scholarly journals Impact of g-Load Shift on Temporal Expression Pattern of Apoptosis-linked Proteins in the Rat Mammary Gland

2019 ◽  
Vol 7 (1) ◽  
pp. 13-21
Author(s):  
Kibrom M. Alula ◽  
James H. Resau ◽  
Osman V. Patel
Keyword(s):  
Mammary Epithelial Cells ◽  
Late Pregnancy ◽  
Mammary Epithelial ◽  
Sprague Dawley ◽  
Sprague Dawley Rats ◽  
Pup Survival ◽  
Associated Proteins ◽  
Rat Mammary Gland ◽  
Tumor Protein P53 ◽  
Temporal Expression Pattern

AbstractAlteration in gravitational load impacts homeorhetic response in rat dams which affects neonatal pup survival. However, the effects of hypergravity (HG) exposure on the abundance of apoptosis-associated proteins in mammary epithelial cells (MECs) have not been characterized. Therefore, we examined whether chronic exposure to HG from midpregnancy alters the abundance of proapoptotic proteins in MECs during the late pregnancy and early lactation. A group of pregnant Sprague Dawley rats were exposed to either HG (2g) or normo-gravity (1g: stationary control [SC]) from days 11 to 20 of gestation (G20). Another set of animals were investigated from day 11 of pregnancy through days 1 and 3 (P1 and P3, respectively) postpartum. Quantitative (pixels [px]/lobule) immunohistochemistry at G20 of Cleaved Caspase-3 (CC-3), Tumor Protein p53 (P53), and vitamin D receptor (VDR) revealed that all the three proteins were increased (p<0.01) in HG rats compared to SC animals. At P1, the HG group had twofold higher (p<0.001) expression of CC-3 relative to the SC group. Approximately, 50% (p<0.001) more VDR was detected in the HG cohorts than SC at P3. These results suggest that a shift in g-load upregulates the expression of key proapoptotic proteins during the pregnancy-to-lactation transition in the rat MECs.

scholarly journals Expression of Prolactin mRNA in Rat Mammary Gland During Pregnancy and Lactation

2000 ◽  
Vol 48 (3) ◽  
pp. 389-395 ◽  
Author(s):  
Toshiki Iwasaka ◽  
Shinobu Umemura ◽  
Kochi Kakimoto ◽  
Haruko Koizumi ◽  
Yoshiyuki R. Osamura
Keyword(s):  
Mammary Gland ◽  
Epithelial Cells ◽  
Mammary Epithelial Cells ◽  
Late Pregnancy ◽  
Mammary Epithelial ◽  
Rt Pcr ◽  
Pregnancy And Lactation ◽  
Rat Mammary Gland ◽  
Pcr Method

We studied the expression of prolactin (PRL) mRNA in the mammary gland of resting, pregnant, lactating, and weanling rats using in situ and solution reverse transcriptase-polymerase chain reaction (RT-PCR). In mid- to late pregnancy and throughout lactation, PRL mRNA was detected in both in situ and solution RT-PCR. These PRL mRNA signals were clearly identified in the cytoplasm of alveolar and ductal mammary epithelial cells by the in situ RT-PCR method. In mid- to late pregnancy, such as at the initiating point of PRL mRNA expression, we confirmed in some cases a lack of PRL mRNA by solution RT-PCR. In addition, in the early weaning phase, no signals were detected by solution RT-PCR. However, slight focal signals were detected in some poorly vacuolated cytoplasm of regressing acinar cells by in situ RT-PCR. These findings suggest that PRL mRNA in rat mammary gland begins in mid- to late pregnancy in parallel with the development of the mammary gland, continues throughout lactation, and declines in the early phase of weaning, with regression of mammary epithelial cells.

scholarly journals Profiling solute-carrier transporters in key metabolic tissues during the postpartum evolution of mammary epithelial cells from nonsecretory to secretory

2019 ◽  
Vol 51 (11) ◽  
pp. 539-552
Author(s):  
Osman V. Patel ◽  
Theresa Casey ◽  
Karen Plaut
Keyword(s):  
Gene Expression ◽  
Adipose Tissue ◽  
Epithelial Cells ◽  
Energy Homeostasis ◽  
Mammary Epithelial Cells ◽  
Expression Patterns ◽  
Late Pregnancy ◽  
Mammary Epithelial ◽  
Sugar Transporters ◽  
Solute Carrier

Modifications in the abundance of solute-carrier (SLC) transcripts in tandem with adjustments in genes-associated with energy homeostasis during the postpartum transition of the mammary epithelial cells (MEC) from nonsecretory to secretory is pivotal for supporting milk synthesis. The goal of this study was to identify differentially expressed SLC genes across key metabolic tissues between late pregnancy and onset of lactation. Total RNA was isolated from the mammary, liver, and adipose tissues collected from rat dams on day 20 of pregnancy (P20) and day 1 of lactation (L1) and gene expression was measured with Rat 230 2.0 Affymetrix GeneChips. LIMMA was utilized to identify the differential gene expression patterns between P20 and L1 tissues. Transcripts engaged in conveying anions, cations, carboxylates, sugars, amino acids, metals, nucleosides, vitamins, and fatty acids were significantly increased ( P < 0.05) in MEC during the P20 to L1 shift. Downregulated ( P < 0.05) genes in the mammary during the physiological transition included GLUT8 and SLC45a3. In the liver, SLC genes encoding for anion, carbonyl, and nucleotide sugar transporters were upregulated ( P < 0.05) at L1. while genes facilitating transportation of anions and hexose were increased ( P < 0.05), from P20 to L1 in the adipose tissue. GLUT1 and GLUT4 in the liver, along with GLUT4 and SGLT2 in the adipose tissue, were repressed ( P < 0.05) at L1. Our results illustrate that MEC exhibit dynamic molecular plasticity during the nonsecretory to secretory transition and increase biosynthetic capacity through a coordinated tissue specific SLC transcriptome modification to facilitate substrate transfer.

scholarly journals Essential role of Orai1 store-operated calcium channels in lactation

2015 ◽  
Vol 112 (18) ◽  
pp. 5827-5832 ◽  
Author(s):  
Felicity M. Davis ◽  
Agnes Janoshazi ◽  
Kyathanahalli S. Janardhan ◽  
Natacha Steinckwich ◽  
Diane M. D’Agostin ◽  
...  
Keyword(s):  
Neural Control ◽  
Mammary Epithelial Cells ◽  
Myoepithelial Cells ◽  
Mammary Epithelial ◽  
Sweat Glands ◽  
Milk Ejection ◽  
Signaling Mechanisms ◽  
Pup Survival ◽  
Imaging Strategy

The nourishment of neonates by nursing is the defining characteristic of mammals. However, despite considerable research into the neural control of lactation, an understanding of the signaling mechanisms underlying the production and expulsion of milk by mammary epithelial cells during lactation remains largely unknown. Here we demonstrate that a store-operated Ca2+ channel subunit, Orai1, is required for both optimal Ca2+ transport into milk and for milk ejection. Using a novel, 3D imaging strategy, we visualized live oxytocin-induced alveolar unit contractions in the mammary gland, and we demonstrated that in this model milk is ejected by way of pulsatile contractions of these alveolar units. In mammary glands of Orai1 knockout mice, these contractions are infrequent and poorly coordinated. We reveal that oxytocin also induces a large transient release of stored Ca2+ in mammary myoepithelial cells followed by slow, irregular Ca2+ oscillations. These oscillations, and not the initial Ca2+ transient, are mediated exclusively by Orai1 and are absolutely required for milk ejection and pup survival, an observation that redefines the signaling processes responsible for milk ejection. These findings clearly demonstrate that Ca2+ is not just a substrate for nutritional enrichment in mammals but is also a master regulator of the spatiotemporal signaling events underpinning mammary alveolar unit contraction. Orai1-dependent Ca2+ oscillations may represent a conserved language in myoepithelial cells of other secretory epithelia, such as sweat glands, potentially shedding light on other Orai1 channelopathies, including anhidrosis (an inability to sweat).

scholarly journals Changes in Gastric Smooth Muscle Cell Contraction during Pregnancy: Effect of Estrogen

2019 ◽  
Vol 2019 ◽  
pp. 1-8 ◽  
Author(s):  
Othman A. Al-Shboul ◽  
Hanan J. Al-Rshoud ◽  
Ahmed N. Al-Dwairi ◽  
Mohammad A. Alqudah ◽  
Mahmoud A. Alfaqih ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Late Pregnancy ◽  
Cyclic Guanosine Monophosphate ◽  
Guanosine Monophosphate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sprague Dawley ◽  
Gastric Smooth Muscle ◽  
Sprague Dawley Rats ◽  
Polymerase Chain ◽  
Synthase Inhibitor

It is well known that pregnancy is associated with frequent gastrointestinal (GI) disorders and symptoms. Moreover, previous reports have shown that estrogen, which changes in levels during pregnancy, participates in the regulation of GI motility and is involved in the pathogenesis of various functional disorders in the stomach. The aim of the current study was to explore the changes in the expression of estrogen receptors (ERs) and examine the effect of estrogen on nitric oxide- (NO-) cyclic guanosine monophosphate (cGMP) pathway and thus relaxation in gastric smooth muscle cells (GSMC) during pregnancy. Single GSMC from early-pregnant and late-pregnant Sprague-Dawley rats were used. Protein and mRNA expression levels of ERs were measured via specifically designed enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR), respectively. NO and cGMP levels were measured via specifically designed ELISA kits. Effect of estrogen on acetylcholine- (ACh-) induced contraction of single GSMC was measured via scanning micrometry in the presence or absence of the NO synthase inhibitor,N-nitro-L-arginine (L-NNA), or guanylyl cyclase inhibitor, 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ). Estrogen increased both NO and cGMP levels and their levels were greater in early compared to late pregnancy. Expression of ERs was greater in early compared to late pregnancy. ACh induced greater contraction of GSMC in late pregnancy compared to early pregnancy. Estrogen inhibited ACh-induced contraction in both periods of pregnancy. Importantly, pretreatment of GSMC with either L-NNA or ODQ abolished estrogen inhibitory action on muscle contraction. In conclusion, GSMC contractile behavior undergoes drastic changes in response to estrogen during pregnancy and this might explain some of the pregnancy-associated gastric disorders.

scholarly journals The Stage-Specific Plasticity of Descending Modulatory Controls in a Rodent Model of Cancer-Induced Bone Pain

Cancers ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 3286
Author(s):  
Mateusz Wojciech Kucharczyk ◽  
Diane Derrien ◽  
Anthony Henry Dickenson ◽  
Kirsty Bannister
Keyword(s):  
Dorsal Horn ◽  
Bone Pain ◽  
Early Stage ◽  
Bone Destruction ◽  
Rodent Model ◽  
Sprague Dawley ◽  
Sprague Dawley Rats ◽  
Rat Mammary Gland ◽  
Descending Control

Pain resulting from metastatic bone disease is a major unmet clinical need. Studying spinal processing in rodent models of cancer pain is desirable since the percept of pain is influenced in part by modulation at the level of the transmission system in the dorsal horn of the spinal cord. Here, a rodent model of cancer-induced bone pain (CIBP) was generated following syngeneic rat mammary gland adenocarcinoma cell injection in the tibia of male Sprague Dawley rats. Disease progression was classified as “early” or “late” stage according to bone destruction. Even though wakeful CIBP rats showed progressive mechanical hypersensitivity, subsequent in vivo electrophysiological measurement of mechanically evoked deep dorsal horn spinal neuronal responses revealed no change. Rather, a dynamic reorganization of spinal neuronal modulation by descending controls was observed, and this was maladaptive only in the early stage of CIBP. Interestingly, this latter observation corresponded with the degree of damage to the primary afferents innervating the cancerous tissue. Plasticity in the modulation of spinal neuronal activity by descending control pathways reveals a novel opportunity for targeting CIBP in a stage-specific manner. Finally, the data herein have translational potential since the descending control pathways measured are present also in humans.

Differential expression of the growth hormone receptor and growth hormone-binding protein in epithelia and stroma of the mouse mammary gland at various physiological stages

1999 ◽  
Vol 161 (1) ◽  
pp. 77-87 ◽  
Author(s):  
YN Ilkbahar ◽  
G Thordarson ◽  
IG Camarillo ◽  
F Talamantes
Keyword(s):  
Growth Hormone ◽  
Mammary Gland ◽  
Epithelial Cells ◽  
Growth Hormone Receptor ◽  
Mammary Epithelial Cells ◽  
Late Pregnancy ◽  
Mammary Glands ◽  
Mouse Mammary Gland ◽  
Mammary Epithelial ◽  
Mrna Levels

Increasing evidence suggests that GH is important in normal mammary gland development. To investigate this further, we studied the distribution and levels of growth hormone receptor (GHR) and GH-binding protein (GHBP) in the mouse mammary gland. At three weeks of age, the epithelial component of the right fourth inguinal mammary gland of female mice was removed. These animals were then either maintained as virgins until they were killed or they were mated. One group of the mated mice was killed on day 18 of pregnancy and the remaining mated animals were allowed to carry their pups until term and were killed on day 6 of lactation. At the time of death, both the intact left and the de-epithelialized right mammary glands were collected from all three groups. Some of the intact glands served as a source of epithelial cells, free of stroma. The mRNA levels for GHR and GHBP were measured in intact glands, epithelia-cleared fat pads, and isolated mammary epithelial cells. GHR and GHBP mRNAs were expressed in both the mammary epithelium and stroma. However, the levels of both GHR and GHBP mRNAs were significantly higher in the stroma as compared with the epithelium component. This increase for both mRNAs was from 3- to 12-fold at each physiological state examined. In the intact gland, both GHR and GHBP transcripts were highest in virgins, declined during late pregnancy, and the lowest levels were found in the lactating gland. GHBP and GHR protein concentrations were also assessed in intact glands and epithelia-free fat pads. Similar to the mRNAs, GHR and GHBP protein levels (means+/-s.e.m.) in intact glands were highest in virgin mice (0.891+/-0.15 pmoles/mg protein and 0.136+/-0.26 pmoles/mg protein respectively), declined during late pregnancy (0. 354+/-0.111 pmoles/mg protein and 0.178+/-0.039 pmoles/mg protein respectively), and were lowest during lactation (0.096+0.037 pmoles/mg protein and 0.017+0.006 pmoles/mg protein respectively). Immunocytochemistry utilizing specific antisera against mouse (m) GHR and mGHBP revealed that the two proteins are localized to both the stroma and parenchyma of mouse mammary glands, with similar patterns of immunostaining throughout the different physiological stages analyzed. GHR immunolocalized to the plasma membrane and cytosol of mammary epithelial cells and adipocytes, whereas the GHBP immunostaining was nuclear and cytosolic. In conclusion, we report here that GHR and GHBP mRNAs and proteins are expressed in both the epithelium and the stroma of mammary glands of virgin, pregnant, and lactating mice. In intact glands, GHR and GHBP proteins, as well as their transcripts are higher in abundance in virgin relative to lactating mice. At all physiological stages, GHR and GHBP mRNA levels are higher in the stroma compared with the parenchyma. These findings indicate that the actions of GH in the mammary gland are both direct through its binding to the epithelia, and indirect by binding to the stroma and stimulation of IGF-I production which, in turn, affects mammary epithelial development.

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