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Crypt cells are involved in kin recognition in larval zebrafish.

Biechl D, Tietje K, Gerlach G, Wullimann MF - Sci Rep (2016)

Bottom Line: Zebrafish larvae imprint on visual and olfactory kin cues at day 5 and 6 postfertilization, respectively, resulting in kin recognition later in life.Then, we tested imprinted and non-imprinted larvae (full siblings) for kin odor detection.We provide the first direct evidence that crypt cells, and likely a subpopulation of microvillous OSNs, but not ciliated OSNs, play a role in detecting a kin odor related signal.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Systemic Neurosciences &Department Biology II, Ludwig-Maximilians-Universität Munich, Grosshadernerstr. 2, 82152 Planegg-Martinsried Germany.

ABSTRACT
Zebrafish larvae imprint on visual and olfactory kin cues at day 5 and 6 postfertilization, respectively, resulting in kin recognition later in life. Exposure to non-kin cues prevents imprinting and kin recognition. Imprinting depends on MHC class II related signals and only larvae sharing MHC class II alleles can imprint on each other. Here, we analyzed which type of olfactory sensory neuron (OSN) detects kin odor. The single teleost olfactory epithelium harbors ciliated OSNs carrying OR and TAAR gene family receptors (mammals: main olfactory epithelium) and microvillous OSNs with V1R and V2R gene family receptors (mammals: vomeronasal organ). Additionally, teleosts exhibit crypt cells which possess microvilli and cilia. We used the activity marker pERK (phosphorylated extracellular signal regulated kinase) after stimulating 9 day old zebrafish larvae with either non-kin conspecific or food odor. While food odor activated both ciliated and microvillous OSNs, only the latter were activated by conspecific odor, crypt cells showed no activation to both stimuli. Then, we tested imprinted and non-imprinted larvae (full siblings) for kin odor detection. We provide the first direct evidence that crypt cells, and likely a subpopulation of microvillous OSNs, but not ciliated OSNs, play a role in detecting a kin odor related signal.

No MeSH data available.


Related in: MedlinePlus

Examples of activated OSN identification and counting.All photographs shown are confocal optical sections. (A–C) Overviews of 9 dpf larval zebrafish cross-sections triple-stained for DAPI, S100 and pERK. (A’-A”’), (B’-B”’), and (C-C”’) show magnified monochromatic pictures of each marker in the olfactory epithelium. Note examples of activated crypt cells in imprinted larvae tested with kin odor (A-A”’) as well as some mOSNs and cOSNs (B-B”’). In non-imprinted larvae tested with kin odor, crypt cells are not activated (C-C”’). (D) shows a DAPI view of the position of the olfactory epithelium relative to eye and olfactory bulb with a corresponding explanatory drawing. Larval brain outline indicates the level of section of (D). Drawing at right bottom gives an overview on the cytoarchitectonic organization of the olfactory epithelium. Abbreviations: ac anterior commissure, CeP cerebellar plate, DT dorsal thalamus, E epiphysis, EmT eminentia thalami, H hypothalamus, Ha habenula, lG lateral glomeruli, MdG mediodorsal glomeruli, MO medulla oblongata, N region of the nucleus of the medial longitudinal fascicle, OB olfactory bulb, oc optic chiasma, ON olfactory nerve, P pallium, Po preoptic region, poc postoptic commissure, PTd dorsal part of posterior tuberculum, PTv ventral part of posterior tuberculum, S subpallium, T tegmentum, TeO tectum opticum TeVe tectal ventricle, Va valvula cerebelli, vg ventral glomeruli, VT ventral thalamus.
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f4: Examples of activated OSN identification and counting.All photographs shown are confocal optical sections. (A–C) Overviews of 9 dpf larval zebrafish cross-sections triple-stained for DAPI, S100 and pERK. (A’-A”’), (B’-B”’), and (C-C”’) show magnified monochromatic pictures of each marker in the olfactory epithelium. Note examples of activated crypt cells in imprinted larvae tested with kin odor (A-A”’) as well as some mOSNs and cOSNs (B-B”’). In non-imprinted larvae tested with kin odor, crypt cells are not activated (C-C”’). (D) shows a DAPI view of the position of the olfactory epithelium relative to eye and olfactory bulb with a corresponding explanatory drawing. Larval brain outline indicates the level of section of (D). Drawing at right bottom gives an overview on the cytoarchitectonic organization of the olfactory epithelium. Abbreviations: ac anterior commissure, CeP cerebellar plate, DT dorsal thalamus, E epiphysis, EmT eminentia thalami, H hypothalamus, Ha habenula, lG lateral glomeruli, MdG mediodorsal glomeruli, MO medulla oblongata, N region of the nucleus of the medial longitudinal fascicle, OB olfactory bulb, oc optic chiasma, ON olfactory nerve, P pallium, Po preoptic region, poc postoptic commissure, PTd dorsal part of posterior tuberculum, PTv ventral part of posterior tuberculum, S subpallium, T tegmentum, TeO tectum opticum TeVe tectal ventricle, Va valvula cerebelli, vg ventral glomeruli, VT ventral thalamus.

Mentions: In this experiment we investigated which OSN type(s) respond to a kin odor produced by full siblings and therefore play a role in olfactory kin recognition and maybe imprinting. To this aim, we stimulated 9 dpf old imprinted (group reared, see Methods) and non-imprinted (isolated reared; see Methods and Fig. 1b) larvae with kin odor for 7 minutes, and compared these two groups with equally reared control groups stimulated with E3 medium. In this experiment, we used the anti-pERK antibody together with an established crypt cell marker, an antibody against the calcium binding protein S10026, in order to examine a possible overlap between these two markers (Fig. 4) and to differentiate S100-positive (S100 +) from S100-negative pERK activated OSNs.


Crypt cells are involved in kin recognition in larval zebrafish.

Biechl D, Tietje K, Gerlach G, Wullimann MF - Sci Rep (2016)

Examples of activated OSN identification and counting.All photographs shown are confocal optical sections. (A–C) Overviews of 9 dpf larval zebrafish cross-sections triple-stained for DAPI, S100 and pERK. (A’-A”’), (B’-B”’), and (C-C”’) show magnified monochromatic pictures of each marker in the olfactory epithelium. Note examples of activated crypt cells in imprinted larvae tested with kin odor (A-A”’) as well as some mOSNs and cOSNs (B-B”’). In non-imprinted larvae tested with kin odor, crypt cells are not activated (C-C”’). (D) shows a DAPI view of the position of the olfactory epithelium relative to eye and olfactory bulb with a corresponding explanatory drawing. Larval brain outline indicates the level of section of (D). Drawing at right bottom gives an overview on the cytoarchitectonic organization of the olfactory epithelium. Abbreviations: ac anterior commissure, CeP cerebellar plate, DT dorsal thalamus, E epiphysis, EmT eminentia thalami, H hypothalamus, Ha habenula, lG lateral glomeruli, MdG mediodorsal glomeruli, MO medulla oblongata, N region of the nucleus of the medial longitudinal fascicle, OB olfactory bulb, oc optic chiasma, ON olfactory nerve, P pallium, Po preoptic region, poc postoptic commissure, PTd dorsal part of posterior tuberculum, PTv ventral part of posterior tuberculum, S subpallium, T tegmentum, TeO tectum opticum TeVe tectal ventricle, Va valvula cerebelli, vg ventral glomeruli, VT ventral thalamus.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4834543&req=5

f4: Examples of activated OSN identification and counting.All photographs shown are confocal optical sections. (A–C) Overviews of 9 dpf larval zebrafish cross-sections triple-stained for DAPI, S100 and pERK. (A’-A”’), (B’-B”’), and (C-C”’) show magnified monochromatic pictures of each marker in the olfactory epithelium. Note examples of activated crypt cells in imprinted larvae tested with kin odor (A-A”’) as well as some mOSNs and cOSNs (B-B”’). In non-imprinted larvae tested with kin odor, crypt cells are not activated (C-C”’). (D) shows a DAPI view of the position of the olfactory epithelium relative to eye and olfactory bulb with a corresponding explanatory drawing. Larval brain outline indicates the level of section of (D). Drawing at right bottom gives an overview on the cytoarchitectonic organization of the olfactory epithelium. Abbreviations: ac anterior commissure, CeP cerebellar plate, DT dorsal thalamus, E epiphysis, EmT eminentia thalami, H hypothalamus, Ha habenula, lG lateral glomeruli, MdG mediodorsal glomeruli, MO medulla oblongata, N region of the nucleus of the medial longitudinal fascicle, OB olfactory bulb, oc optic chiasma, ON olfactory nerve, P pallium, Po preoptic region, poc postoptic commissure, PTd dorsal part of posterior tuberculum, PTv ventral part of posterior tuberculum, S subpallium, T tegmentum, TeO tectum opticum TeVe tectal ventricle, Va valvula cerebelli, vg ventral glomeruli, VT ventral thalamus.
Mentions: In this experiment we investigated which OSN type(s) respond to a kin odor produced by full siblings and therefore play a role in olfactory kin recognition and maybe imprinting. To this aim, we stimulated 9 dpf old imprinted (group reared, see Methods) and non-imprinted (isolated reared; see Methods and Fig. 1b) larvae with kin odor for 7 minutes, and compared these two groups with equally reared control groups stimulated with E3 medium. In this experiment, we used the anti-pERK antibody together with an established crypt cell marker, an antibody against the calcium binding protein S10026, in order to examine a possible overlap between these two markers (Fig. 4) and to differentiate S100-positive (S100 +) from S100-negative pERK activated OSNs.

Bottom Line: Zebrafish larvae imprint on visual and olfactory kin cues at day 5 and 6 postfertilization, respectively, resulting in kin recognition later in life.Then, we tested imprinted and non-imprinted larvae (full siblings) for kin odor detection.We provide the first direct evidence that crypt cells, and likely a subpopulation of microvillous OSNs, but not ciliated OSNs, play a role in detecting a kin odor related signal.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Systemic Neurosciences &Department Biology II, Ludwig-Maximilians-Universität Munich, Grosshadernerstr. 2, 82152 Planegg-Martinsried Germany.

ABSTRACT
Zebrafish larvae imprint on visual and olfactory kin cues at day 5 and 6 postfertilization, respectively, resulting in kin recognition later in life. Exposure to non-kin cues prevents imprinting and kin recognition. Imprinting depends on MHC class II related signals and only larvae sharing MHC class II alleles can imprint on each other. Here, we analyzed which type of olfactory sensory neuron (OSN) detects kin odor. The single teleost olfactory epithelium harbors ciliated OSNs carrying OR and TAAR gene family receptors (mammals: main olfactory epithelium) and microvillous OSNs with V1R and V2R gene family receptors (mammals: vomeronasal organ). Additionally, teleosts exhibit crypt cells which possess microvilli and cilia. We used the activity marker pERK (phosphorylated extracellular signal regulated kinase) after stimulating 9 day old zebrafish larvae with either non-kin conspecific or food odor. While food odor activated both ciliated and microvillous OSNs, only the latter were activated by conspecific odor, crypt cells showed no activation to both stimuli. Then, we tested imprinted and non-imprinted larvae (full siblings) for kin odor detection. We provide the first direct evidence that crypt cells, and likely a subpopulation of microvillous OSNs, but not ciliated OSNs, play a role in detecting a kin odor related signal.

No MeSH data available.


Related in: MedlinePlus