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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

Differential activation of cOSNs, mOSNs and crypt cells by stimulation with different odors.9 day old zebrafish larvae were exposed to either food odor, non-kin larvae odor or E3 Medium (control) (pooled data of Fig. 2). The total number of pERK+activated cOSNs, mOSNs, and crypt cells was counted per larva and statistically analyzed. Box plots show median, upper and lower quartile and whiskers (maximum interquartile range: 1.5). *indicates statistical significance p: ***p < 0.001. (a) cOSNs are strongly activated by food odor. Significantly more pERK+cOSNs were counted in larvae stimulated with food compared to larvae odor (Mann-Whitney U: 4.0, p < 0.001, median (Mdn)food = 54, Mdnlarvae = 2, n = 18) and to control (U < 0.0, p < 0.001, Mdnfood = 54, Mdnctr = 3, nfood = 18, nctr = 20). (b) mOSNs show the highest activation when stimulated with food odor. Significantly more mOSNS were activated by food odor compared to control stimulation (U: 33.5, p < 0.001, Mdnfood = 19, Mdnctr = 4.5, nfood = 18, nctr = 20). pERK + mOSNs stimulated with larvae odor do not differ in numbers compared to controls (U: 116.5, p = 0.062, Mdnlarvae = 7.5, Mdnctr = 4.5, nlarvae = 18, nctr = 20). (c) Crypt cells show no significant difference in pERK+cell numbers due to stimulation with different odors (Kruskall Wallis test: H(2) = 3.197, p = 0.202, nfood = nlarvae odor = 18, nctr = 20).
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f3: Differential activation of cOSNs, mOSNs and crypt cells by stimulation with different odors.9 day old zebrafish larvae were exposed to either food odor, non-kin larvae odor or E3 Medium (control) (pooled data of Fig. 2). The total number of pERK+activated cOSNs, mOSNs, and crypt cells was counted per larva and statistically analyzed. Box plots show median, upper and lower quartile and whiskers (maximum interquartile range: 1.5). *indicates statistical significance p: ***p < 0.001. (a) cOSNs are strongly activated by food odor. Significantly more pERK+cOSNs were counted in larvae stimulated with food compared to larvae odor (Mann-Whitney U: 4.0, p < 0.001, median (Mdn)food = 54, Mdnlarvae = 2, n = 18) and to control (U < 0.0, p < 0.001, Mdnfood = 54, Mdnctr = 3, nfood = 18, nctr = 20). (b) mOSNs show the highest activation when stimulated with food odor. Significantly more mOSNS were activated by food odor compared to control stimulation (U: 33.5, p < 0.001, Mdnfood = 19, Mdnctr = 4.5, nfood = 18, nctr = 20). pERK + mOSNs stimulated with larvae odor do not differ in numbers compared to controls (U: 116.5, p = 0.062, Mdnlarvae = 7.5, Mdnctr = 4.5, nlarvae = 18, nctr = 20). (c) Crypt cells show no significant difference in pERK+cell numbers due to stimulation with different odors (Kruskall Wallis test: H(2) = 3.197, p = 0.202, nfood = nlarvae odor = 18, nctr = 20).

Mentions: Because we observed that stimulus duration did not affect the number of activated OSNs, we plotted pERK activated cells independent of stimulus durations against the two different odors food and non-kin conspecific larvae odor and compared it with controls (Fig. 3). The activation profile of the different OSNs revealed a highly significant activation in response to food odor compared to controls in ciliated and microvillous OSNs (cOSNs, mOSNs) (Fig. 3a,b). Stimulation with non-kin conspecific larvae odor did not show a significant difference in number of activated neurons compared to control stimulation in both mOSNs and cOSNs. In contrast, crypt cells did not show any significant activation in response to both stimuli compared to controls (Fig. 3c). While mOSNs and cOSNs did show a significant activation in response to food but not to non-kin conspecific larvae odor, crypt cells did not respond to either of the stimuli. These results clearly show (a) that within the temporal range tested, stimulus duration has no effect and (b) that pERK is a reliable marker for activated OSNs in zebrafish larvae specific for different odor stimulations.


Crypt cells are involved in kin recognition in larval zebrafish.

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

Differential activation of cOSNs, mOSNs and crypt cells by stimulation with different odors.9 day old zebrafish larvae were exposed to either food odor, non-kin larvae odor or E3 Medium (control) (pooled data of Fig. 2). The total number of pERK+activated cOSNs, mOSNs, and crypt cells was counted per larva and statistically analyzed. Box plots show median, upper and lower quartile and whiskers (maximum interquartile range: 1.5). *indicates statistical significance p: ***p < 0.001. (a) cOSNs are strongly activated by food odor. Significantly more pERK+cOSNs were counted in larvae stimulated with food compared to larvae odor (Mann-Whitney U: 4.0, p < 0.001, median (Mdn)food = 54, Mdnlarvae = 2, n = 18) and to control (U < 0.0, p < 0.001, Mdnfood = 54, Mdnctr = 3, nfood = 18, nctr = 20). (b) mOSNs show the highest activation when stimulated with food odor. Significantly more mOSNS were activated by food odor compared to control stimulation (U: 33.5, p < 0.001, Mdnfood = 19, Mdnctr = 4.5, nfood = 18, nctr = 20). pERK + mOSNs stimulated with larvae odor do not differ in numbers compared to controls (U: 116.5, p = 0.062, Mdnlarvae = 7.5, Mdnctr = 4.5, nlarvae = 18, nctr = 20). (c) Crypt cells show no significant difference in pERK+cell numbers due to stimulation with different odors (Kruskall Wallis test: H(2) = 3.197, p = 0.202, nfood = nlarvae odor = 18, nctr = 20).
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f3: Differential activation of cOSNs, mOSNs and crypt cells by stimulation with different odors.9 day old zebrafish larvae were exposed to either food odor, non-kin larvae odor or E3 Medium (control) (pooled data of Fig. 2). The total number of pERK+activated cOSNs, mOSNs, and crypt cells was counted per larva and statistically analyzed. Box plots show median, upper and lower quartile and whiskers (maximum interquartile range: 1.5). *indicates statistical significance p: ***p < 0.001. (a) cOSNs are strongly activated by food odor. Significantly more pERK+cOSNs were counted in larvae stimulated with food compared to larvae odor (Mann-Whitney U: 4.0, p < 0.001, median (Mdn)food = 54, Mdnlarvae = 2, n = 18) and to control (U < 0.0, p < 0.001, Mdnfood = 54, Mdnctr = 3, nfood = 18, nctr = 20). (b) mOSNs show the highest activation when stimulated with food odor. Significantly more mOSNS were activated by food odor compared to control stimulation (U: 33.5, p < 0.001, Mdnfood = 19, Mdnctr = 4.5, nfood = 18, nctr = 20). pERK + mOSNs stimulated with larvae odor do not differ in numbers compared to controls (U: 116.5, p = 0.062, Mdnlarvae = 7.5, Mdnctr = 4.5, nlarvae = 18, nctr = 20). (c) Crypt cells show no significant difference in pERK+cell numbers due to stimulation with different odors (Kruskall Wallis test: H(2) = 3.197, p = 0.202, nfood = nlarvae odor = 18, nctr = 20).
Mentions: Because we observed that stimulus duration did not affect the number of activated OSNs, we plotted pERK activated cells independent of stimulus durations against the two different odors food and non-kin conspecific larvae odor and compared it with controls (Fig. 3). The activation profile of the different OSNs revealed a highly significant activation in response to food odor compared to controls in ciliated and microvillous OSNs (cOSNs, mOSNs) (Fig. 3a,b). Stimulation with non-kin conspecific larvae odor did not show a significant difference in number of activated neurons compared to control stimulation in both mOSNs and cOSNs. In contrast, crypt cells did not show any significant activation in response to both stimuli compared to controls (Fig. 3c). While mOSNs and cOSNs did show a significant activation in response to food but not to non-kin conspecific larvae odor, crypt cells did not respond to either of the stimuli. These results clearly show (a) that within the temporal range tested, stimulus duration has no effect and (b) that pERK is a reliable marker for activated OSNs in zebrafish larvae specific for different odor stimulations.

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