Regulation of neurotrophic peptide expression in sympathetic neurons: quantitative analysis using radioimmunoassay and real-time quantitative polymerase chain reaction

Girard, Beatrice M.; May, Víctor; Bora, Susan H.; Fina, Frédéric; Braas, Karen M.

doi:10.1016/s0167-0115(02)00191-x

Cited by 51 publications

(81 citation statements)

References 28 publications

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“…Real time PCR to quantify PACAP transcripts was performed as described previously (Girard et al, 2002a;Braas et al, 2004;Girard et al, 2006a). The primers used were identical to those published in our preliminary report (Girard et al, 2006b).…”

Section: Real Time Quantitative Reverse Transcription-polymerase Chaimentioning

confidence: 99%

“…Briefly, 1X buffer was used containing 5 mM MgCl 2 , 200 μM dATP, dGTP, dTTP and dCTP, 0.625 U AmpliTaq Gold and 300 nM of each primer in a final 25 μl reaction. Real-time quantitative PCR was performed on an ABI 7500 Fast Real-Time PCR System (Applied Biosystems, Norwalk, CT) after optimization of cycling parameters which included SYBR Green I melting point analyses (Girard et al, 2002a). Data were analyzed at the termination of each assay using Sequence Detector 1.3 software (Applied Biosystems, Norwalk, CT).…”

Section: Real Time Quantitative Reverse Transcription-polymerase Chaimentioning

confidence: 99%

“…The total RNA quantity for each whole mount preparation was determined using a Nanodrop ND-1000 spectrophotometer (Nanodrop Technologies, Wilmington, DE). 1 μg of RNA per sample was used to synthesize complementary DNA with the Omniscript reverse transcription (QIAGEN Inc., Valencia CA) and oligo-dT primers.Real time PCR to quantify PACAP transcripts was performed as described previously (Girard et al, 2002a;Braas et al, 2004;Girard et al, 2006a). The primers used were identical to those published in our preliminary report (Girard et al, 2006b).…”

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Regulation of neuronal pituitary adenylate cyclase-activating polypeptide expression during culture of guinea-pig cardiac ganglia

et al. 2007

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The trophic neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) increases in many different neuron types following injury; a response postulated to support cell survival and regeneration. In acutely isolated cardiac ganglia, approximately 1% of the cardiac neurons exhibited PACAP immunoreactivity whereas after 72 hours in culture, ~25% of the neurons were PACAP immunoreactive. In contrast, there was no increase in vasoactive intestinal polypeptide (VIP)-immunoreactive (IR) cells. Using a combination of immunocytochemical and molecular techniques, we have quantified PACAP expression, during explant culture of guinea pig cardiac ganglia. Using real time PCR, PACAP transcript levels increased progressively up to 48 hours in culture with no further increase after 72 hours. PACAP transcript levels were reduced by neurturin at 48 hours in culture but not after 24 or 72 hours in culture. In addition, neurturin partially suppressed the percentage of PACAP-IR neurons after 72 hours in culture, an effect mediated by activation of the phosphatidylinositol 3-kinase and mitogen activated protein kinase signaling pathways. The addition of different known regulatory molecules, including CNTF, Il-1β, TNFα, bFGF, TGF βand NGF did not increase the percentage of PACAP-IR neurons after 24 hours in culture; a result indicating that the generation and secretion of these factors did not stimulate PACAP expression. The presence of 20 nM PACAP or 10 μM forskolin increased the percentage of PACAP-IR cardiac neurons in 24 hour cultures, but not in 72 hour cultures. Neither treatment enhanced the number of VIP-IR neurons. The addition of the PAC 1 receptor antagonist, M65 (100 nM) suppressed the 20 nM PACAP-induced increase in percentage of PACAP-IR cells in 24 hour cultures indicating the effect of PACAP was mediated through the PAC 1 receptor. However, 100 nM M65 had no effect on the percentage of PACAP-IR cells in either 24 or 48 hour cultures not treated with exogenous PACAP, suggesting that endogenous release of PACAP likely did not contribute to the enhanced peptide expression. We postulate that the enhanced PACAP expression, which occurs in response to injury is facilitated in the explant cultured cardiac ganglia by the loss of a target-derived inhibitory factor, very likely neurturin. In intact tissues the presence of neurturin would normally suppress PACAP expression. Lastly, our results indicate that many common trophic factors do not enhance PACAP expression in the cultured cardiac neurons. However, the stimulatory role of an, as yet, unidentified factor can not be excluded. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content...

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Section: Real Time Quantitative Reverse Transcription-polymerase Chaimentioning

confidence: 99%

Section: Real Time Quantitative Reverse Transcription-polymerase Chaimentioning

confidence: 99%

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Regulation of neuronal pituitary adenylate cyclase-activating polypeptide expression during culture of guinea-pig cardiac ganglia

et al. 2007

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“…Depolarization of primary cultured sympathetic neurons of the rat SCG elevated Gal mRNA levels which appeared to correlate more closely with Gal peptide production (Girard et al, 2002). The neurotrophic actions of Gal are demonstrated by the upregulation of Gal expression both in DRG neurons and in SCG neurons after nerve injury (Holmberg et al, 2005;Holmes et al, 2005).…”

Section: Discussionmentioning

confidence: 93%

“…(Yang et al, 2009). Many factors, such as leukemia inhibitory factor (LIF), nerve growth factor (NGF), non-neuronal cells, and potassium, have been demonstrated to be involved in the upregulation of Gal in cultured SCG neurons (Klimaschewski, 1997;Girard et al, 2002;Liu et al, 2010). The use of dissociated sympathetic neurons in vitro has provided a considerable amount of information concerning adrenoreceptors which regulate sympathetic transmitter release (Boehm and Huck, 1997).…”

Section: Introductionmentioning

confidence: 99%

Effects of Alpha 1- and Alpha 2-Adrenoreceptor Stimulation on Galanin mRNA Expression in Primary Cultured Superior Cervical Ganglion Neurons

Xing¹,

Chen²,

Liu³

et al. 2011

Biomolecules and Therapeutics

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Abstractan inhibitory modulator of cholinergic and noradrenergic neurotransmission (Miller et al., 1999;Van Hoomissen et al., 2004;Landry et al., 2005;Hobson et al., 2006). Gal is co-localized with norepinephrine (NE) in several regions of the brain, such as locus coeruleus, bed nucleus of the stria terminalis, amygdala, cortex and hippocampus (Hokfelt et al., 1998;Kozicz, 2001;Xu et al., 2001;Holmes et al., 2002;Barrera et al., 2006;Ogren et al., 2006;Simpson et al., 2006). Gal hyperpolarizes noradrenergic locus coeruleus neurons at high concentrations (10 -6 -10 -7 mol/L) and enhances NE-induced hyperpolarization at low concentrations (10 -9 mol/L) (Hokfelt et al., 1998). It has been shown that Gal expression was regulated by NE in the magnocellular neurons of supraoptic nucleus in an ex vivo acute model of rat hypothalamic slices (Melnikova et al., 2006) and in cultured dorsal root ganglion (DRG) neuronsOriginal Article

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Autonomic dysfunction and plasticity in micturition reflexes in human α‐synuclein mice

Hamill

Tompkins

Girard

et al. 2012

Developmental Neurobiology

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Although often overshadowed by the motor dysfunction associated with Parkinson’s disease (PD), autonomic dysfunction including urinary bladder and bowel dysfunctions are often associated with PD and may precede motoric changes; such autonomic dysfunction may permit early detection and intervention. Lower urinary tract symptoms are common in PD patients and result in significant morbidity. The current studies focus on non-motor symptoms in PD using a transgenic mouse model with overexpression of human α-synuclein, the peptide found in high concentrations in Lewy body neuronal inclusions, the histopathologic hallmark of PD. We examined changes in the physiological, molecular, chemical, and electrical properties of neuronal pathways controlling urinary bladder function in transgenic mice. The results of these studies reveal that autonomic dysfunction (i.e., urinary bladder) can precede motor dysfunction. In addition, mice with human α-synuclein overexpression in relevant neuronal populations is associated with alterations in expression of neurotransmitter/ neuromodulatory molecules (PACAP, VIP, substance P, neuronal NOS) within neuronal pathways regulating bladder function as well as with increased NGF expression in the urinary bladder. Changes in the electrical and synaptic properties of neurons in the major pelvic ganglia that provide postganglionic innervation to urogenital tissues were not changed as determined with intracellular recording. The urinary bladder dysfunction observed in transgenic mice likely reflects changes in peripheral (i.e., afferent) and/or central micturition pathways or changes in the urinary bladder. SYN-OE mice provide an opportunity to examine early events underlying the molecular and cellular plasticity of autonomic nervous system pathways underlying synucleinopathies.

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Regulation of neurotrophic peptide expression in sympathetic neurons: quantitative analysis using radioimmunoassay and real-time quantitative polymerase chain reaction

Cited by 51 publications

References 28 publications

Regulation of neuronal pituitary adenylate cyclase-activating polypeptide expression during culture of guinea-pig cardiac ganglia

Regulation of neuronal pituitary adenylate cyclase-activating polypeptide expression during culture of guinea-pig cardiac ganglia

Effects of Alpha 1- and Alpha 2-Adrenoreceptor Stimulation on Galanin mRNA Expression in Primary Cultured Superior Cervical Ganglion Neurons

Autonomic dysfunction and plasticity in micturition reflexes in human α‐synuclein mice

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