Inhibin a is one of the candidate genes that control the ovulation in poultry. To study the genetic effects of inhibin a on apoptosis and proliferation of goose granulosa cells cultured in vitro, two RNA interference ...Inhibin a is one of the candidate genes that control the ovulation in poultry. To study the genetic effects of inhibin a on apoptosis and proliferation of goose granulosa cells cultured in vitro, two RNA interference （RNAi） expression vectors, psiRNA-INHal and psiRNA-INHα2, were constructed to knock down inhibin α gene expression. After 48 h of transfection, the efficiency of these two RNAi expression vectors was examined by fluorescence microscopy. Meanwhile, inhibin protein expression levels, apoptosis indexes （AI） and proliferation indexes （PI） of granulosa cells were analyzed by flow cytometry. In addition, the supernatants were collected to assay the concentrations of estrogen （E2） and progesterone （P） by radioimmunoassay. The results showed that the expression level of inhibin a in the RNAi group were decreased 30%--40% than those in the control groups （P 〈0.05） and the apoptosis indexes and proliferation indexes in the RNAi groups were significantly higher than those in the control groups （P 〈0.05）. However, the E2 concentrations in the RNAi groups were lower than those in the control groups （P 〈0.05）. These results indicate that inhibin a has antagonistic effect on granulosa cell apoptosis.展开更多
[Objective] The aim of this study was to reveal the relationship between inihibin （INH） α precursor gene and seasonal reproduction of goats, and investigate the evolutionary conservation of INHα precursor gene. [ ...[Objective] The aim of this study was to reveal the relationship between inihibin （INH） α precursor gene and seasonal reproduction of goats, and investigate the evolutionary conservation of INHα precursor gene. [ Method] Cloning and sequence analysis of 5＇ flanking region and exon of inihibinα （INHE） precursor gene in twenty ewes between non-seasonal estrous breed （Haimen goats） and seasonal estrous breed （Anhui white goats） was analyzed in this study. [ Result] Compared with Anhui white goats, INHα precursor gene in Haimen goats had three SNP but no amino acid change, while its nucleotide homology was 99.7% and amino acid homology was 100%. The nucleotide homology of INHα precursor gene in goat, cattle, pig, person, chicken, horse, rat and dog ranged from 12.7% to 96.5%. [ Conclusion] INHα precursor gene tends to be highly conserved in species, and any change of nucleotide and amino acid maybe directly influence the function of the whole gene coding and regulation.展开更多
The aims of the study were to determine the distribution of inhibin and its α subunit in some rat tissues by an immunohistochemical method of streptavidin-biotin complex （SABC） and to assess the transport mechanism...The aims of the study were to determine the distribution of inhibin and its α subunit in some rat tissues by an immunohistochemical method of streptavidin-biotin complex （SABC） and to assess the transport mechanism of inhibin by investigating the localization of inhibin and α subunits in the central nervous system （mainly in hypothalamus and pituitary） of ovariectomized rats. Investigations on the extragonadal tissues of ovariectomized rats showed positive expression of inhibin and its α subunit in heart, kidney, spleen, pancreatic gland cells, but no positive reaction sites were seen in lung, liver, submaxillary gland, and adrenal gland. After injection with inhibin α subunit fragment or inhibin extract, positive reaction sites were observed in hypothalamus and pituitary of ovariectomized rats by SABC. Inhibin and its subunit was present in a wide variety of nonreproductive organs and tissues, and its expression was tissue specific, which indicated that inhibin might play a role in regulating tissue function through autocrine/paracrine mechanisms. Inhibin dimer and α subunit could be transported through the BBB by the method of “separation and reconstruction”.展开更多
Aim： To evaluate whether inhibin-B can predict the outcome of a microsurgical epidymal sperm aspiration （MESA） procedure in patients with suspected primary obstructive azoospermia （OA） and if inhibin-B can replac...Aim： To evaluate whether inhibin-B can predict the outcome of a microsurgical epidymal sperm aspiration （MESA） procedure in patients with suspected primary obstructive azoospermia （OA） and if inhibin-B can replace testicular biopsy in the diagnostic work-up of these patients. Methods： Inhibin-B levels and testicular biopsy scores were related to the outcome of MESA in 43 patients with suspected primary OA. MESA was considered to be successful when epididymal sperm could be identified during the procedure. Results： Spermatozoa were present in the epididymal aspirate in 28 out of the 43 patients （65%）. lnhibin-B values were not significantly different in patients with successful or unsuccessful MESA. The modified Johnsen score, however, was significantly lower in patients with unsuccessful MESA （P = 0.003）. A rete testis obstruction or epididymal malfunctioning was found in 15% of patients with suspected primary OA, reflected by unsuccessful MESA despite normal inhibin-B levels and normal testicular histology. Conclusion： Inhibin-B cannot replace testicular biopsy as a diagnostic tool in the work-up of patients with suspected primary OA. Testicular biopsy is useful in identifying patients with spermatogenic arrest, who might have normal inhibin-B values.展开更多
This study aimed to improve egg-laying performance in incubating Magang geese of Guangdong origin and Landaise geese of French origin. In experiment 1, 50 adults, egg-laying Magang geese were inoculated intramuscular...This study aimed to improve egg-laying performance in incubating Magang geese of Guangdong origin and Landaise geese of French origin. In experiment 1, 50 adults, egg-laying Magang geese were inoculated intramuscularly （i.m.） on days 0, 22, and 45 with 1 mL of immunogen containing 1 mg of recombinant chicken inhibin fusion protein. Immunization significantly increased blood antibody titers against inhibin fusion protein, but did not affect the egg-laying performance within 10 days after the first inoculation. From day 15, the egg-laying rate in inhibin-immunized group increased and was significantly higher than the values of control geese from day 40 to 55. However, the reverse was true from day 55 to 75 when more immunized geese developed incubation. In the entire 120 days of the experiment, the immunized geese laid 17.3 eggs in contrast to 16.4 eggs laid by the control geese. From day 30 till the end of the experiment, weight of eggs in the control geese was significantly greater than that in inhibin-immunized birds. In experiment 2, 40 Landaise geese were immunized against inhibin, as described in experiment 1. These geese laid 9.0 eggs on average in contrast to 7.3 eggs laid by nonimmunized control geese over 90 days of egg laying. The above results demonstrated that immunization against recombinant chicken inhibin fusion protein improved egg-laying performance in geese, and the increment was higher in nonincubating geese.展开更多
The aim of this study was to investigate the feasibility of stimulating ovarian fol icle development in order to improve fertility in water buffalo cows by immunization against inhibin. The experiment was carried out ...The aim of this study was to investigate the feasibility of stimulating ovarian fol icle development in order to improve fertility in water buffalo cows by immunization against inhibin. The experiment was carried out in early summer (May) and included 24 multi-parity crossbred Murrah-Swamp buffaloes that were divided into immunized (n=11) and control (n=13) groups. Each immunized cow was administered with a 2-mL immunogen of mineral oil adjuvant containing 2 mg of recombinant inhibinα-subunit fusion protein. The controls were treated with the adjuvant only. Al animals received Ovsynch protocol treatment, starting on the day of the antigen administration, and they were artiifcial y inseminated upon behavioral estrus. As a result, al of the immunized buffaloes generated antibodies against inhibin during the experimental period and had higher plasma concentrations of fol icle-stimulating hormone (FSH), activin, and estradiol (E2) related to estrous expression. A higher proportion of immunized animals expressed estrus behavior than did the controls (72%vs. 30%, P<0.05). On aver-age, inhibin-immunized buffaloes had signiifcantly more large fol icles (≥9 mm in diameter) than the controls (mean±SEM;1.2±0.1 vs. 0.84±0.1, respectively;P<0.05) and a slightly higher mean total number of fol icles (≥2 mm;11.4±0.7 vs. 9.0±1.1, respectively;P=0.09) and smal (2–4 mm) fol icles (8.81±0.6 vs. 6.84±1.0, respectively;P=0.12). A higher percentage of cows ovulated in the immunized group than in the control group (91%(10/11) vs. 54%(7/13), respectively;P<0.05). Moreover, inhibin-immunized cows had slightly larger corpus luteum (CL) than the controls 9 days after ovulation and signiifcantly higher (P<0.01) post-ovulation peak plasma progesterone (P4) concentrations. Immunization against inhibin also mar-ginal y increased the conception rate 42 days after insemination (45.8%vs. 15.4%;P>0.05). These results demonstrate that immunization against inhibin, coupled with the treatment with the Ovsynch protocol, can constitute a new technique to increase fertility in water buffalo cows.展开更多
A total of 40 rats, aged in 30 days. were divided into 4 groups and immunized (intramuscularly injection) with 0 μg(control), 15μg (group 1), 25μg (group 2) or 40 μg (group 3) of inhibin α(1-32) re-combinant expr...A total of 40 rats, aged in 30 days. were divided into 4 groups and immunized (intramuscularly injection) with 0 μg(control), 15μg (group 1), 25μg (group 2) or 40 μg (group 3) of inhibin α(1-32) re-combinant expression plasmid pcINH in combination with liposome. Booster was given without liposome on day 20 after primary immunization. The results showed that 50%(13/26) rats were detected in positive antibody against inhibin. However, the increase of immunization dosage and booster did not promote the ratio of antibody positive rats. The number of matured follicles above 0.8 mm in diameter in the antibody positive rats was 2.3 more than that in the negative rats (P>0. 05). The concentration of blood plasma FSH increased distinctively on day 10 after primary immunization (P<0. 05), but no increase was observed after booster immunization. The 17-β-estradiol levels in blood plasma of rats between the positive and the negative groups had no remarkable differences (P>0.05). These results suggested that recombinant inhibin expression plasmid could stimulate animal body to produce antibody against inhibin.展开更多
基金the National Natural Science Foundation of China (No. 30300253) and Wuhan Chenguang Science and Technology Project (No. 20065004116-25).
文摘Inhibin a is one of the candidate genes that control the ovulation in poultry. To study the genetic effects of inhibin a on apoptosis and proliferation of goose granulosa cells cultured in vitro, two RNA interference （RNAi） expression vectors, psiRNA-INHal and psiRNA-INHα2, were constructed to knock down inhibin α gene expression. After 48 h of transfection, the efficiency of these two RNAi expression vectors was examined by fluorescence microscopy. Meanwhile, inhibin protein expression levels, apoptosis indexes （AI） and proliferation indexes （PI） of granulosa cells were analyzed by flow cytometry. In addition, the supernatants were collected to assay the concentrations of estrogen （E2） and progesterone （P） by radioimmunoassay. The results showed that the expression level of inhibin a in the RNAi group were decreased 30%--40% than those in the control groups （P 〈0.05） and the apoptosis indexes and proliferation indexes in the RNAi groups were significantly higher than those in the control groups （P 〈0.05）. However, the E2 concentrations in the RNAi groups were lower than those in the control groups （P 〈0.05）. These results indicate that inhibin a has antagonistic effect on granulosa cell apoptosis.
基金Supported by Natural Science Foundation of Jiangsu Province(BK2007561)~~
文摘[Objective] The aim of this study was to reveal the relationship between inihibin （INH） α precursor gene and seasonal reproduction of goats, and investigate the evolutionary conservation of INHα precursor gene. [ Method] Cloning and sequence analysis of 5＇ flanking region and exon of inihibinα （INHE） precursor gene in twenty ewes between non-seasonal estrous breed （Haimen goats） and seasonal estrous breed （Anhui white goats） was analyzed in this study. [ Result] Compared with Anhui white goats, INHα precursor gene in Haimen goats had three SNP but no amino acid change, while its nucleotide homology was 99.7% and amino acid homology was 100%. The nucleotide homology of INHα precursor gene in goat, cattle, pig, person, chicken, horse, rat and dog ranged from 12.7% to 96.5%. [ Conclusion] INHα precursor gene tends to be highly conserved in species, and any change of nucleotide and amino acid maybe directly influence the function of the whole gene coding and regulation.
文摘The aims of the study were to determine the distribution of inhibin and its α subunit in some rat tissues by an immunohistochemical method of streptavidin-biotin complex （SABC） and to assess the transport mechanism of inhibin by investigating the localization of inhibin and α subunits in the central nervous system （mainly in hypothalamus and pituitary） of ovariectomized rats. Investigations on the extragonadal tissues of ovariectomized rats showed positive expression of inhibin and its α subunit in heart, kidney, spleen, pancreatic gland cells, but no positive reaction sites were seen in lung, liver, submaxillary gland, and adrenal gland. After injection with inhibin α subunit fragment or inhibin extract, positive reaction sites were observed in hypothalamus and pituitary of ovariectomized rats by SABC. Inhibin and its subunit was present in a wide variety of nonreproductive organs and tissues, and its expression was tissue specific, which indicated that inhibin might play a role in regulating tissue function through autocrine/paracrine mechanisms. Inhibin dimer and α subunit could be transported through the BBB by the method of “separation and reconstruction”.
文摘Aim： To evaluate whether inhibin-B can predict the outcome of a microsurgical epidymal sperm aspiration （MESA） procedure in patients with suspected primary obstructive azoospermia （OA） and if inhibin-B can replace testicular biopsy in the diagnostic work-up of these patients. Methods： Inhibin-B levels and testicular biopsy scores were related to the outcome of MESA in 43 patients with suspected primary OA. MESA was considered to be successful when epididymal sperm could be identified during the procedure. Results： Spermatozoa were present in the epididymal aspirate in 28 out of the 43 patients （65%）. lnhibin-B values were not significantly different in patients with successful or unsuccessful MESA. The modified Johnsen score, however, was significantly lower in patients with unsuccessful MESA （P = 0.003）. A rete testis obstruction or epididymal malfunctioning was found in 15% of patients with suspected primary OA, reflected by unsuccessful MESA despite normal inhibin-B levels and normal testicular histology. Conclusion： Inhibin-B cannot replace testicular biopsy as a diagnostic tool in the work-up of patients with suspected primary OA. Testicular biopsy is useful in identifying patients with spermatogenic arrest, who might have normal inhibin-B values.
文摘This study aimed to improve egg-laying performance in incubating Magang geese of Guangdong origin and Landaise geese of French origin. In experiment 1, 50 adults, egg-laying Magang geese were inoculated intramuscularly （i.m.） on days 0, 22, and 45 with 1 mL of immunogen containing 1 mg of recombinant chicken inhibin fusion protein. Immunization significantly increased blood antibody titers against inhibin fusion protein, but did not affect the egg-laying performance within 10 days after the first inoculation. From day 15, the egg-laying rate in inhibin-immunized group increased and was significantly higher than the values of control geese from day 40 to 55. However, the reverse was true from day 55 to 75 when more immunized geese developed incubation. In the entire 120 days of the experiment, the immunized geese laid 17.3 eggs in contrast to 16.4 eggs laid by the control geese. From day 30 till the end of the experiment, weight of eggs in the control geese was significantly greater than that in inhibin-immunized birds. In experiment 2, 40 Landaise geese were immunized against inhibin, as described in experiment 1. These geese laid 9.0 eggs on average in contrast to 7.3 eggs laid by nonimmunized control geese over 90 days of egg laying. The above results demonstrated that immunization against recombinant chicken inhibin fusion protein improved egg-laying performance in geese, and the increment was higher in nonincubating geese.
基金supported by the National Key Technology R&D Program of China (2011BAD19B02-6)the Open Grant of Guangxi Provincial Key Laboratory of Water Buffalo Genetics, Breeding and Reproduction, China (SNKF-2012-04)
文摘The aim of this study was to investigate the feasibility of stimulating ovarian fol icle development in order to improve fertility in water buffalo cows by immunization against inhibin. The experiment was carried out in early summer (May) and included 24 multi-parity crossbred Murrah-Swamp buffaloes that were divided into immunized (n=11) and control (n=13) groups. Each immunized cow was administered with a 2-mL immunogen of mineral oil adjuvant containing 2 mg of recombinant inhibinα-subunit fusion protein. The controls were treated with the adjuvant only. Al animals received Ovsynch protocol treatment, starting on the day of the antigen administration, and they were artiifcial y inseminated upon behavioral estrus. As a result, al of the immunized buffaloes generated antibodies against inhibin during the experimental period and had higher plasma concentrations of fol icle-stimulating hormone (FSH), activin, and estradiol (E2) related to estrous expression. A higher proportion of immunized animals expressed estrus behavior than did the controls (72%vs. 30%, P<0.05). On aver-age, inhibin-immunized buffaloes had signiifcantly more large fol icles (≥9 mm in diameter) than the controls (mean±SEM;1.2±0.1 vs. 0.84±0.1, respectively;P<0.05) and a slightly higher mean total number of fol icles (≥2 mm;11.4±0.7 vs. 9.0±1.1, respectively;P=0.09) and smal (2–4 mm) fol icles (8.81±0.6 vs. 6.84±1.0, respectively;P=0.12). A higher percentage of cows ovulated in the immunized group than in the control group (91%(10/11) vs. 54%(7/13), respectively;P<0.05). Moreover, inhibin-immunized cows had slightly larger corpus luteum (CL) than the controls 9 days after ovulation and signiifcantly higher (P<0.01) post-ovulation peak plasma progesterone (P4) concentrations. Immunization against inhibin also mar-ginal y increased the conception rate 42 days after insemination (45.8%vs. 15.4%;P>0.05). These results demonstrate that immunization against inhibin, coupled with the treatment with the Ovsynch protocol, can constitute a new technique to increase fertility in water buffalo cows.
基金supported by National Natural Science Foundation of China(30070555).
文摘A total of 40 rats, aged in 30 days. were divided into 4 groups and immunized (intramuscularly injection) with 0 μg(control), 15μg (group 1), 25μg (group 2) or 40 μg (group 3) of inhibin α(1-32) re-combinant expression plasmid pcINH in combination with liposome. Booster was given without liposome on day 20 after primary immunization. The results showed that 50%(13/26) rats were detected in positive antibody against inhibin. However, the increase of immunization dosage and booster did not promote the ratio of antibody positive rats. The number of matured follicles above 0.8 mm in diameter in the antibody positive rats was 2.3 more than that in the negative rats (P>0. 05). The concentration of blood plasma FSH increased distinctively on day 10 after primary immunization (P<0. 05), but no increase was observed after booster immunization. The 17-β-estradiol levels in blood plasma of rats between the positive and the negative groups had no remarkable differences (P>0.05). These results suggested that recombinant inhibin expression plasmid could stimulate animal body to produce antibody against inhibin.
