Cell immunoblot assay study demonstrating the release of PACAP from individual anterior pituitary cells of rats and the effect of PACAP on LH release

Szabó, Enikő; Nemeskéri, Ágnes; Heinzlmann, Andrea; Suzuki, Nobuhiro; Arimura, Akira; Köves, Katalin

doi:10.1016/s0167-0115(02)00186-6

Cited by 26 publications

(15 citation statements)

References 21 publications

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“…Scale bar in the photomicrographs is 500 mm. Cimini et al 1994, Szabó et al 2002, 2004 and has been shown to be suitable for demonstrating hormone release from individual cells in the rat pituitary. As no such immunoassay system has been developed for measuring adenohypophysial hormone levels in the goldfish pituitary, we have previously modified this method to examine if PACAP stimulates Sl or Prl release in dispersed and cultured pituitary cells (Matsuda et al 2008, Azuma et al 2009).…”

Section: Discussionmentioning

confidence: 99%

“…In order to examine the effect of MCH on SL release from goldfish pituitary cells, the cell immunoblot method was carried out according to previous reports (Arita et al 1993, Cimini et al 1994, Szabó et al 2002, 2004 but was modified for use on isolated goldfish pituitary cells (Matsuda et al 2008, Azuma et al 2009). This method allowed for the semiquantitative measurement of SL release from individual cells (Matsuda et al 2008, Azuma et al 2009).…”

Section: Primary Cell Culture and Cell Immunoblot Methodsmentioning

confidence: 99%

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Melanin-concentrating hormone reduces somatolactin release from cultured goldfish pituitary cells

Tanaka¹,

Azuma²,

Nejigaki³

et al. 2009

Journal of Endocrinology

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Melanin-concentrating hormone (MCH)-containing neurons directly innervate the adenohypophysis in the teleost pituitary. We examined immunohistochemically the relationship between MCH-containing nerve fibres or endings and somatolactin (SL)-producing cells in the goldfish pituitary. Nerve fibres or endings with MCH-like immunoreactivity were identified in the neurohypophysis in close proximity to the adenohypophysial cells showing SL-like immunoreactivity. We also examined the effect of MCH on SL release from cultured goldfish pituitary cells and SL synthesis using a cell immunoblot and a real-time PCR method. Treatment of individually dispersed pituitary cells with MCH 10 K7 M for 3 h decreased the area of SL-like immunoreactivity on immunoblots, and MCH-induced reductions in SL release were blocked by treatment with the mammalian MCH receptor (MCHR) antagonist, compound-30, at a concentration of 10 K5 M.Treatment with 10 K7 M MCH for 3 h did not affect sl-a and -b (smtla and -b as given in the Zfin Database) mRNA expression levels. These led us to explore the signal transduction mechanism leading to the inhibition of SL release, for which we examined whether MCH-induced reductions in SL release are mediated by the G i or G q proteincoupled signalling pathway. The MCH-induced reductions in SL release were abolished by treatment with the G i/o protein inhibitors, NF023 (10 K5 M) or pertussis toxin (260 ng/ml), but not by the phospholipase C inhibitor, U-73122 (3!10 K6 M). These results indicate that MCH can potentially function as a hypothalamic factor suppressing SL release via the MCHR, and subsequently through the G i protein to inhibit the adenylate cyclase/cAMP/protein kinase A-signalling pathway in goldfish pituitary cells.

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Section: Discussionmentioning

confidence: 99%

Section: Primary Cell Culture and Cell Immunoblot Methodsmentioning

confidence: 99%

Melanin-concentrating hormone reduces somatolactin release from cultured goldfish pituitary cells

Tanaka¹,

Azuma²,

Nejigaki³

et al. 2009

Journal of Endocrinology

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“…The opaque nature of nitrocellulose membranes makes them unable to be observed directly under a light microscope (Szabo et al 2002;Yamasato and Satoh 2001). Recently, Barry et al (2007) applied a new method for in situ microscopic observation of fungal morphology on nitrocellulose membranes with high-power magnification after treating the membranes with microscope immersion oil.…”

Section: Discussionmentioning

confidence: 99%

Use of nitrocellulose membranes as a scaffold in cell culture

Wang

Deng

et al. 2012

Cytotechnology

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Nitrocellulose membranes, one of the most important and oldest cellulose derivatives, are commonly used for nucleic acid and protein detection in research and diagnostic applications. However, a limited number of studies have explored whether they can act as scaffolds for cell growth. In this study, we investigated this polymeric material for its ability to support the growth of human cells. Eight established cell lines were examined for adherence, growth, spread, and survival on nitrocellulose membranes by optical microscopy after hematoxylin and eosin and/or immunocytochemical staining and by scanning electron microscopy. Apoptosis and leakage of lactate dehydrogenase (LDH) were also assessed. All cells readily adhered to and spread on the surface of nitrocellulose membranes as well as coverslips, and the cells maintained the expression of digestive system-specific genes. No significant change was detected in apoptosis or leakage of LDH from cells grown on nitrocellulose membranes. These results suggested that nitrocellulose membranes have a suitable cytocompatibility towards human cells and that they might be used for tissueengineering scaffolds. Moreover, we demonstrate an additional and underused property of nitrocellulose of specific relevance to microscopic imaging, as it can be rendered virtually transparent, thus the cells growing on such membranes can be observed directly under an optical microscope after staining.

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“…Within this context, however, it should be noted that these responses are highly dependent on the methodological choices. For instance, systemic administration of PACAP could release pituitary hormones, while in vitro studies using static incubation or perfusion of the pituitary cell cultures, had no effect or produced contrasting responses [170,178,179]. It is possible that the hypophysiotropic actions of PACAP in the pituitary gland could be mediated through indirect mechanisms involving paracrine actions.…”

Section: Actions In the Adenohypophysismentioning

confidence: 99%

The Secretin/Pituitary Adenylate Cyclase-Activating Polypeptide/ Vasoactive Intestinal Polypeptide Superfamily in the Central Nervous System

Chu¹,

Lee²,

Siu³

et al. 2006

CNSAMC

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The secretin/PACAP/VIP superfamily contains at least ten brain-gut peptides, including secretin, pituitary adenylate cyclase-activating polypeptide (PACAP), vasoactive intestinal polypeptide (VIP), glucagon, glucagon-like peptide-1 (GLP-1), glucagon like peptide-2 (GLP-2), gastric inhibitory polypeptide (GIP), peptide histidine isoleucine (PHI) or peptide histidine methionine (PHM), and growth hormone-releasing hormone (GHRH). These peptides exhibit a wide tissue distribution in the peripheral systems, indicating their pleiotrophic actions in the body. Meanwhile, their functions in the central nervious system (CNS) have also been consolidated recently. For instance, most of these peptides have shown to serve as neurotransmitters, neuromodulators, neurotrophic factors, and/or neurohormones in the brain, and hence, their potential as novel CNS agents in treating neurological disorders including Autism, Alzheimer's disease, Parkinson's disease and HIV-associated neuronal cell death were recently exploited. In this article, recent progress in research of peptides in this family with particular emphasis on structures, their central functions and potential use in the treatment of neuronal diseases are reviewed.

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Cell immunoblot assay study demonstrating the release of PACAP from individual anterior pituitary cells of rats and the effect of PACAP on LH release

Cited by 26 publications

References 21 publications

Melanin-concentrating hormone reduces somatolactin release from cultured goldfish pituitary cells

Melanin-concentrating hormone reduces somatolactin release from cultured goldfish pituitary cells

Use of nitrocellulose membranes as a scaffold in cell culture

The Secretin/Pituitary Adenylate Cyclase-Activating Polypeptide/ Vasoactive Intestinal Polypeptide Superfamily in the Central Nervous System

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