Ovary - Science topic

Thanks a lot sir !

in mutant mice, for example, the Cre mouse model, I found that granulosa cells get died fast and they didn't grow. I don't know why

Anmi Jose Thanks for your advice. It works for me. Thanks Once again. Highly appreciated. !!

Yes, immune cells play a role in the ovary's defense system. The ovary contains various immune cells, including macrophages, dendritic cells, and lymphocytes, which help protect it from infections and maintain tissue homeostasis. Additionally, immune cells in the ovary also contribute to processes like ovulation and follicle development.

1. Hormonal effects: Dietary fat influences hormone levels, including those crucial for reproductive function such as estrogen and progesterone. Elevated fat intake can stimulate hormone production, affecting ovarian structure and function.

2. Energy storage: Fat is a concentrated energy source, and excess fat intake leads to increased energy reserves stored in tissues, including the ovaries. This surplus energy can contribute to increased ovarian weight.

3. Ovarian follicle development: Adequate nutrition is essential for the development and maturation of ovarian follicles. Excess dietary fat provides necessary energy and nutrients, supporting enhanced follicle development and resulting in larger ovaries.

4. Lipid metabolism: Dietary fat intake influences lipid metabolism and storage. Excessive fat consumption can lead to increased lipid deposition in various tissues, including the ovaries, contributing to increased ovarian weight.

It's crucial to consider diet composition, genetic factors, and individual responses when understanding how dietary fat affects ovarian weight in gilts. Additionally, while increased fat intake may impact ovarian weight, it's essential to maintain a balanced diet to optimize overall health and reproductive performance in animals.

Dear Anaïs Larue ,

Did you fix the problem of gDNA finally?

Testicular differentiation and testosterone secretion by Leydig cells occur at day 36 of pregnancy while the hypothalamic-pituitary-gonadal axis begins to develop when approximately 70 percent (day42) of pregnancy has passed so fetal masculinization and increasing of the fetal plasma androgen concentration is independent of pituitary gonadotropins however, Leydig cells respond to LH from the very early stages. Probably, according to the later activation of the hypothalamic-pituitary-gonadal axis except for testes, other gonads do not have many opportunities to produce hormones

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.

@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.

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

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.

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:

Also please have a look at the following potentially useful article:

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!

I think you can use rats to induce PCOS by exposure to testosterone propionate. I refer you to read this article:

Thank you Carsten Lange for your kind information.

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.

You're welcome P.Kumar Yadav:)

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.

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.

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.

Follow the link

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,

Dear colleague,

I am not working the corona virus.

All best wishes for your continuing successes,

Prof. Otar Shainidze

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 "

Image J software.

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

It is little difficult to give an exact answer without knowing which wavelength you are using for the excitation here. But I assumes (based on the auto-fluorescence observation) this must be either 405 nm or 488 laser lines. If this is the case, I would Highly recommend to control your gain and power during the experiment. If this doesn't help much then try to use alternative laser lines like 561 nm or 640 nm.

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.

this link could be useful:

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.

ATTC (https://www.lgcstandards-atcc.org/Products/Cells_and_Microorganisms/Cell_Lines.aspx?geo_country=es)

Cellosaurus (https://web.expasy.org/cellosaurus/)

Good luck!

Priscila

Interesting question with great insights. Looking forward to the discussion!

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

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

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.

Thank you , this is very helpful

Please take a look at this useful PDF attachment.

It helps you to obtain first-hand experience and forecast in research.

I,ll try it. Wish me luck.

good question

I will do that Hussein A. Abid thanks

Yes, you can.

Greets

Wittwer

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!

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!!

Thank you Dear Dr. Mohammad Tariq Ali Khan

Well see the attached article. It might be of some help.

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

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.

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.

Dear Dr Sujata Magdum,

In attached my paper.

best regards

Thank you very much sir.

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.

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.

CYP11a, CYP21, CYP17, and CYP19,

Thanks for the input. I'm guessing that's what he did.

please use centrifugal spin top filters of 0.22 micron. they will work.

u can get them on (sigma M9160)

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.

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.

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.

Thanks Dear Samer, Do have any reference for that?

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.

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.

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

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.

Laparoscopic ovarian drilling can be of help. Ofcource she may be advice on weight reduction if she is overweight

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

thank you Mr Dhia

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!!

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

You can use repeated Letrozole administration for induction.

Thank u maria and Rubia... 

Will you consider gene mutation?

 Dear Maksim 

greetings

We need to qualify different types of progestagens to draw a solid conclusion about usefulness of these medications

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

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

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

Thank you very much, I'll try some of these protocols. Keep in touch!

Suggest biometry.nci.nih website

Also try the ISUOG / UOG website

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!

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

Organ culture.