• Korean Journal of Veterinary Research
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• v.32 no.3
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• pp.295-308
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• 1992

In order to study on the Sertoli cell, we attempt have been made to measure the average number of each germ cells per Sertoli cell on the 12 stages of cycle in matured korean Jindo dog. The fine structure of Sertoli cell junctional specialization was studied with electron microscope. The results were summarized as follows; 1. The average number of various germ cells associated with Sertoli cell was 9.77 to 13. 80 through stages of cycle and the total average number was 11.62. 2. Sertoli-Sertoli cell junctional specialization was present in seminiferous epilthelium, and Sertoli-spermatid cell junctional specialization rose from stage 8 spermatid, persisted to step 13 spermatid and then disappeared. The structure of Sedoli-spermatid cell juncticnal specialization was not similar to that of Sertoli cxlls. 3. Just after spermiation, free-surface of Sertoli-spermatid cell junctional specialization was replaced by Sertoli cell cytoplasm with tubulobulbar complex at the neiglaboring region observed. 4. The Sertoli cell process was located within the cytoplasm of late stage spermatids. Some membranes of residual body and spermatid cytoplasm partly disappeared, resulting in opening of the cytoplasm of spermatid into that Sertoli cell. This fact suggested that spermatid cytoplasm was partly eliminated.

• Korean Journal of Animal Reproduction
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• v.11 no.3
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• pp.206-217
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• 1987

Testes from the drake and the gander have been examined by the electron microscopy in thin sections in order to examine the spermiogenesis and the structure of spermatozoa. The spermiogenesis can be divided into three stages: early spermatid, nuclear elongation, and matured spermatid. In the early spermatid of the drake, there are thread-like material in the nucleus, a prominent nuclear envelope around the nucleus, and big lumens in the cytoplasm. The shape of the gander's mitochondria in the early spermatid is slender compared to that of the drake, and the inner membrane of the mitochondria is thicker than the outer membrane. The distal centriole of the drake and the gander in the early spermatid is a long hollow cylinder form. In the nuclear elongation stage, elongated nucleus forms two or three cross sections in one spermatid cell and it is surrounded by the amorphous sheath. The nucleus of the matured spermatid is compact and its apical end is covered with acrosome cap and acrosome spine. The axoneme is surrounded by the amorphous material.

• Applied Microscopy
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• v.26 no.1
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• pp.33-45
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• 1996

The spermatogenesis of Korean slug, Incilaria fruhstorferi are observed by electron microscope. The results are as follows: The spermatogenesis of Korean slug, Incilaria fruhstorferi, is processed through the five stages; Spermatogonia, primary spermatocyte, secondary spematocyte, spermatid and spermatozoon. The spermiogenesis, the differentiation of the spermatid, is also processed through the five stages. In stage 1, the numerous and round mitochomdria in the cytoplasm are moved around the nucleus of spermatid. In stage 2, the nucleus of spermatid transformed into the oval shape, and the oval nucleus is surrounded by many rough endoplasmic reticulum. In stage 3, the oval nucleus of spermatid is changed to be curved as an arrow, and then two centrioles appeared behind nucleus. The centriole is sucked into the cytoplasm. and almost all the chromatins are changed into heterochromatins. In stage 4, the nucleus of spermatid are transformed into the oval shape, when the lamella plate chromatin of spermatid form in the nucleoplasm. In stage 5, the oval nucleus is then transformed into the stream-line shape when the lamella plate chromatin of spematid gradually concentrated in the nucleus, and long axoneme ($65{\mu}m$ in length) form from the distal centriole. Two long mitochondria in the middle piece and the main piece of spermatozoon array spirally along a long axoneme, and the mitochondria matrix is gradually filled with electron-dense glycogen particles ($0.1{\mu}m$ in size). The axoneme of spermatozoon consists of typical 9+2 microtubular pattern.

• Korean Journal of Animal Reproduction
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• v.22 no.4
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• pp.395-403
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• 1998

In this study we determined fertilization processes and developmental ability of porcine oocytes following injection of round spermatid in the presence and absence of artificial activation. Electrical stimulation at 2 h before spermatid injection significantly increased the incidence of normal fertilization as compared to those following injection without stimulation or with stimulation immediately after injection. The incidences of two pronuclear formation and apposition were not different in oocytes between following intracytoplasmic spermatid and spermatid nucleus injection. Indirect immunocyto chemistry and laser scanning confocal microscopy study revealed that microtubuels were organized from the oocyte cortex following round spermatid injection, and this seemed to move both male and female pronuclei into the oocyte center. At 6 days following injection blastocoele formation was seen in the eggs following round spermatid (25%) and round spermatid nucleus injection (27%). However, none of oocytes developed to the blastocyst stage at 6 days following sham injection. The average cell numbers of blastocysts at 8 days following injection of spermatid and spermatid nucleus were 87 to 99. These results suggested that either round spermatid or it's nudeus can be used to produce viable embryos by injection into unfertilized oocytes in the pig.

• Reproductive and Developmental Biology
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• v.29 no.4
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• pp.241-245
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• 2005

Porcine oviduct epithelial cells (POEC) are widely used in co-culture experiments to improve early embryonic development, in vitro fertilization in embryo transfer programs for domestic animals and in vitro maturation of immature germ cells. POEC were mechanically isolated and cultured in tissue culture medium 199. Cells grew continuously, and confluent monolayers were formed after 7 days. After forming confluent monolayer of epithelial cells, supernatant was collected as the condition medium for maturing round spermatids in vitro. Round spermatids were also separated mechanically and cultured in the POEC condition medium. In this study we observed that $20\%$ of round spermatid cultured were matured into elongating spermatid after 24 h, and about $10\%$ of round spermatid cultured showed complete elongation (elongated spermatid) within $24\~48$ h of in vitro culture. No further development was observed within $50\~72$ h and transformed cells lost their viability after 72 h. These preliminary findings suggest that the condition medium from POEC may be possible to overcome the round spermatid block by improving the milieu of culture system.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.2
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• pp.193-201
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• 1999

In this study we determined fertilization processes and developmental ability of porcine oocytes following injection of round spermatid in the presence of artificial activation. Electrical stimulation at 3 h before spermatid injection significantly increased the incidence of normal fertilization as compared to those following injection without stimulation or with stimulation immediately after injection. The incidences of two pronuclear formation and apposition were not different in oocytes between following intracytoplasmic spermatid and spermatid nucleus injection. Indirect immunocytochemistry and laser scanning confocal microscopy study revealed that micro tubules were organized from the oocyte cortex following round spermatid injection, and this seemed to move both male and female pronuclei into the oocyte center. Paternal mitochondria which are introduced with spermatid have been observed up to 4-cell. Our study indicated that either round spermatid or it's nucleus can be used to produce viable bovine embryos by injection into unfertilized oocytes.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.531-539
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• 1996

The lectin-binding patterns in the testis of the sexually matured Jindo dog were investigated to study the distribution of glycoconjugates in the seminiferous tubule under light and transmission electron microscopy. Positive reactions to Wheat germ agglutinin(WGA) and Dolichos biflorus agglutinin (DBA) were observed in the Sertoli cell and in the residual body of spermatid with a stronger reaction in the Sertoli cell to the lectins than in the residual body. Strong reactions to Soybean agglutinin(SBA) and Peanut agglutinin(PNA) were observed in the acrosome vesicles of the Golgi- and cap-phase spermatid, while a moderate reaction was observed in the acrosome-phase, maturation-phase spermatid and the residual body. The acrosome area of the spermatid reacted intensively to Griffonia simplicifolia agglutinin( GS-I) when the cell was in the acrosome-phase and maturation-phase, and the same reaction to the GS-I was observed in the residual body. However, the seminiferous tubule did not react to Ulex europeus agglutinin I(UEA-I). The gold-labelling of the Sertoli cells with DBA resulted in positive reactions of the Sertoli cell column and processes when observed under the electron microscopy, while the Golgi-, cap- and acrosome-phase spermatids reacted positively to SBA in the peripheral low-dense area of the acrosome vesicle of spermatid. Based on these results, we concluded that differences in the lectin-binding pattern of the seminiferous tubules were recognized in the Jindo dog compared to other animals.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.327-331
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• 1996

Normal fertilization and pregnancy by round spermatid was achieved from nonobstructive azoospermia patient to be believed untreatable. Therefore, it is suggested that application of round spermatid in human ART program seems to be new treatment of male infertility. Also, it will be needed further research for evaluated fertilization mechanism by round spermatid injection.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.2
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• pp.233-237
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• 1998

We demonstrated that the normal pregnancy and delivery by round spermatid injection (ROSI) into oocytes was achieved from nonobstructive azoospermia patient. In this case, the normal fertilization rate was 50%. All of the two pronuclear stage embryos cleaved and were transferred to the patient's uterus. A singleton pregnancy was achieved and resulted in the birth of normal female infant. This resuJt show that intracytoplasraic injection of round spermatid seems to be new treatment of nonobstmctive azoospermia male infertility. Further research is needed to evaluate the required culture conditions to induce progression of the round spermatid into a more elongated stage.

• Korean Journal of Animal Reproduction
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• v.23 no.4
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• pp.359-364
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• 1999

Recent advances in intracytoplasmic sperm (ICSI) and round spermatid injection (ROSI) would provide exciting opportunities not only for the male infertility but also for studying gamete physiology during fertilization and early development. Furthermore, intracytoplasmic sperm injection could be used to produce transgenic animal (Perry et al., 1999). However, it is not clear in the fertilization processes in mammalian oocytes following intracytoplasmic injection of spermatozoon, isolated sperm head or round spermatid. (omitted)