Objective: Present investigation was aimed to study the Single Nucleotide Variants of the luteinizing hormone beta ($LH{\beta}$) gene and to analyze their association with the semen quality (fresh and post-thawed frozen semen) and luteinizing hormone (LH) concentrations in Murrah buffalo bulls. Methods: Polymerase chain reaction-single stranded conformational polymorphism (PCR-SSCP) and Sanger sequencing method is used to study genetic variability in $LH{\beta}$ gene. LH assay was carried out using enzyme-linked immunosorbent assay method. A fixed general linear model was used to analyze association of single nucleotide polymorphism (SNP) of $LH{\beta}$ gene with semen quality in 109 and LH concentrations in 80 Murrah bulls. Results: $LH{\beta}$ gene was found to be polymorphic. Total six SNPs were identified in $LH{\beta}$ gene g C356090A, g C356113T, g A356701G, g G355869A, g G356330C, and g G356606T. Single Stranded Conformational Polymorphism variants of pattern 2 of exon 1+pattern 2 of exon 2+pattern 1 of exon 3 had highly significant (p<0.01) effect on sperm concentration (million/mL), percent mass motility, acrosome integrity and membrane integrity in fresh and frozen semen whereas significant (p<0.05) effect was observed on percent live spermatozoa. SSCP variants of pattern 2 of exon 1+pattern 2 of exon 2+pattern 1 of exon 3 had highly significant (p<0.01) effect on luteinizing hormone concentrations too. Conclusion: The observed association between SSCP variants of $LH{\beta}$ gene with semen quality parameters and LH concentrations indicated the possibilities of using $LH{\beta}$ as a candidate gene for identification of markers for semen quality traits and LH concentrations in Murrah buffaloes.
Two experiments were conducted to determine whether leptin is a metabolic signal for gonadotropin secretion in ewes. In the first experiment, twenty-eight cyclic Chal ewes were assigned randomly to an energy restricted, no leptin group (ERNL) (60% of maintenance; n = 14) and an energy normal, no leptin group (ENNL) (100% of maintenance; n = 14) for 71 days (6 estrous cycles). Estrus was synchronized with seven consecutive injections of $PGF_{2{\alpha}}$ Biweekly, body weight (BW) and body condition score (BCS) were determined and blood samples were collected to measure plasma leptin concentration. Blood samples were also taken to determine plasma progesterone concentration twice weekly. After each PG injection from the second injection to the end of experiment, four ewes were selected and blood samples were collected at 20 minutes and at hourly intervals for 3 h to detect plasma LH and FSH concentration. In the second experiment, after the ceasing of the estrous cycle caused by energy restriction, six acyclic ewes were selected and randomly allotted to two groups (n = 3) and received the following treatment for four days. Ewes in an energy restricted, leptin group (ERL) were fed with a ration which provided 60% of maintenance energy requirements and intravenously injected with $4{\mu}g$ leptin/kg BW daily. Ewes in an energy excess, no leptin group (EENL) were fed with a ration that provided 180% (120%+60%) of maintenance energy requirements and intravenously injected with 1 ml saline daily. In both groups, blood samples were collected at 20 minutes and at hourly intervals for 3 h before feeding on d 0 and d 5, and for 3 h before and after injections as above on d 2 and d 4 to detect plasma LH and FSH concentration. In the first experiment, BW and BCS from the $2^{nd}$ estrous cycle, and leptin from the $3^{rd}$ estrous cycle to the end of the experiment significantly (p<0.05) decreased. In ERNL ewes, mean plasma concentrations of FSH significantly (p<0.01) decreased from the $4^{th}$ estrous cycle to d 71 and LH pulsatile secretion was suppressed on d 71, so that, mean plasma concentrations of LH (p<0.05), LH pulse frequency (p<0.01) and LH pulse amplitude (p<0.05) significantly decreased. In the second experiment, injection of leptin significantly increased mean circulating concentrations of LH (p<0.05), LH pulse frequency (p<0.01), LH pulse amplitude (p<0.05) and mean circulating concentrations of FSH (p<0.01) and leptin (p<0.01). High energy intake significantly (p<0.05) stimulated pulsatile secretion of LH and leptin secretion (p<0.01), but non-significantly increased plasma FSH concentration. The results of this study indicate that leptin is a metabolic signal for the GnRH-LH/FSH axis in feed-restricted fat-tailed ewes.
Pituitary LH release has been known to be regulated by the hypothalamic gonadotropin releasing hormone (GnRH) and the gonadal steroid hormones. In addition, neurotransmitters and neuropeptides are actively involved in the control of LH secretion. The alteration in LH release might reflect changes in biosynthesis and/or posttranslational processing of LH. However, little is known about the mechanism by which biosynthesis of LH subunits is regulated, especially at the level of transcription. In order to investigate if ovarian steroid hormones regulate the LH subunit gene expression, ${\alpha}\;and\;LH{\beta}$ steady state mRNA levels were determined in anterior pituitaries of ovariectomized rats. Serum LH concentrations and pituitary LH concentrations were increased markedly with time after ovariectomy. ${\alpha}\;and\;LH{\beta}$ subunit mRNA levels after ovariectomy were increased in a parallel manner with serum LH concentrations and pituitary LH contents, the rise in $LH{\beta}$ subunit mRNA levels being more prominent than the rise in ${\alpha}\;subunit$ mRNA. ${\alpha}\;and\;LH{\beta}$ subunit mRNA levels in ovariectomized rats were negatively regulated by the continuous treatment of ovarian steriod hormones for $1{\sim}4\;days$ and $LH{\beta}\;subunit$ mRNA seemed to be more sensitive to negative feedback of estradiol than progesterone. Treatment of estrogen antagonist, LY117018 or progesterone antagonist, RU486 significantly restroed LH subunit mRNA levels as well as LH release which were suppressed by estradiol or progesterone treatment. These results suggest that ovarian steroids negatively regulate the LH synthesis at the pretranslational level by modulating the steady state levels of ${\alpha}\;and\;LH{\beta}\;subunit$ mRNA and $LH{\beta}\;subunit$ mRNA seemed to be more sensitive to negative feedback action of estradiol than progesterone.
This study was carried out to investigate the effect of LH-RH and Gn-RH treatment in Holstein cows with ovarian quiescence and follicular cystic ovaries. The cows with ovarian quiescence and follicular cystic ovaries injected intramuscularly with 100$\mu\textrm{g}$, 200$\mu\textrm{g}$ and 400$\mu\textrm{g}$ of LH-RH and 200$\mu\textrm{g}$ and 400$\mu\textrm{g}$ of Gn-RH respectively. The cows was diagnosed by repeated rectal palpation. The plasma progesterone and estradiol-17$\beta$ concentrations were assayed by radioimmunoassay methods. The resutls of this study were summarized as follows : 1. Ovulations were induced after treatment of LH-RH and Gn-RH. The concentrations of progesterone reached small peak level at luteal phase and estradiol-17$\beta$ reached obvious peak level with the development and maturation of the follicle during the periods of degeneration of the corpus luteum, and normal ovarian cycle activity started subsequently. 2. The cows with ovarian quiescence and follicular cystic ovaries were induced ovulation at 38.9$\pm$5.3 hrs. after treatment of LH-RH in 66.7% cows and at 52.7$\pm$7.9 hrs after treatment of Gn-RH in 60.0% cows respectively. 3. The good ovarian responses were indicated in treatment with 200$\mu\textrm{g}$ to 400$\mu\textrm{g}$ of LH-RH than those treated with 100$\mu\textrm{g}$ in cows with ovarian quiescence, and did not show difference of ovarian responses between treatments with 200$\mu\textrm{g}$ to 400$\mu\textrm{g}$ of Gn-RH in cows with follicular cystic ovaries.
The study was carried out to find out the changes of the sex hormone concentrations in the blood plasma and antrum of follicular and lutein cystic ovaries of Holstein cows. The progesterone, estradiol-17$\beta$, testosterone, FSH and LH from samples of the blood plasma and antrum of cystic ovaries of cows assayed by radioimmunoassay method. The results of this study were summarized as follows : 1. The concentrations fo progesterone and estradiol-17$\beta$ in the blood plasmaat estrous and luteal phase were 0.95$\pm$0.18ng/ml, 11.45$\pm$3.12pg/ml and 4.25$\pm$0.27ng/ml, 6.27$\pm$0.82pg/ml respectively. The concentrations of progesterone and estradiol-17$\beta$ in the antrum fluid of follicles at estrous and luteal phase were 24.8$\pm$4.12ng/ml, 54.3$\pm$7.25pg/ml and 21.7$\pm$3.79ng/ml, 14.3$\pm$2.72pg/ml respectively, and showed significant changes among the estrous and luteal phase and blood plasma and antrum fluid of follicles. 2. The concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the blood plasma of follicular cystic ovareis of cows were 0.85$\pm$0.25ng/ml, 9.23$\pm$2.72pg/ml, 17.12$\pm$3.26pg/ml and 3.78$\pm$1.02mIU/ml respectively. And the concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the antrum fluid of follicular cystic ovareis of cows were 284$\pm$48.21ng/ml, 389$\pm$67.23ng/ml, 12.84$\pm$0.29ng/ml and 1.84$\pm$0.17mIU/ml respectively, and showed significant changes between in the blood plasma and antrum fluid of cystic ovaries. 3. The concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the blood plasma of lutein cystic ovaries of cows were 3.40$\pm$0.78ng/ml, 4.02$\pm$0.42pg/ml, 10.72$\pm$2.74pg/ml and 0.76$\pm$0.12mIU/ml respectively. And the concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the antrum fluid of lutein cystic ovareis of cows were 427$\pm$35.79ng/ml, 0.76$\pm$0.07ng/ml, 3.45$\pm$0.57ng/ml and 0.29$\pm$0.07mIU/ml respectively, and showed significant changes between the blood plasma and antrum fluid of cystic ovaries. 4. Accordingly, the diagnosis of follicular and lutein cystic ovareis of cows from progesterone, estradiol-17$\beta$ and LH levels in the blood plasma and antrum of cystic ovaries of cows is thought to be possible a diagnostic means.
Korea has recently suffered from severe hazes, largely being long-range transported from China but frequently mixed with domestic pollution. It is important to identify the origin of the frequently-occurring hazes, which is however hard to clearly determine in a quantitative term. In this regard, we suggest a possible classification procedure of various hazes into long-range transported haze (LH), Yellow Sand (YS), and urban haze (UH), based on mass loading of fine particles, time lag of PM mass concentrations between two sites aligned with dominant wind direction, backward trajectory of air mass, and the mass ratio of PM2.5 to PM10. The analysis sites are Seoul (SL) and Baengnyeongdo (BN), which are distant about 200 km from each other in the west to east direction. Aerosol concentrations at BN are overall lower than those of SL, indicative of BN being a background site for SL. We found distinct time lag of PM2.5 and PM10 concentrations between BN and SL in case of both LH and YS, but the intensity of YS being stronger than LH. Time scale (e-folding time scale) of LH appears to be longer and more variable than YS, which implies that LH covers much larger spatial scale. In addition, we found linear and significant correlations between ${\tau}_a$ obtained from sunphotometer and ${\tau}_{cal}$ calculated from surface aerosol scattering coefficient for LH episodes, relative to few correlation between those for YS, which might be associated with transported height of YS being much higher than LH. Therefore surface PM concentrations for the YS period are thought to be not representative for vertical integrated amount of aerosol loadings, probably by virtue of decoupled structure of aerosol vertical distribution. Improvement of various hazes classification based on the current result would provide the public as well as researchers with more accurate information of LH, UH, and YS, in terms of temporal scale, size, vertical distribution of aerosols, etc.
The present study investigated the hypophysial responsiveness in terms of GnRH induced LH and FSH release in cycling buffalo during the tropical summer and winter climatic conditions (seasons). Peripheral plasma LH and FSH levels were measured at 1 hour before and 6 hours subsequent to the administration of GnRH (1 ug/kg body weight) or saline on Day 14 of oestrous cycle in 2 groups of buffalo (n = 6 each) during summer and winter seasons. Although GnRH induced LH peak concentrations did not differ during the two seasons, time to attain LH peak concentration was shorter (p < 0.05) and the area under LH peak was 39% higher (p < 0.05) during winter season in comparison to summer season. However, season had no effect on GnRH induced peak FSH concentration, time to attain peak FSH concentration and the area under FSH peak. Pretreatment basal LH and FSH levels did not differ during the two seasons. The present study suggests that the summer season adversely affects the GnRH stimulated release of LH in buffalo.
This study was attempted to investigate the processes of regression of the corpus luteum and uterus after parturition in 2∼3 multiparous Korean native goats. The concentrations of LH, prolactin and progesterone in blood plasma of native goats were measured at 5 day intervals from 10 days prepartum to 35 days postpartum. The pregnancy corpus luteum from goats at Days(D) 1, 10, 20 and 30 days of postpartum were examined by light microscopy. Changes in the uterus fo goats were studies by macroscopic and light microscopic observations during the postpartum period. Mean concentrations of plasma LH were low after parturition and the levels of plasma LH were similar during late gestation and throughout the postpartum period. Mean plasma concentrations of prolactin were 0.30 0.06 and 0.38 0.13ng/ml at Day 5 and Day 10 prepartum, respectively, but PRL levels remained slightly high for 5 weeks after kidding. Mean levels of progesterone in plasma were 0.33 0.05ng/ml on Day 1 postpartum(P<0.01). Through light microscopic survey, pregnancy corpora lutea were quite degeneration by day 10 pospartum. Microscopic changes of CL regression consisted of cytoplasmic eosinophilia and vacuolation, and pyknosis and karyorrhexis of the nucleus of luteal cells. Vascular changes were distended at the periphery ofthe CL. From macroscopic measurements of the uterus, the uteri were returned to their initial non-pregnant stage within a period of 21 dyas after parturition. Following partuition the intercaruncular epithelium was reparied by 20 days. The uterine epithelium was partially recovered in the carucle by 30 days postpartum.
It has been shown that orexin has an inhibitory effect on gonadotropin secretions in non-ruminant animals. The goal of this study was to determine whether orexin affects LH, and FSH secretions in the camel, as a pseudo-ruminant animal, under different dietary energy content. Sixteen castrated camels were randomly divided into 4 groups. Animals in groups 1 and 2 were fed 100% and animals in groups 3 and 4 were fed 50% energy content in their diet for 20 days. After 20 days, animals in groups 1 and 3 received infusions of 1 $\mu{g}$ orexin and groups 2 and 4 received infusions of 2 $\mu{g}$ orexin into their jugular vein. Blood samples were collected from the jugular vein every 20 minutes from 4 h before the first infusion of orexin until 4 h after the last orexin infusion. Lower dietary energy intake and infusions of 2 $\mu{g}$ but not 1 $\mu{g}$ orexin significantly (p<0.01) decreased the mean plasma concentrations and pulse amplitudes of LH of the animals. Infusion of 1 and 2 $\mu{g}$ orexin did not change the secretions of LH of the animals fed NE. Different energy dietary intake and infusion of 1 and 2 $\mu{g}$ orexin did not change the mean plasma concentrations of FSH of the animals in all groups. Infusions of 1 and 2 $\mu{g}$ orexin significantly (p<0.01) decreased the glucose levels of animals fed LE but not in NE fed animals. Additionally, plasma glucose levels of the LE-fed animals in groups 3 and 4 were significantly (p<0.01) lower than those of the animals in groups 1 and 2 fed NE diet. The results of this experiment indicated that orexin may negatively affect LH and FSH in camels with negative energy balance, but not in those with positive energy balance.
Park C. S.;Sung N. D.;Kim C. H.;Jin D. I.;Choi Y. S.;Yi Y. J.
Reproductive and Developmental Biology
/
v.29
no.1
/
pp.25-30
/
2005
This study was carried out to investigate the effects of semen characteristics, frozen-thawed sperm viability and serum FSH, LH, estradiol-17β and testosterone concentrations between breeds and among seasons in boars. In all seasons, Yorkshire boars produced higher semen volume compared with Duroc boars, whereas sperm concentration did not differ significantly between Duroc and Yorkshire boars. Semen volume in spring was higher compared with summer, autumn and winter in both Duroc and Yorkshire boars, but sperm concentration did not differ significantly among seasons. Sperm motility and normal acrosome rate of frozen-thawed sperm produced in spring were higher than those in summer, autumn and winter in both Duroc and Yorkshire boars. Sperm motility of frozen-thawed sperm in Yorkshire boars was higher than that in Duroc boars regardless of seasons. However, normal acrosome rate did not differ significantly between Duroc and Yorkshire boars. Serum FSH concentration in Yorkshire boars was lower than that in Duroc boars in all seasons. However, there were no significant differences on serum FSH concentration of Duroc and Yorkshire boars among seasons. Serum LH and estradiol-17β concentrations did not differ significantly between Duroc and Yorkshire boars. Also, there were no significant differences in serum LH and estradiol-17β concentrations of Duroc and Yorkshire boars among seasons. Serum testosterone concentration in Yorkshire boars was higher than that in Duroc boars in all seasons. In both breeds, serum testosterone concentrations were higher in spring than in summer, autumn and winter. In conclusion, when serum FSH concentrations were low, semen volumes were high, and when serum testosterone concentrations were high, sperm motility and normal acrosome rate of frozen-thawed sperm were high.
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