Expression of the IP3R1 promoter-driven nls-lacZ transgene in Purkinje cell parasagittal arrays of developing mouse cerebellum

J Neurosci Res. 2010 Oct;88(13):2810-25. doi: 10.1002/jnr.22451.

Abstract

The cerebellar Purkinje cell monolayer is organized into heterogeneous Purkinje cell compartments that have different molecular compositions. Here we describe a transgenic mouse line, 1NM13, that shows heterogeneous transgene expression in parasagittal Purkinje cell arrays. The transgene consists of a nuclear localization signal (nls) fused to the beta-galactosidase (lacZ) composite gene driven by the type 1 inositol 1,4,5-trisphosphate receptor (IP(3)R1) gene promoter. IP(3)R1-nls-lacZ transgene expression was detected at a single Purkinje cell level over the surface of a whole-mount X-gal-stained cerebellum because of nuclear accumulation of the nls-lacZ activity. Developing cerebella of 1NM13 mice showed stripe-like X-gal staining patterns of parasagittal Purkinje cell subsets. The X-gal stripe pattern was likely determined by an intrinsic property as early as E15 and showed increasing complexity with cerebellar development. The X-gal stripe pattern was reminiscent of, but not identical to, the stripe pattern of zebrin II immunoreactivity. We designated the symmetrical X-gal-positive (transgene-positive, Tg(+)) Purkinje cell stripes about the midline as vermal Tg1(+), Tg2(a, b)(+) and Tg3(a, b)(+) stripes and hemispheric Tg4(a, b)(+), Tg5(a, b)(+), Tg6(a, b, c)(+), and Tg7(a, b)(+) stripes, where a, b, and c indicate substripes. We also assigned three parafloccular substripes Tg8(a, b, c)(+). The boundaries of X-gal stripes at P5 were consistent with raphes in the Purkinje cell layer through which granule cells migrate, suggesting a possible association of the X-gal stripes with raphe formation. Our results indicate that 1NM13 is a good mouse model with a reproducible and clear marker for the compartmentalization of Purkinje cell arrays.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Cerebellum* / cytology
  • Cerebellum* / embryology
  • Cerebellum* / growth & development
  • Choline / analogs & derivatives
  • Choline / genetics
  • Choline / metabolism
  • Embryo, Mammalian
  • Gene Expression Regulation, Developmental / genetics
  • Gene Expression Regulation, Developmental / physiology*
  • Humans
  • Inositol 1,4,5-Trisphosphate Receptors / genetics*
  • Inositol 1,4,5-Trisphosphate Receptors / metabolism
  • Mice
  • Mice, Transgenic
  • Nerve Tissue Proteins / metabolism
  • Nuclear Localization Signals / genetics*
  • Organ Culture Techniques
  • Promoter Regions, Genetic / genetics
  • Purkinje Cells / physiology*

Substances

  • ITPR1 protein, human
  • Inositol 1,4,5-Trisphosphate Receptors
  • Nerve Tissue Proteins
  • Nuclear Localization Signals
  • zebrin II
  • murexine
  • Choline