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 The effect of branched chain amino acids on proteosynthesis in skeletal muscles of japanese quail during ontogenesis

2011 ◽  
Vol 49 (No. 4) ◽  
pp. 137-143
Author(s):  
J. Antalíková ◽  
M. Baranovská ◽  
J. Jankela
Keyword(s):  
Amino Acids ◽  
Protein Content ◽  
Japanese Quail ◽  
Branched Chain Amino Acids ◽  
Protein Fractions ◽  
Individual Protein ◽  
Japanese Quails ◽  
Branched Chain ◽  
The Individual

We studied the influence of branched chain amino acids on the muscle proteosynthesis of Japanese quail during ontogenesis. We used in vitro incubation of these muscles: musculus extensor metacarpalis radialis (EMR) &ndash; wing muscle, musculus ambiens (MA) &ndash; leg muscle. The incorporation of <sup>14</sup>C-tyrosine into the individual protein fractions was evaluated. Influences of valine, leucine and isoleucine on proteosynthesis on day 14, 28 and 53 of life of Japanese quails were compared. Different patterns of individual protein fractions were detected. During ontogenesis, in the MA the number of fractions remained unchanged while in the EMR it differed. Four fractions with molecular weight 200&ndash;1 000 kDa present on day 14 and 28 were absent on day 53. A new fraction over 200 kDa was detected on day 53. The <sup>14</sup>C-tyrosine incorporation after leucine treatment was enhanced only in the MA of 28&nbsp;days old quails. The protein content in the EMR decreased (50%) in several fractions. The addition of valine had no effect in the MA while in the EMR the protein content decreased in 14 and 28 days old quails. The incorporation of <sup>14</sup>C-tyrosine was decreased by the influence of isoleucine in the EMR of 28 and 53 days old quails, in the MA only in 28 days old birds. We assume that the effect of regulatory amino acids on proteosynthesis depends both on muscle type and on the age of Japanese quail. &nbsp;

scholarly journals Maternal nutrient restriction in late pregnancy programs postnatal metabolism and pituitary development in beef heifers

PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0249924
Author(s):  
John M. Long ◽  
Levi A. Trubenbach ◽  
Kenneth C. Hobbs ◽  
Andrew E. Poletti ◽  
Chelsie B. Steinhauser ◽  
...  
Keyword(s):  
Area Under The Curve ◽  
Late Pregnancy ◽  
Postnatal Growth ◽  
Late Gestation ◽  
Nutrient Restriction ◽  
Postnatal Life ◽  
Beef Heifers ◽  
Maternal Undernutrition ◽  
Ad Libitum

Maternal undernutrition during pregnancy followed by ad libitum access to nutrients during postnatal life induces postnatal metabolic disruptions in multiple species. Therefore, an experiment was conducted to evaluate postnatal growth, metabolism, and development of beef heifers exposed to late gestation maternal nutrient restriction. Pregnancies were generated via transfer of in vitro embryos produced using X-bearing sperm from a single Angus sire. Pregnant dams were randomly assigned to receive either 100% (control; n = 9) or 70% (restricted; n = 9) of their total energy requirements from gestational day 158 to parturition. From post-natal day (PND) 301 until slaughter (PND485), heifers were individually fed ad libitum in a Calan gate facility. Calves from restricted dams were lighter than controls at birth (P<0.05) through PND70 (P<0.05) with no difference in body weight from PND105 through PND485 (P>0.10). To assess pancreatic function, glucose tolerance tests were performed on PND315 and PND482 and a diet effect was seen with glucose area under the curve being greater (P<0.05) in calves born to restricted dams compared to controls. At slaughter, total internal fat was greater (P<0.05) in heifers born to restricted dams, while whole pituitary weight was lighter (P<0.05). Heifers from restricted dams had fewer growth hormone-positive cells (somatotrophs) compared to controls (P<0.05). Results demonstrate an impaired ability to clear peripheral glucose in heifers born to restricted dams leading to increased deposition of internal fat. A reduction in the number of somatotrophs may contribute to the adipogenic phenotype of heifers born to restricted dams due to growth hormone’s known anabolic roles in growth, lipolysis, and pancreatic islet function.

scholarly journals PER2 Mediates CREB-Dependent Light Induction of Per1

2021 ◽  
Author(s):  
Andrea Brenna ◽  
Jürgen A. Ripperger ◽  
Gabriella Saro ◽  
Dominique Glauser ◽  
Zhihong Yang ◽  
...  
Keyword(s):  
Histone Acetyltransferase ◽  
Regulatory Region ◽  
Light Signal ◽  
Physiological Processes ◽  
Evolutionarily Conserved ◽  
The One ◽  
The Individual ◽  
H3 Acetylation

Abstract Light affects many physiological processes in mammals such as entrainment of the circadian clock, regulation of mood, and relaxation of blood vessels. At the molecular level, a stimulus such as light initiates a cascade of kinases that phosphorylate CREB at various sites, including serine 133 (S133). This modification leads CREB to recruit the co-factor CRCT1 and the histone acetyltransferase CBP to stimulate the transcription of genes containing a CRE element in their promoters, such as Period 1 (Per1). However, the details of this pathway are poorly understood. Here we provide evidence that PER2 acts as a co-factor of CREB to facilitate the formation of a transactivation complex on the CRE element of the Per1 gene regulatory region in response to light. Using in vitro and in vivo approaches, we show that PER2 modulates the interaction between CREB and its co-regulator CRTC1 to support complex formation only after a light or forskolin stimulus. Furthermore, the absence of PER2 abolished the interaction between the histone acetyltransferase CBP and CREB. This process was accompanied by a reduction of histone H3 acetylation and decreased recruitment of RNA Pol II to the Per1 gene. Collectively, our data show that PER2 supports the stimulus-dependent induction of the Per1 gene via modulation of the CREB/CRTC1/CBP complex. Remarkably, our results indicate that the molecular mechanism that transduces the light signal to the clock is similar to the one in the filamentous fungus Neurospora crassa to induce frequency (Frq). This suggests an evolutionarily conserved mechanism of this process despite the divergent sequences of the individual components.

scholarly journals IVF affects embryonic development in a sex-biased manner in mice

Reproduction ◽  
2016 ◽  
Vol 151 (4) ◽  
pp. 443-453 ◽  
Author(s):  
Kun Tan ◽  
Zhuqing Wang ◽  
Zhenni Zhang ◽  
Lei An ◽  
Jianhui Tian
Keyword(s):  
Fetal Development ◽  
Late Gestation ◽  
Embryonic And Fetal Development ◽  
Specific Effects ◽  
Males And Females ◽  
Health Complications ◽  
Similar Growth ◽  
Insight Into

Increasing evidence indicates that IVF (IVF includes in vitro fertilization and culture) embryos and babies are associated with a series of health complications, and some of them show sex-dimorphic patterns. Therefore, we hypothesized that IVF procedures have sex-biased or even sex-specific effects on embryonic and fetal development. Here, we demonstrate that IVF-induced side effects show significant sexual dimorphic patterns from the pre-implantation to the prenatal stage. During the pre-implantation stage, female IVF embryos appear to be more vulnerable to IVF-induced effects, including an increased percentage of apoptosis (7.22±1.94 vs 0.71±0.76, P<0.01), and dysregulated expression of representative sex-dimorphic genes (Xist, Hprt, Pgk1 and Hsp70). During the mid-gestation stage, IVF males had a higher survival rate than IVF females at E13.5 (male:female=1.33:1), accompanied with a female-biased pregnancy loss. In addition, while both IVF males and females had reduced placental vasculogenesis/angiogenesis, the compensatory placental overgrowth was more evident in IVF males. During the late-gestation period, IVF fetuses had a higher sex ratio (male:female=1.48:1) at E19.5, and both male and female IVF placentas showed overgrowth. After birth, IVF males grew faster than their in vivo (IVO) counterparts, while IVF females showed a similar growth pattern with IVO females. The present study provides a new insight into understanding IVF-induced health complications during embryonic and fetal development. By understanding and minimizing these sex-biased effects of the IVF process, the health of IVF-conceived babies may be improved in the future.

scholarly journals Interactions and three-dimensional localization of a group of nuclear pore complex proteins.

1994 ◽  
Vol 126 (3) ◽  
pp. 603-617 ◽  
Author(s):  
N Panté ◽  
R Bastos ◽  
I McMorrow ◽  
B Burke ◽  
U Aebi
Keyword(s):  
Nuclear Pore Complex ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Nuclear Pore ◽  
Western Blots ◽  
Cytoplasmic Filaments ◽  
Quick Freezing ◽  
The Individual ◽  
Antibody Labeling

We have used antibodies directed against a number of nuclear pore complex (NPC) proteins to determine their mutual interactions and location within the three-dimensional structure of the NPC. A monoclonal antibody, termed QE5, recognized three NPC polypeptides, p250, NUP153, and p62 on Western blots, and labeled the nuclear envelope of several cultured cell lines by immunofluorescence microscopy. These three polypeptides contained O-linked N-acetylglucosamine residues and were released from the NPC by detergent/high-salt treatment as discrete high molecular weight complexes. p250 was found in association with a novel 75 kD protein, NUP153 was released as a homo-oligomer of about 1 megadalton, and p62 was associated with polypeptides of 58 and 54 kD (previously reported by Finlay, D. R., E. Meier, P. Bradley, J. Horecka, and D. J. Forbes. 1991. J. Cell Biol. 114:169-183). p75, p58, and p54 were not galactosylated in vitro. Xenopus oocyte NEs were labeled with gold-conjugated QE5 and prepared for electron microscopy by quick freezing/freeze drying/rotary metal shadowing. This EM preparation method enabled us to more precisely localize the epitopes of this antibody to the cytoplasmic filaments and the nuclear basket of the NPC. Since QE5 recognizes three O-linked NPC glycoproteins, its labeling was compared with that of the lectin wheat germ agglutinin which recognizes O-linked N-acetylglucosamine moieties. The two probes were found to yield similar, although not identical, distributions of label. To identify the individual proteins with particular NPC components, we have used an anti-peptide antibody against NUP153 and a monospecific anti-p250 polyclonal antibody. Labeling with these two antibodies has documented that NUP153 is a constituent of the nuclear basket with at least one of its epitopes residing in its terminal ring, whereas p250 is a constituent of the cytoplasmic filaments.

scholarly journals 340 Effects of nutrient restriction during mid- to late-gestation on maternal and fetal pancreatic exocrine function in sheep

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 95-95
Author(s):  
Ronald J Trotta ◽  
Taylor M Czech ◽  
Manuel A Vásquez-Hidalgo ◽  
Kimberly A Vonnahme ◽  
Kendall C Swanson
Keyword(s):  
Enzyme Activity ◽  
Protein Content ◽  
Digestive Enzyme ◽  
Total Content ◽  
Pancreatic Exocrine Function ◽  
Late Gestation ◽  
Pancreatic Mass ◽  
Exocrine Function ◽  
Nutrient Restriction ◽  
Pancreatic Exocrine

Abstract To examine the effects of nutrient restriction on fetal and maternal ovine exocrine pancreatic function during mid- to late-gestation, 41 singleton ewes (48.3±0.6 kg BW) were randomly assigned to dietary treatments: 100% (control; CON; n = 20) or 60% of nutrient requirements (restricted; RES; n = 21) from day 50–90 (mid-gestation). At day 90, 14 ewes (CON, n = 7; RES, n = 7) were slaughtered. The remaining ewes were subjected to treatments of nutrient restriction or remained under a control diet from day 90–130 (late-gestation): CON-CON (n = 6), CON-RES (n = 7), RES-CON (n = 7), and RES-RES (n = 7) and were slaughtered at day 130. The pancreas was weighed, subsampled, and assayed for digestive enzyme activity. Enzyme activity was expressed as U/g, U/g protein, or U/pancreas (total content). Total protein content in the pancreas was expressed as g/pancreas. Differences between means were determined using contrasts in the MIXED procedure of SAS. Fetal and maternal pancreatic mass increased (P &lt; 0.04) with day of gestation. Nutrient restriction during mid- (P = 0.01) and late-gestation (P = 0.01) decreased maternal pancreatic mass on d 130. Total fetal pancreatic α-amylase, trypsin, and protein content increased (P &lt; 0.05) with day of gestation. Maternal nutrient restriction during late-gestation tended to decrease (P &lt; 0.08) fetal pancreatic trypsin activity. Nutrient restriction during late-gestation tended to increase (P = 0.07) α-amylase:trypsin in fetal pancreas but tended to have the opposite response in maternal pancreas (P = 0.10). Total maternal pancreatic content of trypsin and protein tended to increase (P &lt; 0.07) as gestation progressed. Nutrient restriction of gestating ewes decreased total content of a-amylase (P = 0.04) and tended to decrease total content of trypsin (P = 0.06) and protein (P = 0.06) in the maternal pancreas on d 90. Nutrient restriction during mid-gestation on d 90 and during late-gestation on d 130 decreased (P = 0.04) maternal α-amylase activity. Maternal nutrient restriction impairs pancreatic exocrine function by reducing maternal and fetal digestive enzyme activity.

Fetal development of in vitro-produced embryos: Possible association with uterine function

2000 ◽  
Vol 77 (E-Suppl) ◽  
pp. 1 ◽  
Author(s):  
C. E. Farin ◽  
P. W. Farin ◽  
P. Blondin ◽  
A. E. Crosier
Keyword(s):  
Fetal Development ◽  
Uterine Function

Rapid, Refined, and Robust Method for Expression, Purification, and Characterization of Recombinant Human Amyloid-beta M1-42

2019 ◽  
Author(s):  
Priya Prakash ◽  
Travis Lantz ◽  
Krupal P. Jethava ◽  
Gaurav Chopra
Keyword(s):  
Amyloid Beta ◽  
Western Blots ◽  
Force Microscopy ◽  
E Coli ◽  
Atomic Force ◽  
Set Up ◽  
Short Time

Amyloid plaques found in the brains of Alzheimer’s disease (AD) patients primarily consists of amyloid beta 1-42 (Ab42). Commercially, Ab42 is synthetized using peptide synthesizers. We describe a robust methodology for expression of recombinant human Ab(M1-42) in Rosetta(DE3)pLysS and BL21(DE3)pLysS competent E. coli with refined and rapid analytical purification techniques. The peptide is isolated and purified from the transformed cells using an optimized set-up for reverse-phase HPLC protocol, using commonly available C18 columns, yielding high amounts of peptide (~15-20 mg per 1 L culture) in a short time. The recombinant Ab(M1-42) forms characteristic aggregates similar to synthetic Ab42 aggregates as verified by western blots and atomic force microscopy to warrant future biological use. Our rapid, refined, and robust technique to purify human Ab(M1-42) can be used to synthesize chemical probes for several downstream in vitro and in vivo assays to facilitate AD research.

Proteomics-Based Characterization of the Effects of MMP14 on the Protein Content of Exosomes from Corneal Fibroblasts

2020 ◽  
Vol 27 (10) ◽  
pp. 979-988
Author(s):  
Kyu-Yeon Han ◽  
Jin-Hong Chang ◽  
Dimitri T. Azar
Keyword(s):  
Protein Content ◽  
Catalytic Domain ◽  
Protein Composition ◽  
Proteomics Analysis ◽  
Knock Out ◽  
Corneal Fibroblast ◽  
Flow Cytometric ◽  
Corneal Fibroblasts ◽  
Liquid Chromatography Tandem Mass

Background: Exosomes secreted by corneal fibroblasts contain matrix metalloproteinase (MMP) 14, which is known to influence pro-MMP2 accumulation on exosomes. Accordingly, we hypothesized that the enzymatic activity of MMP14 may alter the protein content of corneal fibroblast- secreted exosomes. Objective: The aim of this study was to investigate the effects of MMP14 on the composition and biological activity of corneal fibroblast-derived exosomes. Methods: Knock out of the catalytic domain (ΔExon4) of MMP14 in corneal fibroblasts was used to determine the effect of MMP14 expression on the characteristics of fibroblast-secreted exosomes. The amount of secreted proteins and their size distribution were measured using Nano Tracking Analysis. Proteins within exosomes from wild-type (WT) and ΔExon4-deficient fibroblasts were identified by liquid chromatography-tandem mass spectrometry (MS/MS) proteomics analysis. The proteolytic effects of MMP14 were evaluated in vitro via MS identification of eliminated proteins. The biological functions of MMP14-carrying exosomes were investigated via fusion to endothelial cells and flow cytometric assays. Results: Exosomes isolated from WT and ΔExon4-deficient fibroblasts exhibited similar size distributions and morphologies, although WT fibroblasts secreted a greater amount of exosomes. The protein content, however, was higher in ΔExon4-deficient fibroblast-derived exosomes than in WT fibroblast-derived exosomes. Proteomics analysis revealed that WT-derived exosomes included proteins that regulated cell migration, and ΔExon4 fibroblast-derived exosomes contained additional proteins that were cleaved by MMP14. Conclusion: Our findings suggest that MMP14 expression influences the protein composition of exosomes secreted by corneal fibroblasts, and through those biological components, MMP14 in corneal fibroblasts derived-exosomes may regulate corneal angiogenesis.

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