scholarly journals In the porcine placenta, aromatase (CYP19) is specifically expressed in trough-like structures of the trophoblast at the basis and flanks of chorionic folds

2021 ◽  
Vol 77 (07) ◽  
pp. 6550-2021
Author(s):  
PERSHOTAM KHATRI ◽  
GERHARD SCHULER ◽  
ELÇIN ÖZOCAK-BATMAZ ◽  
GÖZDE R. ÖZALP
Keyword(s):  
Complex Geometry ◽  
Cell Types ◽  
Staining Intensity ◽  
Late Gestation ◽  
Weak Staining ◽  
A Minor

Aromatase was localized in the porcine placenta by immunohistochemistry during the first peak of maternal estrogen concentrations (D25; n=3), their nadir around midgestation (D50; n=3) and their second increase in late gestation (D110; n=3). On D25, aromatase was highly expressed throughout the trophoblast. On D50, its expression was restricted to the trophoblast lining the trough-like fossal regions between the bases of chorionic folds. However, immunostaining was detected only in a minor proportion of these locations, with staining intensity being generally only weak to moderate. On D110, distinct to intense immunostaining was found in virtually all trophoblast fossal regions and in similar structures at the flanks of chorionic folds adjacent to the free margins of secondary and tertiary endometrial folds. A weak staining of questionable specificity was occasionally observed in endometrial glands. In other cell types of the utero-placental compartment, aromatase was undetectable. The results suggest that in pigs placental estrogens could be involved in the formation of a more complex geometry of the feto-maternal interface as pregnancy progresses.

scholarly journals Effect of Insect Live Larvae as Environmental Enrichment on Poultry Gut Health: Gut Mucin Composition, Microbiota and Local Immune Response Evaluation

Animals ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 2819
Author(s):  
Elena Colombino ◽  
Ilaria Biasato ◽  
Ilario Ferrocino ◽  
Sara Bellezza Oddon ◽  
Christian Caimi ◽  
...  
Keyword(s):  
Immune Response ◽  
Broiler Chickens ◽  
Environmental Enrichment ◽  
Staining Intensity ◽  
Local Immune Response ◽  
Gut Health ◽  
Insect Larvae ◽  
A Minor ◽  
Dietary Treatments ◽  
Caecal Microbiota

The aim of this study was to evaluate the effect of Hermetia illucens (HI) and Tenebrio molitor (TM) live larvae as environmental enrichment on the mucin composition, local immune response and microbiota of broilers. A total of 180 four-day-old male broiler chickens (Ross 308) were randomly allotted to three dietary treatments (six replicates/treatment; ten animals/replicate): (i) control (C); (ii) C+HI; (iii) C+TM. Live larvae were distributed based on 5% of the expected daily feed intake. At slaughter (39 days of age), samples of duodenum, jejunum and ileum (twelve animals/diet) were submitted to mucin histochemical evaluation. Expression of MUC-2 and cytokines was evaluated by rt-qPCR in jejunum. Mucin staining intensity was not influenced by diet (p > 0.05); however, this varied depending on the intestinal segment (p < 0.001). No significant differences were recorded for IL-4, IL-6 TNF-α, MUC-2 and INF-γ gene expression in jejunum, while IL-2 was lower in the TM group compared to HI and C (p = 0.044). Caecal microbiota showed higher abundance of Clostridium, Saccharibacteria and Victivallaceae in the HI group, while Collinsella was higher in the TM group. The results suggested that live insect larvae did not impair mucin composition or local immune response, and can slightly improve caecal microbiota by enhancing a minor fraction of short chain fatty acid-producing taxa.

scholarly journals Engineering of Adenovirus Vectors Containing Heterologous Peptide Sequences in the C Terminus of Capsid Protein IX

2002 ◽  
Vol 76 (14) ◽  
pp. 6893-6899 ◽  
Author(s):  
Igor P. Dmitriev ◽  
Elena A. Kashentseva ◽  
David T. Curiel
Keyword(s):  
Heparan Sulfate ◽  
Capsid Protein ◽  
Minor Component ◽  
Cell Types ◽  
Binding Motif ◽  
Flag Epitope ◽  
C Terminus ◽  
Protein Ix ◽  
A Minor ◽  
Carboxy Terminal

ABSTRACT The utility of the present generation of adenovirus (Ad) vectors for gene therapy applications could be improved by restricting native viral tropism to selected cell types. In order to achieve modification of Ad tropism, we proposed to exploit a minor component of viral capsid, protein IX (pIX), for genetic incorporation of targeting ligands. Based on the proposed structure of pIX, we hypothesized that its C terminus could be used as a site for incorporation of heterologous peptide sequences. We engineered recombinant Ad vectors containing modified pIX carrying a carboxy-terminal Flag epitope along with a heparan sulfate binding motif consisting of either eight consecutive lysines or a polylysine sequence. Using an anti-Flag antibody, we have shown that modified pIXs are incorporated into virions and display Flag-containing C-terminal sequences on the capsid surface. In addition, both lysine octapeptide and polylysine ligands were accessible for binding to heparin-coated beads. In contrast to virus bearing lysine octapeptide, Ad vector displaying a polylysine was capable of recognizing cellular heparan sulfate receptors. We have demonstrated that incorporation of a polylysine motif into the pIX ectodomain results in a significant augmentation of Ad fiber knob-independent infection of CAR-deficient cell types. Our data suggest that the pIX ectodomain can serve as an alternative to the fiber knob, penton base, and hexon proteins for incorporation of targeting ligands for the purpose of Ad tropism modification.

Histopathology of pituitary tumours

2011 ◽  
pp. 121-127
Author(s):  
Eva Horvath ◽  
Kalman Kovacs
Keyword(s):  
Mammalian Species ◽  
Cell Types ◽  
Pars Intermedia ◽  
Pars Tuberalis ◽  
Posterior Lobe ◽  
Human Pituitary ◽  
Human Pituitary Gland ◽  
Fibre Network ◽  
Pars Anterior ◽  
A Minor

The human pituitary gland consists of two major components: the adenohypophysis comprising the hormone producing cells of the pars anterior, pars intermedia, and pars tuberalis, and the neurohypophysis, also called pars nervosa or posterior lobe (1). In contrast to most mammalian species, the human gland has no anatomically distinct pars intermedia (2). The exclusively proopiomelanocortin (POMC)-producing cells of the pars intermedia are sandwiched between the anterior and posterior lobes in the majority of mammals, whereas in the human they are incorporated within the pars anterior, thereby constituting the pars distalis (3). The pars tuberalis is a minor upward extension of the adenohypophysis attached to the exterior of the lower pituitary stalk. In this chapter we deal only with adenohypophyseal tumours. Histologically, the adenohypophysis consists of a central median (or mucoid) wedge flanked by the two lateral wings. The hormone-producing cell types are distributed in an uneven, but characteristic manner. The cells are arranged within evenly sized acini surrounded by a delicate but well-defined reticulin fibre network giving the pituitary its distinct architecture (4). In the center of the acini is the long-neglected pituitary follicle composed of the agranular nonendocrine folliculo-stellate cells (5).

scholarly journals Abcg2 labels multiple cell types in skeletal muscle and participates in muscle regeneration

2011 ◽  
Vol 195 (1) ◽  
pp. 147-163 ◽  
Author(s):  
Michelle J. Doyle ◽  
Sheng Zhou ◽  
Kathleen Kelly Tanaka ◽  
Addolorata Pisconti ◽  
Nicholas H. Farina ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Satellite Cells ◽  
Muscle Regeneration ◽  
Side Population ◽  
Cell Types ◽  
Lineage Tracing ◽  
Skeletal Muscle Regeneration ◽  
Genetic Lineage ◽  
Multiple Cell ◽  
A Minor

Skeletal muscle contains progenitor cells (satellite cells) that maintain and repair muscle. It also contains muscle side population (SP) cells, which express Abcg2 and may participate in muscle regeneration or may represent a source of satellite cell replenishment. In Abcg2-null mice, the SP fraction is lost in skeletal muscle, although the significance of this loss was previously unknown. We show that cells expressing Abcg2 increased upon injury and that muscle regeneration was impaired in Abcg2-null mice, resulting in fewer centrally nucleated myofibers, reduced myofiber size, and fewer satellite cells. Additionally, using genetic lineage tracing, we demonstrate that the progeny of Abcg2-expressing cells contributed to multiple cell types within the muscle interstitium, primarily endothelial cells. After injury, Abcg2 progeny made a minor contribution to regenerated myofibers. Furthermore, Abcg2-labeled cells increased significantly upon injury and appeared to traffic to muscle from peripheral blood. Together, these data suggest an important role for Abcg2 in positively regulating skeletal muscle regeneration.

Immunocytochemical localization of oestrogen receptors in the endometrium of the ewe

1991 ◽  
Vol 3 (3) ◽  
pp. 321 ◽  
Author(s):  
RA Cherny ◽  
LA Salamonsen ◽  
JK Findlay
Keyword(s):  
Stromal Cells ◽  
Cellular Response ◽  
Individual Cell ◽  
Cell Types ◽  
Staining Intensity ◽  
Steroid Treatment ◽  
Positive Staining ◽  
Cell Type ◽  
Peak Intensity

Immunocytochemistry with monoclonal antibodies to the oestrogen receptor (ER) was used to localize ERs in sections of endometrium obtained from cycling and pregnant Corriedale ewes. Representative tissue from Days 4, 10, 14, 15, 16 and 17 of the cycle (Day 0 = onset of oestrus) and Day 15 of pregnancy was used. ER localization was also examined in tissue obtained from ovariectomized (ovex) ewes with and without subcutaneous implants containing oestrogen, progesterone, or oestrogen and progesterone. ER distribution was examined in caruncular endometrium and intercaruncular endometrium. Staining intensity varied according to cell type, stage of the cycle, steroid treatment and pregnancy. No staining was observed in endothelial cells. In all cases, ER was localized within the nuclei of positive cells. Generally, ER levels were high on Day 4 and declined to negligible values by Day 10 (corresponding to peak progesterone values) except in the deep stroma of caruncular endometrium. Positive staining reappeared in stromal cells of caruncles on Day 13 and in the luminal epithelium of intercaruncular tissue on Day 14. Peak intensity was reached on Day 15 for caruncular tissue and Day 16 for intercaruncular tissue. Ovariectomy did not cause an overall reduction in ER levels, whereas treatment with oestrogen and progesterone had variable effects depending on cell type. Progesterone did not suppress overall ER. In Day 15 pregnant tissue, ER was undetectable in all compartments except deep stroma of caruncles, indicating that factors other than progesterone, perhaps embryonic in origin, were responsible. The observation that individual cell types display differential sensitivities to oestrogen and progesterone as regards their expression of ER is consistent with the role of cell-cell interactions as modulators of cellular response to steroids through the oestrous cycle and in pregnancy.

scholarly journals Reciprocal expression of 17α-hydroxylase-C17,20-lyase and aromatase cytochrome P450 during bovine trophoblast differentiation: a two-cell system drives placental oestrogen synthesis

Reproduction ◽  
2006 ◽  
Vol 131 (4) ◽  
pp. 669-679 ◽  
Author(s):  
G Schuler ◽  
G R Özalp ◽  
B Hoffmann ◽  
N Harada ◽  
P Browne ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Cell Types ◽  
Staining Intensity ◽  
Giant Cells ◽  
Cell System ◽  
Cellular Level ◽  
Specific Staining ◽  
Cytoplasmic Staining ◽  
Distinct Cell ◽  
Trophoblast Giant Cells

No definitive information is yet available on the steroidogenic capacity of the two morphologically distinct cell types forming the bovine trophoblast, the uninucleated trophoblast cells (UTCs) and the trophoblast giant cells (TGCs). Hence, in order to localise 17α-hydroxylase-C17,20-lyase (P450c17) on a cellular level and to monitor its expression as a function of gestational age, placentomes from pregnant (days 80–284; n = 19), prepartal (days 273–282; 24–36 h prior to the onset of labour; n = 3) and parturient cows (n = 5) were immunostained for P450c17 using an antiserum against the recombinant bovine enzyme. At all stages investigated, P450c17 was exclusively found in the UTCs of chorionic villi (CV), where staining was ubiquitous between days 80 and 160, but was largely restricted to primary CV and the branching sites of secondary CV between days 160 and 240. Thereafter, a distinct ubiquitous staining reoccurred in the UTCs of all CV in late pregnant, prepartal and parturient animals. Using an antiserum against human aromatase cytochrome P450 (P450arom), specific cytoplasmic staining was observed in TGCs. In placentomes from pregnant cows, staining intensity was higher in mature compared with immature TGCs and was more pronounced in the trophoblast covering big stem villi compared with the trophoblast at other sites of the villous tree. In placentomes of a parturient cow, specific staining was only found in mature TGCs that survived the normal, but substantial, prepartal decline in TGC numbers. These results clearly showed that bovine UTCs and TGCs exhibit different steroidogenic capacities, constituting a ‘two-cell’ organisation for oestrogen synthesis. P450c17 expression appears to be quickly down-regulated and P450arom is up-regulated when UTCs enter the TGC differentiation pathway.

scholarly journals Promoter DNA Methylation Patterns of Differentiated Cells Are Largely Programmed at the Progenitor Stage

2010 ◽  
Vol 21 (12) ◽  
pp. 2066-2077 ◽  
Author(s):  
Anita L. Sørensen ◽  
Bente Marie Jacobsen ◽  
Andrew H. Reiner ◽  
Ingrid S. Andersen ◽  
Philippe Collas
Keyword(s):  
Dna Methylation ◽  
Cell Types ◽  
Molecular Evidence ◽  
Mesenchymal Progenitors ◽  
H3k27 Methylation ◽  
Differentiated Cells ◽  
Promoter Dna Methylation ◽  
Promoter Dna ◽  
A Minor ◽  
Methylation Patterns

Mesenchymal stem cells (MSCs) isolated from various tissues share common phenotypic and functional properties. However, intrinsic molecular evidence supporting these observations has been lacking. Here, we unravel overlapping genome-wide promoter DNA methylation patterns between MSCs from adipose tissue, bone marrow, and skeletal muscle, whereas hematopoietic progenitors are more epigenetically distant from MSCs as a whole. Commonly hypermethylated genes are enriched in signaling, metabolic, and developmental functions, whereas genes hypermethylated only in MSCs are associated with early development functions. We find that most lineage-specification promoters are DNA hypomethylated and harbor a combination of trimethylated H3K4 and H3K27, whereas early developmental genes are DNA hypermethylated with or without H3K27 methylation. Promoter DNA methylation patterns of differentiated cells are largely established at the progenitor stage; yet, differentiation segregates a minor fraction of the commonly hypermethylated promoters, generating greater epigenetic divergence between differentiated cell types than between their undifferentiated counterparts. We also show an effect of promoter CpG content on methylation dynamics upon differentiation and distinct methylation profiles on transcriptionally active and inactive promoters. We infer that methylation state of lineage-specific promoters in MSCs is not a primary determinant of differentiation capacity. Our results support the view of a common origin of mesenchymal progenitors.

scholarly journals The Spatial and Temporal Origin of Chandelier Cells in Mouse Neocortex

Science ◽  
2012 ◽  
Vol 339 (6115) ◽  
pp. 70-74 ◽  
Author(s):  
Hiroki Taniguchi ◽  
Jiangteng Lu ◽  
Z. Josh Huang
Keyword(s):  
Pyramidal Neurons ◽  
Functional Organization ◽  
Cell Types ◽  
Developmental Trajectory ◽  
Late Gestation ◽  
Homeodomain Protein ◽  
Germinal Zone ◽  
Embryonic Origin ◽  
Bona Fide ◽  
Chandelier Cells

Diverse γ-aminobutyric acid–releasing interneurons regulate the functional organization of cortical circuits and derive from multiple embryonic sources. It remains unclear to what extent embryonic origin influences interneuron specification and cortical integration because of difficulties in tracking defined cell types. Here, we followed the developmental trajectory of chandelier cells (ChCs), the most distinct interneurons that innervate the axon initial segment of pyramidal neurons and control action potential initiation. ChCs mainly derive from the ventral germinal zone of the lateral ventricle during late gestation and require the homeodomain protein Nkx2.1 for their specification. They migrate with stereotyped routes and schedule and achieve specific laminar distribution in the cortex. The developmental specification of this bona fide interneuron type likely contributes to the assembly of a cortical circuit motif.

scholarly journals Immunolocalization of Gla proteins (osteocalcin) in rat tooth germs: comparison between indirect immunofluorescence, peroxidase-antiperoxidase, avidin-biotin-peroxidase complex, and avidin-biotin-gold complex with silver enhancement.

1987 ◽  
Vol 35 (8) ◽  
pp. 825-830 ◽  
Author(s):  
A L Bronckers ◽  
S Gay ◽  
R D Finkelman ◽  
W T Butler
Keyword(s):  
Blood Vessels ◽  
Indirect Immunofluorescence ◽  
Staining Intensity ◽  
Gold Complex ◽  
Silver Enhancement ◽  
Tissue Sections ◽  
Weak Staining ◽  
Mode Of Transport ◽  
Tooth Germs ◽  
Odontoblast Processes

Odontoblasts and osteoblasts synthesize gamma-carboxyglutamatic acid (Gla)-containing proteins which are partially deposited in the mineralizing tissues and partially released into the plasma. Using four immunostaining techniques, we have evaluated the question of whether dentin Gla proteins (DGP) are transported to the mineralization front through the odontoblast processes. Undecalcified sections of rat incisors and molar tooth germs were immunostained with affinity-purified antibodies to DGP using the following methods: indirect immunofluorescence; peroxidase-antiperoxidase (PAP); avidin-biotin-peroxidase complex (ABC-peroxidase); and avidin-biotin-gold complex with silver enhancement (ABC-GSS). The results obtained with these four procedures were compared with respect to the developmental appearance of DGP, staining intensity and presence in odontoblastic processes, predentin, dentin, and blood vessels. Qualitatively, similar results were obtained with the four, with respect to the distribution and developmental appearance of DGP, with two exceptions: indirect immunofluorescence never stained DGP within blood vessels, whereas the other methods occasionally did; and because of its sensitivity, only the ABC-GSS method revealed immunostaining for DGP in odontoblastic processes. All methods revealed weak immunostaining in predentin which was considerably enhanced with hyaluronidase treatment; however, hyaluronidase only moderately increased predentin immunostaining with ABC-GSS. Of these four procedures, ABC-GSS is the most sensitive; however, ABC-GSS appears to detect predominantly antigens at the surface of tissue sections. We conclude that DGP is present in odontoblastic processes but in low amounts; the weak staining was due either to rapid transport of DGP through the process or to the fact that this mode of transport is limited.

scholarly journals Identification of qPCR reference genes suitable for normalising gene expression in the developing mouse embryo

2021 ◽  
Vol 6 ◽  
pp. 197
Author(s):  
John C.W. Hildyard ◽  
Dominic J. Wells ◽  
Richard J. Piercy
Keyword(s):  
Gene Expression ◽  
Mouse Embryo ◽  
Reference Genes ◽  
Developmental Regulation ◽  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Cell Types ◽  
Late Gestation ◽  
Developmental Expression ◽  
Suitable Reference

Background: Progression through mammalian embryogenesis involves many interacting cell types and multiple differentiating cell lineages. Quantitative polymerase chain reaction (qPCR) analysis of gene expression in the developing embryo is a valuable tool for deciphering these processes, but normalisation to stably-expressed reference genes is essential for such analyses. Gene expression patterns change globally and dramatically as embryonic development proceeds, rendering identification of consistently appropriate reference genes challenging. Methods: We have investigated expression stability in mouse embryos from mid to late gestation (E11.5–E18.5), both at the whole-embryo level, and within the head and forelimb specifically, using 15 candidate reference genes (ACTB, 18S, SDHA, GAPDH, HTATSF1, CDC40, RPL13A, CSNK2A2, AP3D1, HPRT1, CYC1, EIF4A, UBC, B2M and PAK1IP1), and four complementary algorithms (geNorm, Normfinder, Bestkeeper and deltaCt). Results: Unexpectedly, all methods suggest that many genes within our candidate panel are acceptable references, though AP3D1, RPL13A and PAK1IP1 are the strongest performing genes overall. HPRT1 and B2M are conversely poor choices, and show strong developmental regulation. We further show that normalisation using our three highest-scoring references can reveal subtle patterns of developmental expression even in genes ostensibly ranked as acceptably stable (CDC40, HTATSF1). Conclusion: AP3D1, RPL13A and PAK1IP1 represent universally suitable reference genes for expression studies in the E11.5-E18.5 mouse embryo.

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