In 1873 Italy, Camillo Golgi created the black reaction technique, which enabled scientists to stain and view the structure of neurons, the specialized cells that compose the nervous system. During the nineteenth century, scientists were studying cells and proposed cell theory, which describes the basic characteristics of cells as fundamental units of life. However, cell theory struggled to explain neurons as they are specialized cells and more complex in structure than cells of other tissues. Prior to Golgi’s black reaction, other neuron staining techniques did not enable scientists to clearly and completely view entire neurons without damaging the tissue and obscuring the form. By enabling scientists to study individual neurons and neural tissues, Golgi’s black reaction enables researchers to better study the nervous system and how it develops.

Julia Barlow Platt studied neural crests in animal embryos and became involved in politics in the US during the nineteenth and twentieth centuries. She researched how body and head segments formed in chicks (Gallus gallus) and spiny dogfish (Squalus acanthias). Platt observed that in the mudpuppy (Necturus maculosus), the coordinated migration of neural crest cells in the embryo produced parts of the nervous system, bones, and connective tissues in the head. Platt's research indicated that the neural crest functioned like a germ layer, it challenged existing theories of what sorts of tissues arose from each of an embryo's germ layers, and it described early developmental stages of the nervous system.

The neuron doctrine is a concept formed during the turn of the twentieth century that describes the properties of neurons, the specialized cells that compose the nervous system. The neuron doctrine was one of two major theories on the composition of the nervous system at the time. Advocates of the neuron doctrine claimed that the nervous system was composed of discrete cellular units. Proponents of the alternative reticular theory, on the other hand, argued that the entire nervous system was a continuous network of cells, without gaps or synapses between the cells. In 1873, physician and reticular theory supporter Camillo Golgi developed a staining technique called the black reaction, a neuron staining technique that allowed for complete visibility of nerve cells, which enabled scientists to view a complete neuron cell and its cellular structures. Later, neuroscientist Santiago Ramón y Cajal used the black reaction to show the existence of synapses, or gaps between neurons, and argued that his evidence supported the neuron doctrine. The confirmation of the neuron doctrine showed that neurons function as discrete and independent cells, not as a single network, within the nervous system.

In the nineteenth century, reticular theory aimed to describe the properties of neurons, the specialized cells which make up the nervous system, but was later disconfirmed by evidence. Reticular theory stated that the nervous system was composed of a continuous network of specialized cells without gaps (synapses), and was first proposed by researcher Joseph von Gerlach in Germany in 1871. Reticular theory played a significant role in developmental neurobiology as it enabled scientists to theorize how the form of neural cells functioned in the context of the broader nervous system, and although disproven, reticular theory contributed to the foundation of the neuron doctrine that informed the modern field of neurobiology.

Camillo Golgi studied the central nervous system during the late nineteenth and early twentieth centuries in Italy, and he developed a staining technique to visualize brain cells. Called the black reaction, Golgi’s staining technique enabled him to see the cellular structure of brain cells, called neurons, with much greater precision. Golgi also used the black reaction to identify structures within animal cells like the internal reticular apparatus that stores, packs, and modifies proteins, later named the Golgi apparatus in his honor. Golgi, along with Santiago Ramón y Cajal, received the Nobel Peace Prize in 1906 for their independent work on the structure of the nervous system. Golgi’s discovery of the black reaction enabled other scientists to better study the structure of the nervous system and its development.

Roger Wolcott Sperry studied the function of the nervous system in the US during the twentieth century. He studied split-brain patterns in cats and humans that result from separating the two hemispheres of the brain by cutting the corpus callosum, the bridge between the two hemispheres of the brain. He found that separating the corpus callosum the two hemispheres of the brain could not communicate and they performed functions as if the other hemisphere did not exist. Sperry studied optic nerve regeneration through which he developed the chemoaffinity hypothesis. The chemoaffinity hypothesis stated that axons, the long fiber-like process of neurons, connect to their target cells through special chemical markers. This challenged the previously accepted resonance principle of neuronal connection. Sperry shared the 1981 Nobel Prize in Physiology or Medicine with David Hubel and Torsten Wiesel.

The syncytial theory of neural development was proposed by Victor Hensen in 1864 to explain the growth and differentiation of the nervous system. This theory has since been discredited, although it held a significant following at the turn of the twentieth century. Neural development was well studied but poorly understood, so Hensen proposed a simple model of development. The syncytial theory predicted that the nervous system was composed of many neurons with shared cytoplasm. These nerves were thought to be present in the embryo from a very early stage and were selected by the function of the target tissue. There were two competing theories to the syncytial theory. Theodor Schwann and Francis Maitland Balfour proposed the sheath cell theory, which states that nerve fibers were the product of secretions by chains of sheath cells. Santiago Ramón y Cajal and Wilhelm His proposed the outgrowth theory of fiber development for individual neurons. The most substantial evidence against the syncytial theory of neural development was produced by Ross Granville Harrison in his studies of the development of nerve fibers.

German embryologist Viktor Hamburger came to the US in 1932 with a fellowship provided by the Rockefeller Foundation. Hamburger started his research in Frank Rattray Lillie's laboratory at the University of Chicago. His two-year work on the development of the central nervous system (CNS) in chick embryos was crystallized in his 1934 paper, "The Effects of Wing Bud Extirpation on the Development of the Central Nervous System in Chick Embryos," published in The Journal of Experimental Zoology. Hamburger was able to use the microsurgical techniques that he had learned from Hans Spemann to show how wing buds influence the development of the CNS in chick embryos. This paper is one of several among Hamburger's important studies on chick embryos and represents the empirical and theoretical cornerstone for his further research on central-peripheral relations in the development of the nervous system.

An important question throughout the history of embryology is whether the formation of a biological structure is predetermined or shaped by its environment. If both intrinsic and environmental controls occur, how exactly do the two processes coordinate in crafting specific forms and functions? When Viktor Hamburger started his PhD study in embryology in the 1920s, few neuroembryologists were investigating how the central neurons innervate peripheral organs. As Hamburger began his research, he had no clue that central-peripheral relations in the development of the central nervous system (CNS) would become one of his major interests for the next seventy-five years. In fact, this research trajectory would lead him to discover programmed cell death as a pivotal mechanism mediating central-peripheral relations, as well as to Nobel-Prize-winning work on nerve growth factors (NGF).