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Master's Dissertation
DOI
10.11606/D.42.2008.tde-03102008-131251
Document
Author
Full name
Claudia Ferreira dos Santos Ruiz Figueiredo
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
Souza, Maria Oliveira de (President)
Abdulkader, Fernando Rodrigues de Moraes
Amorim, José Benedito Oliveira
Title in Portuguese
Papel da proteína quinase C (PKC) na modulação da isoforma 1 do permutador Na+ - H+ (NHE1), em células MDCK.
Keywords in Portuguese
Angiotensina II
Arginina vasopressina
Células MDCK
Isoformas de PKC
NHE1
Phorbol 12-Myristate 13-Acetate
Abstract in Portuguese
O presente trabalho visa contribuir para o esclarecimento da seqüência de eventos intracelulares produzidos pelas PKCs a e e, na modulação do pHi, via NHE1. Os estudos foram realizados em células MDCK e as medidas de pHi efetuadas por microscopia de fluorescência. A expressão das PKCs a e e, bem como do NHE1 foi investigada por western blot, utilizando anticorpos específicos para cada proteína. Os estudos foram realizados na situação controle ou na vigência de PMA ou ANG II, AVP e /ou inibidores específicos para cada receptor hormonal ou isoforma de PKC. Nossos resultados indicam que PMA (10-7 M) estimula a recuperação do pHi, por modular a atividade das PKCs a e e. ANG II e AVP em concentrações fisiológicas estimulam a recuperação do pHi após sobrecarga ácida, concomitante com o aumento da fosforilação da PKC a. Em concentração elevada, ambos hormônios não alteram estes parâmetros. O efeito de ANG II ou de AVP depende da interação de cada hormônio com receptores específicos para modular as vias de sinalização celular envolvidas com o aumento dos níveis de diacilglicerol, cálcio citosólico e AMPc.
Title in English
Role of PKC on exchanger isoform 1 (NHE1), modulation in MDCK cells.
Keywords in English
Angiotensin II
Arginin vasopressin
MDCK cells
NHE1
Phorbol 12-Myristate 13-Acetate
PKC isoforms
Abstract in English
The purpose of this work was to investigate the signaling events of PKCs a e e on the NHE1 activity. The effect of phorbol 12-myristate 13-acetate (PMA), angiotensin II (ANG II) or arginine vasopressin (AVP) on the intracellular pH (pHi) was investigated in MDCK cells by using the fluorescence microscopy and fluorescent probe BCECF/AM. The NHE1 or PKCs a e e expression was examined by western blot and specific antibodies. Our results indicate that PMA (10-7 M) or low concentration of ANG II and AVP induced a significant increase of pH recovery rate and PKC a expression, after intracellular acidification with NH4Cl pulse. ANG II or AVP did not change the PKC a expression. However, in right concentration, both hormones did not change these parameter. In conclusion, the effect of ANG II and AVP on NHE1 activity, depend of specifics membrane receptors and cellular signaling of intracellular calcium, DAG and PKCs a and e.
 
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Publishing Date
2008-10-08
 
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