scholarly journals Profiles of PrKX Expression in Developmental Mouse Embryo and Human Tissues

2005 ◽  
Vol 53 (8) ◽  
pp. 1003-1009 ◽  
Author(s):  
Wei Li ◽  
Zu-Xi Yu ◽  
Robert M. Kotin
Keyword(s):  
Protein Kinase ◽  
Mouse Embryo ◽  
Developmental Stages ◽  
Fetal Brain ◽  
Tissue Type ◽  
Type I ◽  
Human Tissues ◽  
Western Blots ◽  
Protein Levels ◽  
Embryo Sections

Protein kinase × (PrKX), karyotypically located on the human × chromosome, is a type I cAMP-dependent protein kinase. Although a specific role for PrKX has not yet been defined, PrKX gene expression in mouse and human tissues has been profiled only by in situ hybridization and Northern blot analyses and not by protein expression. To determine more precisely the PrKX protein levels, we developed specific anti-PrKX antibodies and examined gestationally staged mouse embryo sections by immunohistochemistry. These results showed that PrKX is ubiquitously distributed and highly expressed in murine central nervous system and heart tissues in early developmental stages and in most organs at later stages but was not detected in either connective tissues or bone. Using Western blots to detect PrKX, total protein extracts from eight different adult or fetal human tissues including brain, heart, kidney, liver, lung, pancreas, spleen, and thymus were analyzed. Although PrKX protein was present in each of the tissues tested, the protein levels varied depending on tissue type and developmental stage. Very low protein levels were found in heart tissues from a 5-month-old fetus and from an adult, whereas PrKX proteins were more abundant in fetal brain, kidney, and liver tissues compared with adult samples of the same tissue type.

scholarly journals Purification of PKC-I, an endogenous protein kinase C inhibitor, and types II and III protein kinase C isoenzymes from human neutrophils

1992 ◽  
Vol 284 (2) ◽  
pp. 399-405 ◽  
Author(s):  
K J Balazovich ◽  
E L McEwen ◽  
M L Lutzke ◽  
L A Boxer ◽  
T White
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Human Neutrophils ◽  
Type I ◽  
Dependent Manner ◽  
Type Ii ◽  
Western Blots ◽  
Type Iii ◽  
Kinase C ◽  
Endogenous Protein

Human neutrophil protein kinase C (PKC) activity is inhibited by an endogenous protein found primarily in the pellet fraction from homogenized specific granules, which was both heat- and proteinase-sensitive [Balazovich, Smolen & Boxer (1986) J. Immunol. 137, 1665-1673]. We now report that two PKC isoenzymes and the endogenous PKC inhibitor, which we named PKC-I, were purified from human neutrophils. A neutrophil soluble fraction that was subjected to DEAE-Sephacel chromatography yielded highly enriched PKC because, by definition, enzymic activity was strictly dependent on Ca2+ and phosphatidylserine. Hydroxyapatite chromatography resolved two peaks of PKC activity. Type II and Type III PKC isoenzymes were each identified on Western blots by using isoenzyme-specific monoclonal antibodies. Unlike rat brain, from which PKC isoenzymes were also purified, Type I PKC was not detected in human neutrophils. Western blots indicated that both Type II and Type III PKC isoenzymes had molecular masses near 80 kDa. In agreement with other reports, PKC was autophosphorylated in vitro. PKC-I, an endogenous neutrophil inhibitor of PKC, was purified to apparent homogeneity by DEAE-Sephacel and S-400 Sephacel chromatography. PKC-I had a molecular mass of 41 kDa. PKC-I inhibited purified PKC activity stimulated by 1,2-diacylglycerols in a concentration-dependent manner, and inhibited PKC-dependent phosphorylation of proteins present in neutrophil cytosol.

The Polycomb group gene Posterior Sex Combs encodes a chromosomal protein

Development ◽  
1993 ◽  
Vol 117 (2) ◽  
pp. 641-655 ◽  
Author(s):  
E.C. Martin ◽  
P.N. Adler
Keyword(s):  
Developmental Stages ◽  
Germ Line ◽  
Polytene Chromosomes ◽  
Polycomb Group ◽  
Mutant Phenotype ◽  
Chromosomal Protein ◽  
Western Blots ◽  
Protein Levels ◽  
Sex Combs ◽  
Drosophila Homolog

The Posterior Sex Combs (Psc) gene of Drosophila has been studied at the molecular level both because it is a Polycomb group (Pc-G) gene and hence required for the maintenance of segmental determination, and because it is the Drosophila homolog of the murine bmi-1 oncogene. Although genetic interactions indicated that Psc functioned as a Pc-G gene, the zygotic mutant phenotype of Psc showed little evidence of segmental transformations. We have examined mutant embryos derived from a mutant maternal germ line and found a stronger mutant phenotype, indicating that the weak zygotic phenotype of Psc is due to maternal rescue. We have found that Psc RNA accumulates in developing oocytes and this maternal RNA is presumably responsible for the maternal rescue. We have studied the expression of the Psc gene at both the RNA and protein levels. On northern blots, we find evidence for two Psc mRNAs and, on western blots, we find evidence for two Psc proteins that are altered either in abundance or size in Psc mutants. The Psc protein accumulates in all regions of the embryo and also in many tissues in a variety of developmental stages. In all cases, it is nuclear, as is its mammalian homolog, the bmi-1 protein. On polytene chromosomes, we find Psc at 45 chromosomal loci where two other Pc-G proteins are present.

Involvement of cyclic adenosine monophosphate-dependent protein kinase isozymes in tissue plasminogen activator secretion by rat Sertoli cells stimulated with follicle-stimulating hormone in vitro

1993 ◽  
Vol 128 (6) ◽  
pp. 555-562 ◽  
Author(s):  
V Laurent-Cadoret ◽  
F Guillou ◽  
Y Combarnous
Keyword(s):  
Protein Kinase ◽  
Plasminogen Activator ◽  
Sertoli Cells ◽  
Tissue Type ◽  
Type I ◽  
Dependent Protein Kinase ◽  
Tissue Type Plasminogen Activator ◽  
Camp Dependent Protein Kinase ◽  
Type Plasminogen Activator ◽  
Dependent Protein

This study was undertaken to investigate, in freshly isolated rat Sertoli cells, the physiological function of the type I and type II cyclic adenosine monophosphate (cAMP)-dependent protein kinase isozymes in tissue-type plasminogen activator secretion and the regulation of this cAMP process by follicle-stimulating hormone (FSH). Follicle-stimulating hormone-induced tissue-type plasminogen activator secretion depends upon intracellular cAMP levels. The changes in cAMP amounts required to activate maximally the tissue-type plasminogen activator secretion are extremely small, a cAMP threshold having to be reached for triggering the tissue-type plasminogen activator output. Intact Sertoli cells were incubated with combinations of cAMP analogs specific for each cAMP-dependent protein kinase type and complementary in their cAMP binding site on the cAMP-dependent protein kinase regulatory subunits: 8-aminohexylamino-cAMP = type 1, site 1; 8-thiomethyl-cAMP = type II, site 1 and N6-benzoyl-cAMP = types I/II, site 2. This allowed us to activate selectively each cAMP-dependent protein kinase type in a synergistic manner and then to evaluate their respective influence in the specific tissue-type plasminogen activator response. We establish that both of the cAMP-dependent protein kinase types are present and functional; the activity of the type I isozyme is preponderant (60%) in the cAMP-dependent tissue-type plasminogen activator secretion. Likewise, when these cAMP analogs were coupled with endogenously generated cAMP by FSH or forskolin, both of the cAMP-dependent protein kinase types were involved in the tissue-type plasminogen activator production. However, only tissue-type plasminogen activator secretion induced by FSH is mediated predominantly by the type I cAMP-dependent protein kinase, although the type II isozyme sustains an appreciable physiological role in the transmission pathway. We suggest some differences in the pattern of action between FSH and forskolin in Sertoli cells.

scholarly journals Post-transcriptional regulation across human tissues

2015 ◽  
Author(s):  
Alexander Franks ◽  
Edoardo Airoldi ◽  
Nikolai Slavov
Keyword(s):  
Transcriptional Regulation ◽  
Cell Types ◽  
Tissue Type ◽  
Mrna Levels ◽  
Human Tissues ◽  
Protein Levels ◽  
Physiological Variability ◽  
Reliable Quantification ◽  
Post Transcriptional Regulation ◽  
Different Cell Types

AbstractTranscriptional and post-transcriptional regulation shape tissue-type-specific proteomes, but their relative contributions remain contested. Estimates of the factors determining protein levels in human tissues do not distinguish between (i) the factors determining the variability between the abundances of different proteins, i.e., mean-level-variability and, (ii) the factors determining the physiological variability of the same protein across different tissue types, i.e., across-tissues variability. We sought to estimate the contribution of transcript levels to these two orthogonal sources of variability, and found that scaled mRNA levels can account for most of the mean-level-variability but not necessarily for across-tissues variability. The reliable quantification of the latter estimate is limited by substantial measurement noise. However, protein-to-mRNA ratios exhibit substantial across-tissues variability that is functionally concerted and reproducible across different datasets, suggesting extensive post-transcriptional regulation. These results caution against estimating protein fold-changes from mRNA fold-changes between different cell-types, and highlight the contribution of post-transcriptional regulation to shaping tissue-type-specific proteomes.

scholarly journals Adiponectin as Well as Compressive Forces Regulate in vitro β-Catenin Expression on Cementoblasts via Mitogen-Activated Protein Kinase Signaling Activation

2021 ◽  
Vol 9 ◽  
Author(s):  
Jiawen Yong ◽  
Julia von Bremen ◽  
Gisela Ruiz-Heiland ◽  
Sabine Ruf
Keyword(s):  
Protein Kinase ◽  
Signaling Pathways ◽  
Mapk Signaling ◽  
Mitogen Activated Protein Kinase ◽  
Western Blots ◽  
Protein Levels ◽  
Mitogen Activated Protein ◽  
Gsk 3Β ◽  
Signaling Activation

We aimed to investigate the molecular effect that adiponectin exerts on cementoblasts especially in the presence of compressive forces. OCCM-30 cells (M. Somerman, NIH, NIDCR, United States) were used. Real-time reverse transcriptase–polymerase chain reaction (RT-PCR) and western blots were employed to verify if the mRNA and protein levels of adiponectin receptors (AdipoRs), mitogen-activated protein kinase (MAPK), and β-catenin signaling were influenced by compressive forces or adiponectin. Moreover, siRNAs targeting P38α, JNK1, ERK1, ERK2, and AdipoRs as well as pharmacological MAPK inhibition were performed. We found that compressive forces increase the expression of AdipoRs. Adiponectin and compression up-regulate P38α,JNK1, ERK1, and ERK2 as well as β-catenin gene expression. Western blots showed that co-stimuli activate the MAPK and β-catenin signaling pathways. MAPK inhibition alters the compression-induced β-catenin activation and the siRNAs targeting AdipoRs, P38α, and JNK1, showing the interaction of single MAPK molecules and β-catenin signaling in response to compression or adiponectin. Silencing by a dominantly negative version of P38α and JNK1 attenuates adiponectin-induced TCF/LEF reporter activation. Together, we found that light compressive forces activate β-catenin and MAPK signaling pathways. Adiponectin regulates β-catenin signaling principally by inactivating the GSK-3β kinase activity. β-Catenin expression was partially inhibited by MAPK blockade, indicating that MAPK plays a crucial role regulating β-catenin during cementogenesis. Moreover, adiponectin modulates GSK-3β and β-catenin mostly through AdipoR1. P38α is a key connector between β-catenin, TCF/LEF transcription, and MAPK signaling pathway.

Exercise training attenuates cardiac inflammation and fibrosis in hypertensive ovariectomized rats

2020 ◽  
Vol 128 (4) ◽  
pp. 1033-1043
Author(s):  
Yi-Yuan Lin ◽  
Yi Hong ◽  
Ming-Cheng Zhou ◽  
Hai-Liang Huang ◽  
Woei-Cherng Shyu ◽  
...  
Keyword(s):  
Angiotensin Ii ◽  
Growth Factor ◽  
Exercise Training ◽  
Estrogen Receptors ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Tissue Growth ◽  
Tissue Type ◽  
Type I ◽  
Protein Levels

This study investigated the effects of exercise training on cardiac inflammatory and cardiac fibrotic pathways in female spontaneously hypertensive rats (SHR), which were divided into a sham-operated sedentary hypertensive group (SHR-S), a sedentary hypertensive ovariectomized group (SHR-O), or a hypertensive ovariectomized group with treadmill exercise training (SHR-OT; 60 min/day, 5 days/wk) for 8 wk. Normotensive female Wistar-Kyoto rats (WKY) served as controls. SOD and catalase (CAT) activities were significantly increased in the SHR-OT group, when compared with the SHR-S or SHR-O groups. The protein levels of estrogen receptor (ER)-α and ER-β became decreased in the SHR-O group, when compared with the WKY or SHR-S groups, but were not changed in the SHR-OT group. The protein level of the angiotensin II type I receptor (AT1R) was increased in the SHR-S group but did not further change in the SHR-O group, whereas it was decreased in the SHR-OT group. The inflammatory-related protein levels of TNF-α, p-NF-κB, cyclooxygenase 2 (COX-2), inducible nitric oxide synthase (iNOS), and IL-6, as well as the fibrotic-related protein levels of transforming growth factor-β (TGF-β), p-Smad2/3, connective tissue growth factor (CTGF), tissue-type plasminogen activator (tPA), matrix metalloproteinase (MMP)-9, and collagen I were increased in the SHR-S group and increased further in the SHR-O group, whereas they were decreased in the SHR-OT group. The coexistence of hypertension and ovariectomy additively increased cardiac inflammatory and fibrotic pathways partially through hypertension-enhanced AT1R and ovariectomy-depressed estrogen receptors. Exercise training appeared to suppress hypertensive ovariectomized heart-induced inflammatory and fibrotic pathways possibly through decreasing AT1R but not through estrogen receptors. NEW & NOTEWORTHY The coexistence of hypertension and ovariectomy appeared to increase cardiac inflammatory and fibrotic pathways likely through hypertension-enhanced angiotensin II type I receptor and ovariectomy-depressed estrogen receptors. Exercise training on a treadmill could prevent hypertensive ovariectomized heart-induced cardiac inflammation and fibrosis via an inflammatory pathway [TNF-α, p-IKK-α/β, p-NF-κB, cyclooxygenase 2 (COX-2), iNOS, and IL-6] and fibrotic pathway [transforming growth factor-β (TGF-β), p-Smad2/3, connective tissue growth factor (CTGF), tissue-type plasminogen activator (tPA), matrix metalloproteinase (MMP)-9, and collagen I] possibly through decreasing angiotensin II type I receptor but not through estrogen receptors.

scholarly journals Development of a Mucus Gland Bioreactor in Loach Paramisgurnus dabryanus

2021 ◽  
Vol 22 (2) ◽  
pp. 687
Author(s):  
Tong Zhou ◽  
Bolan Zhou ◽  
Yasong Zhao ◽  
Qing Li ◽  
Guili Song ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
Grass Carp ◽  
Single Copy ◽  
Type I ◽  
Western Blots ◽  
Paramisgurnus Dabryanus ◽  
Expression Activity ◽  
Transgenic Construct ◽  
Mucus Gland

Most currently available bioreactors have some defects in the expression, activity, or purification of target protein and peptide molecules, whereas the mucus gland of fish can overcome these defects to become a novel bioreactor for the biopharmaceutical industry. In this study, we have evaluated the practicability of developing a mucus gland bioreactor in loach (Paramisgurnus dabryanus). A transgenic construct pT2-krt8-IFN1 was obtained by subcloning the promoter of zebrafish keratin 8 gene and the type I interferon (IFN1) cDNA of grass carp into the SB transposon. The IFN1 expressed in CIK cells exhibited an antiviral activity against the replication of GCRV873 and activated two genes downstream of JAK-STAT signaling pathway. A transgenic loach line was then generated by microinjection of the pT2-krt8-IFN1 plasmids and in vitro synthesized capped SB11 mRNA. Southern blots indicated that a single copy of IFN1 gene was stably integrated into the genome of transgenic loach. The expression of grass carp IFN1 in transgenic loaches was detected with RT-PCR and Western blots. About 0.0825 µg of grass carp IFN1 was detected in 20 µL mucus from transgenic loaches. At a viral titer of 1 × 103 PFU/mL, plaque numbers on plates containing mucus from transgenic loaches reduced by 18% in comparison with those of the control, indicating that mucus of IFN1-transgenic loaches exhibited an antiviral activity. Thus, we have successfully created a mucus gland bioreactor that has great potential for the production of various proteins and peptides.

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