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The Lhx9 homeobox gene controls pineal gland development and prevents postnatal hydrocephalus

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Abstract

Lhx9 is a member of the LIM homeobox gene family. It is expressed during mammalian embryogenesis in the brain including the pineal gland. Deletion of Lhx9 results in sterility due to failure of gonadal development. The current study was initiated to investigate Lhx9 biology in the pineal gland. Lhx9 is highly expressed in the developing pineal gland of the rat with transcript abundance peaking early in development; transcript levels decrease postnatally to nearly undetectable levels in the adult, a temporal pattern that is generally similar to that reported for Lhx9 expression in other brain regions. Studies with C57BL/6J Lhx9 −/− mutant mice revealed marked alterations in brain and pineal development. Specifically, the superficial pineal gland is hypoplastic, being reduced to a small cluster of pinealocytes surrounded by meningeal and vascular tissue. The deep pineal gland and the pineal stalk are also reduced in size. Although the brains of neonatal Lhx9 −/− mutant mice appear normal, severe hydrocephalus develops in about 70 % of the Lhx9 −/− mice at 5–8 weeks of age; these observations are the first to document that deletion of Lhx9 results in hydrocephalus and as such indicate that Lhx9 contributes to the maintenance of normal brain structure. Whereas hydrocephalus is absent in neonatal Lhx9 −/−mutant mice, the neonatal pineal gland in these animals is hypoplastic. Accordingly, it appears that Lhx9 is essential for early development of the mammalian pineal gland and that this effect is not secondary to hydrocephalus.

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Abbreviations

Actb:

Beta-actin

Bsx:

Brain-specific homeobox

Crx:

Cone–rod homeobox

E:

Embryonic day

Gapdh:

Glyceraldehyde-3-phosphate dehydrogenase

LD:

Light–dark schedule

Lhx:

LIM homeobox

Otx:

Orthodenticle homeobox

P:

Postnatal day

Pax:

Paired box

qRT-PCR:

Quantitative real-time reverse transcription PCR

Rax:

Retina and anterior neural fold homeobox

Rn28s1:

28S ribosomal RNA

ZT:

Zeitgeber time

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Acknowledgments

This study was supported by the Intramural Research Program of the Eunice Kennedy Shriver National Institute of Child Health and Human Development of the National Institutes of Health and the Lundbeck Foundation (grant number R108-A10301). We are grateful to Dr. Heiner Westphal’s (NICHD) gift of the Lhx9 −/− mice and for the support provided by Dr. Yangu Zhao (NICHD) in establishing a colony of Lhx9 mutant mice. We recognize the full value of the willingness of Dr. Joseph D. Dougherty, Washington University, to share unpublished observations. The expertise and dedication of Daniel Abebe as applied to the expansion and management of the mutant Lhx9 colony in our facilities, which was essential to the success of this investigation, is greatly valued. The authors declare no competing financial interests.

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Author notes

  1. Fumiyoshi Yamazaki

    Present address: Department of Cell Biology and Anatomy, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan

  2. Cong Fu

    Present address: State Key Laboratory of Theoretical and Computational Chemistry, Institute of Theoretical Chemistry, Jilin University Changchun, Jilin, China

  3. Samuel J. Clokie

    Present address: West Midlands Regional Genetics Laboratories, Birmingham Women’s NHS Foundation Trust, Birmingham, UK

  4. Artem Zykovich

    Present address: Buck Institute for Research on Aging, Novato, CA, USA

Authors and Affiliations

  1. Section on Neuroendocrinology, Program in Developmental Endocrinology and Genetics, The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, 49 Convent Drive, Room 6A82, Bethesda, MD, 20892-4510, USA

    Fumiyoshi Yamazaki, Cong Fu, Samuel J. Clokie, Artem Zykovich, Steven L. Coon & David C. Klein

  2. Department of Neuroscience and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark

    Morten Møller & Martin F. Rath

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Correspondence to David C. Klein.

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Yamazaki, F., Møller, M., Fu, C. et al. The Lhx9 homeobox gene controls pineal gland development and prevents postnatal hydrocephalus. Brain Struct Funct 220, 1497–1509 (2015). https://doi.org/10.1007/s00429-014-0740-x

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