Co-Localisation of Kisspeptin with Galanin or Neurokinin B in Afferents to Mouse GnRH Neurones

I. Kalló, B. Vida, L. Deli, C. S. Molnár, E. Hrabovszky, A. Caraty, P. Ciofi, C. W. Coen, Z. Liposits

Research output: Contribution to journalArticle

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Abstract

The gonadotrophin-releasing hormone (GnRH) secreting neurones, which form the final common pathway for the central regulation of reproduction, are directly targeted by kisspeptin (KP) via the G protein-coupled receptor, GPR54. In these multiple labelling studies, we used ovariectomised mice treated with 17β-oestradiol (OVX+E 2) or vehicle (OVX+oil) to determine: (i) the ultrastructural characteristics of KP-immunoreactive (IR) afferents to GnRH neurones; (ii) their galanin or neurokinin B (NKB) content; and (iii) the co-expression of galanin or NKB with KP in the two major subpopulations of KP neurones located in the rostral periventricular area of the third ventricle (RP3V) and the arcuate nucleus (Arc). Electron microscopic investigation of the neuronal juxtapositions revealed axosomatic and axodendritic synapses; these showed symmetrical or asymmetrical characteristics, suggesting a phenotypic diversity of KP afferents. Heterogeneity of afferents was also demonstrated by differential co-expression of neuropeptides; in OVX+E 2 mice, KP afferents to GnRH neurones showed galanin-immunoreactivity with an incidence of 22.50±2.41% and NKB-immunoreactivity with an incidence of 5.61±2.57%. In OVX+oil animals, galanin-immunoreactivity in the KP afferents showed a major reduction, appearing in only 5.78±1.57%. Analysis for co-localisation of galanin or NKB with KP was extended to the perikaryal level in animal models, which showed the highest KP incidence; these were OVX+E 2 females for the RP3V and OVX+oil females for the ARC. In the RP3V of colchicine-treated OVX+E 2 animals, 87.84±2.65% of KP-IR neurones were galanin positive. In the Arc of the colchicine-treated OVX+oil animals, galanin immunoreactivity was detected in only 12.50±1.92% of the KP expressing neurones. By contrast, the incidence of co-localisation with NKB in the Arc of those animals was 98.09±1.30%. In situ hybridisation histochemistry of sections from OVX+E 2 animals identified galanin message in more than a third of the KP neurones in the RP3V (38.67±11.57%) and in the Arc (42.50±12.52%). These data suggest that GnRH neurones are innervated by chemically heterogeneous KP cell populations, with a small proportion deriving from the Arc group. The presence of galanin within KP axons innervating GnRH neurones and the oestrogen-dependent regulation of that presence add a new dimension to the roles played by galanin in the central regulation of reproduction.

Original languageEnglish
Pages (from-to)464-476
Number of pages13
JournalJournal of Neuroendocrinology
Volume24
Issue number3
DOIs
Publication statusPublished - Mar 2012

Fingerprint

Neurokinin B
Kisspeptins
Galanin
Gonadotropin-Releasing Hormone
Neurons
Incidence
Colchicine
Reproduction
AIDS-Related Complex
Arcuate Nucleus of Hypothalamus
Third Ventricle

Keywords

  • Confocal microscopy
  • Double- and triple-labelling
  • Immunohistochemistry
  • In situ hybridisation
  • Pre-embedding electron microscopy

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism
  • Endocrine and Autonomic Systems
  • Cellular and Molecular Neuroscience

Cite this

Co-Localisation of Kisspeptin with Galanin or Neurokinin B in Afferents to Mouse GnRH Neurones. / Kalló, I.; Vida, B.; Deli, L.; Molnár, C. S.; Hrabovszky, E.; Caraty, A.; Ciofi, P.; Coen, C. W.; Liposits, Z.

In: Journal of Neuroendocrinology, Vol. 24, No. 3, 03.2012, p. 464-476.

Research output: Contribution to journalArticle

Kalló, I. ; Vida, B. ; Deli, L. ; Molnár, C. S. ; Hrabovszky, E. ; Caraty, A. ; Ciofi, P. ; Coen, C. W. ; Liposits, Z. / Co-Localisation of Kisspeptin with Galanin or Neurokinin B in Afferents to Mouse GnRH Neurones. In: Journal of Neuroendocrinology. 2012 ; Vol. 24, No. 3. pp. 464-476.
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T1 - Co-Localisation of Kisspeptin with Galanin or Neurokinin B in Afferents to Mouse GnRH Neurones

AU - Kalló, I.

AU - Vida, B.

AU - Deli, L.

AU - Molnár, C. S.

AU - Hrabovszky, E.

AU - Caraty, A.

AU - Ciofi, P.

AU - Coen, C. W.

AU - Liposits, Z.

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N2 - The gonadotrophin-releasing hormone (GnRH) secreting neurones, which form the final common pathway for the central regulation of reproduction, are directly targeted by kisspeptin (KP) via the G protein-coupled receptor, GPR54. In these multiple labelling studies, we used ovariectomised mice treated with 17β-oestradiol (OVX+E 2) or vehicle (OVX+oil) to determine: (i) the ultrastructural characteristics of KP-immunoreactive (IR) afferents to GnRH neurones; (ii) their galanin or neurokinin B (NKB) content; and (iii) the co-expression of galanin or NKB with KP in the two major subpopulations of KP neurones located in the rostral periventricular area of the third ventricle (RP3V) and the arcuate nucleus (Arc). Electron microscopic investigation of the neuronal juxtapositions revealed axosomatic and axodendritic synapses; these showed symmetrical or asymmetrical characteristics, suggesting a phenotypic diversity of KP afferents. Heterogeneity of afferents was also demonstrated by differential co-expression of neuropeptides; in OVX+E 2 mice, KP afferents to GnRH neurones showed galanin-immunoreactivity with an incidence of 22.50±2.41% and NKB-immunoreactivity with an incidence of 5.61±2.57%. In OVX+oil animals, galanin-immunoreactivity in the KP afferents showed a major reduction, appearing in only 5.78±1.57%. Analysis for co-localisation of galanin or NKB with KP was extended to the perikaryal level in animal models, which showed the highest KP incidence; these were OVX+E 2 females for the RP3V and OVX+oil females for the ARC. In the RP3V of colchicine-treated OVX+E 2 animals, 87.84±2.65% of KP-IR neurones were galanin positive. In the Arc of the colchicine-treated OVX+oil animals, galanin immunoreactivity was detected in only 12.50±1.92% of the KP expressing neurones. By contrast, the incidence of co-localisation with NKB in the Arc of those animals was 98.09±1.30%. In situ hybridisation histochemistry of sections from OVX+E 2 animals identified galanin message in more than a third of the KP neurones in the RP3V (38.67±11.57%) and in the Arc (42.50±12.52%). These data suggest that GnRH neurones are innervated by chemically heterogeneous KP cell populations, with a small proportion deriving from the Arc group. The presence of galanin within KP axons innervating GnRH neurones and the oestrogen-dependent regulation of that presence add a new dimension to the roles played by galanin in the central regulation of reproduction.

AB - The gonadotrophin-releasing hormone (GnRH) secreting neurones, which form the final common pathway for the central regulation of reproduction, are directly targeted by kisspeptin (KP) via the G protein-coupled receptor, GPR54. In these multiple labelling studies, we used ovariectomised mice treated with 17β-oestradiol (OVX+E 2) or vehicle (OVX+oil) to determine: (i) the ultrastructural characteristics of KP-immunoreactive (IR) afferents to GnRH neurones; (ii) their galanin or neurokinin B (NKB) content; and (iii) the co-expression of galanin or NKB with KP in the two major subpopulations of KP neurones located in the rostral periventricular area of the third ventricle (RP3V) and the arcuate nucleus (Arc). Electron microscopic investigation of the neuronal juxtapositions revealed axosomatic and axodendritic synapses; these showed symmetrical or asymmetrical characteristics, suggesting a phenotypic diversity of KP afferents. Heterogeneity of afferents was also demonstrated by differential co-expression of neuropeptides; in OVX+E 2 mice, KP afferents to GnRH neurones showed galanin-immunoreactivity with an incidence of 22.50±2.41% and NKB-immunoreactivity with an incidence of 5.61±2.57%. In OVX+oil animals, galanin-immunoreactivity in the KP afferents showed a major reduction, appearing in only 5.78±1.57%. Analysis for co-localisation of galanin or NKB with KP was extended to the perikaryal level in animal models, which showed the highest KP incidence; these were OVX+E 2 females for the RP3V and OVX+oil females for the ARC. In the RP3V of colchicine-treated OVX+E 2 animals, 87.84±2.65% of KP-IR neurones were galanin positive. In the Arc of the colchicine-treated OVX+oil animals, galanin immunoreactivity was detected in only 12.50±1.92% of the KP expressing neurones. By contrast, the incidence of co-localisation with NKB in the Arc of those animals was 98.09±1.30%. In situ hybridisation histochemistry of sections from OVX+E 2 animals identified galanin message in more than a third of the KP neurones in the RP3V (38.67±11.57%) and in the Arc (42.50±12.52%). These data suggest that GnRH neurones are innervated by chemically heterogeneous KP cell populations, with a small proportion deriving from the Arc group. The presence of galanin within KP axons innervating GnRH neurones and the oestrogen-dependent regulation of that presence add a new dimension to the roles played by galanin in the central regulation of reproduction.

KW - Confocal microscopy

KW - Double- and triple-labelling

KW - Immunohistochemistry

KW - In situ hybridisation

KW - Pre-embedding electron microscopy

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