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Lipopolysaccharide Endotoxin Exposure on Ovarian Follicle Immune Response, Steroidogenesis, Oocyte Competence, and Early Embryonic Development.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Lipopolysaccharide Endotoxin Exposure on Ovarian Follicle Immune Response, Steroidogenesis, Oocyte Competence, and Early Embryonic Development./
Author:	Castro, Briza.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2023,
Description:	116 p.
Notes:	Source: Masters Abstracts International, Volume: 85-03.
Contained By:	Masters Abstracts International85-03.
Subject:	Animal sciences. -
Online resource:	https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=30571066
ISBN:	9798380162920

Lipopolysaccharide Endotoxin Exposure on Ovarian Follicle Immune Response, Steroidogenesis, Oocyte Competence, and Early Embryonic Development.
Castro, Briza.

Lipopolysaccharide Endotoxin Exposure on Ovarian Follicle Immune Response, Steroidogenesis, Oocyte Competence, and Early Embryonic Development. - Ann Arbor : ProQuest Dissertations & Theses, 2023 - 116 p.

Source: Masters Abstracts International, Volume: 85-03.

Thesis (M.S.)--New Mexico State University, 2023.

Reproductive efficiency is necessary for sustainability and profitability of beef cattle operations. Bacterial disease contributes to impaired reproductive efficiency through the release of the lipopolysaccharide (LPS) endotoxin into circulation leading to accumulation within the ovarian follicular environment. Presence of LPS in follicular fluid is a contributor to inadequate reproductive parameters. The deleterious effect of high LPS concentrations on ovarian function following acute bacterial mastitis and metritis has been evaluated in dairy cattle. However, limited information is available regarding impacts of low LPS concentrations resulting from subacute or non-detectable disease states on ovarian physiology. Therefore, the experiments herein investigated the effects of varying low-dose LPS concentrations on intrafollicular cell{A0}growth dynamics, immune responses, and steroidogenesis, as well as, subsequent oocyte competence, and early embryonic development. To evaluate exposure of granulosa cells to LPS, a granulosa cell line (KGN) was cultured with vehicle control or increasing doses of LPS (0.0001, 0.001, 0.01, 0.1, 1, and 10 {phono}{aelig}g/mL LPS) for 48 h in the presence or absence of FSH stimulation. Cell confluency and viability significantly decreased when cultured with 1 and 10 {phono}{aelig}g/mL LPS alone compared with vehicle-treated controls throughout 48 h incubation period. Conversely, no differences were detected among LPS-treated cells when concurrently incubated with FSH when evaluated over 48 h. Granulosa cells cultured with increasing doses of LPS in the presence of FSH demonstrated a linear increase in steroidogenic acute regulatory protein. However, no detectable changes were demonstrated in mRNA gene expression of P450 sidechain cleavage, beta-catenin, or aromatase gene expression. Linear immune responses were observed for increased pro-inflammatory cytokine IL-6, IL-8, and MCP-1 gene expression with increasing LPS doses. However, NF-kB remained unaffected by LPS. A tendency for linear progesterone (P4) decrease was observed in non-FSH stimulated cells, however, FSH-stimulated granulosa cells did not demonstrate linear differences among treatments. Similarly, a linear estradiol (E2) decrease was observed in non-FSH stimulated cells. To further elucidate the effects of low-dose LPS on oocyte competence and embryo development, a subsequent study evaluated bovine oocytes collected from slaughterhouse-derived ovaries and matured with vehicle-control or increasing doses of LPS (0.01, 0.1, and 1 {phono}{aelig}g/mL) for 21 h and evaluated for nuclear maturation. A set of embryos from LPS-matured oocytes were cultured to evaluate cleavage rates at d 3 and blastocyst rates at d 8 along with blastocyst cell counts. A subset of LPS-matured oocytes was fertilized and cultured for time-lapse image capture and analysis of embryo development. Results indicate no significant treatment differences among treatment groups in{A0}percent of oocytes at germinal vesicle (GV), germinal vesicle breakdown (GVBD), meiosis I (MI), or metaphase II (MII). Similarly, treatment differences were not observed in cleavage rates, or blastocyst rates evaluated via traditional microscopy. Treatment with LPS did not affect inner-cell mass or trophectoderm cell counts. Time-lapse embryo evaluation demonstrated no differences among vehicle-treated controls or LPS-matured oocytes in number of zygotes that did not cleave after fertilization or those that cleaved but arrested at the 2-cell stage, 4-cell, prior to morula, or morula stage. However, the percentage of blastocysts derived from oocytes matured in 0.01 {phono}{aelig}g/mL LPS tended to decrease compared to vehicle-treated controls. Comparably, the proportion of embryos that developed to the blastocyst stage was greater in vehicle-treated controls compared with embryos derived from oocytes matured in 0.1 and 1 {phono}{aelig}g/mL LPS. Together, these data elucidate the ability of granulosa cells to respond to varying doses of LPS in a dose-dependent manner by initiating a local immune response within the follicle ultimately leading to increased pro-inflammatory cytokine production. Furthermore, FSH has the ability to protect granulosa cell survival against LPS. Oocytes matured in low LPS doses that subsequently underwent in vitro fertilization, experienced decreased competence to develop to the blastocyst stage when developmental events were evaluated utilizing real time-lapse microscopy. Thus, bacterial endotoxin within the ovarian follicle imparts local immune responses in granulosa cells and the follicular environment, contributing to decreased reproductive efficiency and resulting in impacted oocyte quality and subsequent embryo development.

ISBN: 9798380162920Subjects--Topical Terms:

3174829
Animal sciences.
Subjects--Index Terms:

Embryonic development

Lipopolysaccharide Endotoxin Exposure on Ovarian Follicle Immune Response, Steroidogenesis, Oocyte Competence, and Early Embryonic Development.
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520 $a Reproductive efficiency is necessary for sustainability and profitability of beef cattle operations. Bacterial disease contributes to impaired reproductive efficiency through the release of the lipopolysaccharide (LPS) endotoxin into circulation leading to accumulation within the ovarian follicular environment. Presence of LPS in follicular fluid is a contributor to inadequate reproductive parameters. The deleterious effect of high LPS concentrations on ovarian function following acute bacterial mastitis and metritis has been evaluated in dairy cattle. However, limited information is available regarding impacts of low LPS concentrations resulting from subacute or non-detectable disease states on ovarian physiology. Therefore, the experiments herein investigated the effects of varying low-dose LPS concentrations on intrafollicular cell{A0}growth dynamics, immune responses, and steroidogenesis, as well as, subsequent oocyte competence, and early embryonic development. To evaluate exposure of granulosa cells to LPS, a granulosa cell line (KGN) was cultured with vehicle control or increasing doses of LPS (0.0001, 0.001, 0.01, 0.1, 1, and 10 {phono}{aelig}g/mL LPS) for 48 h in the presence or absence of FSH stimulation. Cell confluency and viability significantly decreased when cultured with 1 and 10 {phono}{aelig}g/mL LPS alone compared with vehicle-treated controls throughout 48 h incubation period. Conversely, no differences were detected among LPS-treated cells when concurrently incubated with FSH when evaluated over 48 h. Granulosa cells cultured with increasing doses of LPS in the presence of FSH demonstrated a linear increase in steroidogenic acute regulatory protein. However, no detectable changes were demonstrated in mRNA gene expression of P450 sidechain cleavage, beta-catenin, or aromatase gene expression. Linear immune responses were observed for increased pro-inflammatory cytokine IL-6, IL-8, and MCP-1 gene expression with increasing LPS doses. However, NF-kB remained unaffected by LPS. A tendency for linear progesterone (P4) decrease was observed in non-FSH stimulated cells, however, FSH-stimulated granulosa cells did not demonstrate linear differences among treatments. Similarly, a linear estradiol (E2) decrease was observed in non-FSH stimulated cells. To further elucidate the effects of low-dose LPS on oocyte competence and embryo development, a subsequent study evaluated bovine oocytes collected from slaughterhouse-derived ovaries and matured with vehicle-control or increasing doses of LPS (0.01, 0.1, and 1 {phono}{aelig}g/mL) for 21 h and evaluated for nuclear maturation. A set of embryos from LPS-matured oocytes were cultured to evaluate cleavage rates at d 3 and blastocyst rates at d 8 along with blastocyst cell counts. A subset of LPS-matured oocytes was fertilized and cultured for time-lapse image capture and analysis of embryo development. Results indicate no significant treatment differences among treatment groups in{A0}percent of oocytes at germinal vesicle (GV), germinal vesicle breakdown (GVBD), meiosis I (MI), or metaphase II (MII). Similarly, treatment differences were not observed in cleavage rates, or blastocyst rates evaluated via traditional microscopy. Treatment with LPS did not affect inner-cell mass or trophectoderm cell counts. Time-lapse embryo evaluation demonstrated no differences among vehicle-treated controls or LPS-matured oocytes in number of zygotes that did not cleave after fertilization or those that cleaved but arrested at the 2-cell stage, 4-cell, prior to morula, or morula stage. However, the percentage of blastocysts derived from oocytes matured in 0.01 {phono}{aelig}g/mL LPS tended to decrease compared to vehicle-treated controls. Comparably, the proportion of embryos that developed to the blastocyst stage was greater in vehicle-treated controls compared with embryos derived from oocytes matured in 0.1 and 1 {phono}{aelig}g/mL LPS. Together, these data elucidate the ability of granulosa cells to respond to varying doses of LPS in a dose-dependent manner by initiating a local immune response within the follicle ultimately leading to increased pro-inflammatory cytokine production. Furthermore, FSH has the ability to protect granulosa cell survival against LPS. Oocytes matured in low LPS doses that subsequently underwent in vitro fertilization, experienced decreased competence to develop to the blastocyst stage when developmental events were evaluated utilizing real time-lapse microscopy. Thus, bacterial endotoxin within the ovarian follicle imparts local immune responses in granulosa cells and the follicular environment, contributing to decreased reproductive efficiency and resulting in impacted oocyte quality and subsequent embryo development.
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