A dual‐fluorescence reporter in the Eomes locus for live imaging and medium‐term lineage tracing

Probst, Simone; Daza, Ray A. M.; Bader, N. G.; Hummel, Jonas; Weiß, Matthias; Tanriver, Yakup; Hevner, Robert F.; Arnold, Sebastian J.

doi:10.1002/dvg.23043

Cited by 6 publications

(9 citation statements)

References 35 publications

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“…XM_006529679.2). Following completion of the RNAscope protocol, sections were immunolabelled using a rabbit anti-RFP polyclonal affinity purified antibody (1: 1,000; Rockland, 600-401-379) [29] and a donkey anti-rabbit Alexa Fluor 555 (1: 400; Invitrogen, A31572).…”

Section: Methodsmentioning

confidence: 99%

Medial Amygdala Kiss1 Neurons Mediate Female Pheromone Stimulation of Luteinizing Hormone in Male Mice

et al. 2018

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Background/Aims: The medial amygdala (MeA) responds to olfactory stimuli and alters reproductive physiology. However, the neuronal circuit that relays signals from the MeA to the reproductive axis remains poorly defined. This study aimed to test whether MeA kisspeptin (MeA^Kiss) neurons in male mice are sensitive to sexually relevant olfactory stimuli and transmit signals to alter reproductive physiology. We also investigated whether MeA^Kiss neurons have the capacity to elaborate glutamate and GABA neurotransmitters and potentially contribute to reproductive axis regulation. Methods: Using female urine as a pheromone stimulus, MeA^Kiss neuronal activity was analysed and serum luteinizing hormone (LH) was measured in male mice. Next, using a chemogenetic approach, MeA^Kiss neurons were bi-directionally modulated to measure the effect on serum LH and evaluate the activation of the preoptic area. Lastly, using in situ hybridization, we identified the proportion of MeA^Kiss neurons that express markers for GABAergic (Vgat) and glutamatergic (Vglut2) neurotransmission. Results: Male mice exposed to female urine showed a two-fold increase in the number of c-Fos-positive MeA^Kiss neurons concomitant with raised LH. Chemogenetic activation of MeA^Kiss neurons significantly increased LH in the absence of urine exposure, whereas inhibition of MeA^Kiss neurons did not alter LH. In situ hybridization revealed that MeA^Kiss neurons are a mixed neuronal population in which 71% express Vgat mRNA, 29% express Vglut2 mRNA, and 6% express both. Conclusions: Our results uncover, for the first time, that MeA^Kiss neurons process sexually relevant olfactory signals to influence reproductive hormone levels in male mice, likely through a complex interplay of neuropeptide and neurotransmitter signalling.

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Section: Methodsmentioning

confidence: 99%

Medial Amygdala Kiss1 Neurons Mediate Female Pheromone Stimulation of Luteinizing Hormone in Male Mice

et al. 2018

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“…(O) The confocal image shows fluorescence signals in the E12.5 neocortex for reporter mice expressing nuclear GFP and cell membrane tdT from the Tbr2 locus. Stable nuclear GFP labels IPC and IPC-derived neurons [ 22 ]. The tdT signal emerges later than GFP and labels IPC-derived neurons [ 22 ].…”

Section: Resultsmentioning

confidence: 99%

“…Stable nuclear GFP labels IPC and IPC-derived neurons [ 22 ]. The tdT signal emerges later than GFP and labels IPC-derived neurons [ 22 ]. The scatter plot demonstrates FACS from E12.5 dorsal telencephalon.…”

Section: Resultsmentioning

confidence: 99%

“…the area from the hem to the corticostriatal sulcus) and a ventrolateral part (i.e. the pallial area from the corticostriatal sulcus to the piriform [22]. The tdT signal emerges later than GFP and labels IPC-derived neurons [22].…”

Section: Plos Geneticsmentioning

confidence: 99%

“…Donson LoxP was verified using Southern blot and PCR. Emx1-Cre, Nkx2.1-Cre, Tbr2-Cre, Ai14 Rosa26 CAG-LSL-tdT , and Tbr2 reporter mice were described [22,24,45,54,55]. Mice containing one of the Cre-alleles and two Donson LoxP alleles were defined as conditional knockouts (Emx1-cKO; Nkx2.1-cKO, Tbr2-cKO).…”

Section: Micementioning

confidence: 99%

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The microcephaly gene Donson is essential for progenitors of cortical glutamatergic and GABAergic neurons

et al. 2021

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Biallelic mutations in DONSON, an essential gene encoding for a replication fork protection factor, were linked to skeletal abnormalities and microcephaly. To better understand DONSON function in corticogenesis, we characterized Donson expression and consequences of conditional Donson deletion in the mouse telencephalon. Donson was widely expressed in the proliferation and differentiation zones of the embryonic dorsal and ventral telencephalon, which was followed by a postnatal expression decrease. Emx1-Cre-mediated Donson deletion in progenitors of cortical glutamatergic neurons caused extensive apoptosis in the early dorsomedial neuroepithelium, thus preventing formation of the neocortex and hippocampus. At the place of the missing lateral neocortex, these mutants exhibited a dorsal extension of an early-generated paleocortex. Targeting cortical neurons at the intermediate progenitor stage using Tbr2-Cre evoked no apparent malformations, whereas Nkx2.1-Cre-mediated Donson deletion in subpallial progenitors ablated 75% of Nkx2.1-derived cortical GABAergic neurons. Thus, the early telencephalic neuroepithelium depends critically on Donson function. Our findings help explain why the neocortex is most severely affected in individuals with DONSON mutations and suggest that DONSON-dependent microcephaly might be associated with so far unrecognized defects in cortical GABAergic neurons. Targeting Donson using an appropriate recombinase is proposed as a feasible strategy to ablate proliferating and nascent cells in experimental research.

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Polarity inversion reorganizes the stem cell compartment of the trophoblast lineage

et al. 2023

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A dual‐fluorescence reporter in the Eomes locus for live imaging and medium‐term lineage tracing

Cited by 6 publications

References 35 publications

Medial Amygdala <b><i>Kiss1</i></b> Neurons Mediate Female Pheromone Stimulation of Luteinizing Hormone in Male Mice

Medial Amygdala <b><i>Kiss1</i></b> Neurons Mediate Female Pheromone Stimulation of Luteinizing Hormone in Male Mice

The microcephaly gene Donson is essential for progenitors of cortical glutamatergic and GABAergic neurons

Polarity inversion reorganizes the stem cell compartment of the trophoblast lineage

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