Xenopus laevis transgenesis by sperm nuclear injection

Stuart J Smith; Lynne Fairclough; Branko V Latinkic; Duncan B Sparrow; Timothy J Mohun

doi:10.1038/nprot.2006.325

Injected nuclei in frog oocytes: fate, enlargement, and chromatin dispersal

Development ◽

10.1242/dev.36.3.523 ◽

1976 ◽

Vol 36 (3) ◽

pp. 523-540

Author(s):

J. B. Gurdon

Keyword(s):

Bovine Serum Albumin ◽

Xenopus Laevis ◽

Serum Albumin ◽

Bovine Serum ◽

Oocyte Nucleus ◽

Oocyte Cytoplasm ◽

Nuclear Injection

A method is described by which nuclei associated with some cytoplasm can be rapidly prepared from a suspension of cells. The method involves the use of lysolecithin and bovine serum albumin. Oocytes of Xenopus laevis were injected with about 200 nuclei prepared from human He La cells by this method. Nuclei were deposited in oocyte cytoplasm, in the oocyte nucleus, or in the dispersed contents of a ruptured oocyte nucleus. Injected He La nuclei enlarge up to several hundred times in volume in the course of a few days. Their enlargement is associated with chromatin dispersion, increased binding of an acidic dye, and with the reduction in size, and eventual disappearance, of nucleoli. The amount of He La nucleus enlargement is much greater when the oocyte nucleus is ruptured. The fate of injected nuclei was followed by the use of HeLa nuclei whose DNA had been previously la belled with [3H] thymidine. La belled DNA does not pass from injected He La nuclei into the oocyte nucleus. Injected nuclei appear not to fuse with each other or with the oocyte nucleus. Nuclei prepared by the above method look morphologically healthy in oocytes cultured in vitro for up to one month after nuclear injection. Nuclei prepared by other methods, such as those involving the use of detergents, undergo deterioration within a few days after injection into oocytes.

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Author response for "Amphibian thalamic nuclear organization during larval development and in the adult frog Xenopus laevis : genoarchitecture and hodological analysis"

10.1002/cne.24899/v2/response1 ◽

2020 ◽

Author(s):

Ruth Morona ◽

Sandra Bandín ◽

Jesús M. López ◽

Nerea Moreno ◽

Agustín González

Keyword(s):

Xenopus Laevis ◽

Larval Development ◽

Nuclear Organization ◽

Author Response ◽

Adult Frog

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Androgen-induced plasticity at a “vocal” neuromuscular synapse

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100167895 ◽

1994 ◽

Vol 52 ◽

pp. 32-33

Author(s):

Darcy B. Kelley ◽

Martha L. Tobias ◽

Mark Ellisman

Keyword(s):

Xenopus Laevis ◽

Motor Neurons ◽

Sexual Differentiation ◽

Molecular Basis ◽

Neuromuscular Synapse ◽

Sexually Dimorphic ◽

Intracellular Protein ◽

Developmental Program ◽

Androgen Action ◽

Clawed Frog

Brain and muscle are sexually differentiated tissues in which masculinization is controlled by the secretion of androgens from the testes. Sensitivity to androgen is conferred by the expression of an intracellular protein, the androgen receptor. A central problem of sexual differentiation is thus to understand the cellular and molecular basis of androgen action. We do not understand how hormone occupancy of a receptor translates into an alteration in the developmental program of the target cell. Our studies on sexual differentiation of brain and muscle in Xenopus laevis are designed to explore the molecular basis of androgen induced sexual differentiation by examining how this hormone controls the masculinization of brain and muscle targets.Our approach to this problem has focused on a highly androgen sensitive, sexually dimorphic neuromuscular system: laryngeal muscles and motor neurons of the clawed frog, Xenopus laevis. We have been studying sex differences at a synapse, the laryngeal neuromuscular junction, which mediates sexually dimorphic vocal behavior in Xenopus laevis frogs.

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