Effect Of Inhibin PCISI Gene Immunization On Reproduction And Reproductive Endocrinology In The Yellow Cattle

Techniques of molecular cloning, gene immunization, enzyme-linked immunoassay(ELISA), radio immunoassay (RIA) and B ultrasonic diagnosis were taken advantage of tostudy the immune response. Effect of inhibin gene immunization on reproduction andreproductive endocrinology in cattle was also carried out. The different immune effectscaused by different purity of vaccine, dosage and adjuvant were analyzed. The maincontents were as follows:1 Immunoreactivitiy of the yellow cattle after inhibin pCISI gene immunizationThe yellow cattle were immunized by the different dosage of highly- or lowly-purifiedinhibin fusion expressing plasmid pCISI (0.75mg, 1.5mg, 2.25mg and 3.0mg) by way ofadjuvant procaine or lipsome. The result showed that: (1) In either P/N value or rate of thepositive antibody against inhibin the two adjuvants such as procaine and lipsome nearly hadthe same extents.(2) The immune ability of group with highly-purified plasmid(OD₂₆₀/OD₂₈₀ between 1.8 and 2.0) was a little higher than that with the lowly-purifiedone(OD₂₆₀/OD₂₈₀ between 1.7 and 1.8 or between 2.0 and 2.1)(P＞0.05). (3) Embryonicnumber was much related to both the P/N value and rate of the positive antibody(P＜0.05).(4)the antibody could be produced on the 10th day after the primary immunization. Except thatdifference between the control and the 0.75mg dosage-group was trivial(P＞0.05), among theother three dosage-groups the difference was significant(P＜0.05) for the two aspects.Whatever P/N value or rate of the antibody against inhibin, dosage of 2.25 mg was the bestof all in the experiment. The difference between 2.25mg dosage-group and 0.75mg one wasvery vivid (P＜0.05), as was group of 2.25mg and 1.5mg. (P＜0.05).2 Effect of inhibin gene immunization on follicular development in yellow cattle.The ability of the follicular development was monitored by B ultrasonic diagnosisduring the estrous phase in the yellow cattle. The results illustrated that: (1) Difference wasnot remarkable for the two given adjuvants whatever in the amount of follicles, rate offollicular development, follicular growth speed or size of the mature follicles. (2) Likewise,purity didn't affect follicular development including the above four aspects. (3) Embryonicnumber examined wasn't linked to both growth speed and size of the two-side follicles. (4)As for antibody level, the result was that the positive group was better than the negativegroup in the amount of the small- and large-sized follicles, double-follicle rate, two-sidefollicular growth speed, but not in the single-follicle rate. (5) As concerned as dosage of theexploited pCISI plasmid, difference was remarkable for all the immunized groups and the control in the follicular amount, singe- and double-follicular rate as well as the follicularsize, with the best of dosage 2.25mg and the worst of dosage0.75mg(P＜0.05). In spite ofthese, 1.5mg was the best for the follicular growth speed compared to the other three doses(P＜0.05).3 Effect of inhibin gene immunization on corpus development in yellow cattle.The two-month period of pregnant corpus luteums on both the left and right sides wereexamined by B ultrasonic machine. The results demonstrated that: (1) For the adjuvant, sizeof the two-side corpus luteum in the procaine group was a little smaller than that in thelipsome group (P＞0.05). (2) Size of the two-side corpus luteum in the highly-purified groupwas slightly higher than that in the lowly-purified one. (3) Both the embryonic number andthe antibody level were not communicated with the size of the left or the right corpusluteum.(4) For the immune dosage, the diameter of the two-side corpus luteum in thecontrol was smaller than that in all the four dosage-groups(P＜0.05). As to the fourimmunized groups, the result also presented that difference was significant between the bestgroup of 2.25mg and the worst group of 0.75mg. The developmental ability of the left andright corpus luteums was similar (P＞0.05).4 Effect of inhibin gene immunization on the early embryonic development in theyellow cattle.Early embryos at the age of two months were also diagnosed by B ultrasonic machine.The result was the following: (1)Amount of the early embryos, twinning rate anddevelopmental parameters of both the coceptus and the embryos in the two-adjuvant groupswas nearly in the same level(P＞0.05). (2) The plasmid purity didn't affected the earlyembryonic development (P＞0.05). (3)Whatever developmental parameters of the leftconceptus or the right one, effect of embryonic number was little(P＞0.05). (4) Effect of theantibody levels is the same as the embryonic number. (P＞0.05). (5) For the fetus number, allthe four immunized groups were higher than the control, but only the dosage of 2.25mg wassignificant (P＜0.05). For the twinning rate, the control was much lower than all the testedgroups (P＜0.05). For the developmental parameters (length, shortness, maximum area of thesection and maximum circle of the section) of the two-side conceptus, the control was largerthan the other four groups (P＞0.05), difference of the length was clear between the group of2.25mg (T₃)and 0.75mg(T₁)(P＜0.05). Besides, difference of the shorness, circle and area ofthe embryos between the four dosage-groups and the control was significant on the rightside(p＜0.05), the same result was between the control and the 0.75mg group(p＜0.05), butthere was neither significance between the four immunized groups and the control nor significance among the four dosage-groups in the left-side embryos.5 Effect of inhibin pCISI gene immunization on reproductive endocrinology of yellowcattle.In order to study effect of inhibin gene immunization on reproductive endocrinology ofthe yellow cattle, level of FSH, E₂ and P₄ were measured by RIA. The result showed that: (1)Content of both progesterone and estradiol was nearly the same in the two adjuvant groupsexcept FSH content was a little higher in the procaine group and the lipsome group(P＞0.05).(2) For purity, during the booster immunization period content of P₄ in the lowly-purifiedgroup was higher than that in the highly-purified group but not for FSH and E₂. (3)As forthe antibody level, content of FSH in the positive group was much higher than that in thecontrol(p＜0.05), and a little higher than that in the control(p＞0.05), content of FSH, E₂ andP₄ was significant between the positive group and the negative group at the state of thebooster immunization but not significant during the primary immunization period. Contentof the above three hormones was almost the same on the stage of primary immunization butnot on the stage of booster immunization between the group with the positive antibody andthe group with the negative antibody. (4)Embryonic number could affect content of bothFSH and P₄ during the booster immunization period and it had nothing with E₂ contentduring the whole period. (5)For the part of the immunized dosage, the difference betweenthe control and 1.5mg group was significant for FSH content on the 45th day of the boosterperiod(P＜0.05), significant for E₂ content between the control and the two dosage-groups of2.25mg and 3.0mg on the 10th day of booster period(P＜0.05), significant for E₂ contentbetween the control and the group of 1.5mg of plasmid(P＜0.05), significant for P₄ contentduring the booster immunization for the control and the immunized groups.On day 10 afterthe booster immunization difference between the two groups was significant (P＜0.05), onday 45 after the booster immunization difference between the two groups was extremelysignificant (P＜0.01).