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

• Reproductive and Developmental Biology
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• v.30 no.2
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• pp.107-113
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• 2006

Development of placenta is a complex process that is critical for the pregnancy and controlled by many factors including cytokines, hormones, growth factors and apoptotic molecules. Recently, it has been shown that progranulin (PGRN) functions in growth of embryo and trophectoderm as well as cell migration. To initiate understanding the role of PGRN in human placental development, we investigated the expression of PGRN mRNA and protein in early and late gestation human placentas, term cytotrophoblast cells and two choriocarcinoma cell lines, JEG-3 and Jar. Reverse transcriptase polymerase chain reaction identified mRNAs derived from the PGRN gene in all samples. Immunoblot analysis showed that PGRN proteins are present in early and late gestation human placentas with decreasing levels over gestation and that PGRN proteins are present in normal and transformed trophoblast cells. Immunohistochemical analysis using paraformaldehyde-fixed tissue sections taken from early and late stages of pregnancy showed that PGRN proteins are present in cytotrophoblast cells, syncytiotrophoblast and extravillous cytotrophoblast cells and that expression pattern of PGRN differed according to the stage of cell differentiation. The results of this study are consistent with the hypothesis that PGRN proteins have critical roles in placental development and suggest that PGRN may function in trophoblast cell growth and differentiation.

• Molecules and Cells
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• v.24 no.1
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• pp.76-82
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• 2007

By combining in situ hybridization (ISH) and immunocytochemistry (IC), microscopic topological localization of mRNAs and proteins can be determined. Although this technique can be applied to a variety of tissues, it is particularly important for use on neuronal cells which are morphologically complex and in which specific mRNAs and proteins are located in distinct subcellular domains such as dendrites and dendritic spines. One common technical problem for combined ISH and IC is that the signal for immunocytochemical localization of proteins often becomes much weaker after conducting ISH. In this manuscript, we report a simplified but robust protocol that allows immunocytochemical localization of proteins after ISH. In this protocol, we fix cultured cortical or hippocampal neurons with 4% paraformaldehyde (PFA), rinse briefly in PBS, and then further fix the cells with $-20^{\circ}C$ methanol. Our method has several major advantages over previously described ones in that (1) it is simple, as it is just consecutive routine fixation procedures, (2) it does not require any special alteration to the fixation procedures such as changes in salt concentration, and (3) it can be used with antibodies that are compatible with either methanol (MeOH-) or PFA-fixed target proteins. To our best knowledge, we are the first to employ this fixation method for fluorescence ISH + IC.

• Journal of the Speleological Society of Korea
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• no.71
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• pp.1-4
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• 2006

Chloromethylnaphthalene is a valuable compound for obtaining of the plant growing stimulator - -napthylacetic acid. Chloromethylation of naphthalene by paraformaldehyde in the presence of glacial acetic acid, phosphoric and hydrochloric acids at temperature 80 - 85C and duration - 6 hours the -chloromethylnaphthalene yield was 55-57%. Using Box-Wilson method for mathematical planning of experiment carried out optimization of its synthesis for purpose increasing -chloromethylnaphthalene yield. Preliminary, one - factor experiments were carried out for selecting independence main parameters influencing on the synthesis. A full factor experiment of 23 with extended matrix of planning was used for optimization. Aiming to increase the -chloromethylnaphthalene yield, the obtained mathematical model was used for program of sharp raising on the reply surface. The received optimal conditions for the -chloromethylnaphthalene synthesis were selected as following: molar ratio of naphthalene parapfsormaldehyde of 1 : 2 temperature - 105C duration of the reaction - 3 hours. The yield of -chloromethylnaphthalene under these optimal conditions was 75%.

• Archives of Pharmacal Research
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• v.25 no.3
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• pp.370-374
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• 2002

Administration of dopamine agonist, apomorphine (2 mg/kg, s.c.), produces cage climbing behavior in mice that exhibit typical dopaminergic stimulation. The present study investigated the pCREB expression level in several brain regions following apomorphine treatment in order to determine whether the increased the dopaminergic activation produced by apomorphine accompanies the changes in pCREB immunoreactivity. A mouse brain was removed at 0min, 10 min, 30 min, 1 h, 2 h, 7 h, and 24 h after apomorphine treatment. The brain tissue was fixed by an intracardiac perfusion with ice-cold 4％ paraformaldehyde in PBS. Immunohistochemical study was conducted using the ABC-DAB method. The data showed that the immunoreactivity of pCREB increased in the striatum, nucleus-accumbens, piriform cortex and the dentate gyrus of the hippocampus of a mouse brain 30 min after the apomorphine treatment. Increased immunoreactivity began to diminish 2 h after the apomorphine treatment in all the brain regions measured. The time course for the pCREB immunoreactivity was similar to the behavioral response induced by the apomorphine treatment. These results suggest that activation of the dopamine receptor is accompanied by an increase in pCREB expression in the mouse brain.

• Journal of Physiology & Pathology in Korean Medicine
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• v.32 no.1
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• pp.30-34
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• 2018

This research was practiced to comparative investigate the distribution of sensory and motor neuron linkaged with Yangji(TE4) by using neural-tracer technology. A total 16 S-D rats were used in the present research. After anesthesia, the rats received micro-injection of $6{\mu}{\ell}$ of cholera toxin B subunit(CTB) into the relation positions of the Yangji(TE4), in the human body for observing the distribution of the linkaged sensory neurons in dorsal root ganglia(DRGs) and motor neurons in the spinal cord(C3~T4) and sympathetic ganglia. 3 days after the micro injection, the rats were anesthetized and transcardially perfused saline and 4% paraformaldehyde, followed by routine section of the DRGs, sympathetic chain ganglia(SCGs) and spinal cord. Marked neurons and nerve fibers were detected by immunohistochemical method and observed by light microscope. The marked neurons were recorded and counted. From this study the distribution of primary sensory and motor neurons linkaged with Yangji(TE4) were concluded as follows. Yangji(TE4) dominated by spinal segments of C5~T1, C6~T4, individually.

• International Journal of Oral Biology
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• v.40 no.4
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• pp.183-187
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• 2015

The present study was aimed to evaluate the influence of glutaraldehyde (GA) concentration on multiple electron microscopic (EM) immunostaining using pre-embedding peroxidase and post-embedding immunogold method. Influence of various concentrations of GA included in the fixative on immuoreactivity was assessed in the multiple immunostaining using antisera against anti-transient receptor potential vanilloid 1 (TRPV1) for peroxidase staining and anti-GABA for immunogold labeling in the rat trigeminal caudal nucleus. Anti-TRPV1 antiserum had specificity in pre-embedding peroxidase staining when tissues were fixed with fixative containing paraformaldehyde (PFA) alone. Immunoreactivity for TRPV1 was specific in tissues fixed with fixative containing 0.5% GA at both perfusion and postfixation steps, though the immunoreactivity was weaker than in tissues fixed with fixative containing PFA alone. Tissues fixed with fixative containing 0.5% GA at the perfusion and postfixation steps showed specific immunogold staining for GABA. The results of the present study indicate that GA concentration is critical for immunoreactivity to antigens such as TRPV1 and GABA. This study also suggests that the appropriate GA concentration is 0.5% for multiple immunostaining with peroxidase labeling for TRPV1 and immunogold labeling for GABA.

• Korean Journal of Veterinary Research
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• v.35 no.1
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• pp.29-37
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• 1995

This study was carried out to investigate the morphological changes of the oxytocin secreting neurons in age-related rat hypothalamus by means of immunohistochemistry. The experimental group was 20 aged female rats (Sprague-Dawley, 25~30 months), and the control group was 10 adult female rats (10 months). All animals were perfused transcardially with 4% paraformaldehyde-lysine-periocdate(PLP), and serial transverse brain sections($30{\mu}m$) were prepared by means of cryotome, and were stained with rabbit anti-oxytocin antisera immunohistochemically, using the free floating method and avidin-viotin peroxidase complex. The results were as followings. 1. The immunostained oxytocin secreting neurons were located at the paraventricular nucleus and supraoptic nucleus of age-related rat hypothalamus chiefly. 2. The numbers of oxytocin secreting neurons decreased at the paraventricular nucleus and supraoptic nucleus of age-related rat hypothalamus(p<0.01) 3. The oxytocin secreting neurons of age-related rat hypothalamus immunostained less than those of the adult rat hypothalamus, and the paraventricular nucleus immunostained greater than supraoptic nucleus in age-related rat. 4. The numbers and expansion of dendrite and axonal varicosities decreased in the age-related rat hypothalamus greatly.

• Korean Journal of Veterinary Research
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• v.56 no.2
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• pp.85-89
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• 2016

We tested a set of conditions for obtaining optimal tissue quality in preparation for histology in samples of mouse brain. C57BL/6J mice were sacrificed and perfused with 4% paraformaldehyde, after which the brains were removed and dehydrated in 30% sucrose solution. The brains were then divided into four groups according to freezing temperature and usage of optimal cutting temperature (OCT) compound. Next, we stained the sectioned brain tissues with Harris hematoxylin and eosin Y and immunohistochemistry was performed for doublecortin. The best quality tissue was obtained at $-25^{\circ}C$ and by not embedding with the OCT compound. When frozen at $-25^{\circ}C$, the embedded tissue was significantly damaged by crystals, while at $-80^{\circ}C$ there were no meaningful differences between qualities of embedded- and non-embedded tissues. Overall, we identified a set of conditions to obtain quality frozen brain sections. Our developed protocol will help resolve matters associated with damage caused to sectioned brain tissue by crystal formation during freezing.

• The Korean Journal of Zoology
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• v.26 no.2
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• pp.107-123
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• 1983

The globus pallidus of normal cats were prepared for electron microscopic study following perfusion with a mixture of 1% paraformaldehyde and 1% glutaraldehyde solution. Neurons of two size categories were identified in 1 $\\mu$m araldite sections and their ultrastructural characteristics were studied in adjacent thin section. 1. Large neurons ($30 \\mum \\times 45 \\mum$ in diameter) had extensive areas of rough surfaced endoplasmic reticulm, abundant perinuclear Golgi complex, numerous mitochondria and lipofusin granule, and had a large spherical nucleus with shallow indentation of nuclear manbrane. Small neurons ($17 \\mum \\times 27 \\mum$ in diameter) had poorly rough surfaced endoplasmic reticulum, moderate number of mitochondria and randomly distributed Golgi complex. The nuclear envelope of this cell frequently showed multiple deep invagination. 2. Three types of axo-somatic synapses were identified on the basis of the size and shape of vesicle in the axon terminal and the symmetrical or asymmetrical thickening at the synaptic site. Type I synaptic terminal shows an even distribution of round and oval synaptic vesicles, and has a symmetrical synaptic thickening. Type II axon terminals reveal mostly round and pleomorphic vesicles and a few vesicles were localized near the presynaptic membrane in pale axoplasm and its synaptic thickening were symmetric. Type III axon terminals contain round vesicles, which were aggregated in the axoplasm, and has a asymmetrical synaptic thickening. 3. The majority of axo-somatic contact with the large and small neurons were type I, and type II and III synapes were rare.