Post mortem single-cell labeling with DiI and immunoelectron microscopy unveil the fine structure of kisspeptin neurons in humans

Szabolcs Takács, Zsuzsanna Bardóczi, Katalin Skrapits, Balázs Göcz, Viktória Váczi, Z. Maglóczky, Iván Szűcs, Gergely Rácz, A. Matolcsy, Waljit S. Dhillo, Masahiko Watanabe, Andrea Kádár, Csaba Fekete, I. Kalló, E. Hrabovszky

Research output: Contribution to journalArticle

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Abstract

Kisspeptin (KP) synthesizing neurons of the hypothalamic infundibular region are critically involved in the central regulation of fertility; these cells regulate pulsatile gonadotropin-releasing hormone (GnRH) secretion and mediate sex steroid feedback signals to GnRH neurons. Fine structural analysis of the human KP system is complicated by the use of post mortem tissues. To gain better insight into the neuroanatomy of the somato-dendritic cellular compartment, we introduced the diolistic labeling of immunohistochemically identified KP neurons using a gene gun loaded with the lipophilic dye, DiI. Confocal microscopic studies of primary dendrites in 100-µm-thick tissue sections established that 79.3% of KP cells were bipolar, 14.1% were tripolar, and 6.6% were unipolar. Primary dendrites branched sparsely, contained numerous appendages (9.1 ± 1.1 spines/100 µm dendrite), and received rich innervation from GABAergic, glutamatergic, and KP-containing terminals. KP neuron synaptology was analyzed with immunoelectron microscopy on perfusion-fixed specimens. KP axons established frequent contacts and classical synapses on unlabeled, and on KP-immunoreactive somata, dendrites, and spines. Synapses were asymmetric and the presynaptic structures contained round and regular synaptic vesicles, in addition to dense-core granules. Although immunofluorescent studies failed to detect vesicular glutamate transporter isoforms in KP axons, ultrastructural characteristics of synaptic terminals suggested use of glutamatergic, in addition to peptidergic, neurotransmission. In summary, immunofluorescent and DiI labeling of KP neurons in thick hypothalamic sections and immunoelectron microscopic studies of KP-immunoreactive neurons in brains perfusion-fixed shortly post mortem allowed us to identify previously unexplored fine structural features of KP neurons in the mediobasal hypothalamus of humans.

Original languageEnglish
Pages (from-to)1-14
Number of pages14
JournalBrain Structure and Function
DOIs
Publication statusAccepted/In press - Jan 29 2018

Fingerprint

Kisspeptins
Immunoelectron Microscopy
Neurons
Dendrites
Gonadotropin-Releasing Hormone
Synapses
Axons
Spine
Vesicular Glutamate Transport Proteins
Perfusion
Neuroanatomy
Synaptic Vesicles
Presynaptic Terminals
Firearms
Carisoprodol
Synaptic Transmission
Hypothalamus
Fertility
Protein Isoforms

Keywords

  • Asymmetric synapse
  • Cell filling
  • Diolistic labeling
  • Electron microscopy
  • Hypothalamus
  • Small clear vesicles

ASJC Scopus subject areas

  • Anatomy
  • Neuroscience(all)
  • Histology

Cite this

Post mortem single-cell labeling with DiI and immunoelectron microscopy unveil the fine structure of kisspeptin neurons in humans. / Takács, Szabolcs; Bardóczi, Zsuzsanna; Skrapits, Katalin; Göcz, Balázs; Váczi, Viktória; Maglóczky, Z.; Szűcs, Iván; Rácz, Gergely; Matolcsy, A.; Dhillo, Waljit S.; Watanabe, Masahiko; Kádár, Andrea; Fekete, Csaba; Kalló, I.; Hrabovszky, E.

In: Brain Structure and Function, 29.01.2018, p. 1-14.

Research output: Contribution to journalArticle

Takács, Szabolcs ; Bardóczi, Zsuzsanna ; Skrapits, Katalin ; Göcz, Balázs ; Váczi, Viktória ; Maglóczky, Z. ; Szűcs, Iván ; Rácz, Gergely ; Matolcsy, A. ; Dhillo, Waljit S. ; Watanabe, Masahiko ; Kádár, Andrea ; Fekete, Csaba ; Kalló, I. ; Hrabovszky, E. / Post mortem single-cell labeling with DiI and immunoelectron microscopy unveil the fine structure of kisspeptin neurons in humans. In: Brain Structure and Function. 2018 ; pp. 1-14.
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AU - Göcz, Balázs

AU - Váczi, Viktória

AU - Maglóczky, Z.

AU - Szűcs, Iván

AU - Rácz, Gergely

AU - Matolcsy, A.

AU - Dhillo, Waljit S.

AU - Watanabe, Masahiko

AU - Kádár, Andrea

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