었으며, 적도절은 핵 표면의 첨체 후 덮개(post-nuclear cap)와 첨체덮개(acrosomal cap) 사이에 위치하고 있었다. 1, 5, 6번의 외측섬유들은 다른 것보다 더 크며, 섬유초 (fibrous sheath)와 종 지주(longitudinal column)은 주편부에서 관찰되어졌으며, 미부에서는 섬유초가 존재하지 않았다. 이상의 결과로 미루어 보아, 정자두부와 꼬리의 형태적인 구조는 정자의 진화 패턴과 종을 분류하는데 유용한 정보를 제공하리라 여겨진다.

• Clinical and Experimental Reproductive Medicine
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• 제28권1호
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• pp.65-72
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• 2001

Objectives: This study was to investigate the fertilization rate after intracytoplasmic sperm injection (ICSI) or partial zona dissection (PZD) of human and hamster sperm into hamster oocyte in in vitro fertilization (IVF). In addition, the possibility of clinical application was evaluated by the comparison of usefulness and difference of these method. Materials and Methods: Hamster immature oocytes were obtained from oviducts superovulated by PMSG and hCG, and hamster sperms were obtained from epididymis. The freezed human sperms were thawed before use. Fertilization were confirmed by two pronuclei, one pronucleus, swollen sperm head or/and two polar bodies at $7{\sim}8$ hour after ICSI or PZD. Results: The fertilization rates after ICSI and PZD of human sperm to hamster oocyte were 3.6%, 64.2%,73.6%, and 55.6% for negative control, positive control, ICSI, and PZD respectively, suggesting that ICSI only showed improved fertilization rate (p<0.01). The fertilization rates after ICSI and PZD of hamster sperm to hamster oocyte were 11.1%, 51.2%, 39.6%, and 72.7% for negative control, positive control, ICSI, and PZD respectively, suggesting that PZD only showed improved fertilization rate (p<0.01). PZD showed significantly higher fertilization rate than ICSI (p<0.05). Conclusions: As for the fertilization rate by ICSI and PZD using hamster oocyte in IVF, ICSI technique was considered to be more useful for human sperm and PZD technique for hamster sperm. Therefore, ICSI technique was considered more appropriate for experimental application using human sperm and hamster oocyte.

• Asian-Australasian Journal of Animal Sciences
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• 제33권9호
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• pp.1411-1420
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• 2020

Objective: The aim of study was to investigate the effects of season and single layer centrifugation (SLC) before cryopreservation on post-thaw bull sperm quality in Thailand. Methods: Semen was collected from 6 bulls (Bos indicus) in summer, rainy season and winter 2014 through 2016. Semen characteristics, sperm morphology, sperm kinematics, viability, chromatin structure and mitochondrial membrane were evaluated. Meteorological data were available from the local meteorological station; Results: Season had an effect on semen characteristics in the raw ejaculate, with higher proportions of normal spermatozoa and lower abnormalities in winter than in the other two seasons. Sperm kinematics, viability, DNA fragmentation index, and mitochondrial membrane potential were not different between seasons. Sperm samples selected by SLC had greater normal morphology and a lower proportion with bent tails than controls and higher values of progressive motility (PRO), beat cross frequency, linearity, straightness, wobble (WOB), and lower values of slow motility, velocity average path (VAP), velocity curved line, and amplitude of lateral head displacement than controls. In addition, SLC-selection had a favorable effect on PRO, VAP, and WOB that differed among seasons. Conclusion: Our results suggested that these bulls were well adapted to their location, with season having an effect on sperm morphology. Moreover, SLC could be used prior to cryopreservation, regardless of season, to enhance normal morphology and kinematics of bull sperm samples without adversely affecting other parameters of sperm quality. However, there was considerable variation among bulls in DNA fragmentation index, mitochondrial membrane potential and sperm viability. In addition, SLC had a positive effect on sperm morphology and sperm kinematics, which could be expected to influence fertility.