Decreased MYPT-1 phosphorylation at Thr696 and Cdc42 protein expression are associated with...
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the serine kinase receptor for TGF-β (TβRI). This response will be considered in the context of responses to endothelin-1 and the options for therapeutically targeting endothelin-1 broadened to include downstream signalling otherwise associated with TGF-β receptor activation. doi:10.1016/j.lfs.2013.12.138 Structure of the precursor of salmon, Oncorhynchus keta, endothelins and phylogenetic analysis Hongyu Wang a , Jiexia Quan a , Tsuyoshi Uchide b , Kaname Saida a,c a National Institute of Advanced Industrial Science and Technology (AIST), Japan b Veterinary Internal Medicine, Department of Small Animal Clinical Sciences, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, Hokkaido, Japan c Graduate School, Shibaura Institute of Technology, Japan E-mail address: [email protected] (K. Saida) Endothelin (ET)-related receptors homologous to mammalian receptors have been cloned from fish, indicating that ET-related ligands may be present in this species. Here we cloned cDNAs encoding preproendothelins (PPETs) from the intestinal cDNAlibrary. Salmon ETs cDNAs encode 200 amino acids, including a 20-amino- acid putativesignal sequence, as well as mature ETs, big ETs, and ET- like sequences. This sequences together with other published PPET sequences were used to analyze the phylogenetic relationship among all ET family genes. doi:10.1016/j.lfs.2013.12.139 Decreased MYPT-1 phosphorylation at Thr696 and Cdc42 protein expression are associated with decreased contractile responses to ET-1 in corpora cavernosa and internal pudendal artery from Goto-Kakizaki diabetic rats Rheure Lopes a , Fernando Carneiro a , Theodora Szasz b , Gisele Bomfim c , Clinton Webb b , Rita Tostes a a University of Sao Paulo, Brazil, Pharmacology, Medical School of Ribeirao Preto, Brazil b Georgia Health Sciences University, USA c Federal University of Mato Grosso, Brazil E-mail address: [email protected] (R. Lopes) Endothelin-1 (ET-1) plays a crucial role in the development of erectile dysfunction (ED). The Goto-Kakizaki (GK) rat is a non-obese type 2 diabetes mellitus model, which displays ED and increased ET- 1 plasma levels. The present study tested the hypothesis that GK rats display increased corpora cavernosa (CC) and internal pudendal artery (IPA) contractions to ET-1 as a contributing mechanism for ED. GK rats demonstrated impaired erectile function represented by decreased ICP/MAP responses after cavernous nerve stimulation. In GK rats contractile responses to ET-1 were decreased in both, CC tissue [Control: 396.00 ± 10 vs GK: 168.00 ± 16.00; Emax, mN/μg of tissue] and in IPA [Control: 25.00 ± 1.75 vs GK: 14.83 ± 1.66; Emax, mN]. Gene expression of prepro-ET-1 [Control: 1.00 vs GK: 0.25 ± 0.04] and ETB receptors [Control: 1.00 vs GK: 0.58 ± 0.09], but not ETA receptors was decreased in CC from GK rats. In GK rats, CC protein expression of ETA receptor [Control: 1.00 vs GK: 4.18 ± 0.58], and phosphorylation of ERK 1/2 [Control: 1.00 vs GK: 1.31 ± 0.09] were increased, whereas ETB receptor expression [Control: 1.00 vs GK: 0.75 ± 0.08], Cdc42 protein expression [Control: 1.00 vs GK: 0.40 ± 0.08] and phosphorylation of MYPT-1 [Control: 1.00 vs GK: 0.36 ± 0.15] were decreased. In conclusion, GK rats display ED and exhibit decreased cavernosal and IPA reactivity to ET-1. Whereas decreased phosphorylation of MYPT-1 and Cdc42 protein expression may account for decreased ET-1 responses, it indicates that ED in GK rats is not associated with augmented CC and IPA reactivity to ET-1. doi:10.1016/j.lfs.2013.12.140 Generation of Edn2-iCre transgenic mice Joseph Cacioppo a , Patrick Lin a , Arnon Gal a , Yongbum Koo a,b , CheMyong Ko a a The Department of Comparative Biosciences, University of Illinois at Urbana-Champaign, Urbana IL, USA b Department of Biotechnology and Biomedical Science, Inje University, Gimhae, South Korea E-mail address: [email protected] (J. Cacioppo) Endothelin-2 (ET-2) is a potent vasoconstrictive peptide. Though similar to ET-1, recent studies suggest that ET-2 may act through distinct pathways, necessitating deeper study. ET-2 may play a role in heart failure, inflammation, macular degeneration, and cancer metas- tasis. It is transiently expressed and tightly regulated in the per- iovulatory ovary, where it may aid ovulation by inducing constriction of the follicular wall. Here, we present a transgenic mouse line that expresses iCre (codon-improved Cre recombinase) under the regula- tion of the promoter of the endothelin-2 (edn2) gene, which was developed as a novel model for characterizing the expression of ET-2 and for conditional deletion of genes in cells where ET-2 is expressed. A vector was generated containing iCre, a polyadenylation signal sequence, and an frt-neomycin-resistance-frt cassette. Two homolo- gous regions of the edn2 gene flanking the ATG start codon were isolated from a BAC (bacterial artificial chromosome) clone and inserted upstream and downstream of the iCre-pA-FNF cassette. Homologous recombination was used to re-insert the cassette into the BAC plasmid. Following purification, the plasmid was inserted into fertilized eggs of C57BL/6 mice through pronuclear injection, and resulting eggs were implanted into pseudopregnant mice. Five Edn2-icre transgene- containing lines of mice were established, and one line was bred with ROSA26 reporter. Offspring were used to localize iCre-expressing cells through X-gal staining. Characterization of the staining pattern revealed that iCre was expressed in granulosa cells of ovulatory follicles, cardiomyocytes, the pituitary, and the liver. We expect this novel mouse model to be a useful tool for future studies on the role of ET-2. doi:10.1016/j.lfs.2013.12.141 The calcitonin gene-related peptide (CGRP) plays beneficial roles in myocardial ischemia elicited by endothelin-1 Satoshi Homma a , Satoshi Sakai a , Ken-ichi Yanagi b , Yumi Miyauchi a , Kazutaka Aonuma a , Takashi Miyauchi a a Department of Medicine, University of Tsukuba, Tsukuba, Japan b Department of Biomedical Sciences, University of Tsukuba, Tsukuba, Japan E-mail address: [email protected] (S. Homma) Purpose: In addition to the adrenergic and cholinergic nerves, the cardiovascular tissues are also innervated by several peptidergic neurons that mediate nonadrenergic noncholinergic (NANC) func- tions. Among such neuropeptides, CGRP is released from capsaicin- sensitive sensory neurons in peripheral organs. CGRP is known as Abstracts e40
Transcript of Decreased MYPT-1 phosphorylation at Thr696 and Cdc42 protein expression are associated with...
the serine kinase receptor for TGF-β (TβRI). This response will beconsidered in the context of responses to endothelin-1 and the optionsfor therapeutically targeting endothelin-1 broadened to includedownstream signalling otherwise associated with TGF-β receptoractivation.
doi:10.1016/j.lfs.2013.12.138
Structure of the precursor of salmon, Oncorhynchus keta,endothelins and phylogenetic analysisHongyu Wanga, Jiexia Quana, Tsuyoshi Uchideb, Kaname Saidaa,c
aNational Institute of Advanced Industrial Science and Technology (AIST),JapanbVeterinary Internal Medicine, Department of Small Animal ClinicalSciences, School of VeterinaryMedicine, Rakuno Gakuen University, Ebetsu,Hokkaido, JapancGraduate School, Shibaura Institute of Technology, JapanE-mail address: [email protected] (K. Saida)
Endothelin (ET)-related receptors homologous to mammalianreceptors have been cloned from fish, indicating that ET-relatedligands may be present in this species. Here we cloned cDNAsencoding preproendothelins (PPETs) from the intestinal cDNAlibrary.Salmon ETs cDNAs encode 200 amino acids, including a 20-amino-acid putativesignal sequence, as well as mature ETs, big ETs, and ET-like sequences. This sequences together with other published PPETsequences were used to analyze the phylogenetic relationship amongall ET family genes.
doi:10.1016/j.lfs.2013.12.139
Decreased MYPT-1 phosphorylation at Thr696 and Cdc42 proteinexpression are associated with decreased contractile responses toET-1 in corpora cavernosa and internal pudendal artery fromGoto-Kakizaki diabetic ratsRheure Lopesa, Fernando Carneiroa, Theodora Szaszb, Gisele Bomfimc,Clinton Webbb, Rita Tostesa
aUniversity of Sao Paulo, Brazil, Pharmacology, Medical School of RibeiraoPreto, BrazilbGeorgia Health Sciences University, USAcFederal University of Mato Grosso, BrazilE-mail address: [email protected] (R. Lopes)
Endothelin-1 (ET-1) plays a crucial role in the development oferectile dysfunction (ED). The Goto-Kakizaki (GK) rat is a non-obesetype 2 diabetes mellitus model, which displays ED and increased ET-1 plasma levels. The present study tested the hypothesis that GK ratsdisplay increased corpora cavernosa (CC) and internal pudendalartery (IPA) contractions to ET-1 as a contributing mechanism for ED.GK rats demonstrated impaired erectile function represented bydecreased ICP/MAP responses after cavernous nerve stimulation. InGK rats contractile responses to ET-1 were decreased in both, CCtissue [Control: 396.00 ± 10 vs GK: 168.00 ± 16.00; Emax, mN/μg oftissue] and in IPA [Control: 25.00 ± 1.75 vs GK: 14.83 ± 1.66; Emax,mN]. Gene expression of prepro-ET-1 [Control: 1.00 vs GK: 0.25± 0.04] and ETB receptors [Control: 1.00 vs GK: 0.58 ± 0.09], but notETA receptors was decreased in CC from GK rats. In GK rats, CCprotein expression of ETA receptor [Control: 1.00 vs GK: 4.18± 0.58], and phosphorylation of ERK 1/2 [Control: 1.00 vs GK: 1.31± 0.09] were increased, whereas ETB receptor expression [Control:1.00 vs GK: 0.75 ± 0.08], Cdc42 protein expression [Control: 1.00 vs
GK: 0.40 ± 0.08] and phosphorylation of MYPT-1 [Control: 1.00 vsGK: 0.36 ± 0.15] were decreased. In conclusion, GK rats display EDand exhibit decreased cavernosal and IPA reactivity to ET-1. Whereasdecreased phosphorylation of MYPT-1 and Cdc42 protein expressionmay account for decreased ET-1 responses, it indicates that ED in GKrats is not associated with augmented CC and IPA reactivity to ET-1.
doi:10.1016/j.lfs.2013.12.140
Generation of Edn2-iCre transgenic miceJoseph Cacioppoa, Patrick Lina, Arnon Gala,Yongbum Kooa,b, CheMyong Koa
aThe Department of Comparative Biosciences, University of Illinois atUrbana-Champaign, Urbana IL, USAbDepartment of Biotechnology and Biomedical Science, Inje University,Gimhae, South KoreaE-mail address: [email protected] (J. Cacioppo)
Endothelin-2 (ET-2) is a potent vasoconstrictive peptide. Thoughsimilar to ET-1, recent studies suggest that ET-2 may act throughdistinct pathways, necessitating deeper study. ET-2 may play a role inheart failure, inflammation, macular degeneration, and cancer metas-tasis. It is transiently expressed and tightly regulated in the per-iovulatory ovary, where it may aid ovulation by inducing constriction ofthe follicular wall. Here, we present a transgenic mouse line thatexpresses iCre (codon-improved Cre recombinase) under the regula-tion of the promoter of the endothelin-2 (edn2) gene, which wasdeveloped as a novel model for characterizing the expression of ET-2and for conditional deletion of genes in cells where ET-2 is expressed. Avector was generated containing iCre, a polyadenylation signalsequence, and an frt-neomycin-resistance-frt cassette. Two homolo-gous regions of the edn2 gene flanking the ATG start codon wereisolated fromaBAC (bacterial artificial chromosome) clone and insertedupstream and downstream of the iCre-pA-FNF cassette. Homologousrecombination was used to re-insert the cassette into the BAC plasmid.Following purification, the plasmid was inserted into fertilized eggs ofC57BL/6 mice through pronuclear injection, and resulting eggs wereimplanted into pseudopregnant mice. Five Edn2-icre transgene-containing lines of mice were established, and one line was bred withROSA26 reporter. Offspring were used to localize iCre-expressing cellsthroughX-gal staining. Characterization of the staining pattern revealedthat iCre was expressed in granulosa cells of ovulatory follicles,cardiomyocytes, the pituitary, and the liver. We expect this novelmouse model to be a useful tool for future studies on the role of ET-2.
doi:10.1016/j.lfs.2013.12.141
The calcitonin gene-related peptide (CGRP) plays beneficial rolesin myocardial ischemia elicited by endothelin-1Satoshi Hommaa, Satoshi Sakaia, Ken-ichi Yanagib, Yumi Miyauchia,Kazutaka Aonumaa, Takashi Miyauchia
aDepartment of Medicine, University of Tsukuba, Tsukuba, JapanbDepartment of Biomedical Sciences, University of Tsukuba, Tsukuba,JapanE-mail address: [email protected] (S. Homma)
Purpose: In addition to the adrenergic and cholinergic nerves, thecardiovascular tissues are also innervated by several peptidergicneurons that mediate nonadrenergic noncholinergic (NANC) func-tions. Among such neuropeptides, CGRP is released from capsaicin-sensitive sensory neurons in peripheral organs. CGRP is known as
Abstractse40
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