Conception Rate and Embryo Development in Guinea Pigs with Synchronized Estrus Induced by Progesterone Implant

Abstract Sire conception rate (SCR) is a field measure of fertility among bulls, but it can be influenced by several factors (Sperm transport, sperm-egg binding, early embryo development, etc). The objective of this study was to evaluate the relationship between SCR, sperm motility, SERPINA5 concentrations, and in vitro embryo development. Measurements were performed in 19 bulls with SCR values ranging from -7.7 to 4.45. For each bull, an aliquot of frozen-thawed semen was used for analyses of total (TMOT) and progressive (PROG) motility. Remaining semen was fixed with 2% formaldehyde, and concentration of SERPINA5 was determined by immunolocalization (antibody SERPINA5/Dylight405; PA5-79976-Invitrogen / ab201798-Abcam). Mean fluorescence intensity was determined in ~200 sperm heads/bull. Approximately 149 oocytes/bull were fertilized in vitro for embryo development analysis (cleavage and blastocyst rates). Statistical procedures were performed in SAS (9.4) using the procedures CORR for correlations (SCR, TMOT, PROG, SERPINA5, cleavage and blastocyst) and GLIMMIX for comparison of “field-fertility” (SCR divided in HIGH or LOW) and “field-embryo-fertility” (LOW-SCR sires were divided based on blastocyst rate (HIGH or LOW) resulting in two classifications; LOW-HIGH≥31% and LOW-LOW≤26%, respectively). There were positive correlations (P < 0.05) between cleavage-blastocyst (r=0.50), SERPINA5-cleavage (r=0.48), and TMOT-PROG (r=0.76). Sire SCR was not associated with SERPINA5, TMOT, PROG, cleavage and blastocyst rate (P > 0.52). Among LOW-SCR sires, LOW-LOW sires (-4.83±0.60) tended to have a better SCR score than LOW-HIGH (-6.18±0.42) sires (P = 0.08), but there were no differences (P > 0.43) between LOW-HIGH, LOW-LOW, and HIGH sires for SERPINA5, TMOT, PROG, and cleavage. In conclusion, some LOW SCR sires have good embryo development indicating a different mechanism for their low SCR; however, these differences in SCR could not be explained by TMOT, PROG, SERPINA5, cleavage and blastocyst. There were, however, positive correlations between cleavage-blastocyst rate, and SERPINA5-cleavage rate.

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63 Effect of sire conception rate on bovine early embryo development

Reproduction Fertility and Development ◽

10.1071/rdv33n2ab63 ◽

2021 ◽

Vol 33 (2) ◽

pp. 138

Author(s):

K. Clark ◽

J. N. Drum ◽

J. A. Rizo ◽

M. S. Ortega

Keyword(s):

Embryo Development ◽

Gradient Centrifugation ◽

Blastocyst Stage ◽

Early Embryo ◽

Conception Rate ◽

Cell Stage ◽

Fluorescent Intensity ◽

Early Embryo Development ◽

High Performing ◽

Sire Conception Rate

Currently, the only measure of sire fertility in the bovine is sire conception rate (SCR), which is determined by Day 70 pregnancy diagnosis and not reflective of early embryo development. Therefore, this study aimed to establish the relationship between SCR and early embryo development. In the first experiment, 65 sires of negative (<−1, n=25), average (−1 to 1, n=19), and high (> +1, n=21) SCR were characterised for their ability to produce embryos using an invitro embryo production (IVP) system. For each sire, 100 cumulus–oocyte complexes (COCs) were used. COCs were matured for 22h, fertilized by co-incubation with sperm selected from density gradient centrifugation for 18h, and then placed in culture medium. A sire of known IVP performance was used as a control in each run. Cleavage and blastocyst rates (BL) were measured on Days 3 and 8 post-insemination, respectively. Photographs were taken on Days 3, 5, and 8 to identify arrest stages of non-blastocyst embryos. Sires were ranked based on their blastocyst rate and grouped into quartiles for statistical analysis. Differences in BL were determined by ANOVA using sire, IVP run, and a sire×IVP run interaction. In addition, the correlation between SCR and BL was determined. All data were analysed using SAS software version 9.4 (SAS Institute Inc.). Mean BL between each quartile was significant (P<0.05), with rates ranging from 8 to 22% and 32 to 62% for the lowest and highest quartile, respectively. There was no correlation (P=0.90) between SCR and BL. Arrest stage was measured by subtracting the number of Day-8 blastocysts from, first, embryos that were morulas on Day 5, and then embryos that were 8- to 16-cell stage embryos on Day 5. This method is based on the assumption that embryos closer to the blastocyst stage on Day 5 are more likely to contribute to the Day 8 blastocyst population. The most frequent arrest stage was the 4- to 6-cell stage (39/52 sires). It has been shown that decreased rates of autophagy are associated with embryonic arrest at the 4- to 8-cell stage in humans, leading us to investigate this mechanism in the second experiment. Select high (n=3) and low (n=4) performing sires identified in experiment 1 were used to generate 4- to 6-cell embryos, and autophagy rates were measured using live immunofluorescence with CYTO-ID autophagy dye (n=20 embryos/sire). The mean fluorescent intensity of each embryo was divided by the number of cells within the embryo. Differences in autophagy between high and low sires were determined by ANOVA using SAS. Interestingly, low-performing sires had a significantly higher autophagy rates than high-performing sires (77.8±3.1 vs. 50.0±3.5). This could indicate that embryos produced with low-performing sires had higher levels of stress than their counterparts. In summary, the effect of sire on embryonic development seems to be independent of the SCR classification. The most common arrest stage observed is the 4- to 6-cell stage, right before embryonic genome activation. Further research is required to elucidate the mechanisms by which sires influence pre-implantation development. This research was supported by USDA-NIFA AFRI Competitive Grant No. 2019-67015-28998.

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INSEMINATION OF THE GUINEA-PIG BY INTRAPERITONEAL INJECTION

Journal of Endocrinology ◽

10.1677/joe.0.0160098 ◽

1957 ◽

Vol 16 (1) ◽

pp. 98-106 ◽

Cited By ~ 25

Author(s):

I. W. ROWLANDS

Keyword(s):

Guinea Pig ◽

Guinea Pigs ◽

Intraperitoneal Injection ◽

Narrow Range ◽

Fertilization Rate ◽

In Utero ◽

Conception Rate ◽

Temporal Relation ◽

Foetal Development

SUMMARY Intraperitoneal injection has been shown to be a method of insemination which produces a high conception rate in guinea-pigs. The proportion that become pregnant increases from 0 to 100% by raising the number of spermatozoa inseminated over the comparatively narrow range of 3·0 × 107-5·0 × 107. The conception rate is high in animals inseminated between 0 and 16 hr after the end of oestrus; all animals (17) inseminated with more than 5·0 × 107 spermatozoa during the first half of this period became pregnant. The incidence of pregnancy is very low in animals inseminated intraperitoneally during oestrus and at 18 hr or more after oestrus. The temporal relation between successful insemination and ovulation is the reverse of that obtaining in mated animals or after intravaginal insemination [Blandau & Young, 1939]. Fertility, determined by an estimate of the fertilization rate, by the number of foetuses in utero and the rate of foetal development is similar to that following mating. Reasons for the failure of fertilization in animals intraperitoneally inseminated during oestrus are discussed.

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Investigation of the relaxant effects of propofol on ovalbumin-induced asthma in guinea pigs

European Journal of Anaesthesiology ◽

10.1017/s0265021506000415 ◽

2006 ◽

pp. 1

Author(s):

I. Bagcivan ◽

O. Cevit ◽

M. K. Yildirim ◽

S. Gursoy ◽

S. Yildirim ◽

...

Keyword(s):

Guinea Pigs

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Hepatocyte Alterations in Guinea Pigs FED Polychlorinated Biphenyls (PCBs), Dibenzodioxins (PCDD), AND DIBENZOFURANS (PCDF)

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100053085 ◽

1982 ◽

Vol 40 ◽

pp. 56-57

Author(s):

J. N. Turner ◽

D. N. Collins

Keyword(s):

Guinea Pigs ◽

Room Temperature ◽

Dose Group ◽

Methyl Cellulose ◽

Uranyl Acetate ◽

Target Organ ◽

Office Building ◽

Lead Citrate ◽

Control Groups ◽

Cooling Fluid

A fire involving an electric service transformer and its cooling fluid, a mixture of PCBs and chlorinated benzenes, contaminated an office building with a fine soot. Chemical analysis showed PCDDs and PCDFs including the highly toxic tetra isomers. Guinea pigs were chosen as an experimental animal to test the soot's toxicity because of their sensitivity to these compounds, and the liver was examined because it is a target organ. The soot was suspended in 0.75% methyl cellulose and administered in a single dose by gavage at levels of 1,10,100, and 500mgm soot/kgm body weight. Each dose group was composed of 6 males and 6 females. Control groups included 12 (6 male, 6 female) animals fed activated carbon in methyl cellulose, 6 males fed methyl cellulose, and 16 males and 10 females untreated. The guinea pigs were sacrificed at 42 days by suffocation in CO2. Liver samples were immediately immersed and minced in 2% gluteraldehyde in cacadylate buffer at pH 7.4 and 4°C. After overnight fixation, samples were postfixed in 1% OsO4 in cacodylate for 1 hr at room temperature, embedded in epon, sectioned and stained with uranyl acetate and lead citrate.

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Glycogen in guinea pig neutrophils: Ultrastructural study of neutrophils at the intradermal site of BCG injection

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100077207 ◽

1983 ◽

Vol 41 ◽

pp. 726-727

Author(s):

Corazon D. Bucana

Keyword(s):

Bone Marrow ◽

Guinea Pig ◽

Electron Density ◽

Peritoneal Cavity ◽

Ultrastructural Study ◽

Guinea Pigs ◽

Random Distribution ◽

Glycogen Content ◽

Polymorphonuclear Neutrophils ◽

Ultrastructural Studies

In the circulating blood of man and guinea pigs, glycogen occurs primarily in polymorphonuclear neutrophils and platelets. The amount of glycogen in neutrophils increases with time after the cells leave the bone marrow, and the distribution of glycogen in neutrophils changes from an apparently random distribution to large clumps when these cells move out of the circulation to the site of inflammation in the peritoneal cavity. The objective of this study was to further investigate changes in glycogen content and distribution in neutrophils. I chose an intradermal site because it allows study of neutrophils at various stages of extravasation.Initially, osmium ferrocyanide and osmium ferricyanide were used to fix glycogen in the neutrophils for ultrastructural studies. My findings confirmed previous reports that showed that glycogen is well preserved by both these fixatives and that osmium ferricyanide protects glycogen from solubilization by uranyl acetate.I found that osmium ferrocyanide similarly protected glycogen. My studies showed, however, that the electron density of mitochondria and other cytoplasmic organelles was lower in samples fixed with osmium ferrocyanide than in samples fixed with osmium ferricyanide.

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Densitometric Identification of Primitive Erythroblasts After Reaction With Diaminobenzidine

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100072083 ◽

1973 ◽

Vol 31 ◽

pp. 328-329

Author(s):

John A. Trotter

Keyword(s):

Red Blood Cells ◽

Analytical Method ◽

Blood Cells ◽

Guinea Pigs ◽

Specific Protein ◽

Visual Examination ◽

Hemoglobin Synthesis ◽

Peroxidatic Activity ◽

Small Density

Hemoglobin is the specific protein of red blood cells. Those cells in which hemoglobin synthesis is initiated are the earliest cells that can presently be considered to be committed to erythropoiesis. In order to identify such early cells electron microscopically, we have made use of the peroxidatic activity of hemoglobin by reacting the marrow of erythropoietically stimulated guinea pigs with diaminobenzidine (DAB). The reaction product appeared as a diffuse and amorphous electron opacity throughout the cytoplasm of reactive cells. The detection of small density increases of such a diffuse nature required an analytical method more sensitive and reliable than the visual examination of micrographs. A procedure was therefore devised for the evaluation of micrographs (negatives) with a densitometer (Weston Photographic Analyzer).

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Annulate lamellae in the Sertoli cells of guinea pig testis after unilateral torsion of the spermatic cord

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100111094 ◽

1984 ◽

Vol 42 ◽

pp. 196-197

Author(s):

J. Chakraborty ◽

A. P. Sinha Hikim ◽

J. S. Jhunjhunwala

Keyword(s):

Sertoli Cells ◽

Guinea Pigs ◽

Spermatic Cord ◽

Present Report ◽

Cell Types ◽

Annulate Lamellae ◽

Spermatogenic Cells ◽

Electron Microscopic ◽

Structural Details ◽

First Time

Although the presence of annulate lamellae was noted in many cell types, including the rat spermatogenic cells, this structure was never reported in the Sertoli cells of any rodent species. The present report is based on a part of our project on the effect of torsion of the spermatic cord to the contralateral testis. This paper describes for the first time, the fine structural details of the annulate lamellae in the Sertoli cells of damaged testis from guinea pigs.One side of the spermatic cord of each of six Hartly strain adult guinea pigs was surgically twisted (540°) under pentobarbital anesthesia (1). Four months after induction of torsion, animals were sacrificed, testes were excised and processed for the light and electron microscopic investigations. In the damaged testis, the majority of seminiferous tubule contained a layer of Sertoli cells with occasional spermatogonia (Fig. 1). Nuclei of these Sertoli cells were highly pleomorphic and contained small chromatinic clumps adjacent to the inner aspect of the nuclear envelope (Fig. 2).

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Alterations in Ultrastructure of Skeletal Muscle From Newborn Guinea Pigs Following in Utero Exposure to Ethanol

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100120126 ◽

1985 ◽

Vol 43 ◽

pp. 686-687

Author(s):

C. Uphoff ◽

C. Nyquist-Battie ◽

T.B. Cole

Keyword(s):

Skeletal Muscle ◽

Guinea Pigs ◽

Muscle Development ◽

In Utero ◽

Ethanol Exposure ◽

Prenatally Exposed ◽

Chronic Alcoholic ◽

Test Kit ◽

Food And Water Intake ◽

Post Intubation

Ultrastructural alterations of skeletal muscle have been observed in adult chronic alcoholic patients. However, no such study has been performed on individuals prenatally exposed to ethanol. In order to determine if ethanol exposure in utero in the latter stages of muscle development was deleterious, skeletal muscle was obtained from newborn guinea pigs treated in the following manner. Six Hartly strain pregnant guinea pigs were randomly assigned to either the ethanol or the pair-intubated groups. Twice daily the 3 ethanol-treated animals were intubated with Ensure (Ross Laboratories) liquid diet containing 30% ethanol (6g/Kg pre-pregnant body weight per day) from day 35 of gestation until parturition at day 70±1 day. Serum ethanol levels were determined at 1 hour post-intubation by the Sigma alcohol test kit. For pair-intubation the Ensure diet contained sucrose substituted isocalorically for ethanol. Both food and water intake were monitored.

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The effect of lead exposure on DCX positive cells in layer II of adult guinea pigs neocortex

Cell Biology International ◽

10.1042/cbi034s018d ◽

2010 ◽

Vol 34 (8) ◽

pp. S18-S18

Author(s):

Kun Xiong ◽

Kai Huang ◽

Lei Shang ◽

Hui Wang ◽

Xiao‑xin Yan ◽

...

Keyword(s):

Lead Exposure ◽

Guinea Pigs

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