Migration Of Gonadotropin-releasing Hormone Neurons Research Articles

Expression of GATA-4 in migrating gonadotropin-releasing neurons of the developing mouse

The hypothalamic gonadotropin-releasing hormone (GnRH) neurons are important regulators of reproductive function. During development, these cells arise in the olfactory placode and migrate to the central nervous system, where they form a diffuse population of neurosecretory cells that mediate central nervous system control of reproduction. Little is known of the mechanisms regulating the differentiation of these cells. Studies of the transcriptional regulation of the GnRH gene have demonstrated the importance of the GATA family of zinc-finger transcription factors in gene expression. Although GATA factors are not highly expressed in mature GnRH-secreting neurons, we report that GATA-4 is highly expressed in migrating GnRH neurons in the developing mouse. We also report that a second DNA binding activity regulating GnRH gene expression at the site of GATA-factor action persists in mature hypothalamus and may also play a role in gene expression in the differentiated GnRH neuron.

Gonadotropin-Releasing Hormone Containing Neurons and Olfactory Fibers During Development: From Lamprey to Mammals

Gonadotropin releasing-hormone (GnRH) regulates the hypothalamo–pituitary–gonadal axis in all vertebrates. The vast majority of GnRH neurons are thought to be derived from progenitor cells in medial olfactory placodes. Several antibodies and lectins that recognize cell surface carbohydrates have been useful for delineating the migratory pathway from the olfactory placodes and vomeronasal organ, through the nasal compartment, and across the cribriform plate into the brain. In rats, α-galactosyl-linked glycoconjugates (immunoreactive with the CC2 monoclonal antibody) are expressed on fibers along the GnRH migration pathway and approximately 10% of the GnRH neuronal population. In lamprey, the α-galactosyl binding lectin, Grifonia simplicifolia-I (GS-1), identifies cells and fibers of the developing olfactory system. In contrast to the CC2 immunoreactive GnRH neurons in rats, the GS-1 does not label a subpopulation of presumptive GnRH neurons in lamprey. Results from these and other experiments suggest that GnRH neurons in developing lamprey do not originate within the olfactory placode, but rather within proliferative zones of the diencephalon. However, the overlap of olfactory- and GnRH-containing fibers from prolarval stages to metamorphosis, suggest that olfactory stimuli may play a major role in the regulation of GnRH secretion in lamprey throughout life. By contrast, olfactory fibers are directly relevant to the migration of GnRH neurons from the olfactory placodes in mammalian species. Primary interactions between olfactory fibers and GnRH neurons are likely transient in mammals, and so in later life olfactory modulation of GnRH secretion is likely to be indirect.

Migration of GnRH-immunoreactive neurons from the olfactory placode to the brain: a study using avian embryonic chimeras

Previous studies suggest that gonadotropin-releasing hormone (GnRH) neurons appear in the olfactory placode and subsequently migrate into the brain during embryonic development. The aim of the present study was to obtain direct evidence for migration of GnRH neurons from the olfactory placode into the brain. Olfactory placodes from quail embryos were transplanted isotopically and isochronically, to replace the unilaterally ablated olfactory placodes of chick embryos. The chimeric embryos were allowed to develop for several days until they reached the embryonic stages when GnRH neurons are seen in the brain in normal embryos. Quail olfactory epithelia were formed in the host chick embryos. Quail olfactory nerves were also formed and reached the olfactory bulb or primordial olfactory bulb. GnRH-immunoreactive cells of quail origin revealed by a triple staining method were observed in the quail olfactory epithelium, quail olfactory nerve, chick olfactory bulb, and chick septo-preoptic area. These results indicate that GnRH neurons originate in the olfactory placode and migrate into the telencephalon including the septo-preoptic area. A migratory route of GnRH neurons was well documented by the use of a quail neuron-specific antibody, QN. The migratory route in the brain is discussed with special reference to the terminal nerve. A GnRH-immunoreactive neuronal group of chick origin appeared in the diencephalon of chimeric embryos. These diencephalic neurons may be of non-placodal origin. FMRFamide-immunoreactive neurons of quail origin were also found in the quail olfactory nerve and the host olfactory bulb, suggesting that FMRFamide neurons also originate in the olfactory placode and migrate into the brain.

Regulation of gonadotropin-releasing hormone neurons by basic fibroblast growth factor

The development of a functional network of gonadotropin-releasing hormone (GnRH) neurons in the central nervous system requires a series of complex regulatory mechanisms, presumably mediated in part by neurotrophic factors. The difficulty in studying factors regulating the development of GnRH neurons stems from their paucity and scattered distribution in the brain; as a result, little was known about the role of neurotrophic factors in the development of the mature GnRH neuronal network. Recent utilization of immortalized GnRH neuronal cell lines (GT1) has enabled us to identify and study specific neurotrophic factors and their functions in vitro. The potent neurotrophic effect of basic fibroblast growth factor (bFGF) and the presence of a high abundance of receptors for bFGF in GT1 cells have led to the hypothesis that bFGF may be an important regulator of GnRH neuron expansion, survival, migration, and connectivity.

Organization of GnRH and FMRF-Amide Systems in Two Primitive Bony Fishes (Order Polypteriformes)

To investigate the evolution of the neural organization of gonadotropin-releasing hormone (GnRH), we have examined GnRH-immunoreactivity in two brachiopterygian fishes (Polypterus palmas and Calamoichthys calabaricus). Distal regions of the terminal nerve (TN) within the medial olfactory nerve contained clusters of GnRH-immunoreactive (ir) perikarya (< 10 microns). More proximal, isolated GnRH-ir neurons were present among TN fascicles as they penetrated the ventral forebrain, and a few ir neurons were observed accompanying GnRH-ir fibers in the rostromedial telencephalon. GnRH-ir neurons were not observed in the preoptic area or ventral hypothalamus. In contrast, a small group of GnRH-ir neurons was localized in the periventricular nucleus of the posterior tuberculum. GnRH-ir fibers were present in widespread areas of the brain, including the olfactory bulb, telencephalon, optic nerve, hypothalamus, thalamus, habenula, optic tectum, tegmentum, pituitary and spinal cord. To further characterize projections of TN neurons, we utilized antiserum to FMRF-amide, a small peptide produced by TN cells in other vertebrates. Perikarya that were FMRF-amide-ir within the TN were similar in distribution to GnRH-TN neurons, and the distribution of FMRF-amide-ir fibers overlapped those of GnRH-ir fibers, thus providing a useful marker for identifying TN projections. An additional population of FMRF-amide-ir neurons was present in the periventricular hypothalamus. Our results suggest that in the polypteriformes, GnRH and FMRF-amide neurons of the TN are similar to those observed in other vertebrates; however, the paucity of GnRH cells in the basal forebrain may be unique to primitive actinopterygians and elasmobranchs, and may result from the lack of migration of GnRH neurons into the forebrain, a phenomenon that likely occurs in all other vertebrate classes. Finally, the identification of GnRH-ir neurons in the posterior tuberculum is consistent with similar, and perhaps homologous, GnRH neurons present in nearly all other vertebrate classes.

Isolation and characterization of the gene responsible for the X chromosome-linked Kallmann syndrome

Kallmann de Morsier Syndrome is defined by the association of an hypogonadism with an anosmia. The hypogonadism is due to a deficiency of GnRH (gonadotropin-releasing hormone). Olfactory bulbs and tracts are under-developed in the patients. Embryological studies have indicated that the migration of GnRH neurons and the axonal extension of olfactory neurons, which both originate in the olfactory epithelium during embryogenesis, were impaired in a fetus affected by X-linked Kallmann Syndrome. By a positional cloning strategy, we have isolated the KAL gene, responsible for the X-linked form of the disease. The gene consists of 14 exons. A highly homologous pseudogene on the Y chromosome has been characterized. The KAL gene encodes a putative secreted protein of 680 amino acids, which contains four fibronectin type III repeats and a four disulphide core motif. The former motif is usually associated with adhesion function. The latter has been described in protein with antiprotease activity. We have isolated the chicken KAL homologue and studied its expression by in situ hybridization during late embryonic development. The gene is expressed in various neuronal populations of the central nervous system, including mitral cells of the olfactory bulbs. We suggested that the KAL protein might be involved in late neuronal differentiation.

The ontogeny of gonadotropin-releasing hormone neurons in the chick.

The origin of gonadotropin-releasing hormone (GnRH) neurons in the olfactory placode and their subsequent migration into the central nervous system (CNS) has been described in several species. The current study was undertaken to delineate the time course and route of GnRH neuronal migration in the chick using antisera generated against both the mammalian form of GnRH (LR1) and a specific chicken GnRH (cGnRH I). Chicks aged E4.5 through day of hatching were processed for GnRH immunocytochemistry. Under the incubation conditions employed in this study, GnRH neurons first appeared on E4.5 in the olfactory epithelium. GnRH neurons exit from the olfactory epithelium and follow the extracranial course of the olfactory nerve to the anterior pole of the developing forebrain. Within the nerve, cells are in close apposition to each other. By E5, cells had reached the surface of the telencephalon. Upon entering the CNS, the GnRH neurons dispersed within the neuropil and continued their migration as individuals. As development progressed, GnRH neurons migrated dorsally and caudally within the brain and reached their adult positions by E12. GnRH axons were detected in the vicinity of GnRH cell bodies with the cGnRH I antiserum as early as E4.5; however, GnRH axons did not reach the median eminence until E14. These results extend earlier reports in mammal and chick on the extracranial origin of GnRH neurons and their path of entry into and dispersion within the CNS.

Intrahypothalamic injection of a cell line secreting gonadotropin-releasing hormone results in cellular differentiation and reversal of hypogonadism in mutant mice.

GT1 is an immortalized cell line that synthesizes and secretes the neurohormone gonadotropin-releasing hormone (GnRH). We have placed these cells into the brains of adult mutant hypogonadal (hpg) mice, which lack a functional GnRH gene, to determine whether such cells could differentiate in situ and support gonadal development. Immunocytochemical detection of GnRH revealed that these cells migrated widely in the central nervous system and elaborated axonal processes which on rare occasion projected to the normal target, the median eminence. Using a battery of antibodies, we demonstrated that these cells could cleave the GnRH precursor and that the amidated decapeptide as well as other cleavage products were present. The presence of biologically active material and its appropriate secretion were further documented by gonadal growth in both males and females. The morphological differentiation of the GT1 cells correlated with the density of cells injected. Those remaining within the injection site and/or forming a tumor retained a simple, rounded or fibroblastic appearance. Those cells that migrated into the host away from such tumors assumed the simple fusiform shape of normal GnRH neurons with dendrites extending from one or both poles. When cell density was drastically reduced a much more complex dendritic arbor was elaborated. These data suggest that such cell lines can be useful in reversing genetic defects and in studying such processes as GnRH neuronal migration, axonal targeting, and cytological differentiation.

Migratory arrest of gonadotropin-releasing hormone neurons in transgenic mice.

Gonadotropin-releasing hormone (GnRH) is important in reproduction, although the mechanism of central hypogonadism in humans remains unclear. Because the GnRH neuron originates from the olfactory placode and migrates to the hypothalamus during development, central hypogonadism in humans could be caused by failure in normal migration of GnRH neurons to the hypothalamus. We report that in transgenic mice expression of the simian virus 40 T antigen, driven by the promoter of human GnRH gene, resulted in central hypogonadism due to an arrest in neuronal migration during development and tumor formation along the migratory pathway. This system appears to be an important animal model of hypogonadotropic hypogonadism in humans. Additionally, olfactory bulb tumors from these animals were dispersed, and a GnRH-secreting neuronal cell line (GN cell line) was established.