scholarly journals Targeted disruption of the aromatase P450 gene (Cyp19) in mice and their ovarian and uterine responses to 17beta-oestradiol

2001 ◽  
Vol 170 (1) ◽  
pp. 99-111 ◽  
Author(s):  
K Toda ◽  
K Takeda ◽  
T Okada ◽  
S Akira ◽  
T Saibara ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Corpus Luteum ◽  
Lipid Droplets ◽  
Smooth Endoplasmic Reticulum ◽  
Ovarian Follicles ◽  
Interstitial Cells ◽  
Aromatase Activity ◽  
Tubular Structures ◽  
Wild Type ◽  
Targeted Disruption

Aromatase P450 (CYP19) is an enzyme catalysing the conversion of androgens into oestrogens. We generated mice lacking aromatase activity (ArKO) by targeted disruption of Cyp19 and report the characteristic features of the ArKO ovaries and uteri as revealed by histological and biochemical analyses. ArKO females were totally infertile but there were as many developing follicles in their ovaries at 8 weeks of age as in wild-type ovaries. Nevertheless, no typical corpus luteum was observed in the ArKO ovaries. Electron microscopy revealed the presence of well-developed smooth endoplasmic reticulum, few lipid droplets and mitochondria with less organized tubular structures in the ArKO luteinized interstitial cells. These ultrastructural features were different from those of the wild-type interstitial cells, where there are many lipid droplets and mitochondria with well-developed tubular structures, characteristic of steroid-producing cells. When ArKO mice were supplemented with 17beta-oestradiol (E(2); 15 microg/mouse) every fourth day from 4 weeks of age for 1 month, increased numbers of follicles were observed in the ovaries as compared with those of untreated ArKO mice, although no typical corpus luteum was detectable. Ultrastructural analysis revealed the disappearance of the accumulated smooth endoplasmic reticulum in the luteinized interstitial cells after E(2 )supplementation. Transcripts of pro-apoptotic genes such as p53 and Bax genes were markedly elevated in the ArKO ovaries as compared with those of wild-type mice. Although E(2) supplementation did not cause suppression of the elevated expression of p53 and Bax mRNAs, it caused marked enhancement of expression levels of lactoferrin and progesterone receptor mRNAs in the uteri as well as increases in uterine wet weight. At 8 months of age, ArKO mice developed haemorrhages in the ovaries, in which follicles were nearly depleted, while age-matched wild-type females still had many ovarian follicles. Furthermore, macrophage-like cells were occasionally observed in the ArKO ovarian follicles. These results suggested that targeted disruption of Cyp19 caused anovulation and precocious depletion of ovarian follicles. Additionally, analysis of mice supplemented with E(2) demonstrated that E(2) apparently supports development of ovarian follicles, although it did not restore the defect in ovulation.

scholarly journals STUDIES ON MICROPEROXISOMES II. A CYTOCHEMICAL METHOD FOR LIGHT AND ELECTRON MICROSCOPY

1972 ◽  
Vol 20 (12) ◽  
pp. 1006-1023 ◽  
Author(s):  
ALEX B. NOVIKOFF ◽  
PHYLLIS M. NOVIKOFF ◽  
CLEVELAND DAVIS ◽  
NELSON QUINTANA
Keyword(s):  
Endoplasmic Reticulum ◽  
Guinea Pig ◽  
Lipid Droplets ◽  
Smooth Endoplasmic Reticulum ◽  
Light And Electron Microscopy ◽  
Distal Tubule ◽  
Striking Difference ◽  
High Concentration ◽  
Electron Microscopic ◽  
Pig Kidney

A modification of the Novikoff-Goldfischer alkaline 3,3'-diaminobenzidine medium for visualizing peroxisomes is described. It makes possible light microscopic as well as electron microscopic studies of a recently described class of peroxisomes, the microperoxisomes. Potassium cyanide (5 x 10–3 M) is included in the medium to inhibit mitochondrial staining, the pH is 9.7 and there is a high concentration of H2O2 (0.05%). Two cell types have been chosen to illustrate the advantages of the new procedure for demonstrating the microperoxisomes: the absorptive cells in the human jejunum and the distal tubule cells in the guinea pig kidney. Suggestive relations of microperoxisomes and lipid are described in the human jejunum. The microperoxisomes are strategically located between smooth endoplasmic reticulum that radiates toward the organelles and contains lipid droplets and "central domains" of highly specialized endoplasmic reticulum which do not show the lipid droplets. The microperoxisomes are also present at the periphery of large lipid-like drops. In the guinea pig kidney tubule there is a striking difference between the thick limb of Henle and distal tubule. The distal tubule has a population of cells with large numbers of microperoxisomes readily visible by light microscopy; these cells are not present in the thick limb of Henle. Other differences between the two are also described.

scholarly journals The yeast lipin orthologue Pah1p is important for biogenesis of lipid droplets

2011 ◽  
Vol 192 (6) ◽  
pp. 1043-1055 ◽  
Author(s):  
Oludotun Adeyo ◽  
Patrick J. Horn ◽  
SungKyung Lee ◽  
Derk D. Binns ◽  
Anita Chandrahas ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Neutral Lipid ◽  
Strong Evidence ◽  
Lipid Droplets ◽  
Neutral Lipids ◽  
Glucose Medium ◽  
Wild Type ◽  
Lipid Levels ◽  
Total Neutral Lipid ◽  
A Site

Lipins are phosphatidate phosphatases that generate diacylglycerol (DAG). In this study, we report that yeast lipin, Pah1p, controls the formation of cytosolic lipid droplets. Disruption of PAH1 resulted in a 63% decrease in droplet number, although total neutral lipid levels did not change. This was accompanied by an accumulation of neutral lipids in the endoplasmic reticulum (ER). The droplet biogenesis defect was not a result of alterations in neutral lipid ratios. No droplets were visible in the absence of both PAH1 and steryl acyltransferases when grown in glucose medium, even though the strain produces as much triacylglycerol as wild type. The requirement of PAH1 for normal droplet formation can be bypassed by a knockout of DGK1. Nem1p, the activator of Pah1p, localizes to a single punctum per cell on the ER that is usually next to a droplet, suggesting that it is a site of droplet assembly. Overall, this study provides strong evidence that DAG generated by Pah1p is important for droplet biogenesis.

Ectopic Thyroid Adenomas at the Base of the Heart of the Dog; Ultrastructural Identification of Dense Tubular Structures in Endoplasmic Reticulum

1971 ◽  
Vol 8 (5-6) ◽  
pp. 421-432 ◽  
Author(s):  
D. C. Thake ◽  
N. F. Cheville ◽  
R. K. Sharp
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Polarity ◽  
Smooth Endoplasmic Reticulum ◽  
Ectopic Thyroid ◽  
Follicular Epithelium ◽  
Tubular Structures ◽  
Normal Thyroid ◽  
Thyroid Adenomas ◽  
Secretory Type ◽  
Ultrastructural Characteristics

Three encapsulated tumors found at the base of the heart of the dog were classified as ectopic thyroid adenomas. The ultrastructural characteristics of normal thyroid follicular epithelium were seen: dense secretory-type granules, cell polarity with apical microvilli, and well-developed endoplasmic reticulum. Long, dense tubules were present within the cisternae of smooth endoplasmic reticulum of neoplastic cells; they were not seen in other tissues. These characteristics distinguish ectopic thyroid tumors from aortic body tumors.

scholarly journals A variant of the pyroantimonate technique suitable for localization of calcium in ovarian tissue.

1979 ◽  
Vol 27 (6) ◽  
pp. 1017-1028 ◽  
Author(s):  
B S Weakley
Keyword(s):  
Endoplasmic Reticulum ◽  
Plasma Membrane ◽  
Lipid Droplets ◽  
Microprobe Analysis ◽  
Smooth Endoplasmic Reticulum ◽  
Ovarian Tissue ◽  
Follicle Cells ◽  
Fixation Time ◽  
X Ray ◽  
Cytoplasmic Processes

Osmium-pyroantimonate solutions for the precipitation of cations are unsuitable for use with delicate mammalian oocytes. A variant of the pyroantimonate technique employing a mixture of pyroantimonate and glutaraldehyde has been found to give successful and repeatable results if a fixation time of 4 hr is used. Calcium-containing antimonate precipitates were localized principally in nuclei, smooth endoplasmic reticulum, Golgi apparatus, mitochondria, and cytoplasmic processes of both oocytes and follicle cells, and along the plasma membrane in small oocytes. Deposits were also concentrated around the periphery of lipid droplets in the follicle cells. The presence of calcium in the precipitates was confirmed by x-ray microprobe analysis.

scholarly journals Ultrastructural analysis of the dorsal body gland of the terrestrial snail Megalobulimus abbreviatus (Becquaert, 1948)

2010 ◽  
Vol 70 (2) ◽  
pp. 341-350 ◽  
Author(s):  
GD Moraes ◽  
M. Achaval ◽  
MM Dal Piva ◽  
MC Faccioni-Heuser ◽  
GF Wassermann ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Lipid Droplets ◽  
Neural Control ◽  
Smooth Endoplasmic Reticulum ◽  
Nerve Endings ◽  
Retrograde Labelling ◽  
Axon Terminals ◽  
Cerebral Ganglia ◽  
Reproductive Gland

The ultrastructure of the reproductive gland, dorsal body (DB), of Megalobulimus abbreviatus was analysed. Electron microscope immunohistochemistry was used to detect FMRFamide-like peptides in the nerve endings within this gland. Nerve backfilling was used in an attempt to identify the neurons involved in this innervation. In M. abbreviatus, the DB has a uniform appearance throughout their supraesophageal and subesophageal portions. Dorsal body cells have several features in common with steroid-secreting gland cells, such as the presence of many lipid droplets, numerous mitochondria with tubular cristae and a developed smooth endoplasmic reticulum cisternae. Throughout the DB in M. abbreviatus numerous axonal endings were seen to be in contact with the DB cells exhibiting a synaptic-like structure. The axon terminals contained numerous electron-dense and scanty electron-lucid vesicles. In addition, the DB nerve endings exhibited FMRFamide immunoreactive vesicles. Injection of neural tracer into the DB yielded retrograde labelling of neurons in the metacerebrum lobe of the cerebral ganglia and in the parietal ganglia of the subesophageal ganglia complex. The possibility that some of these retrograde-labelled neurons might be FMRFamide-like neurons that may represent a neural control to the DB in M. abbreviatus is discussed.

scholarly journals Ultrastructure of rhesus monkey renomedullary interstitial cells

1979 ◽  
Vol 13 (2) ◽  
pp. 75-80 ◽  
Author(s):  
David J. Lewis ◽  
David E. Prentice
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Fine Structure ◽  
Rhesus Monkey ◽  
Rough Endoplasmic Reticulum ◽  
Lipid Droplets ◽  
Stellate Cells ◽  
Interstitial Cells ◽  
Prominent Feature

Summary The fine structure of rhesus monkey renomedullary interstitial cells was studied by electron microscopy. These stellate cells contained variable numbers of lipid droplets, moderate numbers of mitochondria, moderate amounts of rough endoplasmic reticulum, and prominent Golgi zones. In rare instances, apparent release of lipid droplets into the interstitium was observed. The most prominent feature of the interstitial cells was large nuclear pseudoinclusions which were observed in a high proportion of the animals examined.

Histopathologic and Ultrastructural Alterations of White Liver Disease in Sheep Experimentally Depleted of Cobalt

1997 ◽  
Vol 34 (6) ◽  
pp. 575-584 ◽  
Author(s):  
S. Kennedy ◽  
S. McConnell ◽  
H. Anderson ◽  
D. G. Kennedy ◽  
P. B. Young ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Body Weight ◽  
Liver Disease ◽  
Lipid Droplets ◽  
Smooth Endoplasmic Reticulum ◽  
Plasma Concentrations ◽  
Fatty Degeneration ◽  
Vitamin B ◽  
Liver Lesions ◽  
Lipofuscin Granules

Many cobalt-deficient sheep develop liver lesions known as ovine "white liver" disease, but the etiology of these changes is controversial. It has been suggested that cofactors are required for development of liver damage in cobalt-deficient sheep. In this study, one group of lambs ( n=5) was fed a diet low in cobalt (4.5 μg/kg) while a group of control lambs ( n=4) received the same diet after it had been supplemented with cobalt (1000 μg/kg). All cobalt-depleted lambs had reduced growth rate, anorexia, lacrimation, and alopecia, and they eventually became emaciated (mean body weight at end of study: 83% of initial body weight). Plasma concentrations of bilirubin and serum activity of glutamate-oxaloacetate transferase were elevated in these animals, while plasma concentrations of vitamin B12 were reduced (less than 220 pmol/L from day 42). Fatty degeneration of the liver associated with reduced concentrations of vitamin B12 (14.5 pmol/g) was seen in these animals at necropsy at 196 days. Microscopic liver lesions included accumulation of lipid droplets and lipofuscin particles in hepatocytes, dissociation and necrosis of hepatocytes, and sparse infiltration by neutrophils, macrophages, and lymphocytes. Ultrastructural hepatocytic alterations included swelling, condensation and proliferation of mitochondria, hypertrophy of smooth endoplasmic reticulum, vesiculation and loss of arrays of rough endoplasmic reticulum, and accumulation of lipid droplets and lipofuscin granules in cytoplasm of hepatocytes. No liver lesions were seen in control lambs. The results of this study indicate that cofactors are not a prerequisite to development of hepatic damage in cobalt-deficient sheep. Reduced activities of the vitamin B12-dependent enzymes, methylmalonyl CoA mutase and methionine synthase, and lipid peroxidation are of likely pathogenetic importance in the development of the lesions.

Sporogonic development of Haemoproteus meleagridis (Haemosporina: Haemoproteidae) in Culicoides edeni (Diptera: Ceratopogonidae)

1991 ◽  
Vol 69 (7) ◽  
pp. 1880-1888 ◽  
Author(s):  
Carter T. Atkinson
Keyword(s):  
Endoplasmic Reticulum ◽  
Lipid Metabolism ◽  
Outer Surface ◽  
Blood Meal ◽  
Lipid Droplets ◽  
Smooth Endoplasmic Reticulum ◽  
Polar Ring ◽  
Membrane Complex ◽  
Inner Membrane Complex ◽  
Crystalloid Body

Mature oocysts of Haemoproteus meleagridis developed in specimens of Culicoides edeni within 3–6 days after the midges took a blood meal from an infected domestic turkey. Approximately 50 sporozoites budded from the surface of a single sporoblast body. Sporogonic development was similar to that of reptilian, avian, and mammalian haemoproteids that are transmitted by ceratopogonid and tabanid flies, but unlike that of hippoboscid-transmitted species in birds, which form large, slowly developing oocysts with multiple sporoblast bodies. Ultrastructural features of developing oocysts included nuclei with prominent nucleoli, mitochondria, and sandwich-like arrays of crystalloid particles and smooth endoplasmic reticulum (ER) that were present on the outer surface of developing lipid droplets during early stages of sporogony. During budding of sporozoites, crystalloid particles measuring 30–40 nm in diameter were associated with the inner membrane complex of the sporozoite pellicle, arranged in rows between subpellicular microtubules. Mature sporozoites contained an apical complex composed of a polar ring with two anterior apical rings, from two to eight elongate rhoptries, 22 evenly spaced subpellicular microtubules, and a small anterior crystalloid body. Morphological similarities between the ER–crystalloid arrays and the ER–microperoxisome arrays that have been described in cells actively engaged in lipid biosynthesis suggest that the crystalloid body contains enzymes important in lipid metabolism.

scholarly journals Regulation of the yeast triacylglycerol lipases Tgl4p and Tgl5p by the presence/absence of nonpolar lipids

2016 ◽  
Vol 27 (13) ◽  
pp. 2014-2024 ◽  
Author(s):  
Isabella Klein ◽  
Lisa Klug ◽  
Claudia Schmidt ◽  
Martina Zandl ◽  
Martina Korber ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Lipid Droplets ◽  
Lipolytic Activity ◽  
Wild Type ◽  
Yeast Saccharomyces Cerevisiae ◽  
Steryl Esters ◽  
Lysophospholipid Acyltransferases ◽  
Nonpolar Lipids ◽  
The Stability ◽  
In Cells

Tgl3p, Tgl4p, and Tgl5p are the major triacylglycerol lipases of the yeast Saccharomyces cerevisiae. Recently we demonstrated that properties of Tgl3p are regulated by the formation of nonpolar lipids. The present study extends these investigations to the two other yeast triacylglycerol lipases, Tgl4p and Tgl5p. We show that Tgl4p and Tgl5p, which are localized to lipid droplets in wild type, are partially retained in the endoplasmic reticulum in cells lacking triacylglycerols and localize exclusively to the endoplasmic reticulum in a mutant devoid of lipid droplets. In cells lacking steryl esters, the subcellular distribution of Tgl4p and Tgl5p is unaffected, but Tgl5p becomes unstable, whereas the stability of Tgl4p increases. In cells lacking nonpolar lipids, Tgl4p and Tgl5p lose their lipolytic activity but retain their side activity as lysophospholipid acyltransferases. To investigate the regulatory network of yeast triacylglycerol lipases in more detail, we also examined properties of Tgl3p, Tgl4p, and Tgl5p, respectively, in the absence of the other lipases. Surprisingly, lack of two lipases did not affect expression, localization, and stability of the remaining Tgl protein. These results suggest that Tgl3p, Tgl4p, and Tgl5p, although they exhibit similar functions, act as independent entities.

scholarly journals Histomorphometric and immunohistochemical changes in interstitial cells and ovarian follicles of rats treated with metformin during and after induction of permanent estrus

2021 ◽  
Vol 10 (6) ◽  
pp. e5110615536
Author(s):  
Leonardo Augusto Lombardi ◽  
Leandro Sabará Mattos ◽  
Marcio Luis Alves Moura ◽  
Ana Paula Espindula ◽  
Ricardo Santos Simões ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Granulosa Cells ◽  
Caspase 3 ◽  
Ovarian Follicles ◽  
Interstitial Cells ◽  
Nuclear Volume ◽  
Female Rats ◽  
Continuous Illumination ◽  
Ovarian Cysts ◽  
Significant Difference

Objective: To evaluate the histomorphometric and immunohistochemical changes in interstitial cells and ovarian follicles of rats treated with metformin during and after induction of permanent estrus. Methods: Thirty-two adult-female rats with regular estrous cycle were equally divided into four groups: 1) GCtrl - at estrous phase. 2) GPCOS - at permanent-estrous phase. 3) GMet1 - rats and daily treated with metformin (12.5 mg/Kg) during 60 consecutive days, as preventive form and 4) GMet2 - PCOS rats, which remained exposed to 60 days of continuous illumination and treated with metformin. After that, the animals were euthanized, and the ovaries were removed and processed for paraffin embedding. Sections were stained with H.E. for histomorphometry or subjected to immunohistochemistry for Ki-67 and cleaved caspase-3 (Casp-3) detections. Results: The GPCOS showed lack of corpus luteum and several ovarian cysts, as well as interstitial-like cells. The presence of corpus luteum and a significant increase in primary and antral follicles were observed in Mel-treated groups, which also showed a decrease in the number of ovarian cysts and in the area occupied by interstitial-like cells. The presence of corpus luteum along with an increase in the number of primary follicles in the Met2 group were noticed (p<0,01). A significant reduction in number of cysts and in the area occupied by interstitial cells, as well as a decrease in nuclear volume of interstitial cells, were noticed in the Met-treated groups, mainly in the Met2 group. The percentage of cell proliferation was significantly higher in granulosa cells of the Met-treated groups than PCOS group, mainly in the GMet2 (p<0,01), which was similar to the GCtrl group. On the other hand, the percentage of apoptosis (cleaved-caspase-3- positive cells) was significantly higher in the granulosa cells of GPCOS and Met-treated groups than the GCtrl group, but without significant difference, which showed weak cleaved caspase-3 immunoreactivity in those cells. Conclusion: The ovaries of rats treated with metformin showed a decrease in nuclear volume and in the area occupied by interstitial cells, presence of corpus luteum, in addition to a decrease in the number of cysts.

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