Intracellular mechanisms involved in copper-gonadotropin-releasing hormone (Cu-GnRH) complex-induced cAMP/PKA signaling in female rat anterior pituitary cells in vitro.
PBN-AR
Instytucja
Instytut Fizjologii i Żywienia Zwierząt im. Jana Kielanowskiego Polskiej Akademii Nauk
Informacje podstawowe
Główny język publikacji
en
Czasopismo
BRAIN RESEARCH BULLETIN
ISSN
0361-9230
EISSN
1873-2747
Wydawca
PERGAMON-ELSEVIER SCIENCE LTD
DOI
URL
Rok publikacji
2016
Numer zeszytu
Strony od-do
75-82
Numer tomu
120
Link do pełnego tekstu
Liczba arkuszy
Słowa kluczowe
en
Cu-GnRH; Gonadoliberin; Intracellular calcium; PKC; Rat pituitary; cAMP
Streszczenia
Język
en
Treść
The copper-gonadotropin-releasing hormone molecule (Cu-GnRH) is a GnRH analog, which preserves its amino acid sequence, but which contains a Cu(2+) ion stably bound to the nitrogen atoms including that of the imidazole ring of Histidine(2). A previous report indicated that Cu-GnRH was able to activate cAMP/PKA signaling in anterior pituitary cells in vitro, but raised the question of which intracellular mechanism(s) mediated the Cu-GnRH-induced cAMP synthesis in gonadotropes. To investigate this mechanism, in the present study, female rat anterior pituitary cells in vitro were pretreated with 0.1μM antide, a GnRH antagonist; 0.1μM cetrorelix, a GnRH receptor antagonist; 0.1μM PACAP6-38, a PAC-1 receptor antagonist; 2μM GF109203X, a protein kinase C inhibitor; 50mM PMA, a protein kinase C activator; the protein kinase A inhibitors H89 (30μM) and KT5720 (60nM); factors affecting intracellular calcium activity: 2.5mM EGTA; 2μM thapsigargin; 5μM A23187, a Ca(2+) ionophore; or 10μg/ml cycloheximide, a protein synthesis inhibitor. After one of the above pretreatments, cells were incubated in the presence of 0.1μM Cu-GnRH for 0.5, 1, and 3h. Radioimmunoassay analysis of cAMP confirmed the functional link between Cu-GnRH stimulation and cAMP/PKA signal transduction in rat anterior pituitary cells, demonstrating increased intracellular cAMP, which was reduced in the presence of specific PKA inhibitors. The stimulatory effect of Cu-GnRH on cAMP production was partly dependent on GnRH receptor activation. In addition, an indirect and Ca(2+)-dependent mechanism might be involved in intracellular adenylate cyclase stimulation. Neither activation of protein kinase C nor new protein synthesis was involved in the Cu-GnRH-induced increase of cAMP in the rat anterior pituitary primary cultures. Presented data indicate that conformational changes of GnRH molecule resulting from cooper ion coordination affect specific pharmacological properties of Cu-GnRH molecule including specific pattern of intracellular activity induced by complex in anterior pituitary cells in vitro.
Cechy publikacji
ORIGINAL_ARTICLE
Inne
System-identifier
687653
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