• Biomedical Science Letters
• /
• v.4 no.1
• /
• pp.57-63
• /
• 1998

The objectives of this study were to produce polyclonal antibody to adipocyte plasma membrane (APM) proteins isolated from pig, and to investigate its tissue specificity. Plasma membrane proteins from adipocyte, brain, heart, kidney, liver and spleen were isolated using a self-forming Percoll gradient. Sheep (40kg) was immunized three times at three week interval with the purified APM proteins. Blood was taken from non-immunized sheep (NS) and from immunized sheep at 10 (AS-1), 12 (AS-2), and 14 (AS-3) days after the third immunization. Antisera titers and cross-reactivity against other tissues were determined by enzyme-linked immunosorbent assay (ELISA). Antisera reacted strongly to APM proteins showing detectable amounts of antibody at 1：81,000 dilution. And antisera showed much stronger reactivity to APM proteins than any other tissue plasma membrane proteins. Furthermore, tissue specificity of antisera against APM was reconfirmed by immunoblotting using anti-sheep immunoglobulin G-horseradish peroxidase conjugate as a secondary antibody Antisera to APM proteins showed adipocyte specificity compared with other tissues. In conclusion, polyclonal antibody against APM proteins isolated from pig was developed successfully in our laboratory, and these antisera showed tissue specificity with APM.

• Korean Journal of Plant Tissue Culture
• /
• v.25 no.6
• /
• pp.495-500
• /
• 1998

Ultrastructural changes of the isolated and cultured protoplasts from ginseng (Panax ginseng C. A. Meyer) callus were studies with electron microscopy. In the 3-day-cultured protoplasts, the cell organelles such as rough endoplasmic reticulum, ribosome, Golgi complex, mitochondria, proplastid increased in number and observed microtubules. Many vesicles derived from the Golgi complex were evenly distributed in the cytoplasm. Some of such vesicles protruded the outer surface of the plasmalemma, and formed the protuberances. Vacuole derived from endoplasmic reticulum included Golgi vesicles by the invagination of vacuoles. These vacuoles migrated toward the plasmalemma by a fusion process (exocytosis), after fusing the plasmalemma the cell wall materials released from the outer surface of the plasmalemma, and lastly deposited on the plasmalemma. Proplastids containing many starch grains, and microtubules parallel to the plasmalemma were observed near the plasmalemma. Connected fibrils which were observed on the outer surface of the 3-day-cultured protoplast were interpreted as the component of cellulose.

• Journal of Plant Biology
• /
• v.37 no.2
• /
• pp.183-193
• /
• 1994

인삼(Panax ginseng C.A. Meyer)의 뇌두 캘러스로부터 분리한 원형질체와 배양된 원형질체의 미세구조 변화와 원형질막 표면을 투과 및 주사전자현미경으로 조사하였다. 분리 직후의 원형질체에서는 캘러스세포에서보다 작은 액포들이 많이 형성되어 있었다. 또한 활면소포체의 수가 증가하였으며 이들은 원형질막과 평행으로 배열하였다. 활면소포체는 가끔 세포질을 둘러싸서 세포질분리체(cytosegresome)를 형성하였고, 이 구조는 세포질을 분해시킨 후 액포로 변화하기도 하였다. 배양된 원형질체에서는 분리 직후의 원형질체와 비교하여 조면소포체, 딕티오좀, 리보좀, 미토콘드리아, proplastid 및 액포 등의 수가 현저하게 증가하였다. 딕티오좀으로부터 많은 소낭들이 형성되었고 이들은 세포질 전반에 걸쳐 존재하였다. 때로는 소낭들이 원형질막 바깥으로 돌출되어 돌기를 형성하기도 하였다. 배양된 원형질체의 표면에는 섬유소로 구성되어 있을 것으로 추정되는 섬유상 구조들이 형성되어 있었다.

• Journal of Life Science
• /
• v.26 no.8
• /
• pp.875-880
• /
• 2016

The plasma membrane plays a crucial role in relaying signals from the outside environment to the inside of the cells. In eukaryotic cells, the inner leaflets of the plasma membrane are composed mostly of phospholipids, including phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylinositides (PIs). In this study, we tried to analyze the morphological changes induced by EGFP-fused membrane binding proteins, which are targeted to the plasma membrane via specific phospholipids binding. As a result, we found that overexpression of EGFP-P4M-SidM, a specific PI4P binding protein, or EGFP alone, did not induce any morphological changes. On the other hand, overexpression of EGFP-PLCδ1(PH), which is a specific PI(4,5)P₂ binding protein, EGFP-AKT1(PH) which binds to PI(3,4,5)P₃, or EGFP-OSH2(PH)×2 which binds to PI4P and PI(4,5)P₂, could induce the filopodia and lamilapodia formation as well as cell shrinkage. Overexpression of Lact-C2-EGFP which is a specific PS-binding probe, EGFP fused Aplysia phosphodiesterase 4 (ApPDE4) long-form (L(N20)-EGFP) which is localized to the plasma membrane via hydrophobic interaction, or EGFP fused Aplysia PDE4 short-form (S(N-UCR1-2)-EGFP) which is localized to the plasma membrane via electrostatic interaction, could induce cell shrinkage, but not filopodia or lamilapodia formation. Taken together, our data support that the different phospholipid bindings in the plasma membrane could induce different characteristic morphological changes. Thus, we can analyze, characterize, and classify the cellular morphological changes induced by the various phospholipid binding proteins.

• Analytical Science and Technology
• /
• v.26 no.1
• /
• pp.106-112
• /
• 2013

PDE plays an important role in cAMP-mediated cellular signaling within the cells. The proper targeting of each PDE is mediated by unique N-terminal of each PDE isoform. It has been recently reported that supershort-, short- and long-forms of PDE4 in Aplysia were cloned in Aplysia. Long-form of ApPDE4 was localized at plasma membrane and presynaptic terminal in Aplysia sensory neurons. However, it remains elusive which part of ApPDE4 is minimal region for the proper targeting and what are the effects on the cell functions. Here, we identified that N-terminal 13 amino acids of ApPDE4 long-form is minimal regions for the plasma membrane targeting. In addition, overexpression of ApPDE4(N20)-mRFP could induce morphological changes in HEK293T cells. Interestingly, mRFP-$PLC{\delta}1$(PH), which selectively binds to PI4,$5P_2$, could induce morphological changes in similar with that by ApPDE4(N20)-mRFP. These results suggested that binding of ApPDE4(N20) to lipids including PI4,$5P_2$ might be responsible for targeting of ApPDE4 to plasma membrane and morphological changes in HEK293T cells.

• The Korean Journal of Zoology
• /
• v.37 no.1
• /
• pp.120-129
• /
• 1994

생쥐의 배우자간 인식과 수란관과 난자의 상호작용을 연구하기 위하여 수정 전후의 난자와 배아에 형광으로 표지된 5가지 렉틴(UEA-1. LCA, PNA RCA-1, GSL-1)을 처리하여 투명대, 위낭강 원형질 막에서 렉틴의 결합양상을 관찰하였다. 미수정난자 및 수정란의 투명 대에는 UIA-1을 제외한 4가지 렉틴이 결합하였다. PNA RCA-1, GSL-1은 투명대의 의총보다 내층에 강하게 결합되었고 UEA-1과 LCA는 투명대의 전층에서 미약하지만 고르게 결합하였다 배란된 난자의 위난강은 LCA와 PNA에 의해 결합되었으며 UEA-1은 수정 이후부터 GSL-1은 2-세포기 이후부터 위난강에 결합하였다. 난구세포를 제거하고 체외성숙을 일으킨 난자의 위난강에는 5가지 렉틴 모두 결합하지 않았다. 난자와 배아의 원형질막은 PNA. RCA-1, GSL-1에 의해 결합되었고, LCA 및 UEA기은 수정 이후부터 원형질막에 결합하였다. 이상의 결과에서 glucosamine, N-acetylgalactoamine 관기를 함유한 물질이 난구세포에서 생성되어 위난강에 축적되며 배란 후부터 난자의 위난강내 탄수화물관기는 수란관내액의 영향을 받는 것으로 사료된다. 난자 투명대의 탄수화물관기는 내층에 밀집되어 분포하는데 이는 정자의 투명대 관입 조절과 관련된 것으로 사료된다. 수정 후 난자의 원형질막 표면에 fucose와 mannose 등의 탄수화물 잔기가 새로이 노출되며 초기배아 발생동안 위난강에 존재하는 구조물의 렉틴 결합 특성이 유지되었다.

• Journal of Animal Science and Technology
• /
• v.49 no.6
• /
• pp.729-736
• /
• 2007

The objective of this study was to investigate the anti-oxidative effects of pyruvate, taurine and melatonin on sperm characteristics(motility, membrane integrity) and lipid peroxidation(LPO) for in vitro storage of boar semen. Semen was treated with various antioxidants such as pyruvate(1mM), taurine(50mM) and melatonin(100nM) with or without 100uM H2O2. Antioxidant treatments were significantly increased the sperm motility when compare to control group in all incubation periods(P≤0.05). Hypoosmotic swelling test(HOST), membrane integrity was similar to the result of motility. In lipid peroxidation measurement by TBA reactions of spermatozoal plasma membrane, malondialdehyde(MDA) level in control and antioxidant treatments were lower than those of antioxidant plus H2O2 or H2O2 treatment for 3 to 6 h incubation period. Relationships of evaluation methods for sperm viability were investigated by motility, membrane integrity and lipid peroxidation. Among evaluation methods, LPO vs motility and membrane integrity vs LPO were negatively correlated(－0.23~－0.92 and －0.68~－0.85), but membrane integrity vs motility was positively correlated (0.53~0.94) in all treatments. These experiments indicate that supplementation of antioxidant to the semen extender can increase the sperm motility and membrane integrity and decrease the lipid peroxidation of spermatozoal plasma membrane. The HOST might be utilized to evaluate the sperm quality instead of lipid peroxidation or motility.

• Applied Microscopy
• /
• v.4 no.1
• /
• pp.25-33
• /
• 1974

The origin and the function of cytolysomes were studied in the mesophyll cells and the root-tip cells of Glycine max Merr. and Zea mays L. fixed by paraformaldehyde-glutaraldehyde-$OsO_4$. The cytolysome-like structures were found of three main types of configurations: multivesicular, myelin like (multilamllar) and multitubular. More complex and mixed ones were also observed. The origin of these structures seems to be initiated by invaginations or in holdings of the plasmalemma into the cell interior, and that by aggregation and convolution of endoplasmic reticulum in the cytoplasm. Invagination of the plasmalemma were found of two main types of configurations: concentric whorls of lamellar and multivesicular. The structures were also observed within vacuoles and cytoplasm. Since the structurers are widely distributed in the cells and are greatly varied in sizes and shapes. These structures originate from the plasmalemma and the cytoplasm subsequently protrudes into the vacuole, and that seem to play an important role on the formation of the autophagic vacuoles. The possible function and fate of these structures are discussed.

• The Korean Journal of Malacology
• /
• v.9 no.2
• /
• pp.94-102
• /
• 1993

동양달팽이(Nesiohelix samarangae)의 자성생식기 중 정자낭(spermatheca)의 미세구조를 투과전자현미경을 이용하여 관찰하였다. 성체의 정자낭의 내강에서는 정자와 이와 관련된 물질들 또는 상피로부터의 분비물질들이 관찰되었다. 정자낭의 내강을 둘러싸고 있는 벽의 상피는 단층원주상피로서 내강과 연한 세포의 원형질막은 미세융모를 가지고 있었으며 섬모는 가지고 있지 않았다. 이 미세융모들의 기부에서는 고배율 하에서 pinocytotic vesicle들이 관찰되었고 때로 세포의 분비물을 세포 밖으로 방출하는 모습도 관찰되었다. 세포들의 상부세포질에는 많은 수의 mitochondria가 있었고, 때로 분비과립과 lysosomd들이 존재하기도 하였다. 이들보다 아래에는 잘 발달한 골지체와 과립소포체(RER)들이 있었다. 이 부위에서 분비과립들이 많이 형성되어 있는 모습이 세포에 따라 관찰되었고, 그 아래에 즉, 각 세포의 하반부 세포질에 상하로 긴 핵이 관찰되었다. 세포의 기부 원형질막은 상부 세포질 내로 깊게 그리고 수없이 많이 주름을 이루고 있어 인접한 세포와의 경계를 구분할 수 없을 정도였다. 기저막의 아래에는 근육다발과 색소세포들이 결합조직 내에서 관찰되었고, 정자낭의 외막은 큰 액포로 인하여 세포질들이 원형질막 주변으로 밀려나 있는 특이한 세포들로 이루어져 있었다. 정자낭의 기능은 정자를 일시 저장하며 정자낭 상피에서 분비되는 소화액들로 정자를 일차적으로 세포외 소화하고 소화되지 않은 물질들을 흡수하여 이차적으로 세포내소화하는 것으로 생각된다.

• Biomedical Science Letters
• /
• v.6 no.4
• /
• pp.237-244
• /
• 2000

It has been reported that plasma membrane activity of the spermatozoa may be susceptible to be influenced by extracellular osmolality and such membranous changes involve infracellular molecular changes, special regard to the structure of membranous lipids, and the accompanying ion-channel of which are closely related with their fluidity of $Ca^{2+}$ and HCO$^{-}_{3}$. It is of common recognition that a certain kind of sterol acceptor player an important to induce lipid fluctuation of the sperm plasma membrane which have been influenced by BSA administration and came in effect to outflow of cholesterol from the spermatozoa and resulted in changes of ionic fluidity to facilitate adenylyl cyclase, and to induce protein tyrosine phosphorylation by increase of cAMP and activation of PKA. Thus it seems likely that an augmentation of the acrosomal reaction is closely related with protein tyrosine phosphorylation. The following experimental results were obtained in the present study; Under the high osmolality conditions, the spermatozoa motility declined significantly and the structural change of the plasma membrane diminished to confirm that the response degrees to the osmolality depended upon the water transfer volume through the plasma membrane and the changes of cellular volume. Those experimental results suggest that a physiological parameter such as low temperature condition played an important role for presentation of spermatozoa and that inducement of spermatozoa activation for reinforcement of protein tyrosine phosphorylation. On the other hand, it seemed likely that the BSA administration as one of sterol accepters might represent a key role also under the high osmolality condition and their result also suggests that osmolality change, special regard to high osmolality condition may play an important role also in the processes of signal transmission.