IACUC Learning Module - Xenopus aevis

IACUC Learning Module - Xenopus laevis

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Appendix I

Nasco Xenopus Breeding Recommendations

Ovulation

 We recommend that you start the following very soon after receiving your frogs. (If your frogs have been in the laboratory for several weeks, then you should first condition the frogs to breed by following the recommendations in the section on feeding.)

  1. Place the male and female in separate tanks (put female in breeding tank).
  2. Dilute the stock solution of gonadotropin to 500 I.U./ml. For example, a vial containing 2500 I.U. should be diluted with 5 ml of solution. The result will be a solution containing 500 I.U. of chorionic gonadotropin per ml. (Nasco stocks chorionic gonadotropin SA7881L) The solution used to dilute the gonadotropin can be a standard amphibian Ringer's solution* or a 0.9% sodium chloride solution purchased from a drug store. It is important that the diluting solution be sterile. 

    NOTE: Keep the diluted hormone refrigerated. If this is done it should remain active for about 90 days.

     

  3. Inject the male with 150 I.U. of gonadotropin 30-36 hours before the male and female are brought together in the same tank.
  4. 24 hours after the initial injection the male should be given another injection of 150 I.U. At this time the female is given her first injection of 250 I.U.
  5. 8 hours after the female's initial injection give her another injection of 500 I.U.
  6. Occasionally the female will begin dropping her eggs prior to the second injection. If this should happen, give the second injection immediately and place the pair together.
  7. After the female receives her second injection, put the male into the tank with the female.

NOTE: Occasionally an additional injection is necessary for successful ovulation. If there are no eggs by the third day (24 hours after pairing), the female should be given 500 I.U. and the male 150 I.U. of gonadotropin. If the water has become foul, change it when these additional injections are administered.

*Standard Amphibian Ringer's Solution

 To prepare a STOCK of amphibian Ringer's solution weigh out the following chemicals and dissolve them in one liter of distilled water.

Do not use this full strength stock solution.

For use, prepare a 10% solution by adding 10 parts of the stock Ringer's solution to 90 parts of distilled water.

 

NaCl - 6.60 gm
KCl - 0.15 gm
CaCl - 0.15 gm
NaHCO3 - 0.15 gm water - 1 liter

The pH of this stock should be very near 7.8 If the pH needs adjusting, use NaHCO3. Nasco offers frog Ringer's solution (Catalog number SA7855L).

Suggested Injection Schedule

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