OBJECTIVES: The current research was conducted to determine whether hyperpolarization-activated cyclic nucleotide-gated (HCN1-4) channels are expressed in gonadotropin-releasing hormone (GnRH) neurons in the female rat hypothalamus and immortalized GnRH neurons (GT1-7 cells). METHODS: Double-label fluorescence immunohistochemistry was used to colocalize HCN1-4 channels and GnRH in GnRH neurons in the female rat hypothalamus. Reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting, and immunocytochemistry were used to analyze HCN channel gene expression in GT1-7 cells. RESULTS: Double-label fluorescence immunohistochemistry showed that 43% of hypothalamic GnRH neurons immunostained for HCN2 and 90% of GnRH neurons immunostained for HCN3. RT-PCR and Western blot showed expression of all four HCN channel subunits in GT1-7 cells. Double-label immunocytochemistry showed cytoplasmic immunostaining of HCN2 and HCN3 in GT1-7 cells. CONCLUSIONS: This study demonstrates for the first time that HCN channels are expressed in GnRH neurons in the rat hypothalamus and GT1-7 cells. Our research supports the hypothesis that HCN channels may be involved in electrical bursting activity and pulsatile GnRH secretion in endogenous GnRH neurons and GT1-7 cells.
OBJECTIVES: The current research was conducted to determine whether hyperpolarization-activated cyclic nucleotide-gated (HCN1-4) channels are expressed in gonadotropin-releasing hormone (GnRH) neurons in the female rathypothalamus and immortalized GnRH neurons (GT1-7 cells). METHODS: Double-label fluorescence immunohistochemistry was used to colocalize HCN1-4 channels and GnRH in GnRH neurons in the female rathypothalamus. Reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting, and immunocytochemistry were used to analyze HCN channel gene expression in GT1-7 cells. RESULTS: Double-label fluorescence immunohistochemistry showed that 43% of hypothalamic GnRH neurons immunostained for HCN2 and 90% of GnRH neurons immunostained for HCN3. RT-PCR and Western blot showed expression of all four HCN channel subunits in GT1-7 cells. Double-label immunocytochemistry showed cytoplasmic immunostaining of HCN2 and HCN3 in GT1-7 cells. CONCLUSIONS: This study demonstrates for the first time that HCN channels are expressed in GnRH neurons in the rathypothalamus and GT1-7 cells. Our research supports the hypothesis that HCN channels may be involved in electrical bursting activity and pulsatile GnRH secretion in endogenous GnRH neurons and GT1-7 cells.
Authors: Bermary Santos-Vera; Ana Del C Vaquer-Alicea; Cristina E Maria-Rios; Alan Montiel-Ramos; Aynette Ramos-Cardona; Rafael Vázquez-Torres; Priscila Sanabria; Carlos A Jiménez-Rivera Journal: Neurochem Int Date: 2019-02-19 Impact factor: 3.921
Authors: Bermary Santos-Vera; Rafael Vázquez-Torres; Hermes G García Marrero; Juan M Ramos Acevedo; Francisco Arencibia-Albite; María E Vélez-Hernández; Jorge D Miranda; Carlos A Jiménez-Rivera Journal: J Mol Neurosci Date: 2012-12-01 Impact factor: 3.444
Authors: Marc S Stieglitz; Stefanie Fenske; Verena Hammelmann; Elvir Becirovic; Verena Schöttle; James E Delorme; Martha Schöll-Weidinger; Robert Mader; Jan Deussing; David P Wolfer; Mathias W Seeliger; Urs Albrecht; Carsten T Wotjak; Martin Biel; Stylianos Michalakis; Christian Wahl-Schott Journal: Front Mol Neurosci Date: 2018-01-09 Impact factor: 5.639