Embryo Project Encyclopedia

"A Diffusible Agent of Mouse Sarcoma, Producing Hyperplasia of Sympathetic Ganglia and Hyperneurotization of Viscera in the Chick Embryo," by Rita Levi-Montalcini and Viktor Hamburger, appeared in 1953 in the Journal of Experimental Zoology. The paper provided the first evidence that nerve growth factor is a diffusible substance. Nerve growth promoting tumors were implanted into developing embryos to determine whether the tumors stimulated growth by direct contact or by a diffusible substance. The tumors were implanted into different parts of the embryo; one set of experiments implanted the tumor directly in the embryo, while another set of experiments placed the tumor on an exterior membrane. The membrane provided a barrier to direct contact with the nervous system, and allowed some chemical interaction between the tumor and the embryo. The tumor stimulated growth in both orientations, demonstrating that nerve growth factor was a chemical agent. The paper ends with two possible conclusions for the mechanism of nerve growth, the tumor may have been directly stimulating the ganglia by a diffusible signal, or it may have reduced the resistance of the chick tissues to the nerve growth. The term "nerve growth factor" is not explicitly used in this paper, and is referred to as a "diffusible agent" or a "growth promoting agent."

Conrad Hal Waddington's "Experiments on the Development of Chick and Duck Embryos, Cultivated in vitro," published in 1932 in Philosophical Transactions of the Royal Society of London, Series B, compares the differences in the development of birds and amphibians. Previous experiments focused on the self differentiation of individual tissues in birds, but Waddington wanted to study induction in greater detail. The limit to these studies had been the amount of time an embryo could be successfully cultivated ex vivo. Waddington applied in vitro cell culturing techniques to this experiment, as opposed to the chorio-allantoic technique used in many earlier studies. Culturing in vitro consisted of placing the embryo on a clot of adult chicken blood plasma and chick embryo extract in a watch glass. Experiments reported in this paper were divided into three main sections: the development of the embryos in vitro, induction by the endoderm, and induction by the primitive streak.

In "Selective Growth Stimulating Effects of Mouse Sarcoma on the Sensory and Sympathetic Nervous System of the Chick Embryo," Rita Levi-Montalcini and Viktor Hamburger explored the effects of two nerve growth stimulating tumors; mouse sarcomas 180 and 37. This experiment led to the discovery that nerve growth factor was a diffusible chemical and later to discoveries that the compound was a protein. Although this paper was an important step in the discovery of nerve growth factor, the term "nerve growth factor" was not used in this paper. It was instead referred to as a "growth promoting agent." The discovery of nerve growth factor earned Levi-Montalcini and Stanley Cohen, who also discovered epidermal growth factor, the 1986 Nobel Prize in Physiology or Medicine.

The developmental stages of the chick embryo were examined by Viktor Hamburger and Howard L. Hamilton in "A Series of Normal Stages in the Development of the Chick Embryo," published in the Journal of Morphology in 1951. These stages were published to standardize the development of the chick based on varying laboratory conditions and genetic differences. The stages Hamburger and Hamilton assigned were determined by the visible features of the chick embryo. The first stage begins just prior to the primitive streak, with the formation of the embryonic shield, and the final stage, forty-six, ends at the hatching of the chick.

In this paper Viktor Hamburger and Rita Levi-Montalcini collaborated to examine the effects of limb transplantation and explantation on neural development. In 1947 Hamburger invited Levi-Montalcini to his lab at Washington University in St. Louis to examine this question. Independently, each had previously arrived at opposing conclusions based on the same data. Hamburger concluded that limb transplantations caused the ganglia to develop more connections and grow larger while Levi-Montalcini concluded that the ganglia first produce a large amount of neurons, then degenerate the unsuccessful neurons. She concluded that larger ganglia must be due to the increase in successful connections. This joint paper, published in the Journal of Experimental Zoology in 1949, corroborated the findings reported by Levi-Montalcini and established that nerve degeneration is an integral part of development.

Nerve growth factor (NGF) is a signaling protein and growth factor implicated in a wide range of development and maintenance functions. NGF was discovered through a series of experiments in the 1950s on the development of the chick nervous system. Since its discovery, NGF has been found to act in a variety of tissues throughout development and adulthood. It has been implicated in immune function, stress response, nerve maintenance, and in neurodegenerative diseases. It is named for its effect on the critical role it plays in the growth and organization of the nervous system during embryonic development.