Effects of polyunsaturated fatty acids and hormones on synaptogenesis in serum-free medium cultures of mouse fetal hypothalamic cells. 1986

A Tixier-Vidal, and R Picart, and C Loudes, and A F Bauman

The effects of soluble factors on synaptogenesis by mouse fetal hypothalamic cells cultured in chemically defined conditions have been examined using transmission electron microscopy. Hypothalami taken on the 16th day of gestation were mechanically dissociated and cells were seeded in a minimum serum-free medium supplemented or not with the following components: triiodothyronine, corticosterone and a mixture of polyunsaturated fatty acids (arachidonic acid plus docosahexaenoic acid bound to defatted bovine serum albumin). In the minimum serum free medium synapses were found after 10 days in culture. However, the development of synaptic vesicles was very limited, whereas that of the presynaptic and postsynaptic densities was apparently normal. Supplementation of the minimum serum-free medium with triiodothyronine, corticosterone and polyunsaturated fatty acids added simultaneously, permitted a full development of synapses as attested to by the increase in number and the regular shape and diameter of synaptic vesicles as well as by the complexity and diversity of synapse configurations. Among those three factors, polyunsaturated fatty acids clearly played a key role. The ability of synapses formed in culture to respond to potassium evoked depolarization was examined on cultures grown for 12 days in the simultaneous presence of the three above mentioned supplements. Exposure for 3 min to 60 mM potassium chloride induced in synaptic boutons vesicular depletion, apposition of vesicle clusters onto the presynaptic grid, appearance of a rich filamentous network and of some coated vesicles. Return to 3mM potassium chloride induced in 3 min a massive restoration of the population of vesicles which slightly differed from synaptic vesicles in control cultures. These results show that: (1) the formation of synaptic vesicles in this system is regulated by soluble factors among which polyunsaturated fatty acids play a major role, and (2) synapses formed de novo in chemically defined conditions of culture display the same ability to respond to and to recover from potassium evoked depolarization as adult axon terminals. Thus, they offer a suitable model for analysis of the mechanisms involved in membrane traffic in central neurons.

UI MeSH Term Description Entries
D007031 Hypothalamus Ventral part of the DIENCEPHALON extending from the region of the OPTIC CHIASM to the caudal border of the MAMMILLARY BODIES and forming the inferior and lateral walls of the THIRD VENTRICLE. Lamina Terminalis,Preoptico-Hypothalamic Area,Area, Preoptico-Hypothalamic,Areas, Preoptico-Hypothalamic,Preoptico Hypothalamic Area,Preoptico-Hypothalamic Areas
D007797 Laminin Large, noncollagenous glycoprotein with antigenic properties. It is localized in the basement membrane lamina lucida and functions to bind epithelial cells to the basement membrane. Evidence suggests that the protein plays a role in tumor invasion. Merosin,Glycoprotein GP-2,Laminin M,Laminin M Chain,Chain, Laminin M,Glycoprotein GP 2,M Chain, Laminin
D008854 Microscopy, Electron Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen. Electron Microscopy
D011189 Potassium Chloride A white crystal or crystalline powder used in BUFFERS; FERTILIZERS; and EXPLOSIVES. It can be used to replenish ELECTROLYTES and restore WATER-ELECTROLYTE BALANCE in treating HYPOKALEMIA. Slow-K,Chloride, Potassium
D002478 Cells, Cultured Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others. Cultured Cells,Cell, Cultured,Cultured Cell
D003345 Corticosterone An adrenocortical steroid that has modest but significant activities as a mineralocorticoid and a glucocorticoid. (From Goodman and Gilman's The Pharmacological Basis of Therapeutics, 8th ed, p1437)
D005231 Fatty Acids, Unsaturated FATTY ACIDS in which the carbon chain contains one or more double or triple carbon-carbon bonds. Fatty Acids, Polyunsaturated,Polyunsaturated Fatty Acid,Unsaturated Fatty Acid,Polyunsaturated Fatty Acids,Acid, Polyunsaturated Fatty,Acid, Unsaturated Fatty,Acids, Polyunsaturated Fatty,Acids, Unsaturated Fatty,Fatty Acid, Polyunsaturated,Fatty Acid, Unsaturated,Unsaturated Fatty Acids
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013569 Synapses Specialized junctions at which a neuron communicates with a target cell. At classical synapses, a neuron's presynaptic terminal releases a chemical transmitter stored in synaptic vesicles which diffuses across a narrow synaptic cleft and activates receptors on the postsynaptic membrane of the target cell. The target may be a dendrite, cell body, or axon of another neuron, or a specialized region of a muscle or secretory cell. Neurons may also communicate via direct electrical coupling with ELECTRICAL SYNAPSES. Several other non-synaptic chemical or electric signal transmitting processes occur via extracellular mediated interactions. Synapse
D014284 Triiodothyronine A T3 thyroid hormone normally synthesized and secreted by the thyroid gland in much smaller quantities than thyroxine (T4). Most T3 is derived from peripheral monodeiodination of T4 at the 5' position of the outer ring of the iodothyronine nucleus. The hormone finally delivered and used by the tissues is mainly T3. Liothyronine,T3 Thyroid Hormone,3,3',5-Triiodothyronine,Cytomel,Liothyronine Sodium,Thyroid Hormone, T3

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