Science topic

Ovary - Science topic

The reproductive organ (GONADS) in female animals. In vertebrates, the ovary contains two functional parts: the OVARIAN FOLLICLE for the production of female germ cells (OOGENESIS); and the endocrine cells (GRANULOSA CELLS; THECA CELLS; and LUTEAL CELLS) for the production of ESTROGENS and PROGESTERONE.
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The supplier of the HTB-78 line (ovary adenocarcinoma, adherent) recommends culturing on Leibovitz's L-15 Medium in 100% air. Maybe any of You tried to grow on a different medium (DMEM?) in an atmosphere with the addition of 5% CO2?
I will be grateful for all the hints!
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Hi Anna! That medium, the Leibovitz L-15, is designed for culturing cells without needing CO2 control because it uses a different kind of buffer system in place of sodium bicarbonate. There are a ton of mediums that are designed for specific purposes and some of those might support better cell growth, when compared to the traditional DMEM medium. Cells are typically grown in a 5% CO2 atmosphere, which provides a buffer system, and mimic physiological pH conditions. Personally, I always use the standard DMEM formulation for complete medium whenever I am culturing mammalian cells and my cells grow well without any issues. The only time I will consider an alternative formula is if I am experiencing slow growth rates, other noticeable issues, or specific needs (e.g., guiding differentiation) that could be improved by changing the type of medium. You should be just fine using DMEM and 5% CO2, if that is what you would prefer. Then, if you notice slow growth rates and lower than normal viability, you might want to consider switching to the other medium.
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I observe the ovarian development mainly growth stage of the ovarian follicles through light microscopy. So any one can help to me which stain is appropriate to analysis the different stage of ovarian follicles.
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@Omar Hernando Avila-Poveda
Thank you for your suggestion but I am trying to observe the number of dilations in the oviduct which happens due the completion of gonotropic cycles in dipteran insects like mosquitoes. I am not trying to observe tissue sections. Any method for direct staining of ovary for observation under light microscope would be helpful.
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So I desperately need to remove all traces of genomic DNA from my RNA samples. I have tried, what I think are all the possibilities, to remove the DNA, so far without success... My RT - always amplifies.
Here are the different technics I have tried thus far:
- 30 ovaries RNA extraction, Turbo DNase treatment on the QIAGEN RNeasy column
- 30 ovaries RNA extraction, Turbo DNase treatment on the QIAGEN RNeasy column + TurboDNase free rigorous treatment 1uL of DNase
- 30 ovaries RNA extraction, Turbo DNase treatment on the QIAGEN RNeasy column + TurboDNase free rigorous treatment 3uL of DNase
- 30 ovaries RNA extraction, Turbo DNase treatment on the QIAGEN RNeasy column + dilution of cDNA 1/50 + TurboDNase free rigorous treatment 2uL of DNase
- 30 ovaries TRIZOL-Chloroform RNA precipitation (no kit) + TurboDNase free rigorous treatment 2uL of DNase
- 30 ovaries TRIZOL-Chloroform RNA precipitation (no kit) + dilution of cDNA 1/50 + TurboDNase free rigorous treatment 2uL of DNase
- 10 ovaries RNA extraction, Turbo DNase treatment on the QIAGEN RNeasy column
Thanks for any help!
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Pretty sure it is amplification... (see following gel picture: RT- is in the 3rd position). The band has always looked the same no matter the treatment...
And in RTqPCR it also amplified (15-20 cycle vs 10-15 for RT+ samples)
I have tried multiple DNase kits and multiple lots.
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In the discovery of mammalian egg von Baer reported to have seen a yellow fleck inside the follicles; this could correspond to sighting- with the microscopes of that time- to the cumulus o-hutus, which convex face in the antrum could be described as a fleck or point?
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To the best of my knowledge it was Karl Ernst von Baer, who investigated the problem of identifying the structure of the ovum of the dog and found it to be a small yellow spot floating in the follicular fluid. As a result of this work, he published in 1827 the first description of a mammalian egg, Epistola de ovi mammalium et hominis genesi (On the Origin of the Mammalian and Human Ovum).
The yellowish-white point that Karl Ernst von Baer could have seen might be the germinal vesicle possessing nucleolus in the diplotene-arrested oocyte inside the follicle of the dog ovary.
Best
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Answer keeping in view the depletion of the germ cell.
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Its like master sex determination gene which can be either located in Y or X chromosome and some time in duplicated copy of the chromosome. For germ cell depletion, busulfan or other DNA-alkalyting drugs are widely used. The function of these drugs are to bind and alter the functioning of master controlling genes in the gonads such as sox2, sox4, nanosg etc.
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Give your suggestions with publication.
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Dear Pramod Kumar Yadav many thanks for sharing this very interesting technical question with the RG community. Please note that the first article suggested by Shin Murakami is freely available as public full text on RG:
Germ cell depletion from mammalian ovary: Possible involvement of apoptosis and autophagy
Also please have a look at the following potentially useful article:
Fate of the germ cells in mammalian ovary: A review
This review article has also been posted by the authors as public full text on RG, so that you can freely download it as pdf file.
Good luck with your work and please stay safe and healthy!
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Several animal models available for polycystic ovaries. In your experience, which one is more reliable and easy to induce?
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I think you can use rats to induce PCOS by exposure to testosterone propionate. I refer you to read this article:
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I am asking about autophagy inducers and inhibitors for use in mammalian ovaries in vitro as well as in vivo condition.
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Thank you Carsten Lange for your kind information.
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I am asking about a detailed protocol.
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Cut the ovary tissue in small pieces and fix them in karnonovsky for 4-6 hours and after give 0.1 M phosphate buffer changes and stored the tissue in 0.1 M phosphate buffer under 4 degree in freezer until further processing. You can also contact SAIF Faculty in AIIMS for further processing and photography.
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Explain in detail.
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You're welcome P.Kumar Yadav:)
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Our long standing understanding for mammalian ovary is that it has fixed number of germ cells and duty of the ovary is to ovulate high quality ovum for successful fertilization and early embryonic development. However, growing studies suggest that the oocytes can be generated from Oocyte like stem cell/neo-oogenesis/adult oogenesis and fertilized in vitro. Therefore, it is important tom know whether oocytes generated from these sources are as good as the follicular oocytes collected from mammals under natural conditions. Kindly give your opinion on the genomics, proteomics and metabolomics aspects of ocytes generated from Oocyte like stem cell/neo-oogenesis/adult oogenesis.
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Thank you Dr. Ke-Hui Cui M.D., Ph.D. for your personal opinion on introducing a new concepts of Hereditics, Cytohetics and Epicytohetics and considering both egg yolk and egg white as hereditary material. We know that the cytoplasmic inheritance (specially through mitochondrial genome) play important role in determining the fate of the individual (in mammals). I would not comment on your new concept and feel that it is premature to say anything on these new thoughts.
Whatever the research experience I have got on oocyte biology in last 30 years, I believe that the mammalian oocyte is a specialized cells and takes very long time (few oocytes takes very long time from puberty to menopause almost 30 years) to complete (oocyte maturation process; both nuclear maturation as well as cytoplasmic maturation in order to ensure the fully competent oocytes required for successful fertilization, early embryonic development and fate of the individuals).
As you know the mammalian oocytes are microlecithal with little or no yolk. However, mature human sperm are highly compact with no cytoplasm in the head region. As you know that the animal cloning for the first time Prof. Sir Ian Wilmut cloned Dolly and now several mammalian species have successfully been cloned using somatic cell nuclear transfer (SCNT) method. Keeping all these into consideration, I wonder how much these new concepts fit into specially in mammalian oocytes and helpful in oocyte biology of mammals.
Good luck for your new journal "Hereditics" in future.
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Hello - I need to retrieve GV stage oocytes from mice ovaries. I do not know where to get the small tool that is used to mush the ovaries into small pieces. I do not know how to fabricate it either. Does anyone know if it is commercially available? If not, do you have any spares you are willing to part with? Thank you very much in advance.
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Are you trying to isolated the GV stage oocytes for in vitro cultre and analysis? or trying to isolate them and use for biochemical/histological analysis. There is different protocol for different targets. Kindly clarify your purpose.
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Can anyone give suggestions for reducing the autofluorescence/background fluorescence in rat ovary section.
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Thank you Jan Homolak.....
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I transfected the CHO (Chinese hamster ovary) Cells with Plasmid containg Heavy and light chai genes of a mAb, I'ld like to know during which phase of cell growth the mAb will be produced ?
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Hello,
At the transition from lag to log phase during which at this point the cells have enough energy to synthesize DNA and other proteins for themselves which would also include the protein of interest as in this case the mAb.
Best.
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I extract total RNA from two mouse ovaries with Trizol. but I got 56ng/ul total RNA. Could anyone please tell me how many ovary will be used could get more concentration for the total RNA?
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I have preserved my tissue sample (ovary and follicles) in RNA Later and stored at -80 degree celsius. I have tried to extract the RNA using Trizol reagent. I have even washed the tissue with 1X PBS but still the RNA is degraded. I need an advice on this.
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Thanks Bruno Ramos,
We did two 2-min washes with PBS and then we included the tissue in OCT before cutting in a cryostate. Now, we are trying to extract total RNA using three different kits and we observe a highly degraded RNA. It looks like RNA extracted from FFPE, RIN around 2. Did you face simmilar issues before?
Best,
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We would like to investigate the effects of a pesticide on ovary. But if we treated with single dose and take the samples at the 24 hour can we see significant changes at the histology and ultrastructure? Can we evaluate apoptosis and oxidative stress parameters at the 24 hour of pesticide exposure?
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Dear colleague,
I am not working the corona virus.
All best wishes for your continuing successes,
Prof. Otar Shainidze
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The combined oral contraceptive pill is often just called "the pill". It contains artificial versions of female hormones oestrogen and progesterone, which are produced naturally in the ovaries. If sperm reaches an egg (ovum), pregnancy can happen.
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There is a brief item on the internet from five years ago that indicates that the risk for an ectopic pregnancy is increased for women who take oral contraceptives during pregnancy, as follows:
"If you test positive, you should stop taking your birth control pill. Becoming pregnant while on birth control does increase your risk of ectopic pregnancy. An ectopic pregnancy occurs when a fertilized embryo attaches outside the uterus, often in the fallopian tube. Feb 2, 2016 "
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I have done H&E staining of uterus and ovaries of rats, now want to do scoring of the lesions in percentage%, any idea which technique or software I can use??
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Image J software.
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The PGF2α transferred from the uterus to the ovary is thought to occur either by local countercurrent transfer or general systemic transfer. Countercurrent transfer involves the movement of molecules (PGF2α) across the blood vascular system from higher concentrations in the venous effluent (utero-ovarian vein) to an area of lower concentration (ovarian artery). Systemic transfer involves the passage of the molecules through the general circulatory system.
In some species (cow and ewe), PGF2α synthesis from a uterine horn only influences the life span of the CL in the ipsilateral ovary. In other species (sow and perhaps mare), PGF2α synthesis from one horn is sufficient to cause regression of CL in both ovaries.
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it is my understanding that PGF2alpha is synthesized in the uterus (endometrium) and not the uterine horn. it is converted from arachidonic acid in the bovine, ovine and caprine
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Can the consumption of extra fat (2%) in the diet of prepubertal gilts cause an increase only in the weight of the ovaries? The rest of the uterine structures are not affected (no CL, CA..)
Why does this occur?
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But do you know how it can affect? I cannot find literature that relates the increase in dietary fat with the increase in the weight of the ovary
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Is there any in vitro model for Cystic Ovarian Disease available to study the efficacy of various therapeutic options in vitro before it is implemented in COD condition in cows/bovines?
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Thank you Ali Olfati for the interesting paper. I am looking for an in vitro assay to study the ovulation in bovine ovaries collected from a freshly slaughtered animals.
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How to estimate oocyte numbers in the mouse fetal (1dpp) ovary?
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Dear all,
I want to choose a normal ovary cell line and check for its proliferation property when introducing carcinogenic factors. Does anyone have any suggestions for the cell line? Many thanks in advance!
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Hi Le Cong Truc,
When selecting a cell line is important to make sure that it is a good model for the disease you are studding. Normally we do a literature check to see what cell lines are being used for ovary carcinogenesis research.
It is also important to check the specific genetic background of the cell lines to know the organ of origin, differentiation type, mutations, growth rates, and other properties that will make a difference when planning the experiments and interpreting your results. For instance, some cell lines are not ideal to be used in a classical 96 well plate/72h proliferation assay because they don´t grow in low density.
You can check all the cell lines commercially available and its features in ATTC and Cellosaurus.
Good luck!
Priscila
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HEK293 and sf9 are the most used cells for AAV production, but can CHO cells be used for this purpose?
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Interesting question with great insights. Looking forward to the discussion!
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See the image attached below and give me few valuable suggestions regarding gonad intersex. The below image showing 9 months old male zebrafish dissected out viscera to check the gonad status. The fish was exposed to pesticide (secret) at the age of 7 months old. Looking forward to your valuable suggestions.
Thanking you
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Dear Gopi,
If you treated male zebrafish with pesticides, it is quite a normal reaction. Pesticides are widely known endocrine disruption chemical (EDC) which mimic natural estrogens, attach to the sex steroid receptors and also provoke many abnormalities such as intersex gonads. Of course, the histological analysis is crucial in this matter.
best regards,
Hanna
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I am working on toxicity induced by drugs on invivo model(mice) and few compounds that have antioxidant properties that would be used to reduce this toxicity. The organs I would be looking for would be liver, kidney, ovaries, brain, heart. My question is to study antioxidant profile, what would be better to analyse antioxidant enzymes Serum or tissues?
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liver is the best one
but if you know the active gradient and its mode of metabolism or excretion you can choice the organ related
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I want to do sample digestion for AAS or ICP-MS manually to detect some heavy metals, i have uterus and ovaries of rat model, any idea how it can be performed.
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Hi, Abdul,
Sample digestion is very common for AAS and ICP-MS. If you perform the digestion manually ( open digestion), there is a possibility that some of the elements can get lost. It is always preferable to carry out Microwave digestion. Since it is a closed door digestion, no elements will get lost and the digestion process will be more efficient and faster.
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Cause : Usage of Asbestos as insulator . Effect : Mesothelioma cancer ; Lung / peritoneal /larynx and ovary (8) cancer .
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Naser Ahmed Mahmoud Elsawy
Thank you , this is very helpful
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seeds contain rich number of steroids and have an estrogen and progestogen like effect.
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Please take a look at this useful PDF attachment.
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My new work is induction of polycystic ovary
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It helps you to obtain first-hand experience and forecast in research.
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I have extracted total RNA of rat ovaries and uterus by TRIzol reagent. The integrity and purity of extracted RNA was suitable. The cDNA was made using an one step commercial kit. Now, I have problem with RT process , as the received CT is about 36, even for my house kipping gene. I mixed 1 micro-liter of each primer, 3 of cDNA, 10 of cyber green, and 5 of DEPC-treated water. Can anyone tell me what should i do?
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Laurence Stuart Dawkins-Hall
I,ll try it. Wish me luck.
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Antioxidants polycystic ovary , oxidation stress , treatment of PCO
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good question
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genes poly cystic ovary
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I will do that Hussein A. Abid thanks
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Hello, I've been recently trying to standardize the DCFH method for ovaries.
According to the stablished method, ovaries recently dissected have to be incubated immediatly, embeded in tissue-tek, processed on the cryostat and observed by fluorescent microscopy.
Unfortunately the amount of samples I get in a day take many hours to process and I wanted to ask if it was possible to incubate the ovaries with DCFH and store it in tissue tek in a dark place so I could analyze it later or if I could store at -80°C and then incubate the ovary with the DCFH
I haven't found if this is possible or if HAS to be with fresh samples.
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Yes, you can.
Greets
Wittwer
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Hi, I have been using ImageJ to measure the area of fetal ovaries, with Cleaved Caspase 3 and mvh/ddx4 expression for my IF. mvh is the green channel and CC3 is on the red channel.
I haven't had a problem measuring the area of expression of both of these individually (Colour>Split Channels>Adjust>Threshold>Create Selection>Measure, after setting scale and setting measurement as area).
What I need to know is, is there a way to measure the area that is expressing both red and green simultaneously? So that I can quantify the total area +ve expression of germ cell marker and cc3 in the ovary?
n.b. I have also counted the individual number of cells, but I don't think it is possible to manually count those that are CC3+, hense turning to area and density.
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There is a way for quantification different colored labels adding a different program called Ilastik to the analysis process. I faced a similar problem while doing my work and it helped a lot, if you like feel free to reach out to me!
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The protocol using 4% PFA as fixative and subsequently 0.5%Triton X-100 as permeabilizing agent is standardized in the lab and we are able to stain for most other proteins. The antibody for that protein works well for other tissues like ovary. Has anyone faced similar difficulties while staining for membrane proteins and could maybe suggest any changes in protocol?
Thank you.
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Hello! If the membrane protein is outside the cell you could try simply leaving off the Triton and see if that improves the staining. The other thing you could try is ice-cold Methanol fixation, or even Acetone fixation/permeabilization. If that doesn't help, and you have access to a cryostat, you could freeze some tissue in OCT and take a few unfixed cryosections, and post fix with 4% PFA for a very short time. If you get any hints of good staining, at that point you want to then optimize the concentration of primary antibody. Sometimes you need pretty high concentration 1:100 dilution instead of 1:1000. Please keep in mind that many many antibodies sold by suppliers simply do not work. So if you can't get it after a few trials, its not you. Good luck!!
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What is the best chemical in inducing PCO? is Letrozole or Estradiol-Valerate EV?
How can I get non-time consuming method in PCO induction? and what are the doses?
Regards;
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Thank you Dear Dr. Mohammad Tariq Ali Khan
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Hey fellow scientists. Does anyone have experience using RNAscope in P0 ovary fixed frozen tissue? More specifically, which pretreatment should I try?
Thank you for any input and advise!
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Well see the attached article. It might be of some help.
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Hi all,
We are going to house mice for a double-ko investigation. Since this is our first time housing mice, we have no knowledge on how we should optimize breeding.
So here's the info:
One gene is on Chr-X, the other on Chr-11. Both males and females homozygous double-ko should be infertile. The simple ko are supposed to be ok. We are mostly interested in ovaries of the double-ko females, but we'll also check on both simple-ko and wt as controls.
So what strains should we ask to start the colony ? We also want to cryopreserve embryo in the first few weeks.
The collaborator who is going to send the mice proposed this:
Males: Gene1(-/0), Gene2(-/+)
Females: Gene1(-/+), Gene2(-/+)
Do you think that is an appropriate start ?
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In my opinion, If you cross male and female of given pair:
Theoreticallly, Your chances on getting one male double KO and one female double KO in 16 pups if all are alive (you will keep on breeding this pair till you get enough numbers), The details lies here:
(1) 1-/- 2-/- Female Double KO
(2) 1-/0 2-/+ Male
(3) 1-/0 2-/- Male Double KO
(4) 1-/- 2-/+ Female
(5) 1-/- 2-/+ Female
(6) 1-/0 2+/+ Male
(7) 1-/0 2-/+ Male
(8) 1-/- 2+/+ Female
(9) 1-/+ 2-/- Female
(10) 1+/0 2-/+ Male
(11) 1+/0 2-/- Male
(12) 1+/- 2+/- Female
(13) 1-/+ 2-/+ Female
(14) 1+/0 2+/+ Male
(15) 1+/0 2-/+ Male
(16) 1-/+ 2+/+ Female
Once you get a pair of double KOs, and if they are fertile (depending on your genes not affecting male and female fertility), You will be lucky that you can breed among them and get all pups should be double KOs.
Alternatively,
for next breedings, you can may include female mouse number 9 and male number 11 for better results if they are fertile
Hope that helps, let us see if someone comes up with better strategy,
Subhash
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Hi,
I am struggling to find a way to measure oocytes on histological sections with the nucleus while excluding those without the nucleus. Since the measures would involve the whole histological section (i.e. a few hundreds of oocytes to manually be measured) I am trying segmenting and/or thresholding the pictures on imageJ, but without success. The idea underlying this approach is to find a way to automatically detect the "objects" I am specifically interested in.
I've already tried ImageJ Trainable Weka Segmentation and MorphoLibJ.
Here a couple of pictures of ovaries DAPi-stained and what I'd roughly like to obtain. Red and yellow circles for oocytes with and without nucleus, respectively.
Thank you in advance.
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I can advise you to read "Correlation between germinal vesicle and oocyte development in the adult Japanese quail ( Coturnix coturnix japonica)
J. Embryol. exp. Morph. vol 29, 1, 145-157, 1973.
With kind regards, Marc Callebaut.
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Why is polycystic ovary syndrome (PCOS) rising all over the world?
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PCOS is heavily linked to insulin resistance. With the rise of non-communicable diseases globally including type 2 diabetes and metabolic syndrome, the incidence of PCOS is expected to also increase considerably.
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Immunohistochemical protocol in insects specially red cotton bugs
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Dear Dr Sujata Magdum,
In attached my paper.
best regards
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Is it recommended to evaluate the estrogenic potential of a plant extract in female mice without ovariectomy, if we keep a control group without the administration of the extract? Because normally the estrogenic activity is checked in ovariectomized female mice. But if we found the extract efficient, we would be providing it to a female with ovary.
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Thank you very much sir.
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My project is on ovarian biology and to design experiment, I want to know how the in vivo testing could be done
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Drug administration protocols can depend on your formulation. You might want to know about the clearance of a chemical from the GI tract in mice before you give it orally. Once you have given the drug orally it is likely to go to the liver, so any information on liver metabolism, and a so called 'first-pass' effect should be considered. IP can by-pass the liver if needed, but again formulation is a necessary consideration. Dose needs to be determined by experiment, so try a few mice at different doses. "Back off" a bit from doses that cause unwanted toxicity or fatality. Perhaps, above all, have an idea of the effects you might desire to produce, and be observant about any other effects that are produced by any dose you use.
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Many a times we don't get good cellularity to give a clear diagnosis of ovary in cytology. Any advise please?
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I still feel as and when Ovarian lesions are excised, one can prepare a set of good well fixed slides for study and learning from classical lesions.
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As we all, in the field of animal reproduction know that the main response for superovulation in animals is determined by the number of increased numbers and diameters of ovarian follicles. Do we can find out a lab method to assess number of granulosa cells that respond to the external dose of FSH treatment (i.e. correlation between dose of FSH and number of responsive GCs).
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FSH induce Aromatase leading to E2 production so the bedt correlation is the measurement of E2 secretion in culture media. The amount of E2 depends on the GCs responsiveness to FSH.
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Hi everybody,
I am looking for dubious gene(s) that may causing ovary polycystics?? Some genes are highly expressed in people that are tolerating polycystic ovary or infertility.
Sincerely,
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CYP11a, CYP21, CYP17, and CYP19,
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What specific part of the ovary are CHO cells from? I can't find this information in the literature.
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Thanks for the input. I'm guessing that's what he did.
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Im aspirating the follicular fluid from surface follicles of buffalo ovary, centrifuge the fluid and pass it through 40 micron followed by 20 micron and then by 0.22 micron. But 0.22 micron filter get stucked after filtering just 1 ml. Can any one guide me to filter FF easily and in large amount in single go.
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please use centrifugal spin top filters of 0.22 micron. they will work.
u can get them on (sigma M9160)
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I followed Brown- peterson et al, (2011) and atlas of fish histology to classify the maturity stages of ovary and testes of Caspian goby but i am not certain about the type of cells and stages on light micro graphs of Caspian goby testicular and ovarian histology, for example in spawning capable phase for males , two sub phase are recognized by me that are mid and late GE (following pictures) but i do not know it is correct or not
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  • Thanks for your consideration, let me explain i share only two photographs that show spawning capable phase and the type of cells is not my kind of thing in these two pictures i want to know about classifying sub phases with regard to germinal epithelium in these two picture , i took micro graphs in higher magnification (40 x) to identify the type of cells before but i am not sure about my identifications in all pictures (i do not share all of them here)
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Australia's National Health and Medical Research Council (NHMRC) produced a sham review of Water Fluoridation in 2017 by deliberately ignoring over 3000 peer-reviewed scientific papers on Fluoride Toxicology. One important paper the NHMRC suppressed dealt with the mechanism by which Fluoride damages your teeth, with or without metal ions like Aluminium. Fluoride causes increased SATB1, a factor associated with Malignant Cancers and their Metastasis, with many relevant publications relating to Leukemia, Melanoma, Laryngeal and Nasopharyngeal squamous cell carcinoma, cancers of the Bladder, Breast, Cervix, Colon, Kidney, Lung, Ovary, Prostate, Uterus, and Liver.
A couple of papers: Zhang Y, Kim JY, Horst O, Nakano Y, Zhu L, Radlanski RJ, Ho S, Den Besten PK - "Fluorosed mouse ameloblasts have increased SATB1 retention and Gαq activity" PLoS One 9(8):e103994
Fluoride interferes with FoxP3
Zhang G, Zhou B, Han T, Wang M, Du X, Li Q, Wang J. 2012. Decreased percentages of CD4+CD25+ regulatory t cells and foxp3 expression in the spleen of female mice exposed to fluoride. Fluoride 45(4)357-364.
Observed increases in cancer incidence over four decades follow the roll out of Fluoridation and increased Dental Fluorosis. Surely it is time to ban Fluoridation worldwide?
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I agree, none of the articles talks about fluorosis, but the original question is: why if it has been shown that fluoride is toxic, it has not been banned, my answer is yes, Fluoride is toxic, but for produce the effects that are mentioned in the articles on which Dr. Pain is based, the doses are much higher than those used to prevent the caries process.
In some parts of my country, drinking water contains cyanide, but the doses are lower than those that produce toxic effects.
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Greetings all,
I am trying to purify protein from squid ovary (Moisture content, 75%; protein content 19%). However, due to high amount of carbohydrates (8%) it hindering purification in the first step ammonium sulfate precipitation (unable to sediment the protein).
Therefore, I want to separate carbohydrate and proteins from my sample.
Mostly precipitation methods, using solvents like ethanol/acetone has been used to separate these component but functionality of one of the component is affected negatively.
Is there any other method used for separation of carbohydrates and proteins without affecting functionality of both components .
Please give me valuable comments/suggestions to sought out this problem.
Thanks and regards
Avtar
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Dear Khalid,
The link you provided is my previous work about the chemical composition and functional properties of squid ovary.
Thank you for your time.
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I just want to check the expression of a particular mRNA throughout the different tissues (Liver, Kidney, Brain, Muscle, Gills, Intestine, Ovary, Skin, Spleen etc.) in a control/untreated fish. If I go through the Real-time PCR taking all the tissues together, then what method should I follow to meet my quiry?
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Your main issue here is that you need a reference gene (ideally, two or three genes) that you trust to be valid for comparing expression in all your tissues of interest. This is non-trivial, especially for such a broad panel of tissues. I suggest (assuming you haven't done this already) you take a panel of tissues and a selection of candidate reference genes and establish suitable ref genes empirically (via geNorm, normfinder, etc -or ideally multiple assessment algorithms).
Once you have suitable reference genes, it's fine: you can run all your tissues on one plate quite happily (assuming by 'running all the tissues' you mean isolating RNA/synthesising cDNA from each tissue, then loading a different tissue cDNA in each series of wells).
If you have a lot of samples and you're forced into deciding between "all the samples on a plate, for one gene" and "some of the samples on one plate, for multiple genes", choose the former: always choose the former. Each of your samples (for a given genes) will be comparable to each of your other samples (for that gene) provided they are run on the same plate.
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I need to write in my thesis but I did not find in google!
any help would be greatly appreciated.
Masoumeh
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Thanks Dear Samer, Do have any reference for that?
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As you know poly cystic ovaries and infertility is difficult to control in many cases. Sometimes prescribing traditional hormonal drugs and metformin in over weight female with NIDDM is not effective. What is the best therapeutic agents to control these patients?
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PCOS therapy must be phenotype driven. I consider pregnancy wish as a particular case and I prefer letrozole combined or not with low dosis of FSH.
In the presence of biochemical hyperandrogenism the aim is to increase SHBG, inhibit LH pulsatility and diminish free testosterone. I do prefer EE 35ug combined with 2 mg cyproterone acetate. If necessary, I increase CPA
in 25 mg for 10 days each month. In the presence of glucose intolerance or insulin resistance I have experience only with metformin. In the presence of obesity I use a COC containg drospirenone associated with diet, exercise, and orlistat. If possible 1.8 mg day of liraglutide. In the presence of various phenotypes a combined therapy is indicated.For dislipidemia statins should be used.
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I am attempting to isolate ovary from Helicoverpa armigera larvae. I can't find any pictures showing what it looks like during dissection. I know it is possible since there are several articles describing sequencing of gonad transcripts (ovary and testes). Any help? Ideally I would like to see some pictures of what the ovary looks like during dissection and where exactly it is located.
Thanks in advance!
EDIT: Thanks to Muhammad F Chaudhury for providing me the full version of his article titled " Spermatogenesis and Testicular Development of the European Corn Borer, Ostrinia nubilalis (Lepidoptera: Pyraustidae)". In it he describes that in this insect the testis of late-instar larvae are yellow-orange colored, reniform and aprox. 1mm in size.
This information allowed me to identify them in Helicoverpa larvae.
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Thanks to Muhammad F Chaudhury for providing me the full version of his article titled " Spermatogenesis and Testicular Development of the European Corn Borer, Ostrinia nubilalis (Lepidoptera: Pyraustidae)". In it he describes that in this insect the testis of late-instar larvae are yellow-orange colored, reniform and aprox. 1mm in size.
This information allowed me to identify them in Helicoverpa larvae.
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We took in collage that about 14 follicle undergoes follicule phase each month but only follicle which become large than all and due to its secretion of estrogen it, decrease the secreation of fsh but it increase also the no. Of fsh receptor so the it makes intrinsic postive feedback in this large follicle., so the the rest of small follicles will degenerate and only this large follicle will be able to survive(thug life awii), but if that was really the reason for the the ovaries alterntnation in ovum secreaction., why cant one ovary secrete an ovum in 2 successive months?, this may be due to Corpus albican which remains for months till it degenerates (so it. Basically occupy space allowing the other ovary to produce the next month ovum), I really dont know if I am right or wrong, and I havenot searched alot, so if anyone have the answer please till me 😀😀😀
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It is possible that in humans the corpus luteum, via progesterone secretion,
diminishes the local estrogen receptor synthesis. Granulosa cells of the
opposite side have more estrogen receptors and respond better to the FSH
estimulation. I can not remenber the references at this time
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i am isolating EV from culture emdium of amniotic fluid stem cells amd injecting them into damaged ovaries of rats . i was wondering if anyone has tried injecting them into the tail vein and yielded results.
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Yes, there are numerous studies that have injected isolated exosomes/EVs into animal models, and even a couple of studies that have injected them into humans. Many studies have administered exosomes by both IV and intra-tissue (eg intramuscular, intravitreal), with positive results, even cross species in some instances.
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Patient is married from last 5 years and has a child. Now she has difficulty in conceiving the second child. She is on Metformin 500mg twice a day.
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Laparoscopic ovarian drilling can be of help. Ofcource she may be advice on weight reduction if she is overweight
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Bovine infertility 
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Hi,
From my experience supplementation of minerals to infertil dairy cows does only make sense, if you have a proven deficiency - in German herds a rare situation.
Clinical examination should be done to find the reason for the poor BCS. If parasits (fluke e.g.) and other individual diseases can be excluded , the majority of cases you discribed occurs in consequence of subclinical ketosis during a period of time within the first 6 - 8 weeks after parturition.
With regards
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we want to detect alpha estrogen receptor in ovary and uterus of rat with IHC method. But in the result, the negative control stil stained and it he granulosa cell was not stained.
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thank you Mr Dhia
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I want to isolate RNA from 15,5-17,5 dpc ovaries (preferably with Trizol).
How many ovaries I should add in tubes with Trizol? Should I modify the protocol?
The goal is to quantify oocyte-specific genes by qPCR.
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i would homogenize the fresh tissue with trizol in a device like the picture. Then i'd pass it to a centrifuge tube and add chloroform, and so on... http://i.ebayimg.com/00/s/NTcyWDc3MA==/z/ACUAAOxyMZVTkTG2/$_35.JPG
i did this with cartilage and it worked ok.
Another way could be criopulverizing the tissue with liquid nitrogen and a morter. Then add trizol in the morter, mix everything and pass it to the centrifuge tube, then add chloroform.
Everything must be RNAse free or very clean and autoclaved 2x or/and UV irradiated,
Good luck!!
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A 28 years old lady, height 5 feet, weight - 70 kg has been trying to conceive for the last on and a half years. She has a  32-33 days' menstrual cycle. Husband's semen analysis, follicular study and HSG reports are normal. Ultrasound of pelvis reveals evidence of polycystic ovaries with normal sized one ovary and other of 11 cc. She has been put on Tab Metformin 500 mg three times a day and Chirocyst by another Gynaecologist. She gives H/O partial seizures and is on Tab Orcabezapine. The patient uses Ovulation predictor kit to time intercourse. She used it on 14th and 16th day of the present cycle. The menstrual period got overdue by 6 days, yesterday. The patient used the Ovulation predictor kit and the result was positive. Today, she had a normal period. This is the first time that she had a delayed period. The query is, can ovulation predictor kit show positive result just one day prior to the onset of the menstrual period? Do the drugs she is taking interfere with LH levels?
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The specialty of the prescriber and age of the patient suggests possible PCOS in which case I would incline to link metformin to the unexpected changes noticed. Drug use is in that case appropriate and expected to contribute positively in the overall context
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Our research group at Komar University for Science and Technology looking for a suitable compound/chemical to induce cystic ovaries in the rat model, and then treat them with a natural product of our country to see the effect and outcomes.
Now, we need a chemical with fewer side effects to induce cystic ovaries. TQ
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You can use repeated Letrozole administration for induction.
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I am extracting carotenoid pigments from animal tissues like ovary, hepatopancreas etc. of prawn using solvent system Methanol and Dichloromethane. After extraction, can I directly go for HPLC for quantitative analysis or should I need to perform TLC for initial purification? 
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Thank u maria and Rubia... 
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usg shows undetectable to very small uterus with ovaries not clearly visualized with associated blindness.
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Will you consider gene mutation?
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Dear all, according to Grimes DA et al., 2014 ( http://onlinelibrary.wiley.com/doi/10.1002/14651858.CD006134.pub5/abstract;jsessionid=B22DED607EDF16AB77AF3505560F19B1.f04t02 ), treatment of functional ovarian cysts (FOC) with OCP appears no better than watchful waiting. But it's evident that some progestagens (I guess it especially may be refered to 19-norsteroids) have a definite anti-gonadotropic effect. What about this? Does anybody have a reliable information about the positive influence of some OCP or progestagens on FOC? Thanks a lot!
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 Dear Maksim 
greetings
We need to qualify different types of progestagens to draw a solid conclusion about usefulness of these medications
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We obtained several preliminary images from a electron microscope. The sample is Chinese hamster ovary cells being treated by freeze-fracture technique and certain proteins in the membrane are labeled with gold particles (10nm). But we are not really sure if the image is showing a classical mammalian cell membrane surface or not. :I
So hopefully any expert could help me confirm if this is really a mammalian cell membrane. Thanks a lot !
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Hi Chenyang Lan,
In most replicas the transmembrane proteins appear as ~5 nm dots and (since you are trying to label a transmembrane protein) some of  these dots should be nearby the gold label.
While evaporaton of carbon first can improve the immunolabeling efficiency, it can also mask the smaller details of your replica (see also Schlörmann, W., John, M., Steiniger, F., Westermann, M. & Richter, W. Improved antigen retrieval in freeze-fracture cytochemistry by evaporation of carbon as first replication layer. Histochem. Cell Biol. 127, 633–639 (2007). ) thereby making it more difficult to see the transmembrane proteins. It is worth trying the standard 1.2nm Pt - 12-15nm C evaporation (which generally provides quite good details) just to familiarize yourself with how the replicas look. Be aware that with LN2 freezing for knife fractures you have to use tiny amounts of sample and to fracture really at the top of the sample because the quality of freezing decreases rapidly further down.
An alternative culturing and freezing approach is discribed in  Fujimoto, T. & Fujimoto, K. Metal Sandwich Method to Quick-freeze Monolayer Cultured Cells for Freeze-fracture. J. Histochem. Cytochem. 45, 595–598 (1997). this method should give you quite large replicas of (predominantly) the plasma membrane.
best regards!
Rob
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I am carrying out research for automated PCOS diagnosis. So, I need an ultrasound image of the ovary dataset to accomplish my objectives.
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Dear, fellow i am also indeed of ultrasound image scans, but i need the normal ovary scans...and in return if you need scans of effected ovary ultrasound images then please inbox me...thanks
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Any body have the RNAseq data or know the accession number in any database of healthy human ovarian tissue?
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A reasonable start to search for gene expression (from any tissue) could be the gene expression atlas hosted by EBI (https://www.ebi.ac.uk/gxa/home).
This portal hosts (beside others) data from the GTEx project and the FANTOM5 project which characterises gene expression from various tissues.
Hope that helps,
Best,
Peter
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I'm new at this so I would appreciate any protocol for western blot in order to determine ATG7 and LC3I/II in rat ovary. Thanks in advance.
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Thank you very much, I'll try some of these protocols. Keep in touch!
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somebody should help with dataset for ultrasound sound images of the ovary
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Suggest biometry.nci.nih website
Also try the ISUOG / UOG website
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what are the genes which mutated in polycystic ovary syndrome ?
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Hi!
The heterogeneity of Polycystic ovarian syndrome is actually quite intriguing.  Environmental and familial factors contribute to this condition.  If you try to connect the most sound etiology/pathogenesis with the possible cause, youll find that the CYP11a gene which is involved in steroidogenesis.
There is an article that discusses this in detail:
Genetics of Polycystic Ovary Syndrome
Hope this helps!
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CHO-Lec1 cells have a defective N-acetylglucosaminyltransferase gene or it completely lacks it. From the original papers it seems like they have a reduced enzyme activity due to mutations. But in some places I also came across statement that they lack the enzyme (N acetylglucosaminyltransferase )
Stanley P, et al. Selection and characterization of eight phenotypically distinct lines of lectin-resistant Chinese hamster ovary cells. Cell 6: 121-128, 1975. PubMed: 1182798
Stanley P, et al. Chinese hamster ovary cells selected for resistance to the cytotoxicity of phytohemagglutinin are deficient in a UDP-N-acetylglucosamine-- glycoprotein N-acetylglucosaminyltransferase activity. Proc. Natl. Acad. Sci. USA 72: 3323-3327, 1975. PubMed: 1059116
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Hi Nilima,
It is not a complete KO, they only have a point mutation in the gene. The details are available here: http://www.ncbi.nlm.nih.gov/nuccore/U65792
"Glycosylation defect in Lec1 Chinese hamster ovary mutant is due to
a point mutation in N-acetylglucosaminyltransferase I gene"
Best. Norbert
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An isolated ovary is grown by which vitro culture?
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Organ culture.
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I am working on the effect of toxicants on the ovarian production of steroid hormones. Please, can someone recommend the most suitable cell line to be used as a model for this assay?
Thanks.
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Thanks to all.
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Pregnant 21 year old had IA mucinous ovary cancer at week 14th of pregnancy and underwent to oophorectomy + omentectomy+ peritonial biopsy + appendectomy due to her desire to lead pregnancy to term and have baby.
Pregnancy follows with no complications up to now and on February 25th she'll be 39,1 weeks and it will performed a cesarian section.
Our question is about cancer approuch. Is better patient clinical  follow or radical surgery at the same moment of cesarian section?
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Dear Claudio, 
This is a very hard decision. As you know the surgical treatment for ovary cancer is a cytoreduction. So, I think to talk with the oncology team, with the patient and her family pointing all aspects will help you in this hard decision. 
Good luck
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i want to know the equipment to determine that and the methodology for the estimation
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You either need to do the old fashioned grind and bind assays (eg incubate the tissue with radiolabeled oestrad