Neurointermediate Lobe Of Pituitary Research Articles

Chronic naltrexone infusion: Effects on innervation of rat neurointermediate lobe

The rat pituitary neurointermediate lobe contains nerve fibers which are sensitive to the neurotoxic effects of 6-hydroxydopamine and to acute injections of the opiate antagonist, naltrexone. In the present study, naltrexone hydrochloride was administered to adult male Sprague-Dawley rats in a chronic infusion over a period of one week, using Alza osmotic pumps to determine whether a low (10 −5 M) dose could, over a longer period, also induce toxic effects on pituitary innervation. Electron microscopy of pituitary intermediate lobe fibers revealed swollen neurovesicles and membranous structures in nerve terminals, and significantly fewer normal-appearing nerve terminals, after drug infusion. Light microscopic immunostaining of paraffin sections of pituitary glands illustrated some diminution of serotonin (5-HT)-immunofluorescence in the intermediate lobe after naltrexone treatment. Immunostaining of sections from the same animals for tyrosine hydroxylase (TH) revealed intensified fiber staining in the intermediate lobe, with some regions of terminals having a swollen appearance. High pressure liquid chromatographic-electrochemical detection (HPLC-EC) analysis of neurotransmitters in selected brain areas and pituitary indicated a significant increase in norepinephrine levels in the hypothalamus. The observations suggest that intermediate lobe innervation is sensitive to low, continuously infused doses of naltrexone, which acts as a neurotoxin to produce degenerative changes in nerve terminals. The changes appear to be reflected in alterations in staining patterns of neurotransmitters, and may also affect transmitter uptake in damaged terminals.

Effect of Hypertonic Saline on the Corticotropin-Releasing Hormone and Arginine Vasopressin Content of the Rat Pituitary Neurointermediate Lobe.

The changes in the levels of corticotropin-releasing hormone (CRH) and arginine vasopressin (AVP) in the neurointermediate lobe of the pituitary (NIL) following hypertonic saline administration were examined in rats. The plasma osmotic pressure in rats receiving 2% NaCl for 8 days was greatly increased. Plasma AVP concentration in rats receiving 2% NaCl for 8 days were significantly higher than in control rats (566% of the control level). Plasma corticosterone was significantly higher in the saline-treated rats than in controls, whereas plasma ACTH was not significantly different. The pituitary ACTH concentration was much higher in the saline-treated rats than in controls. CRH in the NIL was increased significantly by saline treatment (419% of the control concentration), whereas the CRH in the paraventricular nucleus and median eminence of control and saline-treated rats did not differ significantly. The AVP in the NIL fell greatly in saline treated rats. The extract from both control and saline-treated rats showed a major peak for immunoreactive CRH, with a retention time identical to that of rat CRH. However, the peak was much higher in the extract from saline-treated rats. The immunoreactive AVP peak was greatly reduced in saline-treated rats. These results suggest that hypertonic saline administration increases the CRH in the NIL and causes AVP hypersecretion and/or hyperfunction of magnocellular-NIL CRH might be responsible for pituitary-adrenal stimulation in saline-treated rats.

Differential Effects of Glucocorticoids on Corticotrophin-Releasing Factor in the Rat Pituitary Neurointermediate Lobe and Median Eminence.

We have investigated the effects of glucocorticoid manipulation on corticotrophin-releasing factor (CRF-41) in the neurointermediate lobe (NIL) and median eminence (ME) of the rat using a radioimmunoassay specific for CRF-41. CRF-41 content in the NIL (88+/-7 fmol, mean +/- SEM) was not significantly altered by administration of dexamethasone in drinking water for 12 days (67+/-9 fmol) or adrenalectomy for 12 days (96+/-17 fmol). The saline-stimulated increase in NIL CRF-41 content (154+/-24 fmol) was not affected by dexamethasone (152+/-24 fmol) or adrenalectomy (179+/-21 fmol). In contrast, the content of CRF-41 in the ME declined following dexamethasone treatment (1247+/-92 fmol to 864+/-26 fmol), or adrenalectomy (854+/-78 fmol). Our results provide further evidence that CRF-41 in the NIL and ME can be differentially regulated.

Tissue specific expression of rat peptidylglycine alpha-amidating monooxygenase activity and mRNA.

The tissue specific expression of peptidylglycine alpha-amidating monooxygenase [(PAM) EC 1.14.17.3], an enzyme which catalyzes the formation of amidated bioactive peptides from their glycine-extended precursors, was examined in adult rat. Soluble and membrane-associated PAM enzymatic activities were determined, and the levels and size classes of PAM mRNA were examined by Northern blot analysis. PAM specific activity varied 1000-fold in the tissues examined, with highest levels in heart atrium, pituitary and salivary glands, and hypothalamus. The fraction of total PAM activity that was membrane associated varied from approximately 70% in heart atrium to 10% in neurointermediate pituitary lobe and thyroid gland. Levels of PAM mRNA varied over 300-fold. In the heart atrium, PAM mRNA accounts for more than 0.1% of the mRNA. For many tissues the ratio of total PAM specific activity to PAM mRNA levels was similar; however, PAM activity was higher than expected from mRNA levels in the salivary glands and lower than expected in several tissues, including heart ventricle. Three major size classes of PAM mRNA were identified among the tissues. Use of RNAse H indicated that differences in size were not due to the length of the poly(A) tail. The heart and central nervous system expressed PAM mRNA of the 4.2 kilobase (kb) and 3.8 kb size classes, while the remaining tissues expressed predominantly 3.8 kb and 3.6 kb classes; few tissues contained only one size class of PAM mRNA. The two major forms of PAM mRNA in adult heart atrium differ by the presence or absence of a 315 nucleotide segment in the protein coding region. Using a cDNA probe from within this segment, the 4.2 kb and 3.8 kb size classes of PAM mRNA in the central nervous system appeared to resemble those in the heart atrium. In the remaining tissues, a subset of PAM mRNAs in the 3.8 kb and 3.6 kb size classes hybridized with this probe, suggesting that additional forms of PAM mRNA are present.

Expression of the BRL-3A Insulin-like Growth Factor Binding Protein (rBP-30) in the Rat Central Nervous System

The expression of the BRL 3A insulin-like growth factor binding protein (rBP-30) was characterized in rat brain, hypothalamus, and pituitary tissue and cultured neuronal and astroglial cells. The 27K BP expressed by BRL 3A cells (rBP-30) was found to also be expressed in conditioned media from newborn rat astrocytes and fetal neurons, but not in the medium from the neuroblastoma cell line, B104. Moreover, a polyclonal antibody, anti HEC1, specifically immunoprecipitated the BRL 3A BP from the same conditioned media, as well as from rat cerebrospinal and amniotic fluid and from conditioned medium of cells isolated from the neurointermediate lobe of adult rat pituitary. The same antibody also immunoprecipitated hBP-31 from human CSF. Northern blot analyses showed that rBP-30 mRNA was expressed in adult rat brain and pituitary, fetal brain and liver, and in fetal neurons and newborn astrocytes maintained in culture. We conclude that the BRL 3A BP (rBP-30) is the major insulin-like growth factor binding protein in the rat CNS and may be the rat analog of hBP-31, the predominant BP in human CSF.

Regulation and Secretion of Proopiomelanocortin Peptides from Isolated Perifused Dog Pituitary Pars Intermedia Cells*

The dog pituitary pars intermedia (PI) appears to consist of relative large numbers of ACTH-containing cells in addition to the more abundant alpha MSH-containing cells. Since regulation of PI secretion probably varies across mammalian species, this study was undertaken to identify substances potentially involved in the control of dog PI POMC peptide secretion and to determine if these substances altered the secretion of immunoreactive (IR) ACTH and IR-alpha MSH in a parallel fashion. Pituitary neurointermediate lobes from dogs were collected and dispersed, and the PI cells obtained were perifused. For comparison, rat PI and pars distalis (PD) cells as well as dog PD cells were similarly collected and perifused. Dog PI cells secreted IR-alpha MSH at a basal rate of 125 +/- 59 (mean +/- SD) pg/min.10(5) cells and IR-ACTH at a rate of 40 +/- 9 pg/min.10(5) cells (molar IR-alpha MSH/IR-ACTH = 10). In contrast, secretion rates for IR-alpha MSH and IR-ACTH from perifused rat PI cells were 171 +/- 108 and 3 +/- 2 pg/min.10(5) cells, respectively (molar IR-alpha MSH/IR-ACTH = 179). Using Sephadex G-50 gel filtration chromatography, virtually all of the IR-beta-endorphin secreted by dog PI cells eluted near beta-endorphin (1-31). In addition, all of the IR-alpha MSH secreted by dog PI cells coeluted with synthetic alpha MSH on the G-50 column, but IR-ACTH appeared in two peaks, one eluting near porcine ACTH-(1-39) and another, apparently larger mol wt species. Dopamine and somatostatin were found to inhibit the secretion of IR-alpha MSH and IR-ACTH from perifused dog PI cells in a parallel and dose-dependent fashion. Norepinephrine and epinephrine similarly inhibited POMC peptide secretion, but this effect was blocked by haloperidol, suggesting that it was mediated through a dopamine receptor. CRF stimulated the secretion of both hormones from dog PI, and this effect was abolished by treatment of the cells with either dopamine or somatostatin. Cortisol had no effect on either basal or CRF-stimulated secretion of IR-alpha MSH or IR-ACTH from dog PI cells, but it did inhibit CRF-stimulated IR-ACTH from perifused dog PD. These results suggest that 1) dog PI secretes considerably more IR-ACTH than that in the rat; 2) the probable separate cell sources of IR-alpha MSH and IR-ACTH in dog PI are regulated in an identical fashion; and 3) dopamine, somatostatin, and CRF may function in the physiological or pathophysiological regulation of dog PI.

Immunocytochemical and ultrastructural studies on allografts of the pituitary neurointermediate lobe in the third cerebral ventricle of the rat.

Neurointermediate lobes from adult or 10-day-old rats were implanted by a stereotaxic procedure into the third ventricle of adult male rats, in an area close to the paraventricular nucleus. They were examined, using immunocytochemical and ultrastructural techniques, at times ranging from 1 week to 8 months. All grafts were recovered in a healthy condition although some rejection of the tissue was detected at the 1- and 2-week stages. In the neural lobe, clusters of pituicytes were scattered among the loose network of capillaries, most of which had a fenestrated endothelium. The intermediate lobe remained organized in compact avascular lobules. Axons similar to those projecting into the neurointermediate lobe in situ, but also axons of other types (e.g., somatostatinergic, enkephalinergic) penetrated the grafts. Synapses with melanotrophic cells in the intermediate lobe and neurohaemal contacts in the neural lobe were frequent from 2 1/2 months after transplantation. Immunocytochemical and ultrastructural characteristics indicated intense secretory stimulation of the melanotrophic cells in the early stages. All cells enclosed in a same glandular lobule reacted in a similar manner. In later stages, when re-innervation occurred, the cells recovered their initial characteristics. The overall effect of the re-innervation of the intermediate lobe grafted in this location is inhibitory, as in the lobe in situ.

Co-localization of tyrosine hydroxylase (TH)- and serotonin (5-HT)-immunoreactive innervation in the rat pituitary gland.

Adult male Sprague-Dawley rats were examined for possible co-localization of tyrosine hydroxylase (TH) and serotonin (5-HT) within innervation of the pituitary neurointermediate lobe. Use of sheep antiserum to TH and a secondary antibody coupled to fluorescein isothiocyanate (FITC), followed by rabbit anti-5-HT, then rhodamine-coupled second antibody, produced co-existent staining in some, but not all fibers of both the neural and intermediate lobes, using paraffin embedded tissues. Application of a combination of the two primary antibodies followed by sequential application of secondary antibody also resulted in co-localized antigens in selected fibers. Multiple 'classic' neurotransmitters within the same nerve terminals may modulate selected areas of endocrine tissue to control hormone release.

Corticotropin-releasing factor (CRF) and neurotransmitters modulate melanotropic peptide release from rat neurointermediate pituitary in vitro

Isolated pituitary neurointermediate lobes from adult male Sprague-Dawley rats were incubated for up to 90 minutes in media containing stimulatory or inhibitory agents, to study the potential effects on secretion of alpha-melanocyte stimulating hormone (α-MSH) from the pars intermedia. Dopamine (DA), as expected, inhibited the release of α-MSH, as measured by radioimmunoassay (RIA), while corticotropin-releasing factor (CRF) and serotonin (5-HT) stimulated peptide release to varying degrees over the incubation period. A combination of DA with CRF produced an inhibition of peptide release, suggesting that DA suppresses the stimulatory effects of the hypothalamic peptide. Immune staining of pituitary tissue for beta endorphin or α-MSH, and quantified for staining intensity, supported the specific effects of stimulation or inhibition of the respective treatments. In addition, electron microscopy of incubated tissues demonstrated alterations in secretory vesicles and membranous organelles which were consistent with peptide secretion patterns. Our in vitro findings suggest that neurotransmitters and hypothalamic peptide, which are present together in vivo in the pituitary intermediate lobe, interact at the level of the endocrine cells to modulate pro-opiomelanocortin peptide secretion.

Membrane-associated forms of peptidylglycine alpha-amidating monooxygenase activity in rat pituitary. Tissue specificity.

Membrane-associated peptidylglycine alpha-amidating monooxygenase (PAM) activity was investigated in rat anterior and neurointermediate pituitary tissues and in pituitary AtT-20/D-16v and GH3 cell lines. A substantial fraction of total pituitary PAM activity was found to be membrane-associated. Triton X-100, N-octyl-beta-D-glucopyranoside, and Zwittergent were effective in solubilizing PAM activity from crude pituitary membranes. The distribution of enzyme activity between soluble and membrane-associated forms was tissue-specific. In the anterior pituitary lobe and pituitary cell lines, 40-60% of total PAM activity was membrane-associated while only 10% of the alpha-amidating activity in the neurointermediate lobe was membrane-associated. Soluble and membrane-associated forms of PAM shared nearly identical characteristics with respect to copper and ascorbate requirements, pH optima, and Km values. Upon subcellular fractionation of anterior and neurointermediate pituitary lobe homogenates on Percoll gradients, 12-18% of total PAM activity was found in the rough endoplasmic reticulum/Golgi fractions and 42-60% was localized to secretory granule fractions. For both tissues, membrane-associated PAM activity was enriched in the rough endoplasmic reticulum/Golgi pool, whereas most of the secretory granule-associated enzyme activity was soluble.

Pro-opiomelanocortin-derived peptides in the pig pituitary: α- and γ1-melanocyte-stimulating hormones and their glycine-extended forms

Pro-opiomelanocortin (POMC)-related peptides in extracts of anterior and neurointermediate pituitary lobes from pigs were characterized by gel chromatography, reversed-phase chromatography and radioimmunoassays. The peptide content was ca. 3-fold greater in the anterior lobe compared to the neurointermediate lobe (19.8 nmol POMC/anterior lobe vs 7.0 nmol/neurointermediate lobe). In the neurointermediate lobe 93% of POMC was processed to α-melanocyte-stimulating hormone (α-MSH) and analogs exclusively of low molecular weight. Most of the remaining adrenocorticotropic hormone (ACTH)-related material consisted of the glycine-extended intermediate ACTH-(1–14) and analogs. In contrast only one fourth to one third of the N-terminal part of POMC (N-POMC) was processed to amidated γ-MSH and its C-terminal glycine-extended precursor. The relative amount of amidated γ-MSH was the same as α-MSH and analogs (94%). However, more than 95% of these peptides were of high molecular weight. In the anterior lobe 2.3% of N-POMC was processed and 94% was amidated γ-MSH of only high molecular weight. These results show that γ-MSH and α-MSH are amidated to the same extent and that γ 1-MSH and γ 2-MSH immunoreactivity are present in both the anterior lobe and the neurointermediate lobe. The results suggest that the production of amidated peptides is not regulated by the amidation process itself but at an earlier step (e.g. at the proteolytic cleavage).

Peptide- and transmitter-containing neurons in the mediobasal hypothalamus and their relation to GABAergic systems: possible roles in control of prolactin and growth hormone secretion.

Indirect immunofluorescence histochemistry was used to study the relation among GABAergic, catecholaminergic, cholinergic, and peptidergic neurons in the rat mediobasal hypothalamus. By employing a direct double-labelling procedure using sheep antiserum against glutamic acid decarboxylase (GAD), mouse monoclonal and rabbit antibodies to neurotensin (NT) and rabbit antisera to tyrosine hydroxylase (TH), choline acetyltransferase (ChAT), galanin (GAL), growth hormone-releasing factor (GRF), or somatostatin (SOM), it was demonstrated that GAD-positive fibers and terminals in the external part of the median eminence co-contained immunoreactivity for TH, NT, GAL or GRF, but not for SOM. In the internal part of the median eminence-infundibular stalk, GAD-positive/NT-, GAL-, and GRF-negative and GAD-positive/TH-positive fiber plexa were shown. When a recently developed direct triple-labelling procedure with biotin-conjugated mouse secondary antibodies in conjunction with diethylaminocoumarin (DAMC)-conjugated avidin was employed, presence of GAD/GAL/NT- as well as GAD/GRF/NT-containing varicosities could be demonstrated close to hypophysial portal vessels. In colchicine-pretreated animals, GAD was shown to coexist with TH, NT, or GAL in cell bodies in both the dorsomedial and ventrolateral domains of the arcuate nucleus, but with GRF only in the ventrolateral division. ChAT-positive neurons in the ventrolateral region were also TH-positive. In the ventrolateral arcuate nucleus, triple-labelling followed by elution-restaining showed GAD/NT/GAL/TH-immunoreactivities in the same cells. Similarly, double-labelling with two following elution-restaining steps showed several NT/GAL/GRF/TH-containing cell bodies in this part of the arcuate nucleus. GAD-positive cells in the anterior hypothalamic periventricular area and fibers in the pituitary neurointermediate lobe were also TH-positive. The results demonstrate complex patterns of storage of chemical messengers in neurons of the arcuate nucleus-median eminence complex. Possible neuroendocrine interactions of these systems in the control of prolactin and growth hormone secretion are discussed.

Radioreceptor assay for alpha-MSH using mouse B16 melanoma cells+.

A radioreceptor assay for alpha-MSH is described which is based on cultured mouse B16 melanoma cells and bioactive monoiodinated [Nle4]-alpha-MSH tracer. The assay was used (1) to study the binding characteristics of alpha-MSH to B16 cells, (2) to determine the relative binding activity of MSH peptides, and (3) to measure MSH in tissue extracts. The association of alpha-MSH to B16 cells reached a stable plateau after 3 h at 15 degrees C. At 25 degrees or 37 degrees C, the binding was transient and at 0-1 degree C, the association was very slow. The hormone-receptor complex was relatively stable between 0 degrees and 15 degrees C whereas a 50% dissociation was reached after 90 min at 25 degrees C and after 35 min at 37 degrees C. The mean KD for alpha-MSH of four saturation experiments was 1.3 nM and the number of receptors 9570 per cell. 1,10-Phenanthroline had a stabilizing effect in the binding assay when used at a 0.3 mM concentration. From the MSH peptides tested in the binding assay, some showed similar potencies in three bioassays (tyrosinase, melanin and Anolis skin), whereas others displayed considerably lower bioassay values than expected from the binding data. This shows that binding and bioactivity can be dissociated in some of the MSH peptides. The biological activity of MSH from the neurointermediate lobe of the rat pituitary as measured by its binding to B16 cells corresponds fairly well with RIA results; in the anterior lobe, alpha-MSH values are overestimated because of the large amount of ACTH present.

Neuropeptide y administration acutely increases hypothalamic corticotropin-releasing factor immunoreactivity: Lack of effect in other rat brain regions

The effect of acute central administration of Neuropeptide Y (NPY) to adult male rats on the brain content of corticotropin-releasing factor immunoreactivity (CRF-ir) was investigated. The brain regions studied included frontal cortex, hippocampus, medulla-pons, midbrain-thalamus, cerebellum, neurointermediate lobe of pituitary, median eminence and the remaining hypothalamus. CRF-ir was determined in each of these regions using a radioimmunoassay specific for rat CRF. CRF-ir was found to be significantly increased in the major site of CRF localization in the brain, the hypothalamus, in NPY-treated rats as compared to vehicle-treated controls either 15 minutes (p<0.025) or 45 minutes (p<0.005) post-injection. This increase was localized to the median eminence (p<0.05 after 15 minutes, p<0.01 after 45 minutes). No statistically siginificant differences were noted in any of the other brain regions assessed. Plasma adrenocorticotropin levels were also found to increase following NPY treatment, and effect which became significant after 45 minutes (p<0.05). These data show that NPY can alter the content of hypothalamic CRF and may play a role in its regulation.

Nicotine-induced alterations in brain regional concentrations of native and cryptic Met- and Leu-enkephalin

The distribution of cryptic forms (larger enkephalin-containing peptides) in neostriatum, hypothalamus, spinal cord T 3-L 1 and neurointermediate lobe of pituitary were determined by radioimmunoassay. Optimal conditions for enzymic hydrolysis of the cryptic enkephalins by trypsin and carboxypeptidase B were established. The proportion of total Met- and Leu-enkephalin represented by native pentapeptide varied markedly among these central nervous system regions. Also, the distributions of native and cryptic Met-enkephalin were distinct from that of Leu-enkephalin. Chromatographic separation by HPLC of immunoreactive Met-enkephalin peptides revealed only two peaks corresponding to Met-enkephalin and Met-enkephalin sulfoxide in rather equal amounts. Hydrolysis of cryptic Met-enkephalin also produced only two HPLC-separable peaks of immunoreactive Met-enkephalin, again corresponding to Met-enkephalin and Met-enkephalin sulfoxide. Bioactivity of cryptic striatal Met-enkephalin after hydrolysis was demonstrated by antinociception and catalepsy in rats following its intracerebroven-tricular injection. Repeated short-term administration of nicotine, 0.1 mg/kg IP six times at 30 min intervals, produced significant increases in native and cryptic Met-enkephalin in striatum, consistent with an increase in neuronal release of Met-enkephalin together with increases in synthesis and processing of proenkephalin A in this brain region. This regimen of nicotine also decreased levels of native Met-enkephalin and of both native and cryptic Leu-enkephalin in neurointermediate lobe, consistent with nicotine-induced release of both proenkephalin A- and prodynorphin-derived peptides from neurointermediate lobe.

Evidence of correlations between mast-cell histamine and beta-endorphin (ir) from NIL-pituitary in the homeostasis

This study was designed to characterize physiological events related to a single or repeated experimental anaphylactoid reactions (by Compound 48 80 ) in non-stressed or cold-restrained rats. Acute treatment with Compound 48 80 (1 mg/kg, i.p.) increases Beta-endorphin(ir) content in the neurointermediate lobe (NIL) of rat pituitary. Moreover, repeated treatment with Compound 48 80 showed tolerance effects. These animals, exposed to stressful conditions, exhibited a fully evident paw oedema following carrageenin oedema-test, whereas saline-pretreated rats, under the same experimental conditions, showed reduced local inflammation. Since Compound 48 80 produces characteristic, systemic, anaphylactoid reaction in the rat, with a very high degree of selectivity in its action, our results suggest that mast-cell histamine and Beta-endorphin from NIL pituitary are involved in the body's reactivity to stressful stimuli.

Lithium and haloperidol differentially alter the dynorphin A (1–8) and enkephalin levels in the neurointermediate lobe of rat pituitary

Repeated administration of the antimanic drug lithium (4 mEq/kg/day for 2, 4 or 6 days, i.p.) to rats produced a progressive decline and eventual depletion of dynorphin-A (1–8) (DYN) concentration whereas Met 5-enkephalin (ENK) was only marginally decreased in the neurointermediate lobe of the pituitary (NIL). Administration of a neuroleptic haloperidol neither affected ENK and DYN levels nor influenced lithium-induced changes. The study reveals that lithium produces a preferential perturbation in the dynorphin system relative to the enkephalin system. These results taken together with other evidence, indicate that dynorphin is possibly coreleased with vasopression following lithium administration and provide a pharmacological support to the previously described colocalization and corelease of these endogenous peptides in the NIL.

Colocalization of GABA and tyrosine hydroxylase immunoreactivities in the axons innervating the neurointermediate lobe of the rat pituitary: an ultrastructural immunogold study

Distribution of the γ-aminobutyric acid (GABA)ergic and dopaminergic innervations was studied in the rat neurointermediate lobe using antibodies against GABA and tyrosine hydroxylase. In light microscopy, immunoperoxidase staining revealed similar distribution patterns of the axons reacting with both antibodies. Diffusely scattered in both lobes, they were more concentrated along the marginal zone of the neural lobe. Application of a double, recto-verso, immunogold labelling method in electron microscopy revealed systematic colocalization of GABA and tyrosine hydroxylase (TH) immunoreactivities in the axons innervating the intermediate lobe; in the neural lobe, almost all GABA-immunoreactive axons were also labelled for TH. Thus, GABA and dopamine, hitherto reported to occur in distinct axons, in fact colocalize in the axonal systems which innervate the pituitary neurointermediate lobe. These observations suggest possible interactions (pre- or postsynaptic) of both transmitters in the functional regulation of the intermediate and neural lobes.

IV. Endogenous dopamine and serotonin release from the explanted rat tuberohypophyseal system: Effects of electrical stimulation and neurotensin

Excised blocks of brain tissue encompassing the hypothalamic periventricular nucleus, arcuate nucleus, infundibular stalk and attached pituitary neurointermediate lobe (NIL) were obtained from rats and perifused in vitro. The intact tuberohypophyseal dopaminergic pathway projecting from cell bodies in the hypothalamus to terminations in the pars intermedia and posterior lobes was thus isolated. An electrode placed in the explant delivered electrical pulses at various frequencies to hypothalamic targets. Products released in the immediate vicinity of the NIL were continuously collected and assayed for dopamine and serotonin content. Electrical stimulation of the arcuate nucleus from 1.0 – 10.0 Hz produced a frequency-dependent increase in dopamine, but not serotonin, release from the NIL. Stimulation of the infundibular stalk, however, elevated the release of both neurotransmitters. Addition of neurotensin (0.001 – 1.0 uM) to the bathing media produced a concentration-dependent increase in dopamine, but not serotonin, release. These experiments demonstrate the utility of the tuberohypophyseal explant as a model for use in the study of dopaminergic neuronal function in this neuroendocrine axis.

Immunoreactive dynorphin-A in ovine anterior pituitary and effects of estradiol-17 beta administration.

Anterior lobe (AL) tissue of the ovine pituitary gland contained a form of immunoreactive dynorphin-A (ir-DYN-A) larger than that found in pituitary neurointermediate lobe. Administration of estradiol-17 beta or estradiol-17 beta plus progesterone to ovariectomized sheep decreased AL tissue concentrations of ir-DYN-A but did not affect any LH parameter. Enzymatically dispersed AL cells also contained ir-DYN-A, but specific release during in vitro incubation was too low to be detected even when cells were exposed to gonadotropin-releasing hormone.