생쥐 수정란의 시험관내 배양 및 이식 : Bovine serum albumin 및 sodium lactate 효과의 비교분석

Two-cell embryos were recovered from ICR, Balb/c, and Fl hybrid (CBA X C57Bl/6) females, and cultured in vitro in the Ham's F-I0 medium supplemented with 0.1 % BSA, 20 mM Na-lactate, or 10 mM HEPES. With Fl hybrid embryos, there were significantly higher development in Ham's F-I0 medium supplemented with 0.1% BSA (79.1%) and lower development in that supplemented with 10 mM HEPES. Significant differences between strains were seen also, and preimplantation deveiopment was significantly better with the embryos of the Flhybrid strain and was lower with the embryos of the Balb/c strain. It was concluded that BSA and Na-lactate are the important factors affecting preimplantation development, and embryos of the Fl hybrid strain are less sensitive to culture conditions. After transplantation of blastocysts cultured in vitro, the pregnancy rate of recipient mice and implantation rate of transferred embryos were 60% and 32%, respectively

분리된 토끼 단일 심근세포의 형태 계측학적 및 세포화학적 연구

Isolation of intact single cardiac cells was carried out to investigate the shape, size and structure together with the cytochemical reaction in adult rabbit. Perfusion of the heart with calcium free Tyrode solution containing 0.04% collagenase resulted in 50% viability of single cardiac cells which were confirmed by trypan blue exclusion test and Janus green B stain. Isolated single cardiac cells from the left ventricle and atrium were both rod shape. The average length and width of the cells from the ventricle and the atrium were 132.9 f.1 m and 10.6 f.1 m, 132.0 f.1 m and 12.3 f.1 m respectively. The isolated single cells from the SA and the AV nodes were both oval shape and their long axis and short axis were measured as 33.8 f.1 m and 25.0 f.1 m, and 33.8 f.1 rn and 24.6 f.1 m respectively. All the single cardiac cells contained abundant positive granules after McManus' periodic acid Schiff reaction. The single cells from the SA node and the AV node revealed fibrillar structures in their cytoplasm and the single cells from left ventricle and left atrium showed blue cross striations after Mallory's phosphotungstic acid hematoxylin stain

생쥐 활막세포의 분화에 관한 면역세포화학적 연구

This study was designed to observe the differentiation of synoviocytes in the knee joint of mouse by immunocytochemical technique. The feti were collected on the 15th, 16th, 17th, 18th and 19th day of pregnancy, and the I-day-old and 5-day-old neonates were also collected. The knee joints were fixed in Bourn's solution and processed and finally embedded in paraplast by routine method. The serial sections were stained in H&E. In order to visualize the lysozyme and the Ia antigen in the cell, rabbit anti-lysozyme antibody and monoclonal IgG2a anti-mouse I-Ad were used in the double-bridge peroxidase anti-peroxidase technique. To observe the ultrastructure of the synoviocytes, the junctional complex, the basal lamina and the type of capillaries in the synovial membrane, some tissues were embedded in Epon 812 by the usual method. The joint cavities were formed separately in the posterior portion of the patella and between the femur and tibia in the Ifi-day-old fetus. These were united in the 18-dayold fetus. The synoviocytes, which showed a positive reaction to anti-lysozyme, first appeared in the 18-day-old fetus. These synoviocytes were abundant on the luminal side of the primordial infrapatellar fat pad in the 19-day-old fetus but were absent on the surface of the meniscus and cruciate ligaments. The Ia antigen-positive synoviocytes first appeared in the 5-day-old neonate. In the types of synovial membrane, the fibrous type first appeared in the 17-day-old fetus, the areolar type in the 18-day-old fetus, and the adipose type in the 5-day-old neonate. The synoviocytes did not have the basal lamina at any age. The desmosomal junctions were observed between the synoviocytes in the 8-week-old adult. The capillaries supplying blood in the synovial membrane were the fenestrated type with closing diaphragms, but the basal lamina was incomplete. On the basis of above findings, it is obvious that joint cavities are formed by the fusion of small ones during fetal growth. The lysozyme-positive synoviocytes, which compose the synovial membrane, appear before birth, but the expression of the Ia antigen shows up after birth

자궁유출림프절에서 T 및 B 세포계의 임신기간에 따른 변화에 관한 면역조직화학적 연구

The following experiment was performed to investigate changes of immune status in the uterus draining lymph nodes of pregnant ICR mice. The experimental animals were divided by duration of pregnancy as follows; Group I, virgin control; Group II, pregnancy 2nd day; Group III, pregnancy 5th day; Group IV, pregnancy 8th day; Group V, pregnancy 14th day; Group VI, pregnancy 19th day. The objective of this study was to focus on the changes in both B and T-cell systems. To note changes in the B-cell system, the nunber and distribution pattern of activated B cells were observed, the number of IgM and IgG positive plasma cells was counted" and the distribution pattern of surface IgM and IgG positive cells was observed. For changes in the T cell system, the distribution pattern of L3T4 positive and Lyt-2 positive cells was observed. Histochemical staining for activated B cells by means of membrane alkaline phosphatase activity (mAP) and immunohistochemical staining, such as avidin-biotin-peroxidase complex (ABC) and alkaline phosphatase-anti-alkaline phosphatase (APAAP) techniques for T cell and other B-cell systems were carried out. L3T4 positive cells in the deep cortex and IgM- and IgG-positive plasma cells in the medulla were significantly increased near the term. Lyt-2 positive cells were increased in the medullary cord or corticomedullary junction on the 5th day of pregnancy. The results obtained from the above experiment suggest that the conceptus evokes the immune responses locally as well as systemically and that immune responses against the conceptus involve changes in both the B cell and T cell systems, and especially the the T-cell system in early pregnancy and the B-cell system in late pregnancy may be operating

임신한 생쥐의 자궁유출 임파절에서 면역 글로불린 함유세포 수의 변화

This experiment was performed to detect which IgG subclasses are stimulated by the fetal antigens. The experimental animals were mated syngeneically (female ICR X male ICR) or allogeneically (female ICR X male DDY), and the renal and lumbar lymph nodes draining the uterus were dissected out on the 9th and 18th days of pregnancy. The lymph nodes were stained with methyl green pyronin(MGP) and treated with a immunohistochemical reaction. Then each group was compared with the virgin control group. The following results were obtained: 1. In the group of full term mice which were mated syngeneically, the numbers of the IgG1, IgG2b and IgM containing cells were significantly greater than those of virgin, 2. In the group of mid term mice which were mated allogeneically, the number of the IgG2b containing cells was significantly greater than that of virgin. 3. In the group of full term mice which were mated allogeneically, the numbers of the IgGl, IgG2b, IgG3 and IgM containing cells were significantly greater than those of virgin. 4. In the group of full term mice which were mated allogenecially, the numbers of the IgGl, IgG2b and IgG3 containing cells were significantly greater than those of syngeneic mating. On the basis of the above results, it is suggested that the IgG1, IgG2b and IgG3 were stimulated during pregnancy in mice

Immunohistochemical study of the distribution of sodium-dependent vitamin C transporters in adult rat brain

Sodium-dependent vitamin C transporters (SVCTs) is known to transport the reduced form of ascorbic acid into the cell, whereas the oxidized form of vitamin C (VC) is moved through a facilitative sugar transporter, such as glucose transporter (GLUT). With regard to the distribution of SVCT1 and -2 within the various organs, they were reported to be expressed in different types of cells. Especially in the central nervous system, only SVCT2 mRNA was expressed mainly in neurons and some types of neuroglial cells. However, data on the expression of SVCT proteins in the brain are scant. Therefore, we tried to develop comprehensive data on the distribution of SVCT proteins in adult rat brain by using immunohistochemical techniques for the first time. In our study, SVCT2 immunoreactivities (IRs) were intensely localized in the neurons of cerebral cortex, hippocampus, and Purkinje cells of cerebellum, and much weaker SVCT2 IRs were found in the other brain regions. Judging from double-immunohistochemical data, most of the cells expressing SVCT2 IRs were likely to be neurons or microglia, even though the cells in choroids plexus or ependymal cells around the ventricles also exhibited SVCT2 IRs. Complete mapping of the distribution of SVCT2 IRs was available by using a semiquantitative method. The subcellular localization of SVCT proteins is necessary for understanding the exact role of the protein, so the current overall mapping of SVCT IRs in the rat brain could be the basis for further studies on related subjects