Overexpression of human keratin 16 produces a distinct skin phenotype in transgenic mouse skin

1995 ◽  
Vol 73 (9-10) ◽  
pp. 611-618 ◽  
Author(s):  
Pierre A. Coulombe ◽  
Nicola S. Bravo ◽  
Rudolph D. Paladini ◽  
Diem Nguyen ◽  
Kenzo Takahashi
Keyword(s):  
Wound Healing ◽  
Transgenic Mice ◽  
Transgenic Mouse ◽  
Transgene Expression ◽  
Germ Line ◽  
Mouse Skin ◽  
Transgenic Animals ◽  
Skin Lesions ◽  
Cellular Level ◽  
Outer Root Sheath

Human cytokeratin 16 (K16; 48 kDa) is constitutively expressed in postmitotic keratinocytes in a variety of stratified epithelial tissues, but it is best known for the marked enhancement of its expression in stratified squamous epithelia showing hyperproliferation or abnormal differentiation. Of particular interest to us, K16 is strongly induced at the wound edge after injury to the epidermis, and its accumulation correlates spatially and temporally with the onset of reepithelialization. To examine the properties of K16 in its natural cellular context, we introduced a wild-type human K16 gene into the germ line of transgenic mice. Several transgenic lines were established and characterized. Under most conditions, the human K16 transgene is regulated tissue specifically in the skin of transgenic mice. Animals that feature low levels of transgene expression are indistinguishable from controls during die first 6–8 months of life. In contrast, transgenic animals expressing the transgene at higher levels develop skin lesions at 1 week after birth, coinciding with the emergence of fur. At a cellular level, alterations begin with the reorganization of keratin filaments and are first seen at the level of the hair follicle outer root sheath (ORS), where K16 expression is known to occur constitutively. The lesions then progressively spread to involve the proximal epidermis, with which the ORS is contiguous. Elevated transgene expression is associated with a marked thickening of these two epithelia, along with altered keratinocyte cytoarchitecture and aberrant keratinization but no keratinocyte lysis. The implications of this phenotype for epithelial differentiation, human genodermatoses, and wound healing in skin are discussed.Key words: cytokeratin, skin, skin disease, transgenic mouse, wound healing.

Overexpression of protein kinase C-alpha in the epidermis of transgenic mice results in striking alterations in phorbol ester-induced inflammation and COX-2, MIP-2 and TNF-alpha expression but not tumor promotion

1999 ◽  
Vol 112 (20) ◽  
pp. 3497-3506
Author(s):  
H.Q. Wang ◽  
R.C. Smart
Keyword(s):  
Protein Kinase ◽  
Transgenic Mice ◽  
Normal Mouse ◽  
Mouse Skin ◽  
Tumor Promotion ◽  
Epidermal Differentiation ◽  
Wild Type ◽  
Outer Root Sheath ◽  
Proinflammatory Mediators ◽  
Cox 2

Protein kinase Calpha (PKCalpha) is one of six PKC isoforms expressed in keratinocytes of mouse epidermis. To gain an understanding of the role of epidermal PKCalpha, we have localized its expression to specific cells of normal mouse skin and examined the effect of keratin 5 (K5) promoter directed expression of PKCalpha in transgenic mice. In normal mouse skin, PKCalpha was extensively expressed in the outer root sheath (ORS) keratinocytes of the anagen hair follicle and weakly expressed in keratinocytes of interfollicular epidermis. K5-targeted expression of PKCalpha to epidermal basal keratinocytes and follicular ORS keratinocytes resulted in a tenfold increase in epidermal PKCalpha. K5-PKCalpha mice exhibited no abnormalities in keratinocyte growth and differentiation in the epidermis. However, a single topical treatment with the PKC activator, 12-O-tetradecanoylphorbol-13-acetate (TPA) resulted in a striking inflammatory response characterized by edema and extensive epidermal infiltration of neutrophils that formed intraepidermal microabscesses in the epidermis. Compared to TPA-treated wild-type mice, the epidermis of TPA-treated K5-PKCalpha mice displayed increased expression of cyclooxygenase-2 (COX-2), the neutrophil chemotactic factor macrophage inflammatory protein-2 (MIP-2) mRNA and the proinflammatory cytokine TNFalpha mRNA but not IL-6 or IL-1alpha mRNA. To determine if K5-PKCalpha mice display an altered response to TPA-promotion, 7, 12-dimethylbenz[a]anthracene-initiated K5-PKCalpha mice and wild-type mice were promoted with TPA. No differences in papilloma incidence or multiplicity were observed between K5-PKCalpha mice and wild-type littermates. These results demonstrate that the overexpression of PKCalpha in epidermis increases the expression of specific proinflammatory mediators and induces cutaneous inflammation but has little to no effect on epidermal differentiation, proliferation or TPA tumor promotion.

scholarly journals Overexpression of spermidine/spermine N1-acetyltransferase under the control of mouse metallothionein I promoter in transgenic mice: evidence for a striking post-transcriptional regulation of transgene expression by a polyamine analogue

1999 ◽  
Vol 338 (2) ◽  
pp. 311-316 ◽  
Author(s):  
Suvikki SUPPOLA ◽  
Marko PIETILÄ ◽  
Jyrki J. PARKKINEN ◽  
Veli-Pekka KORHONEN ◽  
Leena ALHONEN ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Transgenic Mice ◽  
Transgenic Mouse ◽  
Cell Injury ◽  
Mrna Level ◽  
Transgenic Animals ◽  
Ultrastructural Changes ◽  
Mouse Line ◽  
Transgenic Mouse Line ◽  
Polyamine Analogue

We recently generated a transgenic mouse line overexpressing spermidine/spermine N1-acetyltransferase (SSAT) gene under its own promoter. The tissue polyamine pools of these animals were profoundly affected and the mice were hairless from early age. We have now generated another transgenic-mouse line overexpressing the SSAT gene under the control of a heavy-metal-inducible mouse metallothionein I (MT) promoter. Even in the absence of heavy metals, changes in the tissue polyamine pools indicated that a marked activation of polyamine catabolism had occurred in the transgenic animals. As with the SSAT transgenic mice generated previously, the mice of the new line (MT-SSAT) suffered permanent hair loss, but this occurred considerably later than in the previous SSAT transgenic animals. Liver was the most affected tissue in the MT–SSAT transgenic animals, revealed by putrescine overaccumulation, significant decrease in spermidine concentration and > 90% reduction in the spermine pool. Even though hepatic SSAT mRNA accumulated to massive levels in non-induced transgenic animals, SSAT activity was only moderately elevated. Administration of ZnSO4 further elevated the level of hepatic SSAT message and induced enzyme activity, but not more than 2- to 3-fold. Treatment of the transgenic animals with the polyamine analogue N1,N11-diethylnorspermine (DENSPM) resulted in an immense induction, more than 40000-fold, of enzyme activity in the liver of transgenic animals, and minor changes in the SSAT mRNA level. Liver spermidine and spermine pools were virtually depleted within 1–2 days in response to the treatment with the analogue. The treatment also resulted in a marked mortality (up to 60%) among the transgenic animals which showed ultrastructural changes in the liver, most notably mitochondrial swelling, one of the earliest signs of cell injury. These results indicated that, even without its own promoter, SSAT is powerfully induced by the polyamine analogue through a mechanism that appears to involve a direct translational and/or heterogenous nuclear RNA processing control. It is likewise significant that overexpression of SSAT renders the animals extremely sensitive to polyamine analogues.

scholarly journals BCR/ABL P210 and P190 cause distinct leukemia in transgenic mice

Blood ◽  
1995 ◽  
Vol 86 (12) ◽  
pp. 4603-4611 ◽  
Author(s):  
JW Voncken ◽  
V Kaartinen ◽  
PK Pattengale ◽  
WT Germeraad ◽  
J Groffen ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Germ Line ◽  
Latency Period ◽  
Chronic Phase ◽  
Transgenic Animals ◽  
Hematopoietic Tissue ◽  
Long Latency ◽  
Short Period ◽  
Kinetics Of ◽  
Overt Disease

DNA constructs encoding BCR/ABL P210 have been introduced into the mouse germ line using microinjection of one-cell fertilized eggs. Kinetics of BCR/ABL P210 expression in transgenic mice were very similar to those of BCR/ABL P190 constructs in transgenic mice. mRNA transcripts were detectable early in embryonic development and also in hematopoietic tissue of adult animals. Expression of BCR/ABL in peripheral blood preceded development of overt disease. P210 founder and progeny transgenic animals, when becoming ill, developed leukemia of B, T-lymphoid, or myeloid origin after a relatively long latency period. In contrast, P190-transgenic mice exclusively developed leukemia of B-cell origin, with a relatively short period of latency. The observed dissimilarities are most likely due to intrinsically different properties of the P190 and P210 oncoproteins and may also involve sequences that control transgene expression. The delayed progression of BCR/ABL P210-associated disease in the transgenic mice is consistent with the apparent indolence of human chronic myeloid leukemia during the chronic phase. We conclude that, in transgenic models, comparable expression of BCR/ABL P210 and BCR/ABL P190 results in clinically distinct conditions.

scholarly journals Increased expression of keratin 16 causes anomalies in cytoarchitecture and keratinization in transgenic mouse skin.

1994 ◽  
Vol 127 (2) ◽  
pp. 505-520 ◽  
Author(s):  
K Takahashi ◽  
J Folmer ◽  
P A Coulombe
Keyword(s):  
Transgenic Mouse ◽  
Mouse Skin ◽  
Ultrastructural Level ◽  
Wild Type ◽  
Wound Edge ◽  
Outer Root Sheath ◽  
Keratin 16 ◽  
Keratin Expression ◽  
Root Sheath ◽  
Keratin Filaments

Injury to epidermis and other stratified epithelia triggers profound but transient changes in the pattern of keratin expression. In postmitotic cells located at the wound edge, a strong induction of K6, K16, and K17 synthesis occurs at the expense of the keratins produced under the normal situation. The functional significance of these alterations in keratin expression is not known. Here, we report that overexpression of a wild-type human K16 gene in a tissue-specific fashion in transgenic mice causes aberrant keratinization of the hair follicle outer root sheath and proximal epidermis, and it leads to hyperproliferation and increased thickness of the living layers (acanthosis), as well as cornified layers (hyperkeratosis). The pathogenesis of lesions in transgenic mouse skin begins with a reorganization of keratin filaments in postmitotic keratinocytes, and it progresses in a transgene level-dependent fashion to include disruption of keratinocyte cytoarchitecture and structural alterations in desmosomes at the cell surface. No evidence of cell lysis could be found at the ultrastructural level. These results demonstrate that the disruption of the normal keratin profile caused by increased K16 expression interferes with the program of terminal differentiation in outer root sheath and epidermis. They further suggest that when present at sufficiently high intracellular levels, K16, along with K6 and K17, appear capable of inducing a reorganization of keratin filaments in the cytoplasm of skin epithelial cells.

A retinoic acid-inducible transgenic marker of sino-atrial development in the mouse heart

Development ◽  
1999 ◽  
Vol 126 (12) ◽  
pp. 2677-2687 ◽  
Author(s):  
J. Xavier-Neto ◽  
C.M. Neville ◽  
M.D. Shapiro ◽  
L. Houghton ◽  
G.F. Wang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Alkaline Phosphatase ◽  
Retinoic Acid ◽  
Transgenic Mice ◽  
Transgene Expression ◽  
Single Pulse ◽  
Transgenic Animals ◽  
Specific Gene ◽  
Mouse Heart ◽  
Pregnant Mice

To study the specification of inflow structures in the heart we generated transgenic animals harboring the human alkaline phosphatase (HAP) gene driven by the proximal 840 bp of a quail SMyHC3 promoter. In transgenic mice, the SMyHC3-HAP reporter was expressed in posterior heart precursors at 8.25 dpc, in sinus venosa and in the atrium at 8.5 and 9.0 dpc, and in the atria from 10.5 dpc onwards. SMyHC3-HAP transgene expression overlapped synthesis and endogenous response to retinoic acid (RA) in the heart, as determined by antibodies directed against a key RA synthetic enzyme and by staining of RAREhsplacZ transgenic animals. A single pulse of all-trans RA administered to pregnant mice at 7.5, but not after 8.5, dpc induced cardiac dismorphology, ranging from complete absence of outflow tract and ventricles to hearts with reduced ventricles expressing both SMyHC3-HAP and ventricular markers. Blockade of RA synthesis with disulfiram inhibited RA-induced transcription and produced hearts lacking the atrial chamber. This study defines a novel marker for atrial-restricted transcription in the developing mouse heart. It also suggests that atrial-specific gene expression is controlled by localized synthesis of RA, and that exclusion of RA from ventricular precursors is essential for correct specification of the ventricles.

scholarly journals Analysis of the Mouse CSF-1 Gene Promoter in a Transgenic Mouse Model1

2003 ◽  
Vol 51 (7) ◽  
pp. 941-949 ◽  
Author(s):  
Sherry L. Abboud ◽  
Maria Bunegin ◽  
Nandini Ghosh-Choudhury ◽  
Kathleen Woodruff
Keyword(s):  
Gene Expression ◽  
Transgenic Mice ◽  
Transgene Expression ◽  
Transcriptional Activity ◽  
Molecular Mechanisms ◽  
Tissue Expression ◽  
Cellular Level ◽  
Liver And Kidney

CSF-1 stimulates monocyte and osteoclast populations. However, the molecular mechanisms involved in regulating CSF-1 gene expression are unclear. To identify regulatory regions that control normal CSF-1 gene expression, a −774/+183-bp fragment of the murine CSF-1 promoter was analyzed in vitro and in vivo. Transcriptional activity was high in cultured osteoblasts that express CSF-1 mRNA compared to ARH-77 B cells that lack CSF-1 gene expression. Transient transfection of osteoblasts with promoter deletion constructs showed that the −774-bp fragment conferred the highest transcriptional activity and contained activator and repressor sequences. To assess the ability of the CSF-1 promoter to confer normal tissue expression of CSF-1, transgenic mice containing the −774/+183-bp region driving the E. coli β-galactosidase (lacZ) reporter gene were generated. β-Gal analysis of whole tissue extracts showed transgene expression in all tissues tested except liver and kidney. At the cellular level, the pattern of β-gal expression in the spleen, thymus, bone, lung, and testes of adult transgenic mice mimicked normal endogenous CSF-1 mRNA expression in non-transgenic littermates detected by in situ hybridization. This region also directed appropriate transgene expression to sites in other tissues known to synthesize CSF-1, with the exception of the liver and kidney. These findings indicate that the −774-bp fragment contains cis-acting elements sufficient to direct CSF-1 gene expression in many tissues. CSF-1 promoter/lacZ mice may be useful for studying the transcriptional mechanisms involved in regulating CSF-1 gene expression in tissues throughout development.

scholarly journals Study into molecular targets of a neuroprotective compound dimebon using a transgenic mice line

2014 ◽  
Vol 60 (3) ◽  
pp. 354-363
Author(s):  
T.A. Shelkovnikova ◽  
A.A. Ustyugov ◽  
V.S. Kokhan ◽  
T.V. Tarasova ◽  
V.K. Medvedeva ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Nervous System ◽  
Transgenic Mice ◽  
Transgene Expression ◽  
Neuroprotective Effect ◽  
Molecular Targets ◽  
Transgenic Animals ◽  
Protein Species ◽  
Ubiquitinated Proteins ◽  
Amyloidogenic Protein

In the present study we have used a transgenic mice overexpressing an amyloidogenic protein, gamma-synuclein, in the nervous system to address the effect of dimebon on proteinopathy progression. Neuroprotective effect of chronic dimebon administration in these mice at organismal level was confirmed by the increased lifespan. Using histological and biochemical approaches we have demonstrated that dimebon reduced the number of amyloid inclusions in spinal cord of transgenic animals and decreased the content of ubiquitinated proteins in detergent-insoluble fractions. These effects are likely to occur at the level of aggregated protein species, since transgene expression was not altered. Thus, pathological protein aggregation serves as one of dimebon targets in neurodegeneration.

scholarly journals Antinuclear Autoantibodies and Lupus Nephritis in Transgenic Mice Expressing Interferon γ in the Epidermis

1997 ◽  
Vol 186 (9) ◽  
pp. 1451-1459 ◽  
Author(s):  
John P. Seery ◽  
Joseph M. Carroll ◽  
Victoria Cattell ◽  
Fiona M. Watt
Keyword(s):  
Transgenic Mice ◽  
Lupus Erythematosus ◽  
Transgenic Animals ◽  
Organ Damage ◽  
Skin Lesions ◽  
Interferon Γ ◽  
Female Mice ◽  
Skin Immune System ◽  
Organ Specific ◽  
Ifn Γ

Systemic lupus erythematosus (SLE) is a potentially fatal non–organ-specific autoimmune disease that predominantly affects women. Features of the disease include inflammatory skin lesions and widespread organ damage caused by deposition of anti-dsDNA autoantibodies. The mechanism and site of production of these autoantibodies is unknown, but there is evidence that interferon (IFN) γ plays a key role. We have used the involucrin promoter to overexpress IFN-γ in the suprabasal layers of transgenic mouse epidermis. There was no evidence of organ-specific autoimmunity, but transgenic animals produced autoantibodies against dsDNA and histones. Autoantibody levels in female mice were significantly higher than in male transgenic mice. Furthermore, there was IgG deposition in the glomeruli of all female mice and histological evidence of severe proliferative glomerulonephritis in a proportion of these animals. Our findings are consistent with a central role for the skin immune system, acting under the influence of IFN-γ, in the pathogenesis of SLE.

scholarly journals Efficient Generation of Mice with Consistent Transgene Expression by FEEST

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Lei Gao ◽  
Yonghua Jiang ◽  
Libing Mu ◽  
Yanbin Liu ◽  
Fengchao Wang ◽  
...  
Keyword(s):  
Mouse Model ◽  
Transgenic Mice ◽  
Transgenic Mouse ◽  
Mouse Models ◽  
Transgene Expression ◽  
Es Cells ◽  
Mouse Line ◽  
Transgenic Mouse Models ◽  
Transgenic Mouse Line ◽  
A Genome

Abstract Transgenic mouse models are widely used in biomedical research; however, current techniques for producing transgenic mice are limited due to the unpredictable nature of transgene expression. Here, we report a novel, highly efficient technique for the generation of transgenic mice with single-copy integration of the transgene and guaranteed expression of the gene-of-interest (GOI). We refer to this technique as functionally enriched ES cell transgenics, or FEEST. ES cells harboring an inducible Cre gene enabled the efficient selection of transgenic ES cell clones using hygromycin before Cre-mediated recombination. Expression of the GOI was confirmed by assaying for the GFP after Cre recombination. As a proof-of-principle, we produced a transgenic mouse line containing Cre-activatable tTA (cl-tTA6). This tTA mouse model was able to induce tumor formation when crossed with a transgenic mouse line containing a doxycycline-inducible oncogene. We also showed that the cl-tTA6 mouse is a valuable tool for faithfully recapitulating the clinical course of tumor development. We showed that FEEST can be easily adapted for other genes by preparing a transgenic mouse model of conditionally activatable EGFR L858R. Thus, FEEST is a technique with the potential to generate transgenic mouse models at a genome-wide scale.

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