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1.
Endocrine ; 57(2): 314-325, 2017 Aug.
Article in English | MEDLINE | ID: mdl-27738887

ABSTRACT

The dopamine D2 receptor is the main dopamine receptor expressed in the human normal pituitary gland. The aim of the current study was to evaluate dopamine D2 receptor expression in the corticotroph cell populations of the anterior lobe and pars intermedia, as well as posterior lobe of the human normal pituitary gland by immunohistochemistry. Human normal pituitary gland samples obtained from routine autopsies were used for the study. In all cases, histology together with immunostaining for adrenocorticotropic hormone, melanocyte-stimulating hormone, prolactin, and neurofilaments were performed and compared to the immunostaining for D2 receptor. D2 receptor was heterogeneously expressed in the majority of the cell populations of the anterior and posterior lobe as well as in the area localized between the anterior and posterior lobe, and arbitrary defined as "intermediate zone". This zone, characterized by the presence of nerve fibers included the residual pars intermedia represented by the colloid-filled cysts lined by the remnant melanotroph cells strongly expressing D2 receptors, and clusters of corticotroph cells, belonging to the anterior lobe but localized within the cysts and adjacent to the posterior lobe, variably expressing D2 receptors. D2 dopamine receptor is expressed in the majority of the cell populations of the human normal pituitary gland, and particularly, in the different corticotroph cell populations localized in the anterior lobe and the intermediate zone of the pituitary gland.


Subject(s)
Corticotrophs/metabolism , Pituitary Gland/cytology , Pituitary Gland/metabolism , Receptors, Dopamine D2/biosynthesis , Adrenocorticotropic Hormone/metabolism , Humans , Immunohistochemistry , Melanocyte-Stimulating Hormones/metabolism , Nerve Fibers/metabolism , Pituitary Gland/innervation , Pituitary Gland, Anterior/cytology , Pituitary Gland, Anterior/innervation , Pituitary Gland, Anterior/metabolism , Pituitary Gland, Intermediate/cytology , Pituitary Gland, Intermediate/innervation , Pituitary Gland, Intermediate/metabolism , Pituitary Gland, Posterior/cytology , Pituitary Gland, Posterior/innervation , Pituitary Gland, Posterior/metabolism , Prolactin/metabolism , Receptors, Dopamine D2/genetics
2.
Acta Histochem ; 118(3): 213-8, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26797189

ABSTRACT

Immunohistochemical techniques were employed to investigate the distribution of amylin-like immunoreactivity in the axolotl (Ambystoma mexicanum) pituitary. Amylin-immunoreactive cells were observed in the pars intermedia, and these cells were found to be immunoreactive for α-melanocyte-stimulating hormone (αMSH) as well. In contrast, αMSH-immunoreactive cells in the pars distalis were immuno-negaitive for amylin. These light microscopic findings were confirmed by immunoelectron microscopy. Amylin-immunoreactive signals were located on the haloes of presumable secretory granules in association with αMSH-immunoreactive signals in the amylin-positive cells. However, in the pars distalis, the αMSH-positive cells did not contain amylin-immunoreactive secretory granules. Western blot analysis of axolotl pituitary extracts revealed the labeling of a protein band at approximately 10.5-kDa by the anti-rat amylin serum, which was not labeled by the anti-αMSH antibody. These findings indicate that amylin secreted from MSH-producing cells in the pars intermedia may modulate MSH secretion in an autocrine fashion and may participate in MSH functions such as fatty homeostasis together with MSH.


Subject(s)
Ambystoma mexicanum/anatomy & histology , Pituitary Gland, Anterior/cytology , Pituitary Gland, Intermediate/cytology , Amphibian Proteins/metabolism , Animals , Cell Shape , Immunohistochemistry , Melanocyte-Stimulating Hormones/metabolism
3.
Am J Physiol Endocrinol Metab ; 307(10): E872-84, 2014 Nov 15.
Article in English | MEDLINE | ID: mdl-25184991

ABSTRACT

Although the importance of kisspeptin in the pituitary is firmly established, the signaling mechanisms for the pituitary actions of kisspeptin are still largely unknown. Somatolactin (SL), a member of the growth hormone (GH)/prolactin (PRL) family, is a pituitary hormone with pleiotropic functions in fish, but its regulation by kisspeptin has not been examined. To investigate the functional role of kisspeptin in SL regulation, expression of two paralogues of goldfish Kiss1 receptors (Kiss1ra and Kiss1rb) were confirmed in immunoidentified SLα but not SLß cells isolated by RT-PCR coupled with laser capture microdissection. In goldfish pituitary cells prepared from neurointermediate lobe (NIL), synthetic goldfish Kiss decapeptides (gKiss1-10 and gKiss2-10) could increase SLα release. Consistent with the lack of Kiss1r expression in SLß cells, SLß release was not altered by kisspeptin stimulation. In parallel experiments, goldfish gKiss1-10 could elevate cyclic adenosine monophosphate (cAMP) production, upregulate protein kinase A (PKA) and protein kinase C (PKC) activities, and trigger a rapid rise in intracellular Ca(2+) levels in goldfish NIL cells. Using a pharmacological approach, cAMP/PKA and phospholipase C (PLC)/PKC pathways and subsequent activation of Ca(2+)/calmodulin (CaM)-dependent cascades were shown to be involved in SLα release induced by gKiss1-10. Apparently, the Ca(2+)-dependent cascades were triggered by extracellular Ca(2+) entry via voltage-sensitive Ca(2+) channels and mobilization of inositol trisphosphate-sensitive intracellular Ca(2+) stores. Our results demonstrate that gKiss1-10 can act directly at the pituitary level to trigger SLα release via a complex network of post-receptor signaling mechanisms.


Subject(s)
Calcium-Calmodulin-Dependent Protein Kinases/metabolism , Cyclic AMP-Dependent Protein Kinases/metabolism , Cyclic AMP/metabolism , Fish Proteins/metabolism , Glycoproteins/metabolism , Kisspeptins/metabolism , Pituitary Gland, Intermediate/metabolism , Pituitary Hormones/metabolism , Protein Kinase C/metabolism , Type C Phospholipases/metabolism , Animals , Goldfish , Pituitary Gland/cytology , Pituitary Gland/metabolism , Pituitary Gland, Intermediate/cytology , Signal Transduction
4.
Mol Cell Endocrinol ; 372(1-2): 49-56, 2013 Jun 15.
Article in English | MEDLINE | ID: mdl-23541636

ABSTRACT

Prolonged exposure to stress elicits profound effects on homeostasis that may lead to cryptogenic disorders such as chronic fatigue syndrome. To investigate the pathophysiology associated with the syndrome, we used a rat continuous stress (CS) model where the pituitary represents one of the most affected organs. Here we found that mRNA for VGF (non-acronymic), a member of the granin family, was induced specifically in the intermediate lobe (IL). This was matched by a concomitant increase at the peptide/protein level assessed by C-terminal antibody. Furthermore, the up-regulation of VGF was confirmed by immunohistochemistry in a subset of melanotrophs. VGF expression was altered in the IL of rats receivingthe dopamine D2 receptor agonist bromocriptine or the antagonist sulpiride. In vitro, dopamine dose-dependently decreased the mRNA levels in cultured melanotrophs. These findings suggest that VGF expression under CS is negatively regulated by dopaminergic neurons projecting from the hypothalamus.


Subject(s)
Dopamine/physiology , Melanotrophs/metabolism , Neuropeptides/genetics , Stress, Physiological , Transcriptional Activation , Animals , Bromocriptine/pharmacology , Cells, Cultured , Dopamine Agonists/pharmacology , Dopamine Antagonists/pharmacology , Dopaminergic Neurons/metabolism , Gene Expression , Hypothalamus/cytology , Hypothalamus/metabolism , Male , Neuropeptides/metabolism , Pituitary Gland, Intermediate/cytology , Pituitary Gland, Intermediate/drug effects , Pituitary Gland, Intermediate/metabolism , Primary Cell Culture , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Signal Transduction , Sulpiride/pharmacology
5.
Gen Comp Endocrinol ; 185: 10-8, 2013 May 01.
Article in English | MEDLINE | ID: mdl-23376532

ABSTRACT

The pars distalis (PD) and the pars intermedia (PI) have the same embryonic origin, but their morphological and functional characteristics diverge during development. The PD is highly vascularized, whereas the highly innervated PI is essentially non-vascularized. Based on our previous finding that vascular endothelial growth factor-A (VEGF-A) is involved in vascularization of the rat PD, attempt was made to generate transgenic Xenopus expressing VEGF-A specifically in the melanotrope cells of the PI as a model system for studying the significance of vascularization or avascularization for the functional differentiation of the pituitary. The PI of the transgenic frogs, examined after metamorphosis, were distinctly vascularized but poorly innervated. The experimentally induced vascularization in the PI resulted in a marked increase in tissue volume and a decrease in the expression of both alpha-melanophore-stimulating hormone (α-MSH) and prohormone convertase 2, a cleavage enzyme essential for generating α-MSH. The transgenic animals had low plasma α-MSH concentrations and displayed incomplete adaptation to a black background. To our knowledge, this is the first report indicating that experimentally induced angiogenesis in the PI may bring about functional as well as structural alterations in this tissue.


Subject(s)
Neovascularization, Physiologic , Pituitary Gland, Intermediate/physiology , Vascular Endothelial Growth Factor A/physiology , Adaptation, Physiological/physiology , Animals , Animals, Genetically Modified , Female , Male , Pituitary Gland/blood supply , Pituitary Gland/cytology , Pituitary Gland, Intermediate/blood supply , Pituitary Gland, Intermediate/cytology , Proprotein Convertase 2/metabolism , Xenopus laevis , alpha-MSH/biosynthesis , alpha-MSH/blood
6.
Mol Endocrinol ; 25(1): 117-27, 2011 Jan.
Article in English | MEDLINE | ID: mdl-21084383

ABSTRACT

The pituitary gland contains six distinct hormone-secreting cell types that are essential for basic physiological processes including fertility and responding to stress. Formation of hormone-secreting cells during development relies on Notch signaling to prevent progenitors from prematurely differentiating. The nature of the signal curtailing Notch signaling in the pituitary is unknown, but a good candidate is the endocytic adaptor protein NUMB. NUMB targets Notch for proteolytic degradation, but it also has a broad range of actions, including stabilizing adherens junctions through interactions with cadherins and influencing cell proliferation by stabilizing expression of the tumor suppressor protein p53. Here, we show that NUMB and its closely related homolog, NUMBLIKE, are expressed in undifferentiated cells during development and later in gonadotropes in the anterior lobe and melanotropes of the intermediate lobe. All four isoforms of NUMB, are detectable in the pituitary, with the shorter forms becoming more prominent after adolescence. Conditionally deleting Numb and Numblike in the intermediate lobe melanotropes with Pomc Cre mice dramatically alters the morphology of cells in the intermediate lobe, coincident with impaired localization of adherens junctions proteins including E-CADHERIN, N-CADHERIN, ß-CATENIN, and α-CATENIN. Strikingly, the border between posterior and intermediate lobes is also disrupted. These mice also have disorganized progenitor cells, marked by SOX2, but proliferation is unaffected. Unexpectedly, Notch activity appears normal in conditional knockout mice. Thus, Numb is critical for maintaining cell-cell interactions in the pituitary intermediate lobe that are essential for proper cell placement.


Subject(s)
Cell Movement , Gene Deletion , Membrane Proteins/genetics , Nerve Tissue Proteins/genetics , Pituitary Gland, Intermediate/cytology , Pituitary Gland, Intermediate/embryology , Pro-Opiomelanocortin/metabolism , Stem Cells/cytology , Animals , Cell Adhesion , Cell Proliferation , Gene Expression Regulation, Developmental , Gonadotrophs/cytology , Gonadotrophs/metabolism , Integrases/metabolism , Intracellular Signaling Peptides and Proteins , Melanotrophs/cytology , Melanotrophs/metabolism , Membrane Proteins/metabolism , Mice , Mice, Knockout , Nerve Tissue Proteins/metabolism , Pituitary Gland, Intermediate/metabolism , Protein Isoforms/genetics , Protein Isoforms/metabolism , Receptors, Notch/metabolism , Stem Cells/metabolism
7.
Neuroendocrinology ; 92(3): 158-67, 2010.
Article in English | MEDLINE | ID: mdl-20551618

ABSTRACT

BACKGROUND/AIMS: Complex intrinsic optical changes (light scattering) are readily observed in the neurointermediate lobe of the mouse pituitary gland following electrical stimulation of the infundibular stalk. Our laboratory has previously identified three distinct phases within the light scattering signal: two rapid responses to action potential stimulation and a long duration recovery. The rapid light scattering signals, restricted to the neurohypophysial portion (posterior pituitary) of the neurointermediate lobe, consist of an E-wave and an S-wave that reflect excitation and secretion, respectively. The E-wave has the approximate shape of the action potential and includes voltage- and current-related components and is independent of Ca(2+) entry. The S-wave is related to Ca(2+) entry and exocytosis. The slow recovery phase of the light scattering signal, which we designated the R-wave, is less well characterized. METHODS: Using high temporal resolution light scattering measurements, we monitored intrinsic optical changes in the neurointermediate lobe of the mouse pituitary gland. Pharmacological interventions during the measurements were employed. RESULTS: The data presented here provide optical and pharmacological evidence suggesting that the R-wave, which comprises signals from the posterior pituitary as well as from the pars intermedia, mirrors volume changes in pars intermedia cells following a train of stimuli applied to the infundibular stalk. These volume changes were blocked by the GABA-receptor antagonists bicuculline and picrotoxin, and were mimicked by direct application of GABA in the absence of electrical stimulation. CONCLUSIONS: These results emphasize the importance of central GABAergic projections into the neurointermediate lobe, and the potential role of GABA in effecting hormone release from the pars intermedia.


Subject(s)
Action Potentials/physiology , Calcium/metabolism , Pituitary Gland, Intermediate/physiology , Receptors, GABA-A/metabolism , Action Potentials/drug effects , Animals , Bicuculline/pharmacology , Female , GABA Antagonists/pharmacology , Mice , Picrotoxin/pharmacology , Pituitary Gland, Intermediate/cytology , Pituitary Gland, Intermediate/drug effects , gamma-Aminobutyric Acid/pharmacology
8.
J Neurophysiol ; 99(3): 1545-53, 2008 Mar.
Article in English | MEDLINE | ID: mdl-18171710

ABSTRACT

We report the first optical recordings of action potentials, in single trials, from one or a few (approximately 1-2 microm) mammalian nerve terminals in an intact in vitro preparation, the mouse neurohypophysis. The measurements used two-photon excitation along the "blue" edge of the two-photon absorption spectrum of di-3-ANEPPDHQ (a fluorescent voltage-sensitive naphthyl styryl-pyridinium dye), and epifluorescence detection, a configuration that is critical for noninvasive recording of electrical activity from intact brains. Single-trial recordings of action potentials exhibited signal-to-noise ratios of approximately 5:1 and fractional fluorescence changes of up to approximately 10%. This method, by virtue of its optical sectioning capability, deep tissue penetration, and efficient epifluorescence detection, offers clear advantages over linear, as well as other nonlinear optical techniques used to monitor voltage changes in localized neuronal regions, and provides an alternative to invasive electrode arrays for studying neuronal systems in vivo.


Subject(s)
Action Potentials/physiology , Fluorescence , Fluorescent Dyes/pharmacology , Neurons/cytology , Presynaptic Terminals/drug effects , Pyridinium Compounds/pharmacology , Action Potentials/drug effects , Animals , Female , In Vitro Techniques , Mice , Microscopy, Electron, Transmission/methods , Neurons/drug effects , Neurons/physiology , Pituitary Gland, Intermediate/cytology , Presynaptic Terminals/ultrastructure , Spectrometry, Fluorescence/methods
9.
Endocrinology ; 148(10): 4649-57, 2007 Oct.
Article in English | MEDLINE | ID: mdl-17615142

ABSTRACT

Cervical stimulation induces two daily rhythmic prolactin surges, nocturnal and diurnal, which persist for several days. We have shown that a bolus injection of oxytocin initiates a similar prolactin rhythm, which persists despite low levels of oxytocin after injection. This suggests that oxytocin may trigger the cervical stimulation-induced rhythmic prolactin surges. To investigate this hypothesis, we infused an oxytocin antagonist that does not cross the blood-brain barrier for 24 h before and after cervical stimulation and measured serum prolactin. We also measured dopaminergic neuronal activity because mathematical modeling predicted that this activity would be low in the presence of the oxytocin antagonist. We thus tested this hypothesis by measuring dopaminergic neuronal activity in the tuberoinfundibular, periventricular hypophyseal, and tuberohypophyseal dopaminergic neurons. Infusion of oxytocin antagonist before cervical stimulation abolished prolactin surges, and infusion of oxytocin antagonist after cervical stimulation abolished the diurnal and significantly decreased the nocturnal surges of prolactin. The rhythmic prolactin surges returned after the clearance of the oxytocin antagonist. Hypothalamic dopaminergic activity was elevated in antiphase with prolactin surges, and the antiphase elevation was abolished by the oxytocin antagonist in the tuberoinfundibular and tuberohypophyseal dopaminergic neurons, consistent with the mathematical model. These findings suggest that oxytocin is a physiologically relevant prolactin-releasing factor. However, the cervical stimulation-induced prolactin surges are maintained even in the absence of oxytocin actions at the lactotroph, which strongly suggests the maintenance of prolactin surges are not dependent upon oxytocin actions at the pituitary gland.


Subject(s)
Cervix Uteri/physiology , Lactotrophs/metabolism , Ovariectomy , Oxytocin/physiology , Prolactin/metabolism , Animals , Circadian Rhythm , Dopamine/metabolism , Electric Stimulation , Female , Median Eminence/cytology , Median Eminence/metabolism , Models, Biological , Neurons/physiology , Ornipressin/analogs & derivatives , Ornipressin/pharmacology , Oxytocin/antagonists & inhibitors , Pituitary Gland, Intermediate/cytology , Pituitary Gland, Intermediate/metabolism , Pituitary Gland, Posterior/cytology , Pituitary Gland, Posterior/metabolism , Rats , Rats, Sprague-Dawley
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