Ultrastructural studies of hamster tracheal epithelium in vivo and in vitro

1980 ◽  
Vol 70 (1) ◽  
pp. 70-81 ◽  
Author(s):  
Johnny L. Carson ◽  
Albert M. Collier ◽  
Shih-Chin S. Hu
Author(s):  
Conly L. Rieder ◽  
S. Bowser ◽  
R. Nowogrodzki ◽  
K. Ross ◽  
G. Sluder

Eggs have long been a favorite material for studying the mechanism of karyokinesis in-vivo and in-vitro. They can be obtained in great numbers and, when fertilized, divide synchronously over many cell cycles. However, they are not considered to be a practical system for ultrastructural studies on the mitotic apparatus (MA) for several reasons, the most obvious of which is that sectioning them is a formidable task: over 1000 ultra-thin sections need to be cut from a single 80-100 μm diameter egg and of these sections only a small percentage will contain the area or structure of interest. Thus it is difficult and time consuming to obtain reliable ultrastructural data concerning the MA of eggs; and when it is obtained it is necessarily based on a small sample size.We have recently developed a procedure which will facilitate many studies concerned with the ultrastructure of the MA in eggs. It is based on the availability of biological HVEM's and on the observation that 0.25 μm thick serial sections can be screened at high resolution for content (after mounting on slot grids and staining with uranyl and lead) by phase contrast light microscopy (LM; Figs 1-2).


1987 ◽  
Vol 13 (2) ◽  
pp. 223-240 ◽  
Author(s):  
Linda N. Curtis ◽  
Johnny L. Carson ◽  
Albert M. Collier ◽  
Todd M. Gambling ◽  
S. S. Hu ◽  
...  

2018 ◽  
Vol 36 (0) ◽  
Author(s):  
N. MOHAMED ◽  
R.M. TAHA ◽  
U.N.A.A. RAZAK ◽  
H. ELIAS

ABSTRACT: An efficient protocol for in vitro flowering was successfully established for Impatiens balsamina cv Dwarf Bush, an important medicinal plant, through tissue culture techniques. Shoot, stem and petiole explants obtained from 4 week-old aseptic seedlings cultured on MS medium supplemented with different concentrations of plant growth regulator (PGR) were used for in vitro flower induction. Gibberellic acid (GA3), benzylaminopurine (BAP) and kinetin (Kin) treatment singly applied in MS media (pH 5.8), could all stimulate flowering at 23-26 oC with photoperiod of 16 hours light and 8 hours dark. It was observed that shoot explants were more responsive than stem explants in floral formation. Regeneration was achieved via direct organogenesis. For shoot explants, the treatment that induced the highest rate of in vitro flowering (7.30 ± 0.16 flowers per plantlet) was 1.0 mg L-1 GA3. Ultrastructural and histological analysis of in vivo and in vitro flowers were done to discover any somaclonal variation. This research described a simple protocol for rapid in vitro flowering that will be very beneficial for further breeding, cytological and molecular biology research.


2020 ◽  
Vol 318 (4) ◽  
pp. L580-L591
Author(s):  
Ajay Kumar ◽  
Kentaro Noda ◽  
Brian Philips ◽  
Murugesan Velayutham ◽  
Donna B. Stolz ◽  
...  

Primary graft dysfunction (PGD) is directly related to ischemia-reperfusion (I/R) injury and a major obstacle in lung transplantation (LTx). Nitrite ([Formula: see text]), which is reduced in vivo to form nitric oxide (NO), has recently emerged as an intrinsic signaling molecule with a prominent role in cytoprotection against I/R injury. Using a murine model, we provide the evidence that nitrite mitigated I/R-induced injury by diminishing infiltration of immune cells in the alveolar space, reducing pulmonary edema, and improving pulmonary function. Ultrastructural studies support severe mitochondrial impairment in the lung undergoing I/R injury, which was significantly protected by nitrite treatment. Nitrite also abrogated the increased pulmonary vascular permeability caused by I/R. In vitro, hypoxia-reoxygenation (H/R) exacerbated cell death in lung epithelial and microvascular endothelial cells. This contributed to mitochondrial dysfunction as characterized by diminished complex I activity and mitochondrial membrane potential but increased mitochondrial reactive oxygen species (mtROS). Pretreatment of cells with nitrite robustly attenuated mtROS production through modulation of complex I activity. These findings illustrate a potential novel mechanism in which nitrite protects the lung against I/R injury by regulating mitochondrial bioenergetics and vascular permeability.


1997 ◽  
Vol 3 (S2) ◽  
pp. 245-246
Author(s):  
A.S. Kaprelyants ◽  
A.A. Kaprelyants ◽  
A.N. Reylan ◽  
R.K. Migunova

The aim of given investigation is to study the effect of cooling upon rat hepatocyte structure using transmission electron microscopic and computer morphometric methods. Ultrastructural and morphometrical characteristics of hepatocytes under liver cooling for various levels under in vivo and in vitro conditions were investigated. Vistar rats of 180-250 g were used in the experiment. Liver cooling (in vivo) was performed by means of original cryoapplicator with different probe temperature (1,2). Liver tissue for transmission electron microscopy was fixed in glutaraldehyde fixator on cocadylate buffer and OsO4. Dehydration was completed on acetone (3). Tissue embedding was done into the mixture of Epon/Araldite epoxy rasin. Ultrathin slices were contrasted by the method of Reinolds. Cell viewing and imaging were accomplished by electron microscope at accelerating power of 75kV.Morphometrical and stereometrical analysis was performed using the “Morpho-Tools” original computer system (c) 1994-1996 A.S. Kaprelyants, A.A. Kaprelyants, A.N. Reylan .


1968 ◽  
Vol 37 (1) ◽  
pp. 123-138 ◽  
Author(s):  
Ramzi S. Cotran ◽  
Morris J. Karnovsky

Peritoneal mesothelium was exposed for 2–60 min to solutions of horseradish peroxidase by incubation in vitro, or after intraperitoneal injection in vivo. Peroxidase was localized, with the electron microscope in the intercellular clefts of the mesothelium, often along their entire lengths, in vesicles adjoining or contiguous with the clefts, and along the peritoneal and basal surfaces of the cell, and also in intracytoplasmic vacuoles. The intercellular junctions of peroxidase-treated mesothelium did not differ from those of controls: open and closed junctions were present in both groups. Intercellular localization was also obtained when the mesothelium was exposed to peroxidase during or after fixation. Although intracellular absorption of peroxidase and its incorporation into larger vacuoles were observed, there was no clearcut evidence of vesicular transport across the mesothelium in these experiments. These findings are consistent with physiologic data which postulate that mesothelial transport can be accounted for, at least in part, by passive diffusion through a system of pores, and they suggest that these pores are located in the intercellular clefts.


1978 ◽  
Vol 87 (4) ◽  
pp. 701-715 ◽  
Author(s):  
F. Peillon ◽  
F. Cesselin ◽  
P. E. Garnier ◽  
A. M. Brandi ◽  
M. Donnadieu ◽  
...  

ABSTRACT To evaluate the in vitro PRL secretion and synthesis by pituitary tumours from acromegalic patients, six somatotrophic and two somatomammotrophic adenomas were collected after surgery and divided into fragments 1 mm3 in size. Fragments were cultured in short-term (1–2–3–4 h) and in long-term organ culture (7 and 14 days). Media were collected for GH and PRL radioimmunoassay and fragments studied by electron microscopy. In two experiments, fragments from a somatotrophic adenoma and from a somatomammotrophic adenoma were cultured for 1 to 4 h and 9 to 16 days with [3H]leucine. In addition the effect of somatostatin (2.5 nmole/ml) upon PRL secretion was studied in short-term incubation. In long-term organ culture PRL concentrations increased in 8 out of 16 media samples collected from the five tumours and one normal pituitary cultured for 14 days, while GH concentrations decreased during the same time in all the experiments. The [3H]PRL/[3H] proteins ratios were 9.7 % on the 9th day of culture and 20.8 % on the 16th day while for GH, the ratios were respectively 40 and 34.7 %. Ultrastructural studies showed, besides GH cells, the presence of PRL secreting cells in the different tumours, after 7 or 14 days of culture. In short-term incubation, PRL concentrations in media increased in most of the samples collected from the three tumours incubated for 1 to 4 h and the [3H]PRL/[3H]proteins ratio was 10.6 % after 1 h and 21 % after 2 h of incubation. A decrease of PRL content (< 50 %) was observed in two of the three tumours incubated with somatostatin. These data indicate that 1) somatotrophic adenomas, as well as somatomammotrophic ones, are able to secrete and synthesize PRL in vitro, 2) PRL synthesis from these adenomas increases during culture, as from normal pituitary, presumably because of the release of PRL-secreting cells from the PRL inhibitory hypothalamic control (PIF), 3) somatostatin may reduce PRL release in vitro from somatotrophic adenomas as reported in some cases in vivo.


1996 ◽  
Vol 8 (7) ◽  
pp. 1029 ◽  
Author(s):  
JK O'Brien ◽  
D Dwarte ◽  
JP Ryan ◽  
WM Maxwell ◽  
G Evans

Development to the blastocyst stage was assessed for oocytes obtained from prepubertal and adult sheep matured and fertilized in vitro. The proportion of cleaved oocytes reaching the blastocyst stage was significantly lower for oocytes derived from prepubertal sheep than for those from adult sheep (7.4% and 24.6% respectively). There were no differences in the metabolism of glucose, glutamine or pyruvate between oocytes matured in vivo and in vitro, or of glucose or pyruvate between oocytes from prepubertal and adult sheep. Glutamine metabolism by mature oocytes from prepubertal sheep was significantly lower than that by oocytes from adult sheep. Ultrastructural studies revealed no differences in the morphology of cytoplasmic organelles of oocytes matured in vitro from prepubertal and adult sheep, but differences in the volume fraction and size of mitochondria and cortical granules were observed. These data suggest that mature oocytes from prepubertal sheep do not possess the developmental potential of their adult-derived counterparts, and this phenomenon may be associated with metabolic and ultrastructural anomalies.


1986 ◽  
Vol 32 (9) ◽  
pp. 1535-1540 ◽  
Author(s):  
Nobuyuki TANAKA ◽  
Tateo SATO ◽  
Kuo-Jeng HSIEH ◽  
Shigetoshi SHIODA

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