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Combinatorial analysis of calcium-binding proteins in larval and adult zebrafish primary olfactory system identifies differential olfactory bulb glomerular projection fields

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Abstract

In the zebrafish (Danio rerio) olfactory epithelium, the calcium-binding proteins (CBPs) calretinin and S100/S100-like protein are mainly expressed in ciliated or crypt olfactory sensory neurons (OSNs), respectively. In contrast parvalbumin and calbindin1 have not been investigated. We present a combinatorial immunohistological analysis of all four CBPs, including their expression in OSNs and their axonal projections to the olfactory bulb in larval and adult zebrafish. A major expression of calretinin and S100 in ciliated and crypt cells, respectively, with some expression of S100 in microvillous cells is confirmed. Parvalbumin and calbindin1 are strongly expressed in ciliated and microvillous cells, but not in crypt cells. Moreover, detailed combinatorial double-label experiments indicate that there are eight subpopulations of zebrafish OSNs: S100-positive crypt cells (negative for all other three CBPs), parvalbumin only, S100 and parvalbumin, parvalbumin and calbindin1, and parvalbumin and calbindin1 and calretinin-positive microvillous OSNs, as well as a major parvalbumin and calbindin1 and calretinin, and minor parvalbumin and calbindin1 and calretinin-only-positive ciliated OSN populations. CBP-positive projections to olfactory bulb are consistent with previous reports of ciliated OSNs projecting to dorsal and ventromedial glomerular fields and microvillous OSNs to ventrolateral glomerular fields. We newly describe parvalbumin-positive fibers to the mediodorsal field which is calretinin free, with its anterior part showing additionally calbindin1-positive fibers, but absence thereof in the posterior part, indicating an origin from microvillous OSNs in both parts. One singular glomerulus (mdG2) exhibits S100 and parvalbumin-positive fibers, apparently originating from all crypt cells plus some microvillous OSNs. Arguments for various olfactory labeled lines are discussed.

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Abbreviations

ac:

Anterior commissure

c:

Cilia

CB(ir):

Calbindin1 (immunoreactive)

CBP(s):

Calcium-binding protein(s)

CeP:

Cerebellar plate (larvae)

Cr:

Crypt OSN

CR(ir):

Calretinin (immunoreactive)

CZ:

Central zone

D/d:

Dendrite

Dp:

Posterior zone of dorsal telencephalic area

DT:

Dorsal thalamus (thalamus)

DZ:

Dorsal zone

EmT:

Eminentia thalami

gl, Gl:

Glomerular layer of olfactory bulb

Glo:

Glomeruli

GPCR:

G-protein-coupled receptor

H:

Hypothalamus

Ha:

Habenula

Had:

Dorsal habenular nucleus

Hav:

Ventral habenular nucleus

icl:

Internal cellular layer of olfactory bulb

lfb:

Lateral forebrain bundle

LG1–4:

Lateral glomeruli 1–4 (larvae)

LOT:

Lateral olfactory tract

m:

Microvilli

MC:

Mitral cell

Mi:

Microvillous OSN

MG1–4:

Medial glomeruli 1–4 (larvae)

MOE:

Main olfactory epithelium

MOS:

Main olfactory system

MOT:

Medial olfactory tract

N:

Region of the nucleus of medial longitudinal fascicle

OB:

Olfactory bulb

oc:

Optic chiasma

OE:

Olfactory epithelium

on:

Optic nerve

ON:

Olfactory nerve

OSN(s):

Olfactory sensory neuron(s)

OR:

Olfactory receptor

P:

Pallium

Po:

Preoptic region

poc:

Postoptic commissure

Pr:

Pretectum (larvae)

PT:

Posterior tuberculum

PV(ir):

Parvalbumin (immunoreactive)

S:

Subpallium

T:

Midbrain tegmentum

Tel:

Telencephalon

TeO:

Optic tectum

Va:

Valvula cerebelli

Vs:

Supracommissural nucleus of ventral telencephalic area

VT:

Ventral thalamus (prethalamus)

Vv:

Ventral nucleus of ventral telencephalic area

a–e:

Dorsal cluster-associated glomeruli 1–5

ap:

Anterior plexus

dc:

Glomeruli of the dorsal cluster

k:

Medial elongated glomerulus

l:

Glomerulus of the lateral chain 1

lc:

Lateral chain

m:

Glomerulus of the lateral chain 2

mdG1–6 :

Mediodorsal posterior glomeruli 1–6

n:

Glomerulus of the lateral chain 3

p:

Glomerulus of the lateral chain 5

q:

Lateroventral posterior glomerulus

u–w:

Glomeruli of the ventral triplet 1–3

vpG:

Ventroposterior glomerulus

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Acknowledgments

We thank the Deutsche Forschungsgemeinschaft (DFG, Bonn) for support within the SPP 1392 (Projects Wu 211/2-1 and Wu 211/1-2) as well as Prof. Dr. Benedikt Grothe and the Graduate School for Systemic Neurosciences (GSN) at the Ludwig-Maximilians-Universität Munich for additional support. We furthermore thank Bea Stiening for technical support and Dr. Olga Alexandrova for help with confocal microscopy.

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  1. Department Biology II, Biocenter, Graduate School of Systemic Neurosciences, Ludwig-Maximilians-Universität Munich, Grosshaderner Str. 2, 82152, Planegg-Martinsried, Germany

    Sigrid Kress, Daniela Biechl & Mario F. Wullimann

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  1. Sigrid Kress
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  2. Daniela Biechl
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  3. Mario F. Wullimann
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Correspondence to Mario F. Wullimann.

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S. Kress and D. Biechl share first authorship.

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Kress, S., Biechl, D. & Wullimann, M.F. Combinatorial analysis of calcium-binding proteins in larval and adult zebrafish primary olfactory system identifies differential olfactory bulb glomerular projection fields. Brain Struct Funct 220, 1951–1970 (2015). https://doi.org/10.1007/s00429-014-0765-1

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