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cariston

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Does anyone know  of or think that cystinuria testing could/would get to the point of ever using stored frozen sperm on testing deceased dogs??     If there were ever to be a test for genetic carriers, would it be possible to have a test performed from frozen for those that wanted to use semen from gone but not forgotten legends and since there was not alot of awareness or testing way back when, would it be possible to perform this type of testing futuristically, or does this sound completely obsurd??     Opinions please?....




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steveoifer

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Reply with quote  #2 
First of all, we need to see affected dogs, not just random samples of healthy dogs. So validation of being C+ would be difficult.

Next, in semen, we only get 1/2 of the genetic value and that just wouldn't do!

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For the betterment of the breed

"Above all, a uniform type should be aimed at by breeders and uniformity of type can only exist in a proportionate ratio in the purity and distinctiveness in any breed"!.........M. Moore
"If breeds did not adhere to a specific shape, form, and colour range, or if breeders disregarded this blueprint, the breed would degenerate to the point that it would hardly resemble the breed at all. Selective breeding does not just create breeds- it preserves them as well." Breeding purebred dogs inherently means accepting limitations on your freedom to just breed anything...Catherine McMillan
" A reinforced consolidation of the American and British standards could be the basis for restoring our breed to the gladiatorial glory of its ancient past, in capability if not in usage".....Norman Howard Carp-Gordon
"I can live with doubt, or not knowing, rather than to have answers that might be wrong"...Richard Feynman
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annagmay

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Reply with quote  #3 
We can use frozen semen to do DNA tests through OptiGen so it is very possible that we will be able to do the same with other DNA tests with frozen semen as they become available.

It is a LOT more economical and convenient to use stored blood for future health tests.  I recommend that everyone pay the extra $35 to have their dog's blood stored with OptiGen when they PRA and CMR DNA test their dog and that they also participate in the CHIC DNA Repository. 

The CHIC DNA Repository will also accept frozen semen.  If anyone goes to use their dog's semen and it is poor quality or not good, please remember it can still be used for research purposes.  The last time we inquired about this, I think they wanted around 2 straws.  To donate semen you will need to make prior arrangements before having it shipped.

Anna 

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steveoifer

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Reply with quote  #4 
At present there is no test at Optigen for cystinuria.

The genetic material in semen from affected dogs would only display half of the material. This could eventually show the marker/markers on the X portion of the semen. But typically one would want the combined genetic information from an affected dog. That can only be had from blood samplings.

If one is using Optigen as a repository for storing blood samplings, then I fully agree with Anna's request, as it might bear fruit in the future.

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For the betterment of the breed

"Above all, a uniform type should be aimed at by breeders and uniformity of type can only exist in a proportionate ratio in the purity and distinctiveness in any breed"!.........M. Moore
"If breeds did not adhere to a specific shape, form, and colour range, or if breeders disregarded this blueprint, the breed would degenerate to the point that it would hardly resemble the breed at all. Selective breeding does not just create breeds- it preserves them as well." Breeding purebred dogs inherently means accepting limitations on your freedom to just breed anything...Catherine McMillan
" A reinforced consolidation of the American and British standards could be the basis for restoring our breed to the gladiatorial glory of its ancient past, in capability if not in usage".....Norman Howard Carp-Gordon
"I can live with doubt, or not knowing, rather than to have answers that might be wrong"...Richard Feynman
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annagmay

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Reply with quote  #5 
OptiGen can do Cystinuria testing on Newfoundlands for $80. 

My post stated that we can currently do PRA and CMR testing through OptiGen using frozen semen,  so it is very likely that when future DNA tests become available we will also be able to use frozen semen for those tests as well. http://www.optigen.com/opt9_shipsubsemen.html   It will depend on the individual lab whether they are willing to extract the DNA from the frozen semen or not to do the DNA tests that they offer and it may require an additional fee.

We can currently use frozen semen for participation in the CHIC DNA Repository.  It is possible that we can also donate frozen semen to the Cystinuria Research Project, but the participants would need to coordinate the details through UPenn.

Frozen semen should never be thrown out or destroyed when someone no longer wants it, but instead it should be donated to the CHIC DNA Repository for research studies.  They need DNA from both healthy dogs and dogs affected with specific health issues.

Anna

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steveoifer

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Reply with quote  #6 
Not in Mastiffs as of yet and I still wonder if half the genetic marker in semen will be as valuable as the whole for use in the genome project?

I suppose if enough positive semen can be DNA'd there might be something that shows up, but if it's polygenic, then the use of the total genetic value  will be needed.

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For the betterment of the breed

"Above all, a uniform type should be aimed at by breeders and uniformity of type can only exist in a proportionate ratio in the purity and distinctiveness in any breed"!.........M. Moore
"If breeds did not adhere to a specific shape, form, and colour range, or if breeders disregarded this blueprint, the breed would degenerate to the point that it would hardly resemble the breed at all. Selective breeding does not just create breeds- it preserves them as well." Breeding purebred dogs inherently means accepting limitations on your freedom to just breed anything...Catherine McMillan
" A reinforced consolidation of the American and British standards could be the basis for restoring our breed to the gladiatorial glory of its ancient past, in capability if not in usage".....Norman Howard Carp-Gordon
"I can live with doubt, or not knowing, rather than to have answers that might be wrong"...Richard Feynman
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cariston

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Reply with quote  #7 
That sounds awesome about the frozen Anna on even saving the low counts for research...


Where can you get the CMR test  done???   Would it valuable for people to start doing that with mastiffs now for information needed in the future?


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HarmonyMastiffs

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Reply with quote  #8 
Quote:
I suppose if enough positive semen can be DNA'd there might be something that shows up, but if it's polygenic, then the use of the total genetic value  will be needed.


Can you break this down Steve so its simpler - I'm having a

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steveoifer

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Reply with quote  #9 

By submitting semen you are giving only one half of the dogs DNA.

Simply put, we all have 46 chromosomes. To make this work, a sperm carries half of the genetic material (23 chromosomes) and an egg has the other half, so if you just submit semen in place of blood, the researcher only has half of the dog's genetic material that was positive!

This may still be useful, but if it took multiple alleles to create the positive, then the subject being examined may not be telling the entire story!


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For the betterment of the breed

"Above all, a uniform type should be aimed at by breeders and uniformity of type can only exist in a proportionate ratio in the purity and distinctiveness in any breed"!.........M. Moore
"If breeds did not adhere to a specific shape, form, and colour range, or if breeders disregarded this blueprint, the breed would degenerate to the point that it would hardly resemble the breed at all. Selective breeding does not just create breeds- it preserves them as well." Breeding purebred dogs inherently means accepting limitations on your freedom to just breed anything...Catherine McMillan
" A reinforced consolidation of the American and British standards could be the basis for restoring our breed to the gladiatorial glory of its ancient past, in capability if not in usage".....Norman Howard Carp-Gordon
"I can live with doubt, or not knowing, rather than to have answers that might be wrong"...Richard Feynman
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HarmonyMastiffs

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Reply with quote  #10 
- I get it!

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annagmay

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Reply with quote  #11 
Everyone that has frozen semen on a dog that they would like to use for DNA health testing should contact the lab doing the health tests for more information and specific directions on whether it can be used or not for the specific test they would like ran.

If anyone has semen on a dog with a specific health problem that is currently being researched and they are willing to donate some of the semen, they should contact the researcher directly to see if they would like the sample.

CMR testing and long-term blood storage along with PRA testing can all be done through OptiGen.  http://www.optigen.com

Every Mastiff being considered for breeding should be CMR and PRA DNA tested.  The PRA can also be clear by parentage since it is a dominant inheritance.  The CMR affected and carrier dogs can be bred regardless of their genotype if the breeding protocol on the DNA test is followed (affected to clear and carrier to clear, etc...) since that is recessively inherited.

Every Mastiff regardless if they are a pet or show dog or if they are healthy or unhealthy or if they are young or old, or if they are being bred or if they are spayed or neutered. etc.....  should have a sample of their blood donated to the CHIC DNA Repository for health research and/or for future DNA health testing.  http://www.offa.org/chicdnabank.html

It would be an added bonus if every Mastiff also had blood stored with OptiGen http://www.optigen.com

Anna


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annagmay

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Reply with quote  #12 
Using semen for DNA Testing and Health Research Projects:

It is true that there are only 23 chromosomes in 1 sperm cell out of the individual dog's 46 chromosomes.  Semen is composed of millions of individual sperm cells in the average dog.  Even semen that is not of the best quality typically has millions of sperm cells. 

When labs distract DNA from semen, they distract all the DNA and they get a representation of all the chromosomes from the donor dog as they are using millions of sperm cells from that dog.  This includes the X & Y chromosomes. 

Each individual Lab will have its own capabilities as far as using Frozen or Fresh Semen for DNA testing and Research projects go.  Currently OptiGen can use Frozen Semen to do all of the DNA tests that they offer on all dogs being tested and UPenn can use Frozen Semen on all of their DNA tests that they offer and both facilities can use Frozen & Fresh Semen to do their Research Projects that involve studying the dogs genes.

If you are serious about donating Frozen Semen for the the Cystinuria Research through UPenn, you need to contact Dr. Paula Henthorn privately to discuss the specifics and verify that your individual dog's Frozen Semen would be useful or not.  PLEASE DO NOT call her unless you are serious  about donating samples and only if you are participating in the research project.  She is very busy and does not need any more distractions unless they will directly benefit the research.

Thank you,

Anna

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annagmay

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Reply with quote  #13 
Currently UPenn only wants frozen semen from Cystinuria Positive dogs that preferably were actual stone formers with documentation to back that information and/or preferably that had a NP urine test ran with a positive test result.

They do not want to waste anyone's valuable frozen semen unless it will be used for gene mapping and they want to verify that it can be used before anyone donates their dog's semen...

Anna

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Reply with quote  #14 
     Semen carries all 46 chromosomes--in a breeding only 23 (1 gene from each pair)genes are in each sperm which  are paired up with the genes from the bitch (1 gene from each of her 26 pairs) in the egg as it is fertilzed--and  in each "pairing" either of the two genes which make up the pair can be used--that's why puppies from the same litter can have very different DNA profiles! 

for example--Think of color inheriance--if a male carries both a brindle gene and a fawn gene he can sire either brindles or "fawns" if bred to a fawn bitch--if his semen only carried 1/2 of his genetic material--you would not be able to get both colors!

So semen does carry ALL of a dog's DNA. Semen is made up of Millions of little single sperms--which are pure DNA.  As Anna pointed out, once a DNA test is produced--you can use frozen semen.

Several dogs long gone have been "Cleared" of the PRA by using some of their frozen semen.

Once a test is produced--we should be able to "back TRack"  by using frozen semen on deceased dogs  to trace carriers and affected dogs..

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cariston

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Reply with quote  #15 
Very interesting, and much more comprehendable as genetics and understanding cystinuria in depth can be very complicated!




Cariston


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steveoifer

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Reply with quote  #16 
Cathy,

Isn't it easier to check the whole, rather than the half multiple times?

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For the betterment of the breed

"Above all, a uniform type should be aimed at by breeders and uniformity of type can only exist in a proportionate ratio in the purity and distinctiveness in any breed"!.........M. Moore
"If breeds did not adhere to a specific shape, form, and colour range, or if breeders disregarded this blueprint, the breed would degenerate to the point that it would hardly resemble the breed at all. Selective breeding does not just create breeds- it preserves them as well." Breeding purebred dogs inherently means accepting limitations on your freedom to just breed anything...Catherine McMillan
" A reinforced consolidation of the American and British standards could be the basis for restoring our breed to the gladiatorial glory of its ancient past, in capability if not in usage".....Norman Howard Carp-Gordon
"I can live with doubt, or not knowing, rather than to have answers that might be wrong"...Richard Feynman
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annagmay

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Reply with quote  #17 
>>>>Isn't it easier to check the whole, rather than the half multiple times?>>>>

I'm not Cathy, but I'll answer anyway.....

No one had asked about easier, it was only asked if it is possible. 

OptiGen would prefer that we send them blood vs semen.  For DNA testing blood is the Gold Standard and recommended choice. 

It is possible to use semen for DNA testing and it is just as accurate & reliable to use.  Many labs have the capability of doing DNA testing with semen.  OptiGen and UPenn happen to have that capability as they have state of the art labs with modern equipment and they have staff that are trained to run the equipment and do the tests....  At this time, OptiGen and UPenn do not charge an extra fee to do DNA tests with semen, but I'm guessing that both would prefer blood.

It is a lot cheaper for individual owners to pay to store blood for future DNA tests.  I used stored blood to do the CMR test on my bitch Patience that had passed away before the test was available.  She was a spayed bitch that had never been bred, but I wanted to be the 1st one to run the test on a Mastiff that had passed away and I was curious and I'm weird when it comes to health testing. 

If I had a stud dog that had passed away and I wanted to run the CMR test on him, but I didn't have stored blood to use, I could use his frozen semen if it was available.  It does not require special shipping to use frozen semen.  A 1/2 a straw in a baggie labeled semen and cushioning to help avoid breakage and an ice pack to keep it cool is all you need.

Anna

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steveoifer

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Reply with quote  #18 
It just stands to reason, with only half of the genetic markers in haploid samples, that a number of tests would be needed in order to determine the entire genetic content.
I would think that this would be wasteful of time & money compared with utilizing the entire set of chromosomes found in blood DNA.
Now, if you can't get affected samples from blood donors, then the next best thing might be from sperm donations, but the better approach, unless I'm off course on this, would be blood!

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For the betterment of the breed

"Above all, a uniform type should be aimed at by breeders and uniformity of type can only exist in a proportionate ratio in the purity and distinctiveness in any breed"!.........M. Moore
"If breeds did not adhere to a specific shape, form, and colour range, or if breeders disregarded this blueprint, the breed would degenerate to the point that it would hardly resemble the breed at all. Selective breeding does not just create breeds- it preserves them as well." Breeding purebred dogs inherently means accepting limitations on your freedom to just breed anything...Catherine McMillan
" A reinforced consolidation of the American and British standards could be the basis for restoring our breed to the gladiatorial glory of its ancient past, in capability if not in usage".....Norman Howard Carp-Gordon
"I can live with doubt, or not knowing, rather than to have answers that might be wrong"...Richard Feynman
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Kiokeemastiffs

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Reply with quote  #19 
For living dogs--blood or buccal swabs.  Blood would make it a test that vets would need to do--if it is set up as a buccal swab (like the long hair test or the AKC DNA test) then owners could do it.

When it comes out--I think it will be a blood test initally.

For a dead dog--either stored blood or semen- A Semen sample would consist of millions of little single sperms  and once blended--all of ther "DNA" tree would be present.

DNA testing has come such a long way in the last 10 years--it takes only a very small amount of sample--which is processed into a "pureed" mixture in the testing solution--and then run through the machine.

So whether it is blood, cheek cells, or semen--it would be blended up and processed into a "slush" solution to be tested. 

I would rather see frozen blood used to test a dead dog--cause once all your frozen semen is gone--the breeding potiential of said dog is gone.

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Cathy (Catie) C. Arney
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prn65

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Reply with quote  #20 
As Cathy said above about using sperm as a sample for DNA testing - a single sample will contain ALL DNA from that dog, both chromosomes inherited from the dam and from the sire.  The test is run on the whole sample (which contains million of individual sperm cells) -all the DNA present is extracted in one tube.

It is possible to extract DNA from a single cell, but for this kind of study, not practical and wouldn't provide any additional information.

Once a potential mutation is identified, then you look at the DNA of the parents and try to track the mutation through different families.

I used to do these kind of studies. SNP chips are an exciting advance in genetics in that they allow you to screen a large number of samples relatively quickly and inexpensively.  What usually happens is that you end up with a number of potential markers, and you have to run more samples to determine which of these markers actually tracks with the disease you are interested in.

The more samples you have, and the more data associated with the samples, the quicker you'll come up with a useful marker that can then be developed into a test like the one to track PRA.

If you don't have enough samples, then the chances of isolating a marker are greatly reduced, due to the random way genes are inherited. Statistics are the backbone of this method, and without sufficient numbers, the results are fuzzy at best.

If sperm is all that is available from an affected dog or one that sired affected dogs, I'd encourage you to submit it (if the researchers want to use it) especially if you've got lots of pedigree information to go with it

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