Midbrain dopaminergic neurons in the mouse: co-localization with Calbindin-D28k and calretinin

doi:10.1016/0306-4522(96)00228-X

Neuroscience

Volume 75, Issue 2, 25 October 1996, Pages 523-533

https://doi.org/10.1016/0306-4522(96)00228-X Get rights and content

Abstract

The calcium-binding proteins Calbindin-D_28k and calretinin are co-localized with dopamine in some of the midbrain dopaminergic neurons in the rat and monkey; the present study sought to examine the pattern of co-localization in the mouse. Double immunofluorescence staining procedures were used for tyrosine hydroxylase (a dopaminergic cell marker) and Calbindin-D_28k or calretinin. Midbrain dopaminergic neurons were examined at four rostrocaudal levels, and the percentage of cells that contained both tyrosine hydroxylase and either of the two calcium-binding proteins was determined in nucleus A8 (retrorubral field), nucleus A9 (substantia nigra pars compacta, pars reticulata and pars lateralis), and nucleus A10 (nucleus paranigralis, ventral tegmental area, interfascicular nucleus, central linear nucleus). The two calcium-binding proteins were similarly distributed in midbrain dopaminergic neurons in the several nuclear groups that comprise nuclei A8, A9 and A10. The calcium-binding proteins were found in the majority (50–100%) of nucleus A10 neurons, whereas in nuclei A8 and A9 (except for the substantia nigra pars lateralis) less than 40% of the cells contained either calcium-binding protein. The pattern of co-localization in the mouse is similar to that reported for the rat and monkey.

The calcium-binding proteins mark the population of midbrain dopaminergic neurons that are less vulnerable to degeneration in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mouse model of Parkinson's disease.

Section snippets

Animals

Three adult male C57BL/6 mice (Taconic Farms, Germantown, NY, U.S.A.), and three adult male FVB mice (Harlan Sprague Dawley, Indianapolis, IN, U.S.A.) were used for immunocytochemical staining (20–25 g). The mice were deeply anesthetized with Nembutal (120 mg/kg, i.p.), and perfused through the ascending aorta with 0.1 M phosphate-buffered saline (PBS; pH 7.4, 2–3 min), and 10% neutral buffered formalin (10 min). The brains were blocked in a coronal plane, and postfixed in formalin for 5–10 h. After

Sections for analysis

Sections from four rostrocaudal levels were chosen for immunocytochemical staining (Fig. 1). Level 1 is at the middle portion of the mammillary nucleus which contains the DA neurons in the substantia nigra pars compacta (SNC), substantia nigra pars reticulata (SNR), and rostral ventral tegmental area (VTA) (Fig. 1A). Level 2 is at the caudal portion of the mammillary nucleus which contains the same regions as in level 1, and also the interfascicular nucleus (IF; Fig. 1B) just medial to the

Characteristics of doaminergic neurons that contain Calbindin-D_28k or calretinin

Most of the TH + Calbindin-D_28k and TH + calretinin double-labeled neurons are medium- to small-sized, multipolar and fusiform-shaped neurons. In contrast, the midbrain DA neurons that lack the calcium-binding proteins Calbindin-D_28k and calretinin, especially in the SNC and VTA, are large-sized multipolar neurons, often greater than 20 μm in the long axis. These neurons are located mainly in the SNC and the dorsal portion of VTA (within PBP), and some at the lateral extent of the CLi. Gerfen et al.8

Conclusions

The calcium-binding proteins Calbindin-D_28k and calretinin are localized within a subpopulation of midbrain DA neurons in the mouse. Midline nuclei, within nucleus A10, and the SNL contain the highest proportion of double-labeled neurons (labeled both for TH and one or both of the calcium-binding proteins). The midline nuclei include IF, PN and Cli. Within nucleus A8, less than half of the DA neurons contain Calbindin-D_28k and/or calretinin, and within nucleus A9 (SNC and SNR), less than 20% of

Acknowledgements

This research was supported by grants from the NIH (NS-30406), American Parkinson Disease Association, and the James Webb Fund of the Dallas Foundation. The authors thank Ms Judy Burdette for secretarial assistance.

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Midbrain dopaminergic neurons in the mouse: co-localization with Calbindin-D28k and calretinin

Abstract

Section snippets

Animals

Sections for analysis

Characteristics of doaminergic neurons that contain Calbindin-D28k or calretinin

Conclusions

Acknowledgements

Neuroscience

Life Sci.

Brain Res.

Neuroscience

TINS

Brain Res.

Neuroscience

Molec. Brain Res.

Brain Res.

Neuroscience

Neuroscience

Brain Res.

Trends Neurosci.

Neuroscience

Neuroscience

Neuroscience

Brain Res.

Brain Res.

Neuroscience

Cell Calcium

Neuroscience

Brain Res. Bull.

Brain Res.

Immunohistochemical localization of calretinin in the rat hindbrain

J. comp. Neurol.

Evidence for the existence of monoamine-containing neurons in the central nervous system. I. Demonstration of monoamines in the cell bodies of brain stem neurons

Acta physiol. scand. Suppl.

Catecholamine innervation of the basal forebrain: IV. Topography of the dopamine projection to the basal forebrain and neostriatum

J. comp. Neurol.

The neostriatal mosaic: compartmental distribution of CaBP and parvalbumin in the basal ganglia of the rat and monkey

Proc. natn. Acad. Sci. U.S.A.

Midbrain dopaminergic neurons in the mouse: co-localization with Calbindin-D_28k and calretinin

Characteristics of doaminergic neurons that contain Calbindin-D_28k or calretinin