Single cell RNA sequencing confirms retinal microglia activation associated with early onset retinal degeneration.
Single cell RNA sequencing confirms retinal microglia activation associated with early onset retinal degeneration.
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DOI：
10.1038/s41598-022-19351-w
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发表时间：
2022-09-10
影响因子：
4.6
通讯作者：
Borooah, Shyamanga
中科院分区：
文献类型：
--
作者：
Kumari, Asha;Ayala-Ramirez, Raul;Zenteno, Juan Carlos;Huffman, Kristyn;Sasik, Roman;Ayyagari, Radha;Borooah, Shyamanga
膜型卷曲相关蛋白（Mfrp）基因突变导致早发性视网膜变性，与视网膜色素变性、小眼症、视盘玻璃疣和中央凹裂相关。在本研究中，使用了先前表征的Mfrp中携带纯合c.498_499insC突变（MfrpKI/KI）的人视网膜变性小鼠模型。携带这种突变的患者在很小的时候就有视网膜变性。该模型显示视网膜下沉积物，并发生早发性光感受器变性。我们在MfrpKI/KI小鼠中观察到大的视网膜下沉积物，其为强CD 68阳性并且与自体荧光斑点共定位。MfrpKI/KI小鼠视网膜小胶质细胞的单细胞RNA测序显示泛巨噬细胞标志物Iba-1和F4/80阳性细胞的数量显著更高，活化标志物（CD 68）的表达增加，而小胶质细胞稳态标志物的表达降低（TMEM 119，P2ry13，P2ry13，Siglech）与野生型小鼠相比，证实了在视网膜免疫染色中观察到的小胶质细胞活化，显示视网膜下区域的小胶质细胞活化。轨迹分析确定了一小群具有活化转录组特征的小胶质细胞，其可以代表表达较高水平APOE的MfrpKI/KI小鼠中的视网膜下小胶质细胞群体。我们使用视网膜冷冻切片的免疫荧光染色验证了这些发现，并发现MfrpKI/KI小鼠中视网膜下Iba-1/ApoE阳性小胶质细胞的数量显著较高，一些视网膜下小胶质细胞也表达低水平的小胶质细胞稳态标记物TMEM 119，证实了小胶质细胞起源。总之，我们证实携带c.498_499insC突变的MfrpKI/KI小鼠在其视网膜中具有显著较高的活化小胶质细胞群体，具有不同的视网膜下小胶质细胞亚群。此外，需要进行研究以确认MfrpKI/KI小鼠中视网膜下小胶质细胞增加的相关性是否是变性的原因。
Mutations in the Membrane-type frizzled related protein (Mfrp) gene results in an early-onset retinal degeneration associated with retinitis pigmentosa, microphthalmia, optic disc drusen and foveal schisis. In the current study, a previously characterized mouse model of human retinal degeneration carrying homozygous c.498_499insC mutations in Mfrp (MfrpKI/KI) was used. Patients carrying this mutation have retinal degeneration at an early age. The model demonstrates subretinal deposits and develops early-onset photoreceptor degeneration. We observed large subretinal deposits in MfrpKI/KI mice which were strongly CD68 positive and co-localized with autofluorescent spots. Single cell RNA sequencing of MfrpKI/KI mice retinal microglia showed a significantly higher number of pan-macrophage marker Iba-1 and F4/80 positive cells with increased expression of activation marker (CD68) and lowered microglial homeostatic markers (TMEM119, P2ry13, P2ry13, Siglech) compared with wild type mice confirming microglial activation as observed in retinal immunostaining showing microglia activation in subretinal region. Trajectory analysis identified a small cluster of microglial cells with activation transcriptomic signatures that could represent a subretinal microglia population in MfrpKI/KI mice expressing higher levels of APOE. We validated these findings using immunofluorescence staining of retinal cryosections and found a significantly higher number of subretinal Iba-1/ApoE positive microglia in MfrpKI/KI mice with some subretinal microglia also expressing lowered levels of microglial homeostatic marker TMEM119, confirming microglial origin. In summary, we confirm that MfrpKI/KI mice carrying the c.498_499insC mutation had a significantly higher population of activated microglia in their retina with distinct subsets of subretinal microglia. Further, studies are required to confirm whether the association of increased subretinal microglia in MfrpKI/KI mice are causal in degeneration.
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影响因子：
9.3
作者：
Lipski DA;Dewispelaere R;Foucart V;Caspers LE;Defrance M;Bruyns C;Willermain F
通讯作者：
Willermain F
影响因子：
9.3
作者：
Noailles A;Fernández-Sánchez L;Lax P;Cuenca N
通讯作者：
Cuenca N
影响因子：
56.9
作者：
Nimmerjahn, A;Kirchhoff, F;Helmchen, F
通讯作者：
Helmchen, F
影响因子：
9.3
作者：
DePaula-Silva, Ana Beatriz;Gorbea, Carlos;Fujinami, Robert S.
通讯作者：
Fujinami, Robert S.
DOI：
10.1073/pnas.1609896113
发表时间：
2016-09-06
影响因子：
11.1
作者：
Chung, Won-Suk;Verghese, Philip B.;Barres, Ben A.
通讯作者：
Barres, Ben A.