Differences in Gene Expression in Preovulatory Follicles after the LH Surge in Dairy Heifers Versus Lactating Cows.

The molecular mechanisms of ovulation and luteinization have not been well established, partially due to lack of a comprehensive understanding of functionally significant genes up-regulated in response to an ovulatory stimulus and the signaling pathways involved. In the present study, transcripts increased in bovine preovulatory follicles following a GnRH-induced LH surge were identified using microarray technology. Increased expression of 368 and 878 genes was detected at 12 (368 genes) and 20 h (878 genes) following GnRH injection. The temporal, cell specific and prostanoid-dependent regulation of selected genes (ADAM10,DBI,CD36,MTSS1,TFG, andRABGAP1) identified from microarray studies and related genes (ADAM17andAREG) of potential significance were also investigated. Expression of mRNA forDBIandCD36was simultaneously up-regulated in theca and granulosa cells (GC) following the LH surge, whereas temporal regulation ofADAM10,MTSS1,TFG, andRABGAP1was distinct in the two cell compartments and increased granulosaTFGandRABGAP1mRNA were prostanoid dependent.AREGmRNA was increased in theca and GCs at 12 and 24 h following GnRH injection.ADAM17mRNA was increased in theca, but reduced in GCs 24 h following GnRH injection. The increasedADAM17andAREGmRNA were prostanoid dependent. ADAM10 and ADAM17 protein were increased specifically in the apex but not the base of preovulatory follicles and the increase in ADAM17 was prostanoid dependent. Results reveal novel information on the regulation of preovulatory gene expression and suggest a potential functional role for ADAM10 and ADAM17 proteins in the region of follicle rupture.

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Comparison of mRNA for IGFs and their binding proteins in the oviduct during the peri-oestrous period between dairy heifers and lactating cows

Reproduction ◽

10.1530/rep-11-0154 ◽

2011 ◽

Vol 142 (3) ◽

pp. 457-465 ◽

Cited By ~ 11

Author(s):

Theerawat Swangchan-Uthai ◽

Siobhan W Walsh ◽

Sarah L H Alexander ◽

Zhangrui Cheng ◽

Mark A Crowe ◽

...

Keyword(s):

Dairy Cows ◽

Early Embryo ◽

Dominant Follicle ◽

Embryo Survival ◽

Lh Surge ◽

Dairy Heifers ◽

Lactating Cows ◽

Holstein Friesian ◽

Igf Binding ◽

Igf Binding Protein

The oviduct provides the environment to support gamete maturation, fertilisation and early embryo development. As there is a high incidence of early embryonic death in lactating dairy cows, this study compared expression of IGF family members in the oviduct between lactating Holstein-Friesian dairy cows (n=16, 81±2.4 days in milk) and nulliparous heifers (n=16, age 1.6±0.07 years) at three stages of the oestrous cycle: A) newly selected dominant follicle in the luteal phase, B) follicular phase before the LH surge and C) pre-ovulatory phase after the LH surge. Expression ofIGF1,IGF2, IGF binding protein 2 (IGFBP2),IGFBP3andIGFBP6mRNA was determined in the ampulla of the oviduct. Oviduct side (ipsilateral or contralateral) with respect to the dominant follicle did not affect gene expression. Expression ofIGF1and all threeIGFBPsincreased significantly between the luteal and the pre-ovulatory phases, with no further significant alteration post-LH surge. Concentrations of circulating IGF1 were higher in heifers than in cows, as was the mRNA expression ofIGF1,IGFBP3andIGFBP6. The pre-LH surge rise inIGFBP2mRNA was only observed in heifers.IGF2expression was not influenced by either age or stage of cycle. These three IGFBPs are generally considered to inhibit IGF action. These results indicate tight regulation of IGF bioavailability in the oviductal environment around oestrus, with pronounced differences between cows and heifers, which are likely to influence early embryonic development. Further studies are required to assess the implications for embryo survival.

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The Increase in Signaling by Kisspeptin Neurons in the Preoptic Area and Associated Changes in Clock Gene Expression That Trigger the LH Surge in Female Rats Are Dependent on the Facilitatory Action of a Noradrenaline Input

Endocrinology ◽

10.1210/en.2015-1323 ◽

2016 ◽

Vol 157 (1) ◽

pp. 323-335 ◽

Cited By ~ 7

Author(s):

Bruna Kalil ◽

Aline B. Ribeiro ◽

Cristiane M. Leite ◽

Ernane T. Uchôa ◽

Ruither O. Carolino ◽

...

Keyword(s):

Gene Expression ◽

Median Eminence ◽

Preoptic Area ◽

Clock Genes ◽

Third Ventricle ◽

Female Rats ◽

Ovariectomized Rats ◽

Lh Surge ◽

Clock Gene Expression

Abstract In rodents, kisspeptin neurons in the rostral periventricular area of the third ventricle (RP3V) of the preoptic area are considered to provide a major stimulatory input to the GnRH neuronal network that is responsible for triggering the preovulatory LH surge. Noradrenaline (NA) is one of the main modulators of GnRH release, and NA fibers are found in close apposition to kisspeptin neurons in the RP3V. Our objective was to interrogate the role of NA signaling in the kisspeptin control of GnRH secretion during the estradiol induced LH surge in ovariectomized rats, using prazosin, an α1-adrenergic receptor antagonist. In control rats, the estradiol-induced LH surge at 17 hours was associated with a significant increase in GnRH and kisspeptin content in the median eminence with the increase in kisspeptin preceding that of GnRH and LH. Prazosin, administered 5 and 3 hours prior to the predicted time of the LH surge truncated the LH surge and abolished the rise in GnRH and kisspeptin in the median eminence. In the preoptic area, prazosin blocked the increases in Kiss1 gene expression and kisspeptin content in association with a disruption in the expression of the clock genes, Per1 and Bmal1. Together these findings demonstrate for the first time that NA modulates kisspeptin synthesis in the RP3V through the activation of α1-adrenergic receptors prior to the initiation of the LH surge and indicate a potential role of α1-adrenergic signaling in the circadian-controlled pathway timing of the preovulatory LH surge.

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Risk factors of S. aureus intramammary infection in pre partum dairy heifers under grazing conditions and molecular characterization of isolates from heifers and cows

Journal of Dairy Research ◽

10.1017/s0022029919001018 ◽

2020 ◽

Vol 87 (1) ◽

pp. 82-88

Author(s):

Cecilia M. Camussone ◽

Ana I. Molineri ◽

Marcelo L. Signorini ◽

Verónica E. Neder ◽

Carlos A. Vitulich ◽

...

Keyword(s):

Risk Factors ◽

Staphylococcus Aureus ◽

Management Practices ◽

Molecular Level ◽

Univariate Analysis ◽

Multivariable Analysis ◽

Cow Milk ◽

Intramammary Infection ◽

Dairy Heifers ◽

Lactating Cows

AbstractThe aims of the research reported here were to identify potential risk factors associated with the presence of Staphylococcus aureus intramammary infection (IMI) in pre partum dairy heifers on 17 dairy farms from three provinces of Argentina and to characterize, at molecular level, isolates from those heifers and lactating cows from two selected herds. A total of 1474 heifers and 4878 lactating cows were studied. The prevalence of Staphylococcus aureus IMI in the heifers, heifers at quarter level and lactating cow mammary quarters was 14.41, 4.82, and 14.65%, respectively. Univariate analysis showed the key variables associated with S. aureus IMI presence in the heifers were: S. aureus IMI prevalence in cows of the lactating herd, the time calves stayed with their dam after birth, the calf rearing system, the place of rearing (own farm or other dairy farm) and fly control on the farm. None of the variables included in the multivariable analysis was associated with the presence of S. aureus IMI in the pre partum heifers, probably due to low variability among management practices used by the farms for rearing the heifer calves. At the molecular level, S. aureus isolates were grouped into three main PFGE clusters and several genotypes within the clusters. Isolates from mammary secretion of pre partum heifers and milk of lactating cows comprised different PFGE clusters in both herds, although two exceptions occurred. The absence of gene fnbpB, which codifies for a virulence factor protein involved in cell invasion by S. aureus, was significantly more frequent in pre partum heifer secretion isolates than in isolates from lactating cow milk. These results suggest that, under these management conditions, isolates from mammary secretions of pre partum heifers do not originate from the milk of lactating cows, but rather other sources to which the heifer is exposed.

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198 PROGESTERONE, ESTROGEN (ER-α AND ER-β), AND OXYTOCIN RECEPTOR GENE EXPRESSION IN CANINE EMBRYOS

Reproduction Fertility and Development ◽

10.1071/rdv25n1ab198 ◽

2013 ◽

Vol 25 (1) ◽

pp. 248

Author(s):

A. A. P. Derussi ◽

A. C. S. Castilho ◽

R. W. A. Souza ◽

R. Volpato ◽

C. R. F. Guaitolini ◽

...

Keyword(s):

Gene Expression ◽

Relative Abundance ◽

Hormone Receptors ◽

Target Genes ◽

Receptor Gene ◽

Rna Extraction ◽

Amplification Efficiency ◽

Mrna Levels ◽

Lh Surge ◽

Total Rna

The aim of this study was to compare the mRNA levels of hormone receptor for progesterone (PR), oestrogen α (ER-α), oestrogen β (ER-β), and oxytocin (OTR) in canine morulae and blastocysts. Ten healthy mature bitches were inseminated based on monitoring vaginal cytology and progesterone concentration. The first insemination was performed on Day 2 after the preovulatory LH surge (progesterone 4 ng mL–1), and the second was performed 48 h later. All females were submitted to ovariohysterectomy (OVH), and the oviduct as well the uterurs were flushed with PBS solution to obtain the embryos. The females were divided into two groups: Group A (n = 5), morulae were collected 8 days after the LH surge and Group B (n = 5), blastocysts were collected 12 days after the LH surge. The pools (n = 10) of embryos (5 embryos/pool) were stored in RNAlater® (Ambion, Life Technologies, USA) at –80°C. The samples were analysed together. The RNA later was removed used PBS calcium free and the total RNA extraction was performed using the Qiagen RNeasy micro-kit (Hildesheim, Germany). Before reverse-transcription (RT) reaction, the total RNA was treated with DNase I Amplification Grade (Invitrogen Life Technologies, Carlsbad, CA, USA). The gene expression of target genes was assessed by real-time RT-qPCR, using SuperScript III for RT and power SYBR Green PCR Master Mix (Applied Biosystems, USA) for cDNA for PCR. The primers for target genes were designed using the software Primer Express® (Applied Biosystems, USA). The gene expression of target genes was normalized by HPRT gene and the relative abundance of mRNA was determined by the ΔΔct method corrected by amplification efficiency using Pffafl’s equation. The means of mRNA relative abundance were compared by t-test. The PR mRNA expression only in blastocysts is similar to the results obtained by Hou et al. (1997) in rat embryos. It is believed that the absence of PR in the early stages of cleavage is due to the indirect action of progesterone by growth factors produced by the maternal reproductive tract (2). Apparently, ER-β action does not occur in the embryo canine phases analysed; however, the action of ER-α seems related to the deployment signal as seen by Hou et al. (1996) in rats. Similarly to findings in the literature, OTR expression decreased in canine embryonic development. This receptor was produced by blastocysts while present in the uterus, which may represent an incidental mechanism to the embryo control of endometrial receptivity, such as also to prevent the development of endometrial luteolytic mechanism. The variation in hormone receptors gene expression in canine embryos can be influencing the transition from morula to blastocyst. In addition, a hormonal influence on these structures can occur in different ways.

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219 FOLLICLE AND HORMONE DYNAMICS IN SINGLE- v. DOUBLE-OVULATING HEIFERS

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab219 ◽

2010 ◽

Vol 22 (1) ◽

pp. 267

Author(s):

M. P. Palhao ◽

M. A. Beg ◽

M. T. Rodrigues ◽

R. R. Araújo ◽

J. H. M. Viana ◽

...

Keyword(s):

Present Experiment ◽

Group Effect ◽

Preovulatory Follicle ◽

Follicle Growth ◽

Blood Samples ◽

Lh Surge ◽

Follicular Wave ◽

Preovulatory Follicles ◽

Main Effect ◽

Plasma Hormone

The present experiment used the Day 4 ablation model for increasing the incidence of double ovulations in heifers. The objective was to compare follicle growth and plasma hormone concentrations associated with single v. double ovulations. Follicles ≥5 mm were ablated at 4 days post-ovulation to induce a prominent FSH surge and a new follicular wave, and 2 injections of PGF2 (12 h apart) were given 2 days later (Day 6) to favor ovulation. Beginning on Day 5, the 3 largest follicles of the induced wave were scanned twice a day until 36 h after the next ovulation. Blood samples were taken at 6-h intervals starting when the largest follicle reached ≥8.0 mm (expected deviation at 8.5 mm; Ginther et al. 1996) and continued until 36 h after the next ovulation. Concentrations of LH and FSH were measured by validated RIA for cattle (LH measured by Ginther et al. 1999; FSH measured by Adams et al. 1992) and concentrations of oestradiol measured by a commercially available RIA kit (Siddiqui et al. 2009). From a total of 31 heifers, 16 (52%) or 15 (48%) developed a single or more than 1 dominant (≥ 10 mm) follicle in the follicular wave after ablation, respectively. For heifers with 2 dominant follicles, the second-largest follicle ovulated in 9 (60%) heifers and the overall double ovulation rate was 29% (9/31). Follicle diameters and plasma hormone concentrations were compared between single ovulators (n = 12) and double ovulators (n = 8). Diameter of the preovulatory follicles did not increase between the LH peak and ovulation in either the single or double ovulations. In double ovulators, the interval from follicle deviation to the peak of the preovulatory LH surge was shorter (1.9 ± 0.2 days v. 2.5 ± 0.2 days; P < 0.02) and the diameter of the largest preovulatory follicle was smaller (12.2 ± 0.5 mm v. 13.3 ± 0.3 mm; P < 0.02) than in single ovulators, respectively. The LH concentrations of the preovulatory surge did not differ between single and double ovulators for 24 h on each side of the peak (main effect of hour only; P < 0.0001). When data were normalized to the LH peak, the peak of the preovulatory FSH and estradiol surges occurred in synchrony with the peak of LH surge for both groups. A group effect (P < 0.0001) for FSH resulted from a lower concentration averaged over hours in double ovulators. Estradiol showed a group by hour interaction (P < 0.008), reflecting greater concentrations in the double ovulators before and at peak. In conclusion, an increased Supported by the Eutheria Foundation, Cross Plains, WI, USA. Submission supported by FAPEMIG.

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Long-term effects of prior diets, dietary transition and pregnancy on adipose gene expression in dairy heifers

PLoS ONE ◽

10.1371/journal.pone.0218723 ◽

2019 ◽

Vol 14 (7) ◽

pp. e0218723

Author(s):

Hilde K. L. Wærp ◽

Sinéad M. Waters ◽

Matthew S. McCabe ◽

Paul Cormican ◽

Ragnar Salte

Keyword(s):

Gene Expression ◽

Long Term Effects ◽

Dairy Heifers ◽

Dietary Transition

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Decreased Inhibin Gene Expression in Preovulatory Follicles Requires Primary Gonadotropin Surges*

Endocrinology ◽

10.1210/endo-124-5-2193 ◽

1989 ◽

Vol 124 (5) ◽

pp. 2193-2199 ◽

Cited By ~ 38

Author(s):

TERESA K WOODRUFF ◽

JoBETH D'AGOSTINO ◽

NEENA B SCHWARTZ ◽

KELLY E MAYO

Keyword(s):

Gene Expression ◽

Preovulatory Follicles

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Energy and lipid metabolism gene expression of D18 embryos in dairy cows is related to dam physiological status

Physiological Genomics ◽

10.1152/physiolgenomics.00091.2013 ◽

2014 ◽

Vol 46 (2) ◽

pp. 39-56 ◽

Cited By ~ 22

Author(s):

D. Valour ◽

S. A. Degrelle ◽

A. A. Ponter ◽

C. Giraud-Delville ◽

E. Campion ◽

...

Keyword(s):

Gene Expression ◽

Lipid Synthesis ◽

Physiological Status ◽

Molecular Characteristics ◽

Morphological Development ◽

Continue Development ◽

Nonesterified Fatty Acids ◽

Lactating Cows ◽

Lipid Metabolism Gene ◽

Acetate Utilization

We analyzed the change in gene expression related to dam physiological status in day (D)18 embryos from growing heifers (GH), early lactating cows (ELC), and late lactating cows (LLC). Dam energy metabolism was characterized by measurement of circulating concentrations of insulin, glucose, IGF-1, nonesterified fatty acids, β-hydroxybutyrate, and urea before embryo flush. The metabolic parameters were related to differential gene expression in the extraembryonic tissues by correlation analysis. Embryo development estimated by measuring the length of the conceptuses and the proportion of expected D18 gastrulating stages was not different between the three groups of females. However, embryo metabolism was greatly affected by dam physiological status when we compared GH with ELC and GH with LLC but to a lesser extent when ELC was compared with LLC. Genes involved in glucose, pyruvate, and acetate utilization were upregulated in GH vs. ELC conceptuses (e.g., SLC2A1, PC, ACSS2, ACSS3). This was also true for the pentose pathway ( PGD, TKT), which is involved in synthesis of ribose precursors of RNA and DNA. The pathways involved in lipid synthesis were also upregulated in GH vs. ELC. Despite similar morphological development, the molecular characteristics of the heifers' embryos were consistently different from those of the cows. Most of these differences were strongly related to metabolic/hormone patterns before insemination and during conceptus free-life. Many biosynthetic pathways appeared to be more active in heifer embryos than in cow embryos, and consequently they seemed to be healthier, and this may be more conducive to continue development.

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