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Zebrin II Is Ectopically Expressed in Microglia in the Cerebellum of Neurogenin 2 Null Mice

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Abstract

Zebrin II/aldolase C expression in the normal cerebellum is restricted to a Purkinje cell subset and is the canonical marker for stripes and zones. This spatial restriction has been confirmed in over 30 species of mammals, birds, fish, etc. In a transgenic mouse model in which the Neurogenin 2 gene has been disrupted (Neurog2−/−), the cerebellum is smaller than normal and Purkinje cell dendrites are disordered, but the basic zone and stripe architecture is preserved. Here, we show that in the Neurog2−/− mouse, in addition to the normal Purkinje cell expression, zebrin II is also expressed in a population of cells with a morphology characteristic of microglia. This identity was confirmed by double immunohistochemistry for zebrin II and the microglial marker, Iba1. The expression of zebrin II in cerebellar microglia is not restricted by zone or stripe or lamina. A second zone and stripe marker, PLCβ4, does not show the same ectopic expression. When microglia are compared in control vs. Neurog2−/− mice, no difference is seen in apparent number or distribution, suggesting that the ectopic zebrin II immunoreactivity in Neurog2−/− cerebellum reflects an ectopic expression rather than the invasion of a new population of microglia from the periphery. This ectopic expression of zebrin II in microglia is unique as it is not seen in numerous other models of cerebellar disruption, such as in Acp2−/− mice and in human pontocerebellar hypoplasia. The upregulation of zebrin II in microglia is thus specific to the disruption of Neurog2 downstream pathways, rather than a generic response to a cerebellar disruption.

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Acknowledgments

These studies were supported by grants from the Children Hospital Research Institute of Manitoba (HM). We are grateful to Carol Schuurmans for providing ngn2 knockout mice, and Marc Del Bigio for providing slides of pontocerebellar hypoplasia and normal control section samples.

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Authors and Affiliations

  1. Department of Human Anatomy and Cell Science, The Children’s Hospital Research Institute of Manitoba (CHRIM), Max Rady College of Medicine, Rady Faculty of Health Sciences, University of Manitoba, Rm 129 BMSB, 745 Bannatyne Avenue, Winnipeg, MB, R3E 0J9, Canada

    Maryam Rahimi-Balaei, Xiaodan Jiao, Shahin Shabanipour & Hassan Marzban

  2. Biological Sciences Platform, Sunnybrook Research Institute, University of Toronto, Toronto, ON, Canada

    Rajiv Dixit & Carol Schuurmans

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  1. Maryam Rahimi-Balaei
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  2. Xiaodan Jiao
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  5. Carol Schuurmans
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Correspondence to Hassan Marzban.

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Electronic Supplementary Material

Fig. Suppl. 1

. A) Acp2−/− and wt sibling cerebellum at P5 and P7 (P5; n = 1 and P7; n = 2) were isolated and extracted total RNA was shipped to the McGill University and Genome Quebec Innovation Centre (MUGQIC). The raw RNA-sequencing data were trimmed and clipped with STAR (Spliced Transcripts Alignment to a Reference) program. RNA-sequencing data analysis indicates that the Neurog2 expression upregulated in Acp2−/− cerebella. B) In order to quantify microglia in cerebellum of the Neurog2−/− and wt, we counted microglia immunostained with Iba1 in six sections through the vermis of P15 animals, capturing 5 areas in each zone (each area = 0.6 mm*0.64 mm). We calculated microglia number to be: 59.69 ± 1.36 per 1mm2 in wild-type cerebella and 55.94 ± 1.28/1mm2 in Neurog2−/− cerebella (mean ± SE). There were no significant differences between wild-type and Neurog2−/− cerebella. (PNG 1.02 mb)

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Rahimi-Balaei, M., Jiao, X., Shabanipour, S. et al. Zebrin II Is Ectopically Expressed in Microglia in the Cerebellum of Neurogenin 2 Null Mice. Cerebellum 18, 56–66 (2019). https://doi.org/10.1007/s12311-018-0944-3

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