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[Preprint]. 2023 Dec 3:2023.12.02.569711.
doi: 10.1101/2023.12.02.569711.

LSD1/KDM1A is essential for neural stem cell differentiation in mice

E C Falkenberry  1 , M Reeves  1 , A Scott  1   2 , D A Myrick  1   2 , C Fallini  2 , G J Bassell  1 , D J Katz  1
Affiliations

LSD1/KDM1A is essential for neural stem cell differentiation in mice

E C Falkenberry et al. bioRxiv. .

Abstract

The proper regulation of neural stem cell differentiation is required for the proper specification of the central nervous system. Here we investigated the function of the H3K4me1/2 demethylase LSD1/KDM1A during neural stem differentiation in mice. Conditional deletion of LSD1 in nestin- positive neural stem cells results in 100% perinatal lethality after birth with severe motor coordination deficits, retarded growth and defects in brain morphology. Despite these severe defects, motor neuron progenitors and the initial motor neuron population are specified normally and motor neurons with normal morphology can be cultured from these mice in vitro. However, motor neurons cultured from mice lacking LSD1 in neural stem cells continue to inappropriately maintain critical neural stem cell proteins. Taken together these results suggest that, as in other mouse stem cell populations, LSD1 is required to deactivate the stem cell program to enable normal neural stem cell differentiation. However, unlike in other mouse stem cell populations, the inappropriate maintenance of the stem cell program during neural stem cell differentiation may compromise neuronal function rather than neuronal specification.

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Figures

Figure 1.
Figure 1.. LSD1 is expressed in neural stem cells.
Immunofluorescence at e12.5 of Lsd1+/+ control (A-K) and Lsd1NSC (L-N) embryos, showing DAPI (A,E,I,L), LSD1 (C,G,J,M), Cre (B,F) and Merge (D,H,K,N). CRE+ NSCs outlined in white (B) are magnified in E-H. Expression of LSD1 in these cells is lost in Lsd1NSC embryos (M).
Figure 2.
Figure 2.. Lsd1NSC animals do not survive past weaning and show stunted growth.
Expected number of mice of each possible genotype (Wt: Wild Type, Het: heterozygous for the Lsd1 floxed allele with and without Cre, Cre-; fl/fl: homozygous for the Lsd1 floxed allele without Cre, Cre+; fl/fl: homozygous for the Lsd1 floxed allele with Cre) versus observed at P0, N=41 from 7 litters (A) and P21, N=79 from 14 litters (B). **Indicates a P<0.01 from chi square analysis. (C,D) Representative images of heterozygous Lsd1+/− control versus Lsd1NSC littermates taken at postnatal day 5 and postnatal day 8, with Lsd1NSC animal engaged in stargazing pose.
Figure 3.
Figure 3.. Specification of motor neurons in Lsd1NSC animals is not affected.
Immunofluorescence at e12.5 showing DAPI (A,E), HB9 (B,F) and OLIG2 (C,G) in Lsd1+/+ control (A-D) versus Lsd1NSC (E-H) embryos, quantified in I and J (Lsd1+/+: N=8, Lsd1NSC: N=5). ns indicates not significant.
Figure 4.
Figure 4.. Lsd1NSC mutant-derived motor neurons inappropriately express stem cell genes.
Motor neurons cultured from e13.5 Lsd1+/+ controls (A) and Lsd1NSC mutants (B) appear morphologically normal. Three examples each of motor neurons cultured from Lsd1NSC mutants (C-K) and Lsd1+/+ controls (L-T), showing the NSC markers NESTIN (C,F,I,L,O,R) and SOX2 (D,G,J,M,P,S) and the neuronal marker TAU (E,H,K,N,Q,T), quantified in U and V. *** = p<.001, calculated using an unpaired t test.
Figure 5.
Figure 5.. Lsd1NSC mutant animals show brain morphology defects in vivo.
H&E histology of (A,B) hippocampus (Hip), (C,D) cerebellum (Cere) and (E,F) CA1 region of the hippocampus (CA1) from Lsd1+/+ control (A,C,E) versus Lsd1NSC (B,D,F) postnatal animals.
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