Fluorescent in situ mRNA detection in the adult mouse cochlea

Because tissue freeze-drying is an excellent way to preserve antigenic conformation, we have tested the feasibility of this technique to reveal nonradioactive in situ hybridization (ISH) of tissue mRNA. We have compared mRNA detection after different methods of tissue preservation, freeze-drying, cryosectioning, and formaldehyde or methanol fixation. Our results show that nonradioactive ISH is more sensitive for tissues preserved by freeze-drying than for other tissue preparations. We have demonstrated that freeze-drying allows combination of ISH and immunohistochemistry for simultaneous detection of mRNA and antigen because with this technique of tissue preservation ISH does not affect the sensitivity or the amount of the detected antigens. This work underscores the fact that tissue freeze-drying is an easy, convenient, and reliable technique for both ISH and immunohistochemistry and achieves excellent structural conditions for nonradioactive detection.

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

Journal of Histochemistry & Cytochemistry ◽

10.1177/002215540305100407 ◽

2003 ◽

Vol 51 (4) ◽

pp. 455-469 ◽

Cited By ~ 14

Author(s):

Marjo Aitola ◽

Christine M. Sadek ◽

Jan-Åke Gustafsson ◽

Markku Pelto-Huikko

Keyword(s):

Adult Mouse ◽

Coiled Coil ◽

Proliferating Cells ◽

Helix Loop Helix ◽

Aryl Hydrocarbon ◽

Spatiotemporal Expression ◽

Mouse Tissues ◽

Murine Homologue

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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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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“In-bone” Utricle Cultures—A Simplified, Atraumatic Technique for In Situ Cultures of the Adult Mouse (Mus musculus) Utricle

Otology & Neurotology ◽

10.1097/mao.0b013e31827ca330 ◽

2013 ◽

Vol 34 (2) ◽

pp. 353-359 ◽

Cited By ~ 2

Author(s):

Henry C. Ou ◽

Vincent Lin ◽

Edwin W. Rubel

Keyword(s):

Mus Musculus ◽

Adult Mouse

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In Situ Hybridization to Embryo Whole Mounts and Tissue Sections: mRNA Detection and Application to Developmental Studies

In Situ Hybridization ◽

10.1002/9783527615070.ch5 ◽

2008 ◽

pp. 91-121 ◽

Cited By ~ 3

Author(s):

T. Jowett ◽

M. Mancera ◽

A. Amores ◽

Y-L. Yan

Keyword(s):

In Situ Hybridization ◽

Developmental Studies ◽

Mrna Detection ◽

Tissue Sections ◽

Whole Mounts

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TEX101 is shed from the surface of sperm located in the caput epididymidis of the mouse

Zygote ◽

10.1017/s0967199405003394 ◽

2005 ◽

Vol 13 (4) ◽

pp. 325-333 ◽

Cited By ~ 23

Author(s):

Takeshi Takayama ◽

Takuya Mishima ◽

Miki Mori ◽

Tomoko Ishikawa ◽

Takami Takizawa ◽

...

Keyword(s):

Adult Mouse ◽

Epididymal Sperm ◽

Cell Surfaces ◽

Testicular Sperm ◽

Additional Information ◽

Reactive Protein ◽

Surface Molecules ◽

Specific Manner ◽

Caput Epididymidis

It is generally believed that cell-to-cell cross-talk and signal transduction are mediated by cell surface molecules that play diverse and important regulatory roles in spermatogenesis and fertilization. Recently, we identified a novel plasma membrane-associated protein, TES101-reactive protein (TES101RP, or TEX101), on mouse testicular germ cells. In this study, we investigate Tex101 mRNA expression in the adult mouse testis using in situ hybridization, and we examine the fate of TEX101 during sperm transport by immunohistochemical and Western blot analyses. Tex101 mRNA was expressed in a stage-specific manner in spermatocytes and in step 1–9 spermatids of the testis, but not in spermatogonia. Although the TEX101 protein remained on the cell surfaces of step 10–16 spermatids and testicular sperm, it was shed from epididymal sperm located in the caput epididymidis. The results of this study provide additional information on germ cell-specific TEX101 expression during spermatogenesis and post-testicular sperm maturation.

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