Calretinin immunoreactivity in the developing thalamus of the rat: a marker of early generated thalamic cells

doi:10.1016/S0306-4522(97)00443-0

Neuroscience

Volume 83, Issue 4, 12 January 1998, Pages 1203-1214

https://doi.org/10.1016/S0306-4522(97)00443-0 Get rights and content

Abstract

The present work was aimed to study the immunocytochemical localization of the calcium-binding protein, calretinin, in the rat thalamus from embryonic day 14 to the third postnatal week. In the adult rat thalamus, calretinin immunoreactivity is intensely expressed in some intralaminar and midline nuclei, as well as in selected regions of the reticular nucleus. At embryonic day 14, calretinin was expressed by immature and migrating neurons and fibres laterally to the neuroepithelium of the diencephalic vesicle in the region identified as reticular neuroepithelium. At embryonic day 16, immunoreactive neurons were present in the primordium of the reticular nucleus and in the region of the reticular thalamic migration, where neurons showed the morphology of migratory cells. At the end of embryonic development and in the first postnatal week, calretinin-positive neurons were observed in selected region of the reticular nucleus and it was intensely expressed in some intralaminar and midline nuclei. Bands of immunopositive fibres were also observed crossing the thalamus. During the second postnatal week, the immunolabelling in the reuniens, rhomboid, paraventricular and central medial thalamic nuclei remains very intense while a decrease of immunoreactivity in mediodorsal, centrolateral and laterodorsal nuclei was observed. The immunostaining of fibres, particularly evident in the perinatal period, progressively decreased and it was no longer visible by the end of the second postnatal week when the distribution and intensity of calretinin immunostaining was similar to that observed in the adult rat thalamus.

The present findings indicate that the immunolocalization of calretinin can be used to identify subsets of thalamic neuronal population during pre- and postnatal maturation allowing also the detection of the migratory pattern of early generated reticular thalamic neurons.

Section snippets

Experimental procedures

Wistar albino rats (Wistar, Charles River), ranging from embryonic day (E)14 to postnatal day (P)21 were used in this study. The results observed in these animals were compared with two 40-day-old rats. All the experiments were undertaken in accordance with guidelines established in the Principles of Laboratory Animal Care (NIH Publication no. 86–23, revised 1985).

Sixteen embryos (E14, E16, E18, E19) were obtained from chloral hydrate (4%; 1 ml/100 g body weight, i.p.)-anaesthetized pregnant

Embryonic day 14

At E14, the portion of the diencephalic vesicle which gives rise to the thalamus was almost exclusively composed by neuroepithelium, although a zone of differentiating cells was already present. These cells flanked a small neuroepithelial protuberance and concavity (Fig. 1A,B) designated by Altman and Bayer[3]as reticular neuroepithelium because they are destined to produce Rt neurons. Calretinin at this age was expressed in some postmitotic neurons and fibre tracts laterally to the

Discussion

The present findings demonstrate that calretinin immunoreactivity appears early during thalamic development (E14) and is expressed by immature and migrating neurons, laterally to the neuroepithelium of the diencephalic vescicle. At E16, calretinin-IR neurons were present in the Rtp and some of them showed the characteristic morphology of migrating cells. At the end of embryonic development and in the first postnatal days, calretinin was intensely expressed in some intralaminar and midline

Acknowledgements

This work was partially supported by the Paolo Zorzi Association for Neuroscience.

References (46)

L Anglister et al.
Localization of voltage-sensitive calcium channels along developing neurites: their possible role in regulating neurite elongation
Devl Biol.
(1982)
K.G Baimbridge et al.
Calcium-binding proteins in the nervous system
Trends Neurosci.
(1992)
M.R Celio
Calbindin and parvalbumin in the rat nervous system
Neuroscience
(1990)
C Frassoni et al.
Postnatal development of calbindin and parvalbumin immunoreactivity in the thalamus of the rat
Devl Brain Res.
(1991)
J.C Gander et al.
Inhibition of the proliferative cycle and apoptotic events in Wdr cells after down-regulation of the calcium-binding protein calretinin using antisense oligodeoxynnucleotides
Expl Brain Res.
(1996)
M.J Hubbard et al.
Calbindin-28KDa and calbindin-30KDa (calretinin) are localized in the particulate fraction of rat brain
Fedn Eur. biochem. Socs Lett.
(1995)
E Király et al.
Ontogeny of calretinin in chick dorsal root ganglion neurons
Devl Brain Res.
(1992)
E Király et al.
In vitro detection of calretinin immunoreactivity in chicken embryo dorsal root ganglion neurons: a possible developmental marker
Devl Brain Res.
(1993)
H Komuro et al.
Intracellular Ca²⁺ fluctuations modulate the rate of neuronal migration
Neuron
(1996)
J Kuznicki et al.
The expression of calretinin in transfected PC12 cells provides no protection against Ca²⁺ overload or trophic factor deprivation
Biochim. biophys. Acta mol. cell. Res.
(1996)

V Mockel et al.

Vulnerability to excitotoxic stimuli of cultured rat hippocampal neurons containing the calcium binding proteins calretinin and calbindin D28K

Brain Res.

(1994)

L Puelles et al.

Prenatal development of calbindin immunoreactivity in the dorsal thalamus of the rat

Neuroscience

(1992)

A Résibois et al.

Calretinin in rat brain: an immunohistochemical study

Neuroscience

(1992)

J.H Rogers

Immunoreactivity for calretinin and other calcium-binding proteins in cerebellum

Neuroscience

(1989)

B Schwaller et al.

Characterization of a polyclonal antiserum against the purified human recombinant calcium-binding protein calretinin

Cell Calcium

(1993)

D.M Vogt Weisenhorn et al.

Localization of calretinin in cells of layer I (Cajal–Retzius cells) of the developing cortex of the rat

Devl Brain Res.

(1994)

L Winsky et al.

Calretinin distribution in the thalamus of the rat: immunohistochemical and in situ hybridization histochemical analyses

Neuroscience

(1992)

T Yamaguchi et al.

The effect of insulin and insulin-like growth factor-1 on the expression of calretinin and calbindin D28K in rat embryonic neurons in culture

Neurochem. Int.

(1995)

L.C Abbot et al.

Development of calretinin-immunoreactive unipolar brush-like cells and afferent pathway to embryonic and early postnatal mouse cerebellum

Anat. Embryol.

(1995)

J Altman et al.

Development of the diencephalon in the rat. VI. Re-evalutation of the embryonic development of the thalamus on the basis of thymidine-radiographic datings

J. comp. Neurol.

(1979)

J Altman et al.

Development of the rat thalamus: III. Time and site of origin and settling pattern of neurons of the reticular nucleus

J. comp. Neurol.

(1988)

J Altman et al.

Development of the rat thalamus: I. Mosaic organization of the thalamic neuroepithelium

J. comp. Neurol.

(1988)

J Altman et al.

Development of the rat thalamus. VI. The posterior lobule of the thalamic neuroepithelium and the time and site of origin and settling pattern of neurons of the lateral geniculate and lateral posterior nuclei

J. comp. Neurol.

(1989)

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It seems to be very similar to what we observed at E40 and E50 in the guinea pig AM. Frassoni et al. (1998) also detected thick CR-ir fibers crossing the developing thalamus of the rat, but only during perinatal and first postnatal weeks. Later during the postnatal development, these fibers decreased and disappeared in adulthood.
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Calretinin immunoreactivity in the developing thalamus of the rat: a marker of early generated thalamic cells

Abstract

Section snippets

Experimental procedures

Embryonic day 14

Discussion

Acknowledgements

Devl Biol.

Trends Neurosci.

Neuroscience

Devl Brain Res.

Expl Brain Res.

Fedn Eur. biochem. Socs Lett.

Devl Brain Res.

Devl Brain Res.

Neuron

Biochim. biophys. Acta mol. cell. Res.

Brain Res.

Neuroscience

Neuroscience

Neuroscience

Cell Calcium

Devl Brain Res.

Neuroscience

Neurochem. Int.

Development of calretinin-immunoreactive unipolar brush-like cells and afferent pathway to embryonic and early postnatal mouse cerebellum

Anat. Embryol.

Development of the diencephalon in the rat. VI. Re-evalutation of the embryonic development of the thalamus on the basis of thymidine-radiographic datings

J. comp. Neurol.

Development of the rat thalamus: III. Time and site of origin and settling pattern of neurons of the reticular nucleus

J. comp. Neurol.

Development of the rat thalamus: I. Mosaic organization of the thalamic neuroepithelium

J. comp. Neurol.

Development of the rat thalamus. VI. The posterior lobule of the thalamic neuroepithelium and the time and site of origin and settling pattern of neurons of the lateral geniculate and lateral posterior nuclei

J. comp. Neurol.