Reproductive function is essential to the survival of all species. In mammals and other vertebrates, the control of reproduction relies on the hypothalamic-pituitary-gonadal axis, with the primary driving force provided by hypothalamic GnRH neurons. In the median eminence, the decapeptide GnRH are released in a unique pattern from GnRH neuroterminals into the portal capillary system as part of reproductive cycle. During aging, the biological rhythms of GnRH release are altered in a species-specific manner, with a reduction of GnRH pulsatility and surge in aging female rats resulting in reproductive senescence, which happens much earlier than gonadal failure in rats. Relatively few studies have focused on regulation of GnRH release at the neuroterminal level in the median eminence during reproductive aging. Therefore, the aims of this dissertation are to 1) Study the regulation of GnRH secretion at the neuroterminal level, focusing on glutamate transmission; 2) Ascertain the interaction between GnRH neuroterminals and their surrounding microenvironment focused on glial cells and the portal capillary system in the median eminence; and 3) Analyze age and hormone effects on GnRH neuroterminals and their microenvironment. An aging ovariectomized female rat model was used to study the effects of age and hormones on GnRH neuroterminal system. Fluorescence microscopy, confocal microscopy and transmission electron microscopy were used in conjunction with several imaging analysis tools. I mastered the use of cryo-embedding multi-probe immunogold labeling electron microscopy, which was essential to visualize and quantify the ultrastructral changes in GnRH neuroterminals. I combined the serial electron microscopy with cryo-embedding immunogold electron microscopy preparation and developed a new technique to examine biological markers with a three-dimensional perspective at the cellular level. Results from a series of four research projects showed: 1) There is a novel glutermatergic pathway in GnRH neuroterminals, which may regulate GnRH secretion; 2) There are dramatic age related morphological changes in the GnRH neuroterminal /glia/ portal capillary system of the median eminence that may be involved in reproductive senescence and other neuroendocrine system impairments with age; 3) Serial electron microscopy combined with immunogold labeling technique is a useful method to study the regulation of neuronal signaling pathway. Although my studies were performed on a rat model, it seems reasonable to predict that some of these changes in the median eminence with age may apply to other species, including humans, relevant to some of the menopausal symptoms in women. / text

This dissertation presents experimental research investigating the behavior of particles on two different types of anisotropically curved liquid interfaces: cylinders and catenoids. The results are compared to the behavior predicted by theoretical models. Several types of liquids and many types of particles were examined. The size scale of the surfaces ranges from microns to millimeters, with nanometer and micron sized particles. Semi-cylinders, a few hundred microns in diameter, were made by creating a line of liquid on a surface. Three different fluids were used to create the semi-cylinders: Gallium, ionic liquids, and molten polystyrene (PS). Particle behavior on semi-cylinder liquid interfaces made from these materials was observed. Scanning electron microscopy (SEM) and optical microscopy were used to determine the location and assembly (related to particle attraction) of the particles on the surfaces of the fluids. PS semi-cylinders with silica particles were found to be the most promising experimental route, as PS will flow when heated above its Tg and will solidify when cooled to room temperature. As a solid, the PS surface is easily analyzed. Scanning force microscopy (SFM) was used on the PS semi-cylinders to image the deformation to the interface surrounding the particle, and a quadropolar deformation was found. PS catenoids, a few microns tall, were also investigated. The catenoids were produced by placing thin PS films heated above their Tg between two electrodes, separated from the surface of the film by a small air gap. A voltage was applied across the electrodes to create an electric field that produced electrohydrodynamic instabilities on the surface of the film that led to the formation of catenoids of molten PS that spanned the electrode gap. Semi-catenoids, several mm long, were also made from an ionic liquid by using chemically patterned wafers. SEM and optical microscopy were used to determine the particle location on the catenoid surfaces. The PS catenoids were found to be the most promising experimental system, and particles were observed to locate preferentially along the edges of the catenoid, instead of around the center as predicted.