Evidence for nodose ganglion as the source of innervation to the anterior lobe of the pituitary gland.

Recent studies have provided convincing evidence for the presence of peptidergic nerve fibers in the pituitary anterior lobe in several animal species. This study was aimed at elucidating the origin of this innervation by neuroanatomical tracing, denervation experiments, and immunohistochemistry. Immunohistochemistry against substance P and growth-associated protein 43 revealed a dense fiber plexus within the anterior lobe, and these markers were mostly colocalized. Retrograde tracing with Fluorogold from the pituitary gland stained neurons in the hypothalamus, superior cervical ganglia and the nodose ganglia. None of the Fluorogold-labelled neurons in the hypothalamus or superior cervical ganglion were substance P-immunoreactive, while many of the neuronal cell bodies in the nodose ganglion exhibited substance P immunoreactivity. There were no Fluorogold-labelled neurons in the trigeminal, otic or cervical dorsal root ganglia. Surgical transection of the pituitary stalk or bilateral removal of the superior cervical ganglion did not abolish the anterior lobe nerve fibers, and anterograde tracing with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindo-carbocyanine perchlorate from the pituitary stalk failed to stain any nerve fibers within the anterior lobe. Our findings suggest that the nodose ganglion neurons likely innervate the pituitary anterior lobe, while neither hypothalamus nor sympathetic superior cervical ganglion may be a source of this innervation. By showing a distinct neuronal system in the pituitary anterior lobe our findings (i) support the previous functional studies demonstrating a distinct regulation of the morphology of the anterior lobe innervation by hormonal changes, and (ii) suggest that the innervation of the pituitary anterior lobe is a part of the visceral innervation by the vagus nerve rather than a part of the other intracranial innervation. These findings provide a neuroanatomic basis for the reported observations about the neural regulation of the pituitary anterior lobe.

Inducible expression of tryptophan hydroxylase without serotonin synthesis in hypothalamic dopaminergic neurons.

In the present study we have further studied the previous findings that rat hypothalamic dopaminergic neuronal cell groups may express tryptophan hydroxylase (TpH), the serotonin synthesizing enzyme, without a detectable serotonin synthesis. Chemical and mechanical neuronal injuries, namely colchicine treatment and axonal transection, respectively, were performed, and distributions of neurons exhibiting immunoreactivity for TpH and/or tyrosine hydroxylase (TH), the dopamine synthesizing enzyme, were analyzed throughout the hypothalamic periventricular and arcuate nuclei. After colchicine treatment there was a statistically significant 87% (P = 0,01) increase in the number of TpH expressing neurons, while TH expression remained essentially similar. Axonal transection resulted also in a statistically significant 131% (P < 0,01) increase in the number of TpH expressing neurons, while TH expression was not significantly altered. All TpH expression coexisted with TH expression, and the induction of TpH expression by neuronal injuries occurred evenly throughout the rostrocaudal length of the territory studied. A possible serotonin synthesis by TpH was examined by giving drugs that increase brain serotonin synthesis, but no immunohistochemically detectable serotonin synthesis could be found in any of the TpH expressing neurons. Finally the possibility was studied that the relative shortage of the cofactor tetrahydrobiopterin would limit serotonin synthesis. However, an administration of tetrahydrobiopterin did not result in detectable serotonin synthesis in these neurons. Taken together these results suggest that dopaminergic neurons in the hypothalamic periventricular and arcuate nuclei are able to express TpH, this expression is induced after neuronal injury, and this induction occurs similarly throughout the territories studied. TpH expression occurs independently of TH expression, and the newly expressed TpH appears not to synthesize serotonin, regardless of pharmacological pretreatments. Thus, our findings (i) support the idea that neurons may possess inducible expression of nonfunctional transmitter-synthesizing enzymes, in this case TpH, and (ii) suggest that expression of an enzyme synthesizing a certain transmitter may not necessarily imply the corresponding transmitter phenotype.

Expression of nitric oxide synthase in hypothalamic nuclei following axonal injury or colchicine treatment.

Nitric oxide (NO) has recently gained much attention due to its apparently double-edged role in neuronal injury. This study was aimed at elucidating neuronal nitric oxide synthase (nNOS) expression in the brain after two types of injury, namely axonal transection and colchicine treatment. The neurosecretory hypothalamo-pituitary pathway served as a model for the reaction of central neurons to these two types of injury. Axonal transection, i.e., pituitary stalk section, resulted in a qualitative increase in NOS content in the supraoptic and paraventricular nuclei. In these nuclei, there was also an increase in the number of NOS-expressing neurons after the operation. Surprisingly, in the periventricular nucleus, a strong decrease in the number of NOS-positive magnocellular neurons was observed in the anterior part of the nucleus. Intracerebroventricular injection of colchicine resulted in an increase in the cell count in the paraventricular nucleus, while the other nuclei remained unchanged. Our results suggest that axonal injury results in an increase in nNOS expression in the major neurosecretory nuclei, while the periventricular nucleus shows the opposite reaction. Colchicine treatment has an effect similar to that of axotomy in the major neurosecretory nuclei, suggesting that an increase in NOS expression may be induced by interrupted axonal transport. In the periventricular nucleus, the decrease in the number of NOS-containing neurons suggests differences among hypothalamic NOS-containing neuron groups in response to neuronal injury.

Peptide GEGLSS-like immunoreactivity in the rat central nervous system.

A rabbit antiserum was raised against the N-terminal fragment peptide, GEGLSS (Gly-Glu-Gly-Leu-Ser-Ser) of bovine neuropeptide AF (NPAF, A18Famide). NPAF is an octadecapeptide isolated from the bovine brain together with neuropeptide FF (NPFF). GEGLSS-like immunoreactivity was localized with immunofluorescence technique in colchicine-treated rats in neuronal cell bodies of the supraoptic (SON) and paraventricular (PVN) hypothalamic nuclei. A few neurons were also observed in the retrochiasmatic part of the SON. GEGLSS-like immunoreactivity was also localized to nerve terminals of the posterior pituitary. No GEGLSS-ir neuronal cell bodies were observed in the medial hypothalamus, in an area that contains NPFF-ir neurons. GEGLSS immunoreactivity was also seen in the fibers and terminals of nucleus of the solitary tract. We injected a retrograde tracer, fluorogold, to the posterior pituitary gland and visualized GEGLSS-ir neuronal cell bodies double-labeled with the tracer in SON, PVN, and SOR. The pituitary stalk transsection totally abolished the GEGLSS-ir structures from the posterior pituitary. Our results suggest that GEGLSS immunoreactivity in the rat brain has a more limited distribution than NPFF immunoreactivity. GEGLSS immunoreactivity was partially colocalized with arginine-vasopressin and oxytocin in neuronal cell bodies in the SON and PVN. Considering the fact that the known rat NPFF-NPAF precursor does not contain GEGLSS structure, the detected GEGLSS immunoreactivity may be derived from a previously unknown precursor.