scholarly journals PSII-35 Melatonin supplementation and restricted nutrition do not affect chorionic somatomammotropin (CSH) concentration in ovine placenta from mid- to late- gestation

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 244-244
Author(s):  
Hanaa Mansour ◽  
Caleb O Lemley ◽  
Russell Anthony ◽  
Kendall C Swanson ◽  
Anna T Grazul-Bilska ◽  
...  
Keyword(s):  
Histological Analysis ◽  
Late Gestation ◽  
Fetal Membrane ◽  
Nutrient Restriction ◽  
Binucleate Cells ◽  
Tissue Sections ◽  
Vasoactive Factors ◽  
Ovine Placenta ◽  
Chorionic Somatomammotropin ◽  
Texas Red

Abstract Melatonin plays a role as a vasodilator. Vasoactive and angiogenic factors are expressed by placental binucleate cells (BNC) and produce chorionic somatomammotropin (CSH), known to impact fetal and placental growth. We hypothesized that melatonin supplementation and restricted nutrition from mid- to late-gestation would alter CSH concentration and some characteristics of BNC in placenta. At day 50 of gestation, ewes carrying singletons were randomly assigned to a 2 × 2 factorial design and were fed either an adequate (ADQ; 100% NRC; n = 15) or restricted (RES; 60% NRC; n = 15) diet supplemented with 0 (CON, n = 14) or 5 mg of melatonin (MEL; n = 16). Placentomes were collected on day 130 of gestation and preserved in formalin for histological analysis. Cotyledon (COT) were snap frozen for western immunoblotting analyses. Tissue sections were stained using biotinylated Dolichos Biflurus (DBA; a marker of fetal membrane) lectin and fluorescein labeled Texas red-avidin and fluorescein labeled Griffonia Simplifolica (BS) lectin (a marker of BNC). The number, area, and diameter of BNC in COT were determined by image analysis. For immunoblotting, protein was extracted from COT in SDS phosphate buffer, loaded equally, and separated on 12.5% polyacrylamide gels. Protein was transferred to PVDF membranes and incubated with rabbit anti-CSH. Bands were visualized and imaged. Data were analyzed using Proc Mixed procedure of SAS. Melatonin supplementation and restricted nutrition did not affect BNC number, area, or diameter, or CSH protein expression. While we reject our hypothesis that melatonin supplementation and nutrient restriction would alter the CSH concentration and BNC characteristics in COT, we continue to evaluate if the BNC produce angiogenic or vasoactive factors that may influence placental and mammary gland functions in sheep.

Effects of Maternal Nutrient Restriction During Gestation on LH Production in the Female Bovine Fetus

2021 ◽  
Vol 99 (Supplement_2) ◽  
pp. 25-26
Author(s):  
Sterling H Fahey ◽  
Sarah West ◽  
John M Long ◽  
Carey Satterfield ◽  
Rodolfo C Cardoso
Keyword(s):  
Protein Content ◽  
Fetal Development ◽  
Monozygotic Twins ◽  
Late Gestation ◽  
Nutrient Restriction ◽  
Western Blots ◽  
Physiological Processes ◽  
Individual Potential ◽  
The Individual

Abstract Gestational nutrient restriction causes epigenetic and phenotypic changes that affect multiple physiological processes in the offspring. Gonadotropes, the cells in the anterior pituitary that secrete luteinizing hormone (LH) and follicle-stimulating hormone (FSH), are particularly sensitive to nutritional changes during fetal development. Our objective herein was to investigate the effects of gestational nutrient restriction on LH protein content and number of gonadotropes in the fetal bovine pituitary. We hypothesized that moderate nutrient restriction during mid to late gestation decreases pituitary LH production, which is associated with a reduced number of gonadotropes. Embryos were produced in vitro with X-bearing semen from a single sire then split to generate monozygotic twins. Each identical twin was transferred to a virgin dam yielding four sets of female twins. At gestational d 158, the dams were randomly assigned into two groups, one fed 100% NRC requirements (control) and the other fed 70% of NRC requirements (restricted) during the last trimester of gestation, ensuring each pair of twins had one twin in each group. At gestational d 265, the fetuses (n = 4/group) were euthanized by barbiturate overdose, and the pituitaries were collected. Western blots were performed using an ovine LH-specific antibody (Dr. A.F. Parlow, NIDDK). The total LH protein content in the pituitary tended to be decreased in the restricted fetuses compared to controls (P < 0.10). However, immunohistochemistry analysis of the pituitary did not reveal any significant changes in the total number of LH-positive cells (control = 460±23 cells/0.5 mm2; restricted = 496±45 cells/0.5 mm2, P = 0.58). In conclusion, while maternal nutrient restriction during gestation resulted in a trend of reduced LH content in the fetal pituitary, immunohistological findings suggest that these changes are likely related to the individual potential of each gonadotrope to produce LH, rather than alterations in cell differentiation during fetal development.

scholarly journals Timing of nutrient restriction and programming of fetal adipose tissue development

2004 ◽  
Vol 63 (3) ◽  
pp. 397-403 ◽  
Author(s):  
Michael E. Symonds ◽  
Sarah Pearce ◽  
Jayson Bispham ◽  
David S. Gardner ◽  
Terence Stephenson
Keyword(s):  
Adipose Tissue ◽  
Uncoupling Protein ◽  
Tissue Growth ◽  
Fat Deposition ◽  
Late Gestation ◽  
Nutrient Restriction ◽  
Adult Obesity ◽  
Uncoupling Protein 1 ◽  
Voltage Dependent Anion Channel ◽  
Brown Adipose

It is apparent from epidemiological studies that the timing of maternal nutrient restriction has a major influence on outcome in terms of predisposing the resulting offspring to adult obesity. The present review will consider the extent to which maternal age, parity and nutritional restriction at defined stages of gestation can have important effects on fat deposition and endocrine sensitivity of adipose tissue in the offspring. For example, in 1-year-old sheep the offspring of juvenile mothers have substantially reduced fat deposition compared with those born to adult mothers. Offspring of primiparous adult mothers, however, show increased adiposity compared with those born to multiparous mothers. These offspring of multiparous ewes show retained abundance of the brown adipose tissue-specific uncoupling protein 1 at 1 month of age. A stimulated rate of metabolism in brown fat of these offspring may act to reduce adipose tissue deposition in later life. In terms of defined windows of development that can programme adipose tissue growth, maternal nutrient restriction targetted over the period of maximal placental growth results in increased adiposity at term in conjunction with enhanced abundance of mRNA for the insulin-like growth factor-I and -II receptors. In contrast, nutrient restriction in late gestation, coincident with the period of maximal fetal growth, has no major effect on adiposity but results in greater abundance of specific mitochondrial proteins, i.e. voltage-dependent anion channel and/or uncoupling protein 2. These adaptations may increase the predisposal of these offspring to adult obesity. Increasing maternal nutrition in late gestation, however, can result in proportionately less fetal adipose tissue deposition in conjunction with enhanced abundance of uncoupling protein 1.

scholarly journals Immunocytochemical localization of protein antigens in large sections of tissues embedded in water-soluble embedding media.

1986 ◽  
Vol 34 (4) ◽  
pp. 535-538 ◽  
Author(s):  
T M Duello ◽  
F C Gumkowski
Keyword(s):  
Water Soluble ◽  
Placental Lactogen ◽  
Protein Antigens ◽  
Ph Change ◽  
Binucleate Cells ◽  
Tissue Sections ◽  
Pre Treatment ◽  
Ethanol Pretreatment ◽  
Large Sections ◽  
Absolute Etoh

JB4 and Immunobed are water-soluble embedding media used for embedding large blocks of tissue. Immunobed was specifically designed for immunocytochemistry because ethanol extraction of an additive in the monomer of the resin is reported to render tissue sections permeable to immunoglobulins. We have modified the manufacturer's protocol to accomplish localization of two protein antigens in tissues embedded in either JB4 or Immunobed. Luteinizing hormone-beta (LH beta) was localized in sections of rat and bovine pituitary tissues and bovine placental lactogen (bPL) was localized in sections of placentomes from bovine placentas. Sections received one of the following pre-treatments: absolute EtOH; NaHCO3 buffer, pH 6-10; EtOH followed by NaHCO3 buffer; one of several enzymes; EtOH followed by enzyme; NaHCO3 buffer followed by enzyme. Anti-LH beta stained only pituitary gonadotrophs and anti-bPL stained only placental binucleate cells, as assessed by absorption controls. Pre-treatment with enzyme was required for staining of sections, but an alkaline pH change (NaHCO3) had little or no effect. Ethanol pretreatment had little or no effect alone or in conjunction with NaHCO3 or enzyme. Sections were sufficiently thin (1.5 micron) to afford resolution of structure, but suitably large (approximately 2 cm2) to minimize problems of sampling.

Hormone Ontogeny in the Ovine Fetus. II. Ovine Chorionic Somatomammotropin in Mid and Late Gestation in the Fetal and Maternal Circulations*

Endocrinology ◽  
1979 ◽  
Vol 104 (6) ◽  
pp. 1828-1833 ◽  
Author(s):  
P. D. GLUCKMAN ◽  
S. L. KAPLAN ◽  
A. M. RUDOLPH ◽  
M. M GRUMBACH
Keyword(s):  
Late Gestation ◽  
Ovine Fetus ◽  
Chorionic Somatomammotropin

The Effects of Cortisol on the Binucleate Cell Population in the Ovine Placenta During Late Gestation

Placenta ◽  
2002 ◽  
Vol 23 (6) ◽  
pp. 451-458 ◽  
Author(s):  
J.W. Ward ◽  
F.B.P. Wooding ◽  
A.L. Fowden
Keyword(s):  
Cell Population ◽  
Late Gestation ◽  
Binucleate Cell ◽  
Ovine Placenta
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