Regulation and Function of C-Type Natriuretic Peptide (CNP) in Gonadotrope-Derived Cell Lines
Samantha M Mirczuk,
Andrew J Lessey,
Alice R Catterick,
Rebecca M Perrett,
Christopher J Scudder,
Jordan E Read,
Victoria J Lipscomb,
Stijn J Niessen,
Andrew J Childs,
Craig A McArdle,
Imelda M McGonnell,
Robert C Fowkes
Affiliations
Samantha M Mirczuk
Endocrine Signalling Group, Royal Veterinary College, University of London, Royal College Street, London NW1 0TU, UK
Andrew J Lessey
Endocrine Signalling Group, Royal Veterinary College, University of London, Royal College Street, London NW1 0TU, UK
Alice R Catterick
Endocrine Signalling Group, Royal Veterinary College, University of London, Royal College Street, London NW1 0TU, UK
Rebecca M Perrett
Laboratories for Integrative Neuroscience and Endocrinology, Department of Clinical Sciences at South Bristol, University of Bristol, Whitson Street, Bristol BS13NY, UK
Christopher J Scudder
Endocrine Signalling Group, Royal Veterinary College, University of London, Royal College Street, London NW1 0TU, UK
Jordan E Read
Endocrine Signalling Group, Royal Veterinary College, University of London, Royal College Street, London NW1 0TU, UK
Victoria J Lipscomb
Clinical Science and Services, Royal Veterinary College, Hertfordshire AL9 7TA, UK
Stijn J Niessen
Clinical Science and Services, Royal Veterinary College, Hertfordshire AL9 7TA, UK
Andrew J Childs
Comparative Biomedical Sciences, Royal Veterinary College, University of London, Royal College Street, London NW1 0TU, UK
Craig A McArdle
Laboratories for Integrative Neuroscience and Endocrinology, Department of Clinical Sciences at South Bristol, University of Bristol, Whitson Street, Bristol BS13NY, UK
Imelda M McGonnell
Comparative Biomedical Sciences, Royal Veterinary College, University of London, Royal College Street, London NW1 0TU, UK
Robert C Fowkes
Endocrine Signalling Group, Royal Veterinary College, University of London, Royal College Street, London NW1 0TU, UK
C-type natriuretic peptide (CNP) is the most conserved member of the mammalian natriuretic peptide family, and is implicated in the endocrine regulation of growth, metabolism and reproduction. CNP is expressed throughout the body, but is particularly abundant in the central nervous system and anterior pituitary gland. Pituitary gonadotropes are regulated by pulsatile release of gonadotropin releasing hormone (GnRH) from the hypothalamus, to control reproductive function. GnRH and CNP reciprocally regulate their respective signalling pathways in αT3-1 gonadotrope cells, but effects of pulsatile GnRH stimulation on CNP expression has not been explored. Here, we examine the sensitivity of the natriuretic peptide system in LβT2 and αT3-1 gonadotrope cell lines to continuous and pulsatile GnRH stimulation, and investigate putative CNP target genes in gonadotropes. Multiplex RT-qPCR assays confirmed that primary mouse pituitary tissue express Nppc, Npr2 (encoding CNP and guanylyl cyclase B (GC-B), respectively) and Furin (a CNP processing enzyme), but failed to express transcripts for Nppa or Nppb (encoding ANP and BNP, respectively). Pulsatile, but not continuous, GnRH stimulation of LβT2 cells caused significant increases in Nppc and Npr2 expression within 4 h, but failed to alter natriuretic peptide gene expression in αT3-1 cells. CNP enhanced expression of cJun, Egr1, Nr5a1 and Nr0b1, within 8 h in LβT2 cells, but inhibited Nr5a1 expression in αT3-1 cells. Collectively, these data show the gonadotrope natriuretic peptide system is sensitive to pulsatile GnRH signalling, and gonadotrope transcription factors are putative CNP-target genes. Such findings represent additional mechanisms by which CNP may regulate reproductive function.
