Aint/Tacc3 Is Highly Expressed in Proliferating Mouse Tissues During Development, Spermatogenesis, and Oogenesis

Aint was originally identified on the basis of its interaction in vitro with the aryl hydrocarbon nuclear receptor translocator (Arnt). Arnt is a common heterodimerization partner in the basic helix-loop–helix (bHLH)-PER-ARNT-SIM (PAS) protein family and is involved in diverse biological functions. These include xenobiotic metabolism, hypoxic response, and circadian rhythm. In addition, Arnt has a crucial role during development. Aint is a member of a growing family of transforming acidic coiled-coil (TACC) proteins and is the murine homologue of human TACC3. Here we report the spatiotemporal expression of Tacc3 mRNA and protein in embryonic, postnatally developing, and adult mouse tissues using in situ hybridization and immunocytochemistry. Tacc3 mRNA was highly expressed in proliferating cells of several organs during murine development. However, the only adult tissues expressing high levels were testis and ovary. Immunocytochemistry revealed that Tacc3 is a nuclear protein. Our results suggest that Tacc3 has an important role in murine development, spermatogenesis, and oogenesis.

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MAP 4: occurrence in mouse tissues.

The Journal of Cell Biology ◽

10.1083/jcb.99.4.1309 ◽

1984 ◽

Vol 99 (4) ◽

pp. 1309-1315 ◽

Cited By ~ 62

Author(s):

L M Parysek ◽

C F Asnes ◽

J B Olmsted

Keyword(s):

Tissue Specificity ◽

Adult Mouse ◽

Neuroblastoma Cell ◽

Neuroblastoma Cells ◽

Polyclonal Antiserum ◽

The Other ◽

Mouse Tissues ◽

Protein Map ◽

Microtubule Protein

A polyclonal antiserum to a microtubule-associated protein (MAP) from mouse neuroblastoma cells (MAP 4) was used to examine the distribution of this protein in mouse tissues. Immunoblots of neuroblastoma cell microtubule protein preparations demonstrated that the antiserum reacted with a triplet of proteins at 215,000-240,000 mol wt. Antibodies affinity purified from any of the bands showed cross-reaction with the other bands, indicating these polypeptides were all immunologically related. Antibodies specific to MAP 4 decorated microtubules in cultured murine cells fixed with glutaraldehyde, and diffuse staining was seen following treatment of cells with nocodazole. The antiserum reacted with MAP 4 in extracts of brain, heart, liver, and lung from adult mouse; the triplet in brain was more closely spaced than in the other tissues or neuroblastoma cells. In kidney, spleen, and stomach, only a single band (band 4) was labeled; this band was immunologically related to the triplet and was also present in all tissues positive for the triplet. Skeletal muscle, sperm, and peripheral blood contained no reactive polypeptides. After taxol-induced polymerization, the MAP 4 triplet was preferentially associated with the microtubule pellet whereas band 4 remained in the supernatant. These data indicate that there is tissue specificity in the distribution of MAP 4, and that some tissues contain a polypeptide related to MAP 4 (band 4) that does not bind to microtubules in vitro.

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Biosynthesis and in vivo localization of the decapentaplegic-Vg-related protein, DVR-6 (bone morphogenetic protein-6).

The Journal of Cell Biology ◽

10.1083/jcb.120.2.493 ◽

1993 ◽

Vol 120 (2) ◽

pp. 493-502 ◽

Cited By ~ 90

Author(s):

N A Wall ◽

M Blessing ◽

C V Wright ◽

B L Hogan

Keyword(s):

Translational Regulation ◽

Adult Mouse ◽

Protein Localization ◽

Cell Types ◽

Morphogenetic Protein ◽

Mouse Tissues ◽

Bronchiolar Epithelium ◽

Bone Morphogenetic Protein 6

DVR-6 (BMP-6 or Vgr-1) is a member of the TGF-beta superfamily of polypeptide signaling molecules. In situ hybridization studies have previously shown that DVR-6 RNA is expressed in a variety of cell types in the mouse embryo, but no information has been available on protein localization and biosynthesis. We have produced a polyclonal antibody to the proregion of DVR-6 and used it to localize the protein in whole mount and sectioned embryonic, newborn, and adult mouse tissues. DVR-6 protein is expressed in the mouse nervous system beginning at 9.5 days postcoitum (d.p.c.) and continues through adulthood. A variety of epithelial tissues also produce DVR-6 protein, including the suprabasal layer of the skin, bronchiolar epithelium, and the cornea. Additionally, a stably transfected cell line, BMGE+H/D6c4, is used to study the biosynthesis of DVR-6 protein and evidence is presented for translational regulation of DVR-6 expression.

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Expression of laminin isoforms in mouse myogenic cells in vitro and in vivo

Journal of Cell Science ◽

10.1242/jcs.108.12.3795 ◽

1995 ◽

Vol 108 (12) ◽

pp. 3795-3805 ◽

Cited By ~ 7

Author(s):

F. Schuler ◽

L.M. Sorokin

Keyword(s):

In Situ Hybridization ◽

Basement Membranes ◽

Chain Gene ◽

Muscle Fibres ◽

Myogenic Cells ◽

Mouse Tissues ◽

Laminin 1

The expression of laminin-1 (previously EHS laminin) and laminin-2 (previously merosin) isoforms by myogenic cells was examined in vitro and in vivo. No laminin alpha 2 chainspecific antibodies react with mouse tissues, 50 rat monoclonal antibodies were raised against the mouse laminin alpha 2 chain: their characterization is described here. Myoblasts and myotubes from myogenic cell lines and primary myogenic cultures express laminin beta 1 and gamma 1 chains and form a complex with a 380 kDa alpha chain identified as laminin alpha 2 by immunofluorescence, immunoprecipitation and PCR. PCR from C2C12 myoblasts and myotubes for the laminin alpha 2 chain gene (LamA2) provided cDNA sequences which were used to investigate the in vivo expression of mouse LamA2 mRNA in embryonic tissues by in situ hybridization. Comparisons were made with specific probes for the laminin alpha 1 chain gene (LamA1). LamA2 but not LamA1 mRNA was expressed in myogenic tissues of 14- and 17-day-old mouse embryos, while the laminin alpha 2 polypeptide was localized in adjacent basement membranes in the muscle fibres. In situ hybridization also revealed strong expression of the LamA2 mRNA in the dermis, indicating that laminin alpha 2 is expressed other than by myogenic cells in vivo. Immunofluorescence studies localized laminin alpha 2 in basement membranes of basal keratinocytes and the epithelial cells of hair follicles, providing new insight into basement membrane assembly during embryogenesis. In vitro cell attachment assays revealed that C2C12 and primary myoblasts adhere to laminin-1 and -2 isoforms in a similar manner except that myoblast spreading was significantly faster on laminin-2. Taken together, the data suggest that laminins 1 and 2 play distinct roles in myogenesis.

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Hydroxylated Chalcones as Aryl Hydrocarbon Receptor Agonists: Structure-Activity Effects

Toxicological Sciences ◽

10.1093/toxsci/kfaa179 ◽

2020 ◽

Author(s):

Hyejin Park ◽

Un-Ho Jin ◽

Keshav Karki ◽

Clinton Allred ◽

Laurie A Davidson ◽

...

Keyword(s):

Aryl Hydrocarbon Receptor ◽

Adult Mouse ◽

Vitro Assay ◽

Aryl Hydrocarbon ◽

Caco2 Cells ◽

Structure Activity ◽

Dependent Activity ◽

Maximal Induction ◽

Dna Complex

Abstract Hydroxylated chalcones are phytochemicals which are biosynthetic precursors of flavonoids and their 1,3-diaryl-prop-2-en-1-one structure is used as a scaffold for drug development. In this study, the structure-dependent activation of aryl hydrocarbon receptor (AhR)-responsive CYP1A1, CYP1B1 and UGT1A1 genes was investigated in Caco2 colon cancer cells and in non-transformed young adult mouse colonocytes (YAMC) cells. The effects of a series of di- and trihydroxychalcones as AhR agonists was structure-dependent with maximal induction of CYP1A1, CYP1B1 and UGT1A1 in Caco2 cells observed for compounds containing 2,2′-dihydroxy substituents and this included 2,2′-dihydroxy-, 2,2′,4′-trihydroxy- and 2,2′,5′-trihydroxychalcones. In contrast, 2′,4,5′-, 2′3′,4′-, 2′,4,4′-trihydroxy, and 2′,3-, 2′,4-, 2′,4′-, and 2′,5-dihydroxychalcones exhibited low to non-detectable AhR activity in Caco2 cells. In addition, all of the hydroxychalcones exhibited minimal to non-detectable activity in YAMC cells, whereas 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induced CYP1A1, CYP1B1 and UGT1A1 in Caco2 and YAMC cells. The activity of AhR-active chalcones was confirmed by determining their effects in AhR-deficient, Caco2 cells. In addition, 2,2′-dihydroxychalcone induced CYP1A1 protein and formation of an AhR-DNA complex in an in vitro assay. Simulation and modeling studies of hydroxylated chalcones confirmed their interactions with the AhR ligand binding domain and was consistent with their structure-dependent activity as AhR ligands. Thus, this study identifies hydroxylated chalcones as AhR agonists with potential for these phytochemicals to impact AhR mediated colonic pathways.

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In Situ Analysis of Cellular Proliferation in Canine, Feline and Equine Tumors by Immunohistochemistry: A Comparison of Bromodeoxyuridine, Proliferating Cell Nuclear Antigen, and Interchromatin-Associated Antigen Immunostaining Techniques

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063879400600409 ◽

1994 ◽

Vol 6 (4) ◽

pp. 453-457 ◽

Cited By ~ 11

Author(s):

Alain Pierre Théon ◽

Loretta Metzger ◽

Stephen Griffey

Keyword(s):

Cell Proliferation ◽

Proliferating Cell Nuclear Antigen ◽

Cellular Proliferation ◽

Nuclear Antigen ◽

Brdu Incorporation ◽

Immunohistochemical Detection ◽

Proliferating Cells ◽

Proliferating Cell

Cell proliferation in canine, feline, and equine tumors was evaluated using immunohistochemical detection of in vitro 5–bromodeoxyuridine (BrdU) incorporation, proliferating cell nuclear antigen (PCNA), and interchromatin-associated antigen (p105). Ten tumors in each species were analyzed. The tumor proliferative fraction (PF) was defined as the percentage of labeled nuclei for 5,000 tumor nuclei counted. Immunoreactivity was observed with all techniques in all species. A good correlation was observed between the proliferative fractions measured with the BrdU (PFBrdU) and PCNA (PFPCNA) techniques ( rs = 0.523, P = 0.0026). There was no correlation between the PFs measured with the BrdU (PFBrdU) and p105 (PFP105) techniques. Using the median values obtained from the different approaches as cutoff points to define slowly and rapidly proliferating tumors, there was an 80% agreement ( P = 0.009) between PFBrdU and PFPCNA and no agreement between PFBrdU and PFP105 The results of this study indicate that both BrdU and PCNA labeling methods can be used reliably for identifying proliferating cells in animal tumors. In addition, PCNA could be used to replace the BrdU method to assess tumor proliferative fraction because it does not require pretreatment of tissues.

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Polymerization pathway of mammalian nonmuscle myosin 2s

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1808800115 ◽

2018 ◽

Vol 115 (30) ◽

pp. E7101-E7108 ◽

Cited By ~ 7

Author(s):

Xiong Liu ◽

Shi Shu ◽

Edward D. Korn

Keyword(s):

Coiled Coil ◽

Polymerization Process ◽

Light Chains ◽

Biological Functions ◽

Nonmuscle Myosin ◽

Heavy Chains ◽

Filament Dynamics ◽

Bipolar Filament

The three mammalian nonmuscle myosin 2 (NM2) monomers, like all class 2 myosin monomers, are hexamers of two identical heavy (long) chains and two pairs of light (short) chains bound to the heavy chains. The heavy chains have an N-terminal globular motor domain (head) with actin-activated ATPase activity, a lever arm (neck) to which the two light chains bind, and a coiled-coil helical tail. Monomers polymerize into bipolar filaments, with globular heads at each end separated by a bare zone, by antiparallel association of their coiled-coil tails. NM2 filaments are highly dynamic in situ, frequently disassembling and reassembling at different locations within the cell where they are essential for multiple biological functions. Therefore, it is important to understand the mechanisms of filament polymerization and depolymerization. Monomers can exist in two states: folded and unfolded. It has been thought that unfolded monomers form antiparallel dimers that assemble into bipolar filaments. We now show that polymerization in vitro proceeds from folded monomers to folded antiparallel dimers to folded antiparallel tetramers that unfold forming antiparallel bipolar tetramers. Folded dimers and tetramers then associate with the unfolded tetramer and unfold, forming a mature bipolar filament consisting of multiple unfolded tetramers with an entwined bare zone. We also demonstrate that depolymerization is essentially the reverse of the polymerization process. These results will advance our understanding of NM2 filament dynamics in situ.

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The pro-apoptotic protein death-associated protein 3 (DAP3) interacts with the glucocorticoid receptor and affects the receptor function

Biochemical Journal ◽

10.1042/bj3490885 ◽

2000 ◽

Vol 349 (3) ◽

pp. 885-893 ◽

Cited By ~ 24

Author(s):

Sanna M. HULKKO ◽

Hideki WAKUI ◽

Johanna ZILLIACUS

Keyword(s):

Glucocorticoid Receptor ◽

Transcriptional Activation ◽

Heat Shock Protein 90 ◽

Dominant Negative ◽

Dependent Manner ◽

Interaction Domain ◽

Helix Loop Helix ◽

Aryl Hydrocarbon ◽

Apoptotic Protein

The yeast two-hybrid system was used to isolate cDNAs encoding proteins that interact with the glucocorticoid receptor (GR) ligand-binding domain in a ligand-dependent manner. One isolated cDNA encoded a fragment of death-associated protein 3 (DAP3), which has been implicated as a positive mediator of apoptosis. In vitro experiments showed that the full-length DAP3 also interacted with GR. The main interaction domain was mapped to the N-terminal region of DAP3 that had previously been shown to function in a dominant-negative fashion, protecting cells from apoptosis. Co-transfection experiments in COS-7 cells showed that DAP3 had a stimulatory effect on the ligand-induced transcriptional activation by GR and also increased the steroid-sensitivity. Furthermore, DAP3 formed a complex with several other nuclear receptors and some basic helix–loop–helix/Per–Arnt–Sim proteins, as well as with heat-shock protein 90 (hsp90) (Arnt is the aryl-hydrocarbon-receptor nuclear translocator, and Per and Sim are the Drosophila proteins Period and Single-minded). The results suggest that DAP3 could have an important role in GR action, possibly by modulating the cytoplasmic GR–hsp90 complex. Since glucocorticoids can induce apoptosis, the pro-apoptotic DAP3 protein may be involved in this function of GR.

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Paraspeckles are subpopulation-specific nuclear bodies that are not essential in mice

The Journal of Cell Biology ◽

10.1083/jcb.201011110 ◽

2011 ◽

Vol 193 (1) ◽

pp. 31-39 ◽

Cited By ~ 183

Author(s):

Shinichi Nakagawa ◽

Takao Naganuma ◽

Go Shioi ◽

Tetsuro Hirose

Keyword(s):

Adult Mouse ◽

Expression Patterns ◽

Physiological Role ◽

Growth Conditions ◽

Nuclear Bodies ◽

Cellular Functions ◽

Mouse Tissues

Nuclei of higher organisms are well structured and have multiple, distinct nuclear compartments or nuclear bodies. Paraspeckles are recently identified mammal-specific nuclear bodies ubiquitously found in most cells cultured in vitro. To investigate the physiological role of paraspeckles, we examined the in vivo expression patterns of two long noncoding RNAs, NEAT1_1 and NEAT1_2, which are essential for the architectural integrity of nuclear bodies. Unexpectedly, these genes were only strongly expressed in a particular subpopulation of cells in adult mouse tissues, and prominent paraspeckle formation was observed only in the cells highly expressing NEAT1_2. To further investigate the cellular functions of paraspeckles, we created an animal model lacking NEAT1 by gene targeting. These knockout mice were viable and fertile under laboratory growth conditions, showing no apparent phenotypes except for the disappearance of paraspeckles. We propose that paraspeckles are nonessential, subpopulation-specific nuclear bodies formed secondary to particular environmental triggers.

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