• International Journal of Oral Biology
• /
• 제41권3호
• /
• pp.133-139
• /
• 2016

The ultrastructural parameters related to synaptic release of endings which are presynaptic to tooth pulp afferent terminals (p-endings) were analyzed to understand the underlying mechanism for presynaptic modulation of tooth pulp afferents. Tooth pulp afferents were labelled by applying wheat-germ agglutinin conjugated horseradish peroxidase to the rat right lower incisor, whereafter electron microscopic morphometric analysis with serial section and reconstruction of p-endings in the trigeminal oral nucleus was performed. The results obtained from 15 p-endings presynaptic to 11 labeled tooth pulp afferent terminals were as follows. P-endings contained pleomorphic vesicles and made symmetrical synaptic contacts with labeled terminals. The p-endings showed small synaptic release-related ultrastructural parameters: volume, $0.82{\pm}0.45{\mu}m^3$ ($mean{\pm}SD$); surface area, $4.50{\pm}1.76{\mu}m^2$; mitochondrial volume, $0.15{\pm}0.07{\mu}m^3$; total apposed surface area, $0.69{\pm}0.24{\mu}m^2$; active zone area, $0.10{\pm}0.04{\mu}m^2$; total vesicle number, $1045{\pm}668.86$; and vesicle density, $1677{\pm}684/{\mu}m^2$. The volume of the p-endings showed strong positive correlation with the following parameters: surface area (r=0.97, P<0.01), mitochondrial volume (r=0.56, P<0.05), and total vesicle number (r=0.73, P<0.05). However, the volume of p-endings did not positively correlate or was very weakly correlated with the apposed surface area (r=-0.12, P=0.675) and active zone area (r=0.46, P=0.084). These results show that some synaptic release-related ultrastructural parameters of p-endings on the tooth pulp afferent terminals follow the "size principle" of Pierce and Mendell (1993) in the trigeminal nucleus oralis, but other parameters do not. Our findings may demonstrate a characteristic feature of synaptic release associated with p-endings.

• Asian-Australasian Journal of Animal Sciences
• /
• 제21권4호
• /
• pp.532-537
• /
• 2008

In order to investigate the factors affecting the culture of mouse preantral follicles in vitro, we examined the effect of culture media, protein supplements, and culture period on their growth. The oocyte diameter (initial size: $55.6{\pm}2.5{\mu}m$) was progressively increased during culture, and the maximum size ($72.0{\pm}2.4{\mu}m$) was reached on day 10 of the in vitro culture. The chromatin configuration in the germinal vesicle (GV) oocyte progressively shifted from a non-surrounded nucleolus (NSN) to a surrounded nucleolus (SN). On day 10 of the culture, most of the oocytes progressed to the SN pattern. The survival and metaphase II rates of the oocytes in alpha-minimal essential medium (alpha-MEM) were significantly higher (p<0.05) than those in Waymouth and tissue culture medium (TCM)-199. As a protein source, fetal bovine serum (FBS) was more suitable for the culture of mouse preantral follicles as compared to human follicular fluid (hFF) and bovine serum albumin (BSA); the optimal concentration of FBS was 5%. These results suggest that in a culture of mouse preantral follicles, alpha-MEM and 5% FBS are an optimal medium and a protein source, respectively; further, the 10 days of culture is required for the complete growth of oocytes in this culture system.

• Journal of Pharmaceutical Investigation
• /
• 제22권2호
• /
• pp.115-124
• /
• 1992

The characteristics and stabilities of phosphatidylcholine liposomes covalently coupled with immunoglobulin fragments prepared by the REV method were investigated by the dynamic light scattering, absorbance and calcein release. Using a sulfhydryl-reactive phospholipid derivative of N-[4$({\rho}-maleimido-phenyl)$ butyl] phosphatidylethanolamine (MPB-PE), Fab' antibody fragments were covalently combined with preformed large unilamellar vesicles (LUV), Coupling ratio was $250\;{\mu}g$ of $Fab'/{\mu}mol$ of phospholipid in vesicles, From dynamic light scattering, it was found that the size of the vesicles increases as the ratio of cholesterol to lipid increases, but that apparently, the size of liposomes was not sensitive to the existence of Fab' fragments. Regardless of inserting Fab' fragments, the absorbance of liposomes decreased as the amounts of bile salt (BS) added. At very low BS concentrations, BS/lipid aggregates would be formed in the outer vesicles monolayer, while, at the high BS concentrations, mixed micelles would be preferred. The vesicles incorporated with Fab' fragments, however, are more resistant to the bile salts than the MPB-PE vesicle are. The absorbance of vacant liposomes and calcein release resulted in that the Fab' vesicles and MPB-PE vesicles by the REV method are very stable, but that those by the sonication method sufferred the significant change of turbidities.

• Animal cells and systems
• /
• 제3권4호
• /
• pp.375-383
• /
• 1999

Gonadal development, gametogenesis, reproductive cycle, and first sexual maturity of Reishia clavigera were investigated monthly from July 1998 to June 1999 through cytological and histological observations. R. clavigera had separate sexes, and was an internal fertilizer. The ma1e penis was located near the two tentacles. The ovary and testis were composed of a great number of oogenic lobules and spermatogenic tubules, respectively. The size of ripe oocyte ranged from 130 to 140 ${\mu}$m in diameter. The peripheral cytoplasm of the germinal vesicle of the ripe oocyte in many cases were surrounded by smaller yolk granules, while the eccentric cytoplasm was occupied with larger ones. The reproductive cycle of R. clavigera could be classified into five successive stages: early active, late active, ripe, spawning, and recovery. Spawning of females occurred from early July to August when the seawater reached above 24.8$^{\circ}C$. Spawning of males occurred from early June to August in the water above 22.8$^{\circ}C$. Minimum size for sexual maturity of both sexes was above 10.0 mm in shell height. Each egg capsule was a cylinder or spindle in shape, 4-6 mm in length and 1-2 mm in width. Colors of newly spawned egg capsules showed yellowish white or pale yellow, while those with veliger larvae showed pale black, and released larvae or dead egg capsules showed black violet. The fecundity in an egg capsule ranged from 70 to 91 eggs (mean＝80.28 eggs).

• Animal cells and systems
• /
• 제2권4호
• /
• pp.501-506
• /
• 1998

In the goby Micropercops swinhonis, the development of its egg's enveloping layers could be divided into 4 stages. In the earliest developmental period, stage I, there is a simple oocyte surrounded by a layer of squamous follicular cells. Stage II corresponds to the yolk vesicle stage of vitellogenesis. Here the initial follicular layer has become bilaminar with the retention of its outer squamous cell layer and the acquisition of an inner cuboidal cell layer just over the zona radiata. The number and size of the cuboidal cells increases throughout this stage. Stage III corresponds to the yolk granule stage of true vitellogenesis. Here the cuboidal cells begin to be replaced by columnar cells. As the oocyte grows, the columnar cells increase in size. The columnar cells produce cytoplasmic neutral mucins and by the end of this stage their cytoplasm has been filled with this mucin. In stage IV a single layer of squamous cells still remained as the outer follicular layer of the oocyte. The secretory activity of the inner follicular layers' columnar cells has ceased and they had lost their cell wall integrity and ended as a series of bullet-shaped, neutral mucin deposits.

• 한국동물학회지
• /
• 제26권2호
• /
• pp.107-123
• /
• 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.

• 한국발생생물학회지:발생과생식
• /
• 제25권2호
• /
• pp.75-82
• /
• 2021

We studied steroid metabolites produced from red-spotted grouper ovarian follicles during maturation. Oocytes with 350-500 ㎛ diameter were in vitro incubated in the presence of [³H] 17α-hydroxyprogesterone as a precursor. Steroid metabolites were extracted from incubated media and oocytes. The extracts were separated and identified using thin layer chromatography, high performance liquid chromatography and gas chromatography-mass spectrometry. The identified metabolites were androstenedione (A₄), testosterone (T) and estrone (E₁). The metabolites of A₄ was dominant in all size of oocytes and it was the highest in 480 ㎛ diameter oocytes. The metabolites of two progestins, 17α,20β-dihydroxy-4-pregnen-3-one and 17α,20α-dihydroxy-4-pregnen-3-one were detected in the oocytes less than 480 ㎛ diameter although they were not identified definitely. In the oocytes of 480 ㎛ diameter, metabolite of progestin was the highest, and germinal vesicle (GV) was still in the middle of cytoplasm. In the oocytes of 500 ㎛ diameter, GV was began to migrate and the major metabolites were A₄ and E₁. The metabolite of E₁ was detected in all size of oocytes and it was higher than that of E₂. These results suggest that oocytes of 480 ㎛ diameter are the transitional stage involving steroidogenic shift to final oocyte maturation and potential function of E₁ during maturation process.

• Applied Microscopy
• /
• 제25권2호
• /
• pp.29-38
• /
• 1995

The fine structure of the hemocytes in Wolf spider, Pardosa astrigera, is discribed and compared with that of similar cells in other spider species and insects. Five hemocyte types are identified in the hemolymph: prohemocyte, plasmatocyte, granulocyte, spherulocyte and adipohemocyte. Prohemocytes are small with a relatively undifferentiated cytoplasm. The nucleus is comparatively large and has a perinuclear space. Plasmatocytes and granulocytes are pinocytotic function in the hemolymph of the body. The plasmatocytes have some coated pits on the plasma membrane and well developed Golgi complex, The granulocytes appear sequence of events in the formation of coated vesicle from a coated pit on its plasma membrane. Golgi complex become well expressed and give rise to small secretory vesicles which fuse to large bodies. The spherulocytes are larger in cell size than other hemocytes. Their cytoplasm is filled with spherules. The spherules contain the floccurent materials and the helical structured materials, which are 220nm in length and 80nm in width. The adipohemocytes are oval shaped and have a number of lipid droplets.

• Bulletin of the Korean Chemical Society
• /
• 제34권3호
• /
• pp.743-748
• /
• 2013

The temperature dependence of the partition coefficients of a neuropeptide, substance P (SP), in isotropic acidic bicelles was investigated by using a pulsed field gradient nuclear magnetic resonance diffusion technique. The addition of negatively charged dimyristoylphosphatidylserine to the neutral bicelle changed the SP partitioning a little, which implies that the hydrophobic interaction between the hydrophobic residues of SP and the acyl chains of lipid molecules is the major interaction while the electrostatic interaction is minor in SP binding in a lipid membrane. From the temperature dependence of the partition coefficients, thermodynamic functions were calculated. The partitioning of SP into the acidic bicelles is enthalpy-driven, as it is for small unilamellar vesicles and dodecylphosphocholine micelles, while peptide partitioning into a large unilamellar vesicle is entropy-driven. This may mean that the size of lipid membranes is a more important factor for peptide binding than the surface curvature and surface charge density.

• Applied Microscopy
• /
• 제19권2호
• /
• pp.1-16
• /
• 1989

To investigate the research of the tumor seeker using liposomes with negatively stained. The fine structure and size in liposomes composed of PC, DSPC, DAPC and SM phospholipid were observed. And the yield of vesicle affected with sonication by mechanical force were determined. The results were as follows. 1. The stain of 2% UA are obtained a good resolution from electron microscopic observation to compare the negatively liposomes with PTA, AM and UA solution. 2. The fine structure of liposomes except DAPC, PC and CH alone samples could be observed from EM experiment of liposomes composed of PC, DSPC, DAPC and SM phospholipid. 3. The results of experiment from 10, 20, 40 and 60 times sonication with 30sec, prepared 0.1mM NTA-SM-liposomes were obtained unilamella vesicles from groups of multilamella vesicles of phospholipid. 4. About 50nm diameter liposomes obtained through membration filtration step to prepare homogenized liposomes.