Mouse chick chimera: a new model to study the in ovo developmental potentialities of mammalian somites

Chimeras were prepared by transplanting somites from 9-day post-coitum mouse embryos or somitic dermomyotomes from 10-day post-coitum mouse embryos into 2-day-old chick embryos at different axial levels. Mouse somitic cells then differentiated in ovo in dermis, cartilage and skeletal muscle as they normally do in the course of development and were able to migrate into chick host limb. To trace the behavior of somitic myogenic stem cells more closely, somites arising from mice bearing a transgene of the desmin gene linked to a reporter gene coding for Escherichia coli beta-galactosidase (lacZ) were grafted in ovo. Interestingly, the transgene was rapidly expressed in myotomal muscles derived from implants. In the limb muscle mass, positive cells were found several days after implantation. Activation of desmin nls lacZ also occurred in in vitro cultures of somite-derived cells. Our experimental method facilitates investigation of the mechanisms of mammalian development, allowing the normal fate of implanted mouse cells to be studied and providing suitable conditions for identification of descendants of genetically modified cells.

Download Full-text

EFFECTS OF ANAEROBIOSIS ON THE RATES OF MULTIPLICATION OF MAMMALIAN CELLS CULTURED IN VITRO

Canadian Journal of Biochemistry and Physiology ◽

10.1139/y60-108 ◽

1960 ◽

Vol 38 (1) ◽

pp. 871-878 ◽

Cited By ~ 1

Author(s):

Samuel Dales

Keyword(s):

Mammalian Cells ◽

Glucose Utilization ◽

Lactic Acid Production ◽

Cell Types ◽

In Vitro Cultures ◽

Cell Multiplication ◽

Embryonic Mouse ◽

Mouse Cells ◽

Oxidation Of Glucose

To test the effects of anaerobiosis on the rate of multiplication and carbohydrate metabolism of mammalian cells in vitro, cultures of a 'permanent' line, Earle's L strain cells, and of freshly explanted embryonic mouse cells were propagated in the presence and absence of oxygen. Contrary to the findings of several other investigators, our results show that the multiplication of both cell types was depressed by anaerobiosis. Anaerobiosis for at least 7 days, did not, however, bring about unbalanced growth in L cells, nor did it affect their capability to divide rapidly soon after they were returned to aerobic conditions. From the rates of glucose utilization, lactic acid production, and cell multiplication it was estimated that the rate of division in the two cell types studied was proportional to the energy which could be released from either glycolysis or complete oxidation of glucose.

Download Full-text

Teratogenic activity of methadone hydrochloride in mouse and chick embryos

Development ◽

10.1242/dev.30.2.449 ◽

1973 ◽

Vol 30 (2) ◽

pp. 449-458

Author(s):

A. Jurand

Keyword(s):

Spinal Cord ◽

Central Nervous System ◽

Nervous System ◽

Neural Plate ◽

Mouse Embryos ◽

Chick Embryos ◽

In Ovo ◽

Cervical Area ◽

Extensive Necrosis

Teratogenic activity of methadone HCl (Physeptone, Burroughs Wellcome and Co.) was tested on inbred JBT/Jd and outbred Q strain mouse embryos and on chick embryos. 22–24 mg/kg injected subcutaneously on the 9th day of pregnancy caused by the 13th day exencephaly in 56 out of 479 JBT/Jd embryos but after 32 mg/kg only in 1 out of 220 of the Q strain. Some affected JBT/Jd embryos showed also rachischisis in the cervical area. The second abnormality shown by the embryos of both strains is Z-shaped kinkage of the spinal cord. In explanted chick embryos cultured in vitro as well as in embryos treated in ovo methadone causes non-closure of the neural tube with extensive necrosis of the neural plate cells in the cephalic region. The results of this study indicate that methadone, which is a neutropic drug, has an embryotoxic activity directed against the developing central nervous system.

Download Full-text

Induction and abrogation of unresponsiveness in nude mouse cells.

Journal of Experimental Medicine ◽

10.1084/jem.144.6.1458 ◽

1976 ◽

Vol 144 (6) ◽

pp. 1458-1464 ◽

Cited By ~ 4

Author(s):

R Bösing-Schneider ◽

M Haug

Keyword(s):

T Cells ◽

T Cell ◽

Nude Mouse ◽

Nude Mice ◽

In Vitro Cultures ◽

Spleen Cells ◽

Mouse Cells ◽

In Vivo Administration

Nude mice were injected with antigen and T cells at different times to induce unresponsiveness to SRBC. Spleen cells derived from these mice were tested in vitro for the capability to produce antibody-forming cells against sheep erythrocytes in the presence of a T-cell-replacing factor. It was found that priming with antigen alone did not result in paralysis but a later injection of thymus-derived lymphocytes together with antigen results in unresponsiveness of these cells in vitro, provided there was an interval of several days between the in vivo administration of thymus lymphocytes and the explantations of cells to in vitro cultures.

Download Full-text

The development of monosomy 19 mouse embryos

Development ◽

10.1242/dev.69.1.223 ◽

1982 ◽

Vol 69 (1) ◽

pp. 223-236

Author(s):

Terry Magnuson ◽

Sandra Smith ◽

Charles J. Epstein

Keyword(s):

Inbred Strains ◽

Mouse Embryos ◽

Specific Gene ◽

Mammalian Development ◽

Two Dimensional Gel Electrophoresis ◽

Autosomal Trisomy ◽

Morula Stage ◽

A Cell

In general, autosomal monosomy is lethal much earlier in mammalian development than autosomal trisomy. In an attempt to understand why monosomy is so deleterious, we have begun to characterize the development of mouse embryos monosomic for chromosome 19. A dramatic loss of monosomy 19 embryos was found to occur between days 3 and 4 of development. This loss occurred both in vivo and in vitro and with intact blastocysts or isolated inner cell masses. Experiments with inbred strains showed that this loss was not due to the expression of recessive lethal genes. While monosomic embryos were found to have fewer cells than normal and trisomic litter-mates beginning at the early morula stage, the ability to form blastocystsis not interfered with. Electron microscopy revealed no difference in the cellular ultrastructure of monosomic when compared with diploid embryos. Furthermore, two-dimensional gel electrophoresis did not reveal any differences in the proteins synthesized by monosomic, trisomic or diploid litter-mates when examined at day 3 ofdevelopment. These results indicate a lack of gross genomic disturbances in monosomic embryos. When monosomy ↔ diploid chimaeras were made, viable monosomic cells were found in day-9 post-implantation embryos, well past the lethal period. Thus, in chimaeric embryos, the normal cells appear to be able to provide whatever is lacking, suggesting that monosomy 19 is not a cell lethal. Instead, death may be due to a dosage alterationin specific gene products needed during early development.

Download Full-text

EFFECTS OF ANAEROBIOSIS ON THE RATES OF MULTIPLICATION OF MAMMALIAN CELLS CULTURED IN VITRO

Canadian Journal of Biochemistry and Physiology ◽

10.1139/o60-108 ◽

1960 ◽

Vol 38 (8) ◽

pp. 871-878 ◽

Cited By ~ 20

Author(s):

Samuel Dales

Keyword(s):

Mammalian Cells ◽

Glucose Utilization ◽

Lactic Acid Production ◽

Cell Types ◽

In Vitro Cultures ◽

Cell Multiplication ◽

Embryonic Mouse ◽

Mouse Cells ◽

Oxidation Of Glucose

Download Full-text

Some Structural Features of Metaphase Chromosomes of Mice and Men

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100060969 ◽

1967 ◽

Vol 25 ◽

pp. 190-191

Author(s):

Godfrey C. Hoskins ◽

Betty B. Hoskins

Keyword(s):

Electron Microscopy ◽

Electron Micrographs ◽

Structural Features ◽

Metaphase Chromosomes ◽

The Future ◽

Of Mice And Men ◽

Mouse Cells ◽

Human And Mouse

Metaphase chromosomes from human and mouse cells in vitro are isolated by micrurgy, fixed, and placed on grids for electron microscopy. Interpretations of electron micrographs by current methods indicate the following structural features.Chromosomal spindle fibrils about 200Å thick form fascicles about 600Å thick, wrapped by dense spiraling fibrils (DSF) less than 100Å thick as they near the kinomere. Such a fascicle joins the future daughter kinomere of each metaphase chromatid with those of adjacent non-homologous chromatids to either side. Thus, four fascicles (SF, 1-4) attach to each metaphase kinomere (K). It is thought that fascicles extend from the kinomere poleward, fray out to let chromosomal fibrils act as traction fibrils against polar fibrils, then regroup to join the adjacent kinomere.

Download Full-text

Human Adenovirus Uptake by Preirnplantation Mouse Embryos

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100094607 ◽

1972 ◽

Vol 30 ◽

pp. 268-269

Author(s):

D. G. Chase ◽

W. Winters ◽

L. Piko

Keyword(s):

Hela Cells ◽

Human Adenovirus ◽

Mouse Embryos ◽

Equilibrium Density ◽

Cell Stage ◽

Virus Attachment ◽

Preimplantation Mouse Embryos ◽

Embryo Culture Medium ◽

Preimplantation Mouse ◽

Mouse Cells

Although the outlines of human adenovirus entry and uncoating in HeLa cells has been clarified in recent electron microscope studies, several details remain unclear or controversial. Furthermore, morphological features of early interactions of human adenovirus with non-permissive mouse cells have not been extensively documented. In the course of studies on the effects of human adenoviruses type 5 (AD-5) and type 12 on cultured preimplantation mouse embryos we have examined virus attachment, entry and uncoating. Here we present the ultrastructural findings for AD-5.AD-5 was grown in HeLa cells and purified by successive velocity gradient and equilibrium density gradient centrifugations in CsCl. After dialysis against PBS, virus was sedimented and resuspended in embryo culture medium. Embryos were placed in culture at the 2-cell stage in Brinster's medium.

Download Full-text