Changes in expression of the lysosomal membrane glycoprotein, LAMP-1 in interdigital regions during embryonic mouse limb development, in vivo and in vitro

The glutamine analogue, 6-diazo-5-oxo-L-norleucine (DON), has been shown to inhibit biosynthesis of purines and glycosaminoglycans, presumably by blocking the glutaminedependent steps in the biosynthetic pathways. The teratogenic potential of DON on the developing mouse limb-bud in vivo and in vitro was studied in an attempt to discriminate whether DON is exerting its teratogenic effect by interfering with glycosaminoglycan orpurine metabolism. A single intramuscular injection of DON (0·5 mg/kg) to ICR/DUB mice on day 10 of gestation resulted in 76% resorption, while fetuses surviving to day 17 exhibited growth retardation, median cleft lip, and limb malformations. Concurrent administration of Lglutamine (250 mg/kg) provided no protection against resorption or malformations, while 5-aminoimidazolecarboxamide (AIC, 250 mg/kg) decreased the resorption rate to 34% without significantly altering the incidence of malformations. Injection of DON alone on day 11 resulted in 87% of fetuses exhibiting limb malformations, with only 2% resorption. Concurrent injection of AIC decreased the frequency of limb malformations to 32%. L-Glutamine, D-glucosamine, or inosinic acid were without any protective effect in vivo. DON (5 μg/ml medium) added in vitro to organ cultures of day 11 mouse limb-buds caused all limbs to evidence cartilage abnormalities. In this system, either L-glutamine or D-glucosamine (0·5 mg/ml medium) provided protection against DON effects while AIC (0·5 mg/ml medium) offered no protection in vitro. These data suggest that DON exerts its effects in vivo by interfering with purine metabolism while in vitro its teratogenic action may be interruption of glycosaminoglycan biosynthesis. This may reflect upon the relative importance of growth and differentiation to limb development in vivo and in vitro. These data infer that limb development in vitro relies more on the differentiative process (differentiation of cartilage) than on growth, whereas limb development in vivo is dependent, at this stage, to a greater extent on growth for normal phenotypic expression.

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Effects of cytosine arabinoside on in vivo and in vitro mouse limb development

In Vitro ◽

10.1007/bf02615104 ◽

1977 ◽

Vol 13 (7) ◽

pp. 434-442 ◽

Cited By ~ 3

Author(s):

Jeanne M. Manson ◽

Michael L. Dourson ◽

Carl C. Smith

Keyword(s):

Cytosine Arabinoside ◽

Limb Development ◽

Mouse Limb

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The ulnar-mammary syndrome gene, Tbx3, is a direct target of the retinoic acid signaling pathway, which regulates its expression during mouse limb development

Molecular Biology of the Cell ◽

10.1091/mbc.e11-09-0790 ◽

2012 ◽

Vol 23 (12) ◽

pp. 2362-2372 ◽

Cited By ~ 14

Author(s):

Reyna Deeya Ballim ◽

Cathy Mendelsohn ◽

Virginia E. Papaioannou ◽

Sharon Prince

Keyword(s):

Retinoic Acid ◽

Signaling Pathway ◽

Limb Development ◽

Postnatal Life ◽

Functional Protein ◽

Embryonic Limb ◽

Mouse Limb ◽

Direct Target

TBX3, a member of the T-box transcription factor gene family, is a transcriptional repressor that is required for the development of the heart, limbs, and mammary glands. Mutations in TBX3 that result in reduced functional protein lead to ulnar-mammary syndrome, a developmental disorder characterized by limb, mammary gland, tooth, and genital abnormalities. Increased levels of TBX3 have been shown to contribute to the oncogenic process, and TBX3 is overexpressed in several cancers, including breast cancer, liver cancer, and melanoma. Despite its important role in development and postnatal life, little is known about the signaling pathways that modulate TBX3 expression. Here we show, using in vitro and in vivo assays, that retinoic acid (RA) activates endogenous TBX3 expression, which is mediated by an RA–receptor complex directly binding and activating the TBX3 promoter, and we provide evidence that this regulation may be functionally relevant in mouse embryonic limb development. Our data identify TBX3 as a direct target of the RA signaling pathway and extend our understanding of the role and regulation of TBX3 in limb development.

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Hst-1 (FGF-4) antisense oligonucleotides block murine limb development.

The Journal of Cell Biology ◽

10.1083/jcb.130.4.997 ◽

1995 ◽

Vol 130 (4) ◽

pp. 997-1003 ◽

Cited By ~ 18

Author(s):

T Ochiya ◽

H Sakamoto ◽

M Tsukamoto ◽

T Sugimura ◽

M Terada

Keyword(s):

Organ Culture ◽

Limb Development ◽

Culture System ◽

Inhibitory Effect ◽

Defined Medium ◽

Limb Bud ◽

Embryonic Mouse ◽

Organ Culture System ◽

Mouse Limb

The initiation of limb development depends on the site specific proliferation of the mesenchyme by the signals from the apical ectodermal ridge (AER) in embryonic mouse. We have previously reported that the local expression of Hst-1/Fgf-4 transcripts in AER of the mouse limb bud is developmentally regulated, expressed at 11 and 12 days post coitus (p.c.) embryo. In an effort to further understand the role of Hst-1/FGF-4 in mouse limb development, an antisense oligodeoxynucleotides (ODNs) study was performed. We first established a novel organ culture system to study mouse limb development in vitro. This system allows mouse limb bud at 9.5-10-d p.c. embryo, when placed on a sheet of extracellular matrix in a defined medium, to differentiate into a limb at 12.5-d p.c. embryo within 4.5 d. Using this organ culture system, we have shown that exposure of 9.5-10-d p.c. embryonal limb bud explants to antisense ODNs of Hst-1/FGF-4 blocks limb development. In contrast, sense and scrambled ODNs have no inhibitory effect on limb outgrowth, suggesting that Hst-1/FGF-4 may work as a potent inducing factor for mouse limb development.

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Sorting of mannose 6-phosphate receptors and lysosomal membrane proteins in endocytic vesicles.

The Journal of Cell Biology ◽

10.1083/jcb.107.6.2491 ◽

1988 ◽

Vol 107 (6) ◽

pp. 2491-2501 ◽

Cited By ~ 127

Author(s):

H J Geuze ◽

W Stoorvogel ◽

G J Strous ◽

J W Slot ◽

J E Bleekemolen ◽

...

Keyword(s):

Lysosomal Membrane ◽

Membrane Glycoprotein ◽

Horse Radish Peroxidase ◽

Western Blots ◽

Lysosomal Membrane Proteins ◽

Mannose 6 Phosphate ◽

Lysosomal Membrane Glycoprotein ◽

Endocytic Vesicles ◽

Kinetics Of ◽

Combined Data

The intracellular distributions of the cation-independent mannose 6-phosphate receptor (MPR) and a 120-kD lysosomal membrane glycoprotein (lgp120) were studied in rat hepatoma cells. Using quantitative immunogold cytochemistry we found 10% of the cell's MPR located at the cell surface. In contrast, lgp120 was not detectable at the plasma membrane. Intracellularly, MPR mainly occurred in the trans-Golgi reticulum (TGR) and endosomes. lgp120, on the other hand, was confined to endosomes and lysosomes. MPR was present in both endosomal tubules and vacuoles, whereas lgp120 was confined to the endosomal vacuoles. In cells incubated for 5-60 min with the endocytic tracer cationized ferritin, four categories of endocytic vacuoles could be discerned, i.e., vacuoles designated MPR+/lgp120-, MPR+/lgp120+, MPR-/lgp120+, and vacuoles nonimmunolabeled for MPR and lgp120. Tracer first reached MPR+/lgp120-, then MPR+/lgp120+, and finally MPR-/lgp120+ vacuoles, which are assumed to represent lysosomes. To study the kinetics of appearance of endocytic tracers in MPR-and/or lgp120-containing pools in greater detail, cells were allowed to endocytose horse-radish peroxidase (HRP) for 5-90 min. The reduction in detectability of MPR and lgp120 antigenicity on Western blots, due to treatment of cell homogenates with 3'3-diaminobenzidine, was followed in time. We found that HRP reached the entire accessible pool of MPR almost immediately after internalization of the tracer, while prolonged periods of time were required for HRP to maximally access lgp120. The combined data suggest that MPR+/lgp120+ vacuoles are endocytic vacuoles, intermediate between MPR+/lgp120-endosomes and MPR-/lgp120+ lysosomes, and represent the site where MPR is sorted from lgp120 destined for lysosomes. We propose that MPR is sorted from lgp120 by selective lateral distribution of the receptor into the tubules of this compartment, resulting in the retention of lgp120 in the vacuoles and the net transport of lgp120 to lysosomes.

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Mechanism of Aloe Vera extract protection against UVA: shelter of lysosomal membrane avoids photodamage

Photochemical & Photobiological Sciences ◽

10.1039/c5pp00409h ◽

2016 ◽

Vol 15 (3) ◽

pp. 334-350 ◽

Cited By ~ 19

Author(s):

Daniela Rodrigues ◽

Ana Cláudia Viotto ◽

Robert Checchia ◽

Andreza Gomide ◽

Divinomar Severino ◽

...

Keyword(s):

Aloe Vera ◽

Lysosomal Membrane

Aloe Vera extract exhibited remarkable ability of reducing both in vitro and in vivo photodamage, even though it does not have anti-radical properties.

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Nerve-dependent accumulation of myosin light chain 3 in developing limb musculature

Development ◽

10.1242/dev.101.4.673 ◽

1987 ◽

Vol 101 (4) ◽

pp. 673-684

Author(s):

P.A. Merrifield ◽

I.R. Konigsberg

Keyword(s):

Neural Tube ◽

Light Chain ◽

Limb Development ◽

Myosin Light Chain ◽

Chorioallantoic Membrane ◽

In Ovo ◽

Fast Myosin ◽

Limb Musculature

Myosin alkali light chain accumulation in developing quail limb musculature has been analysed on immunoblots using a monoclonal antibody which recognizes an epitope common to fast myosin light chain 1 (MLC1f) and fast myosin light chain 3 (MLC3f). The limb muscle of early embryos (i.e. up to day 10 in ovo) has a MLC profile similar to that observed in myotubes cultured in vitro; although MLC1f is abundant, MLC3f cannot be detected. MLC3f is first detected in 11-day embryos. To determine whether this alteration in MLC3f accumulation is nerve or hormone dependent, limb buds with and without neural tube were cultured as grafts on the chorioallantoic membrane of chick hosts. Although differentiated muscle develops in both aneural and innervated grafts, innervated grafts contain approximately three times as much myosin as aneural grafts. More significantly, although aneural grafts reproducibly accumulate normal levels of MLC1f, they fail to accumulate detectable levels of MLC3f. In contrast, innervated grafts accumulate both MLC1f and MLC3f, suggesting that the presence of neural tube in the graft promotes the maturation, as well as the growth, of muscle tissue. This is the first positive demonstration that innervation is necessary for the accumulation of MLC3f that occurs during normal limb development in vivo.

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Vitamin A alters the internal viscosity of fragments of limb-bud mesenchyme

Development ◽

10.1242/dev.59.1.325 ◽

1980 ◽

Vol 59 (1) ◽

pp. 325-339

Author(s):

T. E. Kwasigroch ◽

D. M. Kochhar

Keyword(s):

Retinoic Acid ◽

Vitamin A ◽

Mesenchymal Cells ◽

Limb Bud ◽

Mouse Limb ◽

Vitamin A Acid ◽

Internal Viscosity ◽

Fusion Experiments

Two techniques were used to examine the effect of vitamin A compounds (vitamin A acid = retinoic acid and vitamin A acetate) upon the relative strengths of adhesion among mouse limb-bud mesenchymal cells. Treatment with retinoic acid in vivo and with vitamin A acetate in vitro reduced the rate at which the fragments of mesenchyme rounded-up when cultured on a non-adhesive substratum, but these compounds did not alter the behavior of tissues tested in fragment-fusion experiments. These conflicting results indicate that the two tests measure different activities of cells and suggest that treatment with vitamin A alters the property(ies) of cells which regulate the internal viscosity of tissues.

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A new in vitro system for studying secondary palate development

Development ◽

10.1242/dev.34.2.485 ◽

1975 ◽

Vol 34 (2) ◽

pp. 485-495

Author(s):

L. Brinkley ◽

G. Basehoar ◽

A. Branch ◽

J. Avery

Keyword(s):

Gas Permeation ◽

Present System ◽

Embryonic Mouse ◽

Fluid Medium ◽

In Vitro System ◽

Palate Development ◽

Fixed Orientation ◽

The Brain

An in vitro system was devised which supports palate development in partially dissected embryonic mouse heads. The heads were suspended in the culture chamber so that they were not held in a fixed orientation and were constantly surrounded with a fluid medium. Under these circumstances the developing palate must effect closure without the aid of gravitational forces. The culture medium was constantly circulated, gassed with 95% O2, 5% CO2 using hollow fiber gas permeation devices, and kept at 34°C. Swiss-Webster mouse embryos of 12 days 12–18 h (ca. 48 h prior to expected in vivo closure) or 13 days 8–14 h (ca. 24 h prior to closure) were used to test the ability of the system to support palatal development. Embryonic heads were dissected in one of two ways before culture: brain and tongue removed, or brain, tongue and mandible removed. After 24 h in culture, preparations of either age with only the brain and tongue removed had made substantially greater progress than their counterparts with the brain, tongue and mandible removed. With only the brain and tongue removed, the palatal shelves were contacting, adhered or fused in 67 % of the older embryos, whereas most of the embryos of the same age cultured with the brain, tongue and mandible removed had shelves that were not fully elevated and still separated by a moderate gap. Thus for maximal progress in the present system, the oral cavity must be intact except for the tongue.

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CD44v10: An Antimetastatic Membrane Glycoprotein for Pancreatic Cancer

The International Journal of Biological Markers ◽

10.1177/172460080301800206 ◽

2003 ◽

Vol 18 (2) ◽

pp. 130-138 ◽

Cited By ~ 5

Author(s):

F. Navaglia ◽

P. Fogar ◽

E. Greco ◽

D. Basso ◽

A.L. Stefani ◽

...

Keyword(s):

Pancreatic Cancer ◽

Membrane Glycoprotein ◽

Rt Pcr ◽

In Vivo Experiments ◽

Microtiter Plates ◽

Pancreatic Metastases ◽

Metastatic Behavior ◽

Mrna Analysis

Aims The aims of this study were 1) to investigate the mRNA pattern of CD44 variants in three primary (MIA PaCa 2, PANC-1, PSN-1) and two metastatic (CAPAN-1, SUIT-2) pancreatic cancer (PC) cell lines; 2) to ascertain whether the genetic transfer of CD44s and CD44v10 modifies the adhesion of PC cells to the extracellular matrix (ECM) in vitro and their metastatic behavior in vivo. Methods CD44 mRNA analysis was done by means of RT-PCR. Adhesion to ECM the was assessed using coated microtiter plates. For the study of CD44v10 insertion in the CAPAN-1 line, liposome-mediated DNA transfer was used. SCID mice were employed for in vivo experiments. Results CD44v10 mRNA was not expressed by the CAPAN-1 nor by four of the six SUIT-2-derived clones. The stable expression of CD44v10 by modified CAPAN-1 significantly enhanced fibronectin adhesion. Mice without either liver or pancreatic metastases were more frequently found among the animals injected with modified (CD44v10 expressing) than with non-modified CAPAN-1. Conclusions 1) It is possible to differentiate between metastatic and non-metastatic PC cells on the basis of CD44v10 expression; 2) CD44v10 seems to be involved in mediating fibronectin adhesion in vitro and in counteracting metastases in vivo.

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