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AU672559B2 - A method of inducing superovulation in cattle - Google Patents
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AU672559B2 - A method of inducing superovulation in cattle - Google Patents

A method of inducing superovulation in cattle Download PDF

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Publication number
AU672559B2
AU672559B2 AU31706/95A AU3170695A AU672559B2 AU 672559 B2 AU672559 B2 AU 672559B2 AU 31706/95 A AU31706/95 A AU 31706/95A AU 3170695 A AU3170695 A AU 3170695A AU 672559 B2 AU672559 B2 AU 672559B2
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Australia
Prior art keywords
inhibin
cow
superovulation
follicle
cattle
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Ceased
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AU31706/95A
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AU3170695A (en
Inventor
Takako Ito
Masato Konishi
Toshiro Takedomi
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National Federation of Agricultural Cooperative Associations
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National Federation of Agricultural Cooperative Associations
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00Medicinal preparations containing peptides
    • A61K38/16Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22Hormones
    • A61K38/24Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00Medicinal preparations containing antigens or antibodies
    • A61K39/0005Vertebrate antigens

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Endocrinology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Reproductive Health (AREA)
  • Zoology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)

Description

-I-i 1 I
I
AUSTRALIA
Patents Act 1990 NATIONAL FEDERATION OF AGRICULTURAL COOPERATIVE
ASSOCIATIONS
ORIGINAL
B
a e e v .er 0 e ovn o S•o uvo u a o *M 6 COMPLETE SPECIFICATION STANDARD PATENT Invention Title: A method for inducing superovulation in cattle The following statement is a ftll description of this invention including the best method of performing it known to us:-
I
i' I- .1dq_
.WW
*1 7 BACKGROUND OF THE INVENTION The present invention relates to a method for inducing superovulation in cattle. More particularly, the present invention relates to a method for introducing superovulation at a higher and steady level to collect normal embryos efficiently. The present invention relates also to a method for reproducing cattle in which a significantly large number of ova or embryos can be collected when a cow having an increased number of ovarian follicles responsive to follicle-stimulating hormone to be injected during superovulation treatment is produced and subjected to the superovulation treatment.
0004 0000 on.
Description of the Related Prior Art Animals are classified into two categories, the monotocous animals cattle, horse and human being) which bring forth only one young per liter and the polytocous animals pig and dog) which bring forth a plurality of youngs per liter. It is considered 20 that the number of ovulations is controlled by hormones derived from hypothalamus-pituitary gland-gonads.
To collect many embryos as possible from cattle which is a monotocous animal, the superovulation technique for inducing ovulation of multiple ova has -2 been used. Usual method for superovulation in cattle comprises injection of an exogenous hormone iporcine follicle-stimulating hormone (FSH) or equine chorionic gonadotropin (eCG; formerly known as pregnant mare serum gonadotropin). That is, superovulation-inducing method in the embryo transfer technique has come into wide use as a technique for efficient bovine reproduction. The superovulation treatment in cattle can be induced by injecting follicle-stimulating hormone on day 9 to day 13 of estrous cycle to develop a large number of ovarian follicles, and by injecting prostaglandin F2a 48 to hours after the injection of follicle-stimulating hormone to induce luteolysis of the existing corpus lutea. The luteolysis results in ovulation by removing the inhibitory effect of progesterone on the action of hypothalamus and pituitary gland.
i o0'° On the other hand, it has been reported that immunological inhibition of inhibin which suppresses the secretion of follicle-stimulating hormone from pituitary 20 gland in cattle accelerates the secretion of folliclestimulating hormone from pituitary gland and consequently permits development of ovarian follicles and ovulation of multiple ova.
tilt Inhibin is a peptide hormone which is secreted in testis or ovarian follicles and suppresses the production and secretion of FSH in the pituitary gland.
In the 1980's, the rapid development of genetic engineering gradually revealed the whole aspect and i i 1
I
3 action of inhibin whose existence and structure had not been elucidated. It is being revealed that in mature cows, inhibin suppresses FSH secretion from the pituitary gland specifically. If inhibitory control of inhibin is possible in cows, secretion of endogenous FSH is accelerated. Therefore, it is expected that ovulation of multiple ova can be induced in every estrous cycle. In recent years, energetic attempts have been made in sheep and cattle to suppress inhibin by immunization and thereby increase the number of ova ovulated.
However, when standard superovulation method comprising injection of FSH or eCG is used, there is a i large variation in the number of ovarian follicles responsive to the hormone among treatments or cows.
This variation is one of the big problems in the art because it is difficult to collect embryos at a higher 4 and steady level with an excellent quality from superior-donor cows which have excellent genetical nature. Consequently, this variation increases the production cost per embryo.
Therefore, a considerable economical effect in the technique for bovine embryo transfer can be certainly obtained if cows which response at a higher and steady level to superovulation treatment are produced by elucidating the relationship between the responsiveness to superovulation and the state of p 4 ovarian follicles or hor.nones involved in follicular development, and carrying out a certain treatment.
It has been reported that there is a positive correlation between the number of ovarian follicles with of 4 to 6 mm in diameter at the initiation of superovulation treatment and the number of ova ovulated by the superovulation (Romero, Albert, Brink, Z.
and Seider, G.E. Jr., Theriogenology, 35, 265 (abstract) 1991). It has also been reported that the number of ova ovulated is decreased when the superovulation treatment is carried out in the luteal phase in which functional dominant follicles are present (Guilbaut, Grasso, Lussier, Rouillier, and Matton, J.
Reprod. Fertil., 91, 81-89, 1991; Huhtinen, Rainio Analto, Bredbacka, P. and Maki-Tanila, A., Theriogenology, 37, 457-463, 1992). In addition, it has been reported that inhibin secretion is increased with follicular development (Martin, Fogwell, R.L. and sq...
Ireland Biology of Reproduction, 44, 693-700, 20 1991). Recently, the present inventors found that the number of recruited follicles was increased by vaccinating a cow against inhibin. Although a manner in which dominant follicles affect on the response to superovulation treatment has not yet been elucidated, the above-mentioned reports suggest the following causal sequence: the presence of the dominant follicles the S" acceleration of inhibin secretion the decrease of the number of the recruited follicles the reduction of the response to superovulation treatment.
SUMMARY OF THE INVENTION From a new viewpoint, the present inventors found that the number of ova ovulated is significantly increased when a mature cow vaccinated against inhibin to have an increased number of ovarian follicles with a diameter of 4 to 6 mm is subjected to conventional superovulation treatment. When this technique is employed, the secretion of endogenous folliclestimulating hormone is accelerated. Therefore, no antibody against follicle-stimulating hormone is produced by repetition of the treatment, so that there is no fear of reduction of the effect.
15 BRIEF DESCRIPTION OF THE DRAWING Fig. 1 shows a change of antibody titer for inhibin caused by vaccination against inhibin.
4 DESCRIPTION OF THE PREFERRED EMBODIMENTS 44 .The present invention is illustrated below in detail with an example.
C
r t .i Example 1 The following experiments were carried out for S' counting ova ovulated by cows subjected to superovulation treatment after immunization against inhibin.
t O b .W 6 1. Experimental method Experiment 1 and experiment 2 were carried out.
Experiment 1 Five non-multiparous cows (Holstein) were used for confirming changes of the antibody titer for inhibin and the number of recruited follicles caused by vaccination against inhibin.
Experiment 2 Using 30 cows (Holstein and Japanese Black), the influence of vaccination against inhibin on the result of the superovulation treatment was investigated.
Five non-multiparous Holsteins and 9 multiparous Japanese Blacks were vaccinated against inhibin and then subjected to superovulation by the use of FSH by a usual method. As controls, 5 non-multiparous Holsteins and 11 multiparous Japanese Blacks were subjected to superovulation treatment without vaccination against inhibin.
The above groups were compared in the number of ova collected.
20 Antigen A product obtained by conjugating rabbit serum albumin to a polypeptice synthesized by copying the amino acid sequence from 1 to 26 (numbering from the Nterminal end) positions of the alpha-subunit of porcine 25 inhibin.
Test animals Non-multiparous Holsteins and multiparous Japanese Blacks, which had normal estrous cycles.
0 00 6 oo 000 oooe 00o0 DOO o 0 o 000000 a aoo 1 00 0 .o o 0o o 0 o00 0 0 000oo o aouo 0 kj9 0 0 D 00 0 o 00 0 0 00 0 006 0 0 o 0 0 0 Il
-U
-7- Immunization method Vaccination was employed. Primary injection was carried out as follows: a mixture of 1 mg of the peptide (corresponding to a fragment of inhibin) and complete Freund's adjuvant was injected to each test animal subcutaneously at 3 to 4 places. At 8 weeks and weeks after the primary injection, booster injections of the peptide was given at 0.5 mg as described above injection.
Follicle-stimulating hormone Extract from pituitary gland.
Luteolytic chemical Cloprostenol.
Superovulation treatment The superovulation treatment was started by FSH-injection at day 10 to day 12 of estrous cycle. The B; PSH was intramuscularly injected 6 times at 12-hour intervals in the following reducing dose: 6, 6, 5, 5, 3 .00.
and 3 mg. At 48 hours after the start of the FSH
S
r 20 injections, 750 pg of Cloprostenol was intramuscularly injected. Animals exhibiting estrus were artificially inseminated twice throughout estrus.
Determination of the number of ova ovulated, and embryos collected i 25 At day 7 to day 8 of rut after the superovulation treatment, embryos were collected nonsurgically from uterus. The number of the embryos
I
8 -8collected, the number of normal embryos and the percentage of the normal embryos were determined.
2. Results In the present study, vaccination against inhibin successfully induced elevated antibody titers against inhibin. When the antibody titers were elevated, the num er of recruited follicles (5 to 10 mm in diameter) showed a tendency to increase. When the number of recruited follicles was thus increased with an increase of the antibody titers for inhibin, the superovulation in the cows increased the number of ova and embryos collected without decrease of the percentage of normal embryos.
[Change of the number of recruited follicles during 15 vaccination against inhibin] t 4 4 4 4 4 0.
1 00 0 0 00 0 0 *00 00 0 a 0' 00.U C0 0 00 0 0 0 0 0 00 -00 00 00 000 0 Table 1 The number of follicles of 5-10 mm diameter Weeks after initiation 036 10 118 2279 Treated of036 10 118 2279 co vaccination Treatment Primary First Second injection booster booster 530 2 62 8 1 6 7 1 534 3 4 2 5 5 8 6 8 13 536 4 6 3 12 2 6 5 7 4 537 4 8 4 9 3 10 11 13 8 539 4 7 4 14 3 9 9 10 Mean S.D. 3.2±1.0 6.2±1.3 3. 0±0. 99.5±3.1 12.8±1.3 7.8±1.6 7.6±2.2 7.8±4.0 8.0±3.3 p From the results shown in Table 1, it can be seen that the number of recruited follicles with a diameter of 5 to 10 mm showed a tendency to increase after the primary injection and the boosters.
[Improvement of response to superovulation treatment by immunization against inhibin] Table 2 Results of superovulation induction and embryo collection Results of superovulation and ova collection Number Number Number P Percentage Group of of ova/ of normal of normal 'of norma! cows embryos embryo embryo collected (b/a 100) (b/alo) Control 14 5.9±4.1P 3.
7 ±3.5 P 62.7 Experiment 16 12.9±6.8P 8.9±4.0P 69.0 The superscript p indicates that there is a significant difference (p 0.01) in the same column (Student ttest).
mean S.D.
The superovulation treatment induced an increased number of both embryos collected and normal embryos in the inhibin-immunized group as compared to the untreated group (control).
ii r~r 4 e a *4, a4 (4 I. I i4 t 41 li L L t -I t '44' 444* S4 I 6 I

Claims (7)

1. A method for inducing superovulation in cattle, which comprises vaccinating a cow against inhibin present in its body to produce antibody therein, and then injecting a follicle-stimulating hormone exogenously.
2. A method according to Claim 1, wherein the follicle-stimulating hormone is follicle-stimulating hormone (FSH) or equine chorionic gonadotropin (eCG).
3. A method according to Claim 1, wherein the cow \O is a milk cow or beef cattle, which has normal estrous cycles after puberty.
4. A method according to Claim 1, wherein the vaccination against inhibin is carried out by using as antigen a product obtained by attaching albumin to a \S polypeptice synthesized by copying the amino acid sequence from 1 to 26 (numbering from the N-terminal end) positions of the alpha-subunit of porcine or bovine inhibin, and injecting a mixture of the antigen and an adjuvant to the cow several times subcutaneously.
5. A method according to Claim 1, wherein the vaccination against inhibin is carried out by injection 04a 0 S" of the antigen.
S6. A method for ovulation of a significantly large number of ova in cattle, which comprises S 25 subjecting a cow vaccinated against inhibin to usual superovulation treatment by injecting follicle- stimulating hormone to the cow on day 9 to day 13 of h 1, aanrr*rr~ rs~~ul--i---rrar raui--~a 12 luteal phase or after 7 to 12 days of residence of a corpus luteum hormone drug in the cow, injecting luteolytic preparation or removing the corpus luteum hormone preparation 2 or 3 days after the injection, and thereby inducing rut.
7. A method according to Claim 6, the antigen used for the vaccination is bovine or porcine natural (extracted) inhibin, a synthesized inhibin fragment, or copied inhibin. Dated this 14th day of September 1995 NATIONAL FEDERATION OF AGRICULTURAL COOPERATIVE ASSOCIATIONS Patent Attorney for the applicants F B RICE CO a o 7 o ao+ a O 0 o a o 9 a 0 o oo 0 at a e a 00 9 O 6 0 9 09 0 Ot 9 1 i i i, M- Ic ABSTRACT A method for inducing superovulation in cattle which comprises vaccinating a cow against inhibin, and then injectIng thereto follicle-stimulating hormone to cause ovulation of a large number of developed ovarian follicles. According to this method, normal embryos can be efficiently collected by inducing superovulation in cows at a higher and steady level. o 4.. 4 04 4o 4 *C 4 4 4C C+. tt e>
AU31706/95A 1994-12-09 1995-09-15 A method of inducing superovulation in cattle Ceased AU672559B2 (en)

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JP6-306274 1994-12-09
JP6306274A JP2899534B2 (en) 1994-12-09 1994-12-09 Method of inducing superovulation in cattle

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AU672559B2 true AU672559B2 (en) 1996-10-03

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2768627B1 (en) * 1997-09-19 2000-03-24 Agronomique Inst Nat Rech VETERINARY MEDICINAL PRODUCT CONTAINING MODIFIED EQUINE CHORIOGONADOTROPIN TO INDUCE OVULATION IN THE MARE AND STIMULATE THE PRODUCTION OF TESTOSTERONE IN THE STANDARD
KR101349009B1 (en) * 2012-05-03 2014-01-21 대한민국 In vivo production method of bovine embyos by emporary undernutrition
BR102014005376A2 (en) * 2014-03-07 2016-02-10 Ouro Fino Saúde Animal Ltda inhibin alpha antigen, inhibin alpha coding gene, fusion protein encoding gene, process of obtaining inhibin alpha antigen, antigenic composition, use of inhibin alpha antigen, and use of antigenic composition
CN111134084A (en) * 2020-01-06 2020-05-12 浙江省农业科学院 Method for optimizing timing insemination effect of replacement gilts

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU7055291A (en) * 1990-01-08 1991-08-05 Stefan Heiden Drug for improving the reactions of the ovary and the production of ova and embryos in domestic mammals in connection with biotechnological embryo transfer

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* Cited by examiner, † Cited by third party
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US4864019A (en) * 1985-11-08 1989-09-05 The Salk Institute For Biological Studies Antibodies to inhibin and conjugates produced therefrom
JPH01500121A (en) * 1986-06-24 1989-01-19 ザ・サルク・インステチュート・フォー・バイオロジカル・スタディーズ sheep inhibin

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU7055291A (en) * 1990-01-08 1991-08-05 Stefan Heiden Drug for improving the reactions of the ovary and the production of ova and embryos in domestic mammals in connection with biotechnological embryo transfer

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CA2158116A1 (en) 1996-06-10
JP2899534B2 (en) 1999-06-02
JPH08154957A (en) 1996-06-18
FR2727866A1 (en) 1996-06-14
AU3170695A (en) 1996-06-27

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