A synonymous single nucleotide polymorphism (g.36417726C > A) in the Lama2 gene influencing fat deposition is associated with post-partum anestrus interval in Murrah buffalo.

Postpartum absence of estrus exhibition known as postpartum anestrus interval (PPAI) for more than 90 days after calving is a concerning issue for dairy buffalo farmers' economy. The PPAI duration is influenced by both management practices and animal genetics. Investigating genetic markers associated with PPAI is crucial for incorporating them into marker-assisted selection programs. Towards this goal, our study focused on exploring potential genetic markers from early postpartum adipose tissue gene networks. We successfully identified 24 Single Nucleotide Polymorphisms (SNPs) within 9 candidate genes. In our initial analysis involving 100 buffaloes, we detected a significant association (P = 0.02267) between a specific synonymous SNP within the Lama2 gene (g.36417726C > A) and PPAI. This finding was subsequently validated (P = 0.02937) in a larger cohort of 415 buffaloes, where the SNP explained 1.36 % of the genetic variance. Intriguingly, buffaloes with the CC genotype of this SNP exhibited a PPAI that was 12.71 ± 3.21 days longer compared to buffaloes with AA and CA genotypes. To gain insight into the functional relevance of this SNP, a computational analysis was performed which indicated that the C allele of the SNP (g.36417726C > A) increased the stability of LAMA2 mRNA compared to the A allele. This computational prediction was corroborated by observing a significant increase (P = 0.01798) in Lama2 gene expression (greater than 8-fold) and higher fat percentage (P < 0.05) in adipose tissue of CC genotypes (48.78 ± 1.87 %) compared to AA genotypes (33.59 ± 4.5 %). Furthermore, we noted a significant (P < 0.05) upregulation of C/ebpß, Pparγ, Fasn, C/ebpα, and Pnpla2 genes, along with the downregulation of Bmp2 and Ptch1 in CC genotypes as opposed to AA genotypes. This observation suggests the involvement of the Pparγ-mediated pathway in both adipogenesis and lipolysis within CC genotypes. In summary, our comprehensive analysis involving association and functional validation underscores the potential of the SNP (g.36417726C > A) within the Lama2 gene as a promising genetic marker against extended PPAI in Murrah buffalo.

Seasonal absence of supplementary corpora lutea in pregnant mares and the relationship with pregnancy loss.

Two experiments were conducted in the Northern (UK) and Southern (Brazil) hemispheres to determine the effect of season (month of conception) on the development of supplementary CL (SCL) and the relationship with pregnancy loss. In experiment 1, 199 pregnancies were followed between Day 14 and term, to determine the number of SCL and pregnancy viability (Northern Hemisphere). From the 199 pregnancies, 178 were obtained from inseminations during the breeding season (March-September), while the rest, 21 pregnancies resulted from conceptions in the non-breeding season (October to February). Pregnancies conceived in the breeding season were more likely (P < 0.01) to have at least 1 SCL (75.8 %, 135/178) than pregnancies from the non-breeding season (33 %, 7/21). However, the pregnancy loss between Days 35 and 120 of pregnancy in mares with no SCL was similar (3.5 %, 2/57; P >0.1) than from mares with SCL (1.4 %, 2/142). In Experiment 2 (Southern Hemisphere), three groups of recipients were used based on their ovarian activity at the time of embryo transfer: Anestrus (n = 8), transitional (n = 7) and cyclic (n = 7) recipient mares. While all transitional and cyclic mares developed at least 1 SCL, only 50 % of anestrous recipients (4/8) developed SCL by 120 of gestation. In conclusion, the development of SCL in pregnant mares is influenced by the time of season of conception, therefore it appears to be regulated by the photoperiod and the endogenous seasonal variation in gonadotropin concentrations. Mares with no SCL were not at increased risk of pregnancy loss.

Progesterone Device Use Improves Ovum Pick-Up Efficiency in Acyclic Donors.

The aim of this study was to evaluate follicular dynamics and ovum pick-up (OPU) efficacy in untreated mares or mares treated with an intravaginal progesterone (P4) device during seasonal anestrus (acyclic) and during the breeding season (cyclic). Six mares (mean age = 5 years), were recruited into an ovum pick-up scheme that was performed every 14 days with and without the P4 device, during the acyclic and cyclic phases. Aspirations amounted to seven procedures with or without the P4 device during each phase. Five ultrasound assessments were performed at each interval between the OPUs. Data on follicular number and diameter as well as the numbers of recovered and the percentage of recovered oocytes were also collected. The number of follicles from mares in the acyclic phase was higher (P < .005) regardless of the treatment. However, the follicular diameter was smaller for the P4 group (P < .005) from the 2nd to the 5th evaluation post-OPU procedure. The percentage of oocytes recovered during the acyclic phase was higher for mares treated with the P4 device (P < .005). The P4 device resulted in follicles with smaller diameters and facilitated OPU efficacy.

Differentially expressed transcripts study during pregnancy and postpartum anestrus of yak (Bos grunniens).

Background: Yak is the main livestock species in the plateau area, and its reproductive performance is low, usually two years or three years. A very few of yaks recover within a certain period of time after delivery and smoothly enter the next estrous cycle, while most of them enter the postpartum anestrus and show no estrus performance. However, the key biological factors and influencing mechanisms that cause postpartum anestrus in yaks are not clear. Objective: To study the expression of differential transcripts in ovaries of yak during pregnancy and postpartum anestrus. Methods: Each three yaks in pregnancy and anestrus under natural grazing conditions in Haiyan County, Qinghai Province were selected and slaughtered, and their ovaries were collected and sent to Biomarker Technologies. Oxford Nanopore Technologies single-molecule real-time electrical signal sequencing technology was used to perform full-length transcriptome sequencing. Astalavista software was used to identify the types of alternative splicing events in yak estrus and pregnancy, and TAPIS pipeline was used to identify alternative polyadenylation. Results: The results showed that there were 1751 differentially expressed transcripts (DETs) between pregnancy and anestrus in yak, of which 808 were upregulated and 943 were downregulated. GO analysis showed that the biological processes of DETs were mainly reproductive, reproductive and rhythmic processes. KEGG analysis showed that the DET cell junction-related adhesion junction protein (ß-catenin) and amino terminal kinase (JNK) were involved in FAs (local adhesion). Phosphatidylinositol-3-kinase (PI3K) is involved in the PI3K/AKT/mTOR signaling pathway. Circadian rhythm output cycle failure (Clock) and brain and muscle tissue aromatic hydrocarbon receptor nuclear transporter-like protein 1 (Bmal1) are involved in circadian rhythm signaling pathway. Conclusion: This study found that ß-catenin, JNK, PI3K, Clock and Bmal1 were closely related to postpartum anestrus in yak.

Could the different estrus periods significantly affect body temperature on clinical examination in bitches?

Clinical examination is essential to make a differential diagnosis list, and body temperature measurement is an indispensable part of the clinical examination. Throughout the estrus cycle, body temperature may fluctuate high enough to affect the clinical examination results. Aim of this study, whether there were any differences between the different estrus periods, body parts (rectal, intravaginal and corneal), and methods (contact thermography and infrared thermography) in regard to temperature in bitches. The animal material of this study consisted of 61 healthy, non-pregnant, non-sportive bitches (2.78 ± 0.22 mean years of age; mean weight 23.81 ± 1.61kg) of different breeds. Corneal temperature measurement results were found the least constant and significant differences were observed between different estrus periods except in the anestrus period. Rectal temperature (RT) measurement results were the most constant. However, in ANOVA analysis, significant differences were observed between the different estrus periods, and the difference between the anestrus and diestrus periods was very close to 0.5 °C. Although, in the Bland-Altman plot analysis, the agreement was observed only between rectal and intravaginal temperature (p: 0.572, r2: 0.70), the limits of agreement were not clinically acceptable. In conclusion, neither corneal temperature measurement with infrared technology nor intravaginal temperature measurement should not be an alternative to RT measurement, and RT measurement results should evaluate carefully considering the estrus period.

Kisspeptin and lactational anestrus: Current understanding and future prospects.

Lactational anestrus, characterized by the suppression of pulsatile gonadotropin-releasing hormone (GnRH)/luteinizing hormone (LH) release, would be a strategic adaptation to ensure survival by avoiding pregnancy during lactation in mammals. In the present article, we first provide a current understanding of the central regulation of reproduction in mammals, i.e., a fundamental role of arcuate kisspeptin neurons in mammalian reproduction by driving GnRH/LH pulses. Second, we discuss the central mechanism inhibiting arcuate Kiss1 (encoding kisspeptin) expression and GnRH/LH pulses during lactation with a focus on suckling stimulus, negative energy balance due to milk production, and the role of circulating estrogen in rats. We also discuss upper regulators that control arcuate kisspeptin neurons in rats during the early and late lactation periods based on the findings obtained by a lactating rat model. Finally, we discuss potential reproductive technology for the improvement of reproductive performance in milking cows.

Reproductive performance of hair ewes and rams implanted with melatonin previous to the anestrus season in northwest Mexico.

Ovine reproductive behavior depends on annual photoperiodic cycle and its impact on endogenous melatonin secretion. In this regard, exogenous melatonin administration previous to the physiological anestrus period could modify the reproductive performance of sheep in the northwest of Mexico. Two independent studies were performed to evaluate such hypothesis in hair sheep implanted with melatonin prior to the anestrus season in latitudes 24° and 25° of Mexico. Study 1 involved 15 rams assigned to one of three treatments receiving 0mg (n=5), 18mg (n=5), or 36mg (n=5) of melatonin subcutaneously. Study variables were measured monthly since implantation (d0) and included testosterone concentration, scrotal circumference, mass motility, individual motility, and sperm concentration. Study 2 included 50 ewes assigned to one of two treatments receiving 0mg (n=25) or 18mg (n=25) of melatonin subcutaneously. In ewes, progesterone concentration and the frequency of females in anestrus were measured during the implantation (-30d), as well as at the beginning (0d) and at the end (45d) of the mating period, while pregnancy rate was determined by ultrasonography 45d after. Continuous variables were analyzed using a mixed effects model considering treatment, time, and the treatment by time interaction as fixed effects. Animal nested within treatment was the random effect. Binary variables were analyzed using the chi-square test. In males, melatonin improved testosterone and sperm concentrations (P<0.05), while in females, a 28% higher pregnancy rate was observed in implanted ewes (P<0.05). Therefore, melatonin enhanced reproductive parameters in both sexes and its exogenous administration previous to the anestrus season in northwest Mexico could be more effective in rams.

Group or individual housing does not reduce socio-sexual and reproductive responses in anestrous goats during the first contact with the photo-stimulated buck.

Herein, we examined the effect of the type of housing (individual vs group housing) of anestrus female goats on plasma cortisol concentrations and socio-sexual behaviors during first contact with photo-stimulated male goats and determined the impact on sexual and reproductive responses after 15 d of contact with photo-stimulated males. Therefore, after weaning, 10 females each were individually and group-housed, respectively. Ten bucks were rendered sexually active by exposure to artificially long days (16 h of light and 8 h of darkness per day) for 2.5 mo. During the naturally increasing photoperiod, 15-mo-old females were exposed and maintained with males. On day 0, during the first contact with males, individually- and group-housed females exhibited similar plasma cortisol concentrations (22.6 vs 27.4 ng/mL, respectively). Likewise, socio-sexual behaviors did not differ between examined groups on day 0. Moreover, the interval from first contact with males to the first estrus, short and normal ovulatory cycles, ovulation, fertility, and prolificacy rates were similar between differently housed females. Furthermore, mounting attempts did not differ between males in contact with either female group. In conclusion, individually- and group-housed anestrus females displayed elevated and similar plasma cortisol concentrations during first contact with photo-stimulated males and similar socio-sexual behaviors and reproductive responses when exposed to photo-stimulated males.

The post-ovulatory rise in progesterone is lower and the persistence of oestrous behaviour longer during the first compared with the second cycle of the breeding season in mares.

Mares are seasonally polyoestrous breeders. Therefore, the first ovulation of the season, following winter anoestrus, is the only cycle in which mares ovulate without the presence of an old CL from the previous cycle. The objective of this study was to compare the length of oestrous behaviour, and plasma progesterone concentrations during the early post-ovulatory period between mares after the first and second ovulation of the breeding season. Overall, 38 mares and 167 oestrous periods were used in the study. From those, 11 mares were used during the first and subsequent oestrous period to measure and compare the post-ovulatory rise in progesterone concentration, whereas all the mares were used to compare the length of the post-ovulatory oestrous behaviour between the first and subsequent cycles of the breeding season. The persistence of the post-ovulatory oestrus was longer (p < .001) following the first ovulation of the year (median of 52 h) compared with the subsequent ovulations (median of 36 h for second and later ovulations groups; n = 38 mares). The progesterone concentration at any of the four 8 h-intervals analysed (28, 36, 76 and 84 h post-ovulation) was lower (p < .01) following the first versus the second ovulation of the year. By 36 h post-ovulation the progesterone concentration of mares at the second ovulation of the year had passed the threshold of 2 ng/ml (2.1 ± 0.33 ng/ml), whereas in the first cycle it was 1.2 ± 0.13 ng/ml. In conclusion, mares had lower progesterone concentrations in their peripheral circulation and longer persistence of oestrous behaviour following the first ovulation of the year compared with the second and subsequent ovulatory periods of the breeding season.

Current status of assisted reproductive technologies in buffaloes.

Buffaloes are raised by small farm holders primarily as source of draft power owing to its resistance to hot climate, disease, and stress conditions. Over the years, transformation of these animals from draft to dairy was deliberately carried out through genetic improvement program leading to the development of buffalo-based enterprises. Buffalo production is now getting more attention and interest from buffalo raisers due to its socioeconomic impact as well as its contribution to propelling the livestock industry in many developing countries. Reproduction of buffaloes, however, is confronted with huge challenge and concern as being generally less efficient to reproduce compared with cattle due to both intrinsic and extrinsic factors such as poor estrus manifestation, silent heat, marked seasonal infertility, postpartum anestrus, long calving interval, delayed puberty, inherently low number of primordial follicles in their ovaries, high incidence of atresia, and apoptosis. Assisted reproductive technologies (ARTs) are major interventions for the efficient utilization of follicle reserve in buffaloes. The present review focuses on estrus and ovulation synchronization for fixed time artificial insemination, in vitro embryo production, intracytoplasmic sperm injection, cryopreservation of oocytes and embryos, somatic cell nuclear transfer, the factors affecting utilization in various ARTs, and future perspectives in buffaloes.

Maximizing the reproductive performances of anestrus dairy buffalo cows using GnRH analogue-loaded chitosan nanoparticles during the low breeding season.

The present study was designed to evaluate the efficiency of GnRH-loaded chitosan-TPP nanoparticles (GnRH-CsNPs) to improve the reproductive performance of anestrus buffaloes during the summer. Egyptian buffaloes (n = 50) were synchronized for estrus either with the ovsynch protocol (OVSs, n = 20) or with the modified ovsynch protocol (OVSm, n = 30) using GnRH-CsNPs. Sera samples were collected on days 0, 7, and 10 (insemination day); and days15 and 30 post insemination for estradiol (E2) and progesterone (P4) determination. The follicles were monitored on day 0 and day 10. CLs diameter and pregnancy diagnosis were scanned on day 30 post insemination. Using a half dose of GnRH as GnRH-CsNPs in the OVSm improved (P < 0.05) the number of total and large follicles, and the total surface area of subordinate and large follicles compared to the OVSs protocol. The OVSm increased (P < 0.05) the largest follicles diameter on day 10 and CLs diameter on day 30 post insemination compared to the OVSs protocol. E2 concentration was greater (P < 0.05) on insemination day, and P4 concentrations were higher (P < 0.05) on days 15 and 30 post insemination in the OVSm than in the OVSs. The estrus induction rate was 80 % higher (P < 0.05) in the OVSm and cows exhibited estrus within 17.54 h earlier than the OVSs (50 % within 42 h), respectively. The conception rate increased in OVSm compared to OVSs (75 vs. 40 %, respectively). In conclusion, nanofabrication of GnRH allowed to reduce hormonal dose to 50 % without any adverse effects on fertility, and improved ovarian activity and reproductive performance of anestrus buffaloes during the low breeding season.

Anti-Müllerian hormone concentrations in queens throughout the estrous cycle.

Studies in cats and dogs have proven the usefulness of anti-Müllerian hormone (AMH) as a diagnostic tool to determine the castration status or to diagnose ovarian remnant syndrome. Yet the secretion pattern of AMH over the estrous cycle in queens has not been investigated so far. Seven healthy sexually intact female cats were examined daily for signs of estrous behavior over a trial period of 4 months. Five queens showed regular estrous behavior, 1 queen was mated in her first heat and 1 queen never showed any signs of heat. To distinguish between inter-estrus and metestrus progesterone levels were determined. Serum samples for AMH and progesterone measurement were collected from the regular cycling queens in late anestrus, at several times during heat, inter-estrus and metestrus, from the mated queen during her first heat and during pregnancy, and in the acycling queen at various times during the trial period. The measured AMH values in anestrus were significantly (P < 0.05) higher than in heat (P < 0.001), metestrus (P = 0.12) and inter-estrus (P = 0.449). In anestrus the median AMH levels were 10.26 ng/ml (range 4.96 to 22.90 ng/ml), in heat 5.97 ng/ml (range 3.32- 22.96 ng/ml), in inter-estrus 10.47 (range 3.35-22.96 ng/ml) and in metestrus 6.38 ng/ml (range 4.50-10.75 ng/ml. The pregnant cat showed median AMH concentrations of 6.47 ng/ml (range 5.60-9.80 ng/ml) during her pregnancy. The acycling queen had solely low AMH values with a median concentration of 0.39 ng/ml. In conclusion there were high variations of the AMH levels among and within the individual cats and between heat cycles in the single cat. Remarkable high AMH concentrations were measured in the younger queens of the study in their first estrous cycles and also in anestrus, when less ovarian activity is expected. Further studies are necessary to emphasize the reasons for these high AMH concentrations especially in young queens.

Sexually active male goats stimulate the endocrine and sexual activities of other males in seasonal sexual rest through the "buck-to-buck effect".

Male goats rendered sexually active by exposure to a photoperiodic treatment are more efficient than untreated goats in stimulating LH secretion and ovulation in seasonally anestrous goats. This phenomenon is called the "male effect." Here, we determined whether sexually active bucks are able to stimulate the endocrine and sexual activities of other bucks in seasonal sexual rest through the phenomenon that we called the "buck-to-buck effect." We used bucks rendered sexually active (SA) during sexual rest by exposure to 2.5 mo of artificial long days (16 h of light per d) and untreated, sexually inactive (SI) bucks. In Experiment 1, we determined the short-term (21 d) LH and testosterone responses of sexually inactive bucks joined with a SA or SI buck. In Experiment 2, we determined the long-term (60 d) testosterone and sexual behavior responses of sexually inactive bucks joined with 2 SA or SI bucks. In Experiment 3, we determined the efficacy of bucks initially exposed to the buck-to-buck effect, the SABB bucks, to thereafter induce a "classical" male effect in seasonally anestrous goats. In Experiments 1 and 2, there was an interaction between time and groups in LH and testosterone plasma concentrations (P < 0.01). In Experiment 1, plasma LH concentrations were greater in bucks joined with a SA buck than in those joined with an SI buck (P < 0.05). In Experiments 1 and 2, testosterone concentrations were greater in bucks joined with SA bucks than in those joined with SI bucks (P < 0.05). In addition, in Experiment 2, the sexually inactive bucks joined with SA bucks displayed more nudging than those joined with SI bucks (P < 0.001). In Experiment 3, kidding rates did not differ between females joined with SA (34 of 40: 85%) or SABB bucks (32 of 40: 80%; P > 0.05). We concluded that the endocrine and sexual activities of bucks during sexual rest can be stimulated by SA bucks. In addition, SABB bucks are able to stimulate the reproductive activity of seasonally anestrous goats.

A single nucleotide polymorphism of the thyrotropin releasing hormone degrading ectoenzyme (TRHDE) gene is associated with post-partum anestrus in Murrah buffalo.

Thyrotropin releasing hormone degrading enzyme (TRHDE) gene is implicated in Thyrotropin releasing hormone (TRH) mediated prolactin secretion. It has been shown that the prolactin secretion alters the Gonadotropin-releasinghormone(GnRH) mediated estrous cycle. Therefore, TRHDE may also regulate postpartum anestrus. Earlier studies reported the role of non-synonymous single nucleotide polymorphism (SNPs) in various pathophysiological conditions by altering the structure and function of the proteins. Hence, in the present study, we identified SNPs in the putative promoter, first exon, middle exon and 3'-UTR containing the last exon of TRHDE gene and determined their association with postpartum anestrus (PPA) in Murrah buffaloes. We found one non synonymous SNP (G > C at 118095875 bp on chromosome 4) in the first exon of TRHDE and performed its association analysis in a population sample of 50 extreme PPA (residual PPAI: 123.06 ± 12.98 days) and 50 normal (residual PPAI: -80.46 ± 3.19 days) buffaloes. The residual PPAI value was the observed PPAI adjusted for the effect of 38 non-genetic factors. The analysis showed a significant (P < 0.004167) association of this SNP with PPA in buffaloes. Molecular dynamics simulations (MDS) also supported that the C allele altering Glutamine to Histidine at the amino acid 148 of TRHDE could enhance the stability and rigidity of TRHDE protein, which may lower its activity, increase TRH and prolactin, and reduce GnRH in PPA buffaloes. The MDS analysis further strengthens the association of the SNP (G > C) in the TRHDE gene with PPA condition in Murrah buffaloes. However, further investigation is needed to prove the MDS observations.

A customized long acting formulation of the kisspeptin analog C6 triggers ovulation in anestrus ewe.

The modulation of the kisspeptin system holds promise as a treatment for human reproductive disorders and for managing livestock breeding. The design of analogs has overcome some unfavorable properties of the endogenous ligands. However, for applications requiring a prolongation of drug activity, such as ovulation induction in the ewe during the non-breeding season, additional improvement is required. To this aim, we designed and tested three formulations containing the kisspeptin analog C6. Two were based on polymeric nanoparticles (NP1 and NP2) and the third was based on hydrogels composed of a mixture of cyclodextrin polymers and dextran grafted with alkyl side chains (MD/pCD). Only the MD/pCD formulation prolonged C6 activity, as shown by monitoring luteinizing hormone (LH) plasma concentration (elevation duration 23.4 ± 6.1, 13.7 ± 4.7 and 12.0 ± 2.4 h for MD/pCD, NP1 and NP2, respectively). When compared with the free C6 (15 nmol/ewe), the formulated (MD/pCD) doses of 10, 15 and 30 nmol/ewe, but not the 90 nmol/ewe dose, provided a more gradual release of C6 as shown by an attenuated LH release during the first 6 h post-treatment. When tested during the non-breeding season without progestogen priming, only, the formulated 30 nmol/ewe dose triggered ovulation (50% of ewes). Hence, we showed that a formulation with an adapted action time would improve the efficacy of C6 with respect to inducing ovulation during the non-breeding season. This result suggests that formulations containing a kisspeptin analog might find applications in the management of livestock reproduction but also point to the possibility of their use for the treatment of some human reproductive pathologies.

Meprin A1 subunit beta gene polymorphism is associated with the length of post-partum anestrus interval in Murrah buffaloes.

Postpartum anestrus interval (PPAI) is the interval between parturition and the first postpartum estrus exhibition in animals. Appearance of both normal and PPA buffaloes under the same farm conditions indicates the role of possible genetic predisposition to PPA. To identify the genetic and non-genetic factors associated with PPA in buffaloes, we collected data on PPAI and other 38 non-genetic variables from 575 Murrah buffaloes in the field conditions and identified the PPA associated non-genetic factors in our previous study. To explore the genetic factors associated with the unexplained variation in PPAI residuals, the present study identified 41 single nucleotide polymorphisms (SNPs) in 13 candidate genes using Sanger sequencing. Exploration of their association with the PPAI residuals of 50 extreme PPA and 50 normal buffaloes identified the significant (P < 0.01) association of the SNP (g.37219977A>G) in the 3'-UTR region of the Meprin A 1 subunit beta (Mep1b) gene with PPAI, which was further validated (P = 0.058) in a large population sample (n = 417). Bioinformatics analysis of the 3'-UTR region has identified three miRNA, bta-miR-2420, bta-miR-2325b and bta-miR-453 that could regulate Igf-1 in the plasma of animals with different genotypes (GG, AG and AA). The higher Igf-1 levels in the GG genotypes than that of AA and AG genotypes of this SNP (g.37219977A>G) further suggest the association of Mep1b gene with PPA condition in Murrah buffaloes. As a result of this study, we propose that buffaloes with protective alleles at this SNP be selected to improve the herd's reproductive efficiency.

The Influence of Intravaginal Gestagens Treatment on the Morphological Features and Endometrial Steroid Hormone Receptors Content during Anestrus Type II in Dairy Cattle.

BACKGROUND: Gestagens are the most widely used therapy in anestrus type II. The aim of this research is to evaluate the effectiveness of the vaginal progesterone inserts therapy in anestrus type II in cows. METHODS: The study was conducted on 33 cows. Progesterone (PR) and estrogen (ER) receptors expression in endometrium was assessed on a molecular level based on mRNA tissue expression. Additionally, blood 17ß-estradiol and progesterone levels were evaluated. RESULTS: A decrease in mRNA expression of A and B PR and ER α was noted in treated and untreated animals. In the treated group, an increase of ERß mRNA expression was observed, while a decreased was found in untreated animals. There was increased PR, ERα and ß expression in endometrial tissue in treated cows, and decreased expression of these factors in untreated cows. In the treated group, recurrence of ovarian cyclicity was noted in 52% of animals and pregnancy was obtained in 34.8% of them, while in the untreated group, recurrence did not occur. In the control group, spontaneous recurrence of ovarian cyclicity was not observed. An increase of PR expression was correlated with increased proliferation of endometrial cells. CONCLUSIONS: It seems likely that the endometrium is well developed and ready for placentation after removing the exogenous source of progesterone and preventing the recurrence of cyclicity of ovaries.

Transcriptome and proteome analysis of pregnancy and postpartum anoestrus ovaries in yak.

BACKGROUND: Domestic yaks are the most important livestock species on the Qinghai-Tibetan Plateau. Adult female yaks normally breed in the warm season (July to September) and enter anestrous in the cold season (November to April). Nevertheless, it is unclear how ovarian activity is regulated at the molecular level. OBJECTIVES: The peculiarities of yak reproduction were assessed to explore the molecular mechanism of postpartum anestrus ovaries in yaks after pregnancy and parturition. METHODS: Sixty female yaks with calves were observed under natural grazing in Haiyan County, Qinghai Province. Three yak ovaries in pregnancy and postpartum anestrus were collected. RNA sequencing and quantitative proteomics were employed to analyze the pregnancy and postpartum ovaries after hypothermia to identify the genes and proteins related to the postpartum ovarian cycle. RESULTS: The results revealed 841 differentially expressed genes during the postpartum hypoestrus cycle; 347 were up-regulated and 494 genes were down-regulated. Fifty-seven differential proteins were screened: 38 were up-regulated and 19 were down-regulated. The differential genes and proteins were related to the yak reproduction process, rhythm process, progesterone-mediated oocyte maturation, PI3K/AKT signaling pathway, and MAPK signaling pathway categories. CONCLUSIONS: Transcriptome and proteomic sequencing approaches were used to investigate postpartum anestrus and pregnancy ovaries in yaks. The results confirmed that BHLHE40, SF1IX1, FBPX1, HSPCA, LHCGR, BMP15, and ET-1R could affect postpartum hypoestrus and control the state of estrus.

MTNR1A gene polymorphisms and reproductive recovery after seasonal anoestrus in different Mediterranean sheep breeds.

The aims of this study were 1) to investigate the effect of MTNR1A gene polymorphisms on reproductive performance in ewes of one Italian and two Slovenian dairy sheep breeds (Sarda, Istrian Premenka and Boska, respectively) which were located at different latitudes, and 2) to highlight if the different season of the male placement with females that was utilized in the different breeding systems in Sardinia (Italy) and Slovenia resulted in different effects of these polymorphisms on reproductive functions. Reproductively mature ewes (n = 100) from each breed were utilized to conduct the study. To evaluate the reproductive efficiency, lambing dates and number of lambs born were recorded per ewe; additionally, the duration in days from ram placement with ewes to lambing (DRPEL), litter size and the fertility rate were determined based on lambing dates. In each breed, there were eight nucleotide variations within the MTNR1A gene exon II, two of which (g.17355358 and g.17355171), respectively, resulted in a valine to isoleucine, and alanine to aspartic acid substitution, in amino acid sequence. The SNPs at position g.17355452 and g.17355458 were determined to have effects on reproductive performance. Genotypes C/C and C/T at g.17355452 in Bovska and Sarda and genotype A/A at g.17355458 in Istrian Pramenka were associated with a greater fertility and a lesser duration in days from ram placement with ewes to lambing. These findings confirmed that the nucleotide sequences of the MTNR1A gene could affect reproductive functions of Mediterranean sheep.

Transcriptome Analysis of Neuroendocrine Regulation of Ovine Hypothalamus-Pituitary-Ovary Axis during Ovine Anestrus and the Breeding Season.

Most sheep are seasonal estrus, and they breed in autumn when the days get shorter. Seasonal estrus is an important factor that affects the productivity and fertility of sheep. The key point to solve this problem is to explore the regulation mechanism of estrus in sheep. Therefore, in this study, transcriptomic sequencing technology was used to identify differentially expressed mRNAs in the hypothalamus, pituitary and ovary of Small Tail Han sheep (year-round estrus) and tan sheep (seasonal estrus) among luteal, proestrus and estrus stages. There were 256,923,304,156 mRNAs being identified in the hypothalamus, pituitary and ovary, respectively. Functional analysis showed that the photosensor, leucine and isoleucine biosynthesis pathways were enriched significantly. It is speculated that photoperiod may initiate estrus by stimulating the corresponding pathways in hypothalamus. ODC1, PRLH, CRYBB2, SMAD5, OPN1SW, TPH1 are believed to be key genes involved in the estrogen process. In conclusion, this study expanded the database of indigenous sheep breeds, and also provided new candidate genes for future genetic and molecular studies on the seasonal estrus trait in sheep.