Exploring Brain Genoarchitecture by Single and Double Chromogenic In Situ Hybridization (ISH) and Immunohistochemistry (IHC) on Cryostat, Paraffin, or Floating Sections

Ferrán, José Luis; Ayad, Abdelmalik; Merchán, Paloma; Morales-Delgado, Nicanor; Sánchez-Arrones, Luisa; Alonso, Antonia; SandovalSandoval, Juan Eugenio; Bardet, Sylvia M.; Corral-San-Miguel, Rubén; Sánchez-Guardado, Luis Óscar; Hidalgo‐Sánchez, Matías; Martı́nez-de-la-Torre, Margaret; Puelles, Luis

doi:10.1007/978-1-4939-2303-8_5

Cited by 26 publications

(26 citation statements)

References 9 publications

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“…Other data shown are from our laboratory. The procedures we follow for ISH performed on floating sections, with immunoreaction as a counterstain or not, were published in Ferran et al (). Methodological data for transgenic Azin2‐LacZ material were reported in Martinez‐de‐la‐Torre, Lambertos, Peñafiel, and Puelles ().…”

Section: Some Molecular Markersmentioning

confidence: 99%

Concentric ring topology of mammalian cortical sectors and relevance for patterning studies

Puelles

Alonso

Garcı́a-Calero

et al. 2019

J of Comparative Neurology

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158

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Models aiming to explain causally the evolutionary or ontogenetic emergence of the pallial isocortex and its regional/areal heterogeneity in mammals use simple or complex assumptions about the pallial structure present in basal mammals and nonmammals. The question arises: how complex is the pattern that needs to be accounted for in causal models? This topic is also paramount for comparative purposes, since some topological relationships may be explained as being ancestral, rather than newly emerged. The mouse pallium is apt to be reexamined in this context, due to the breadth of available molecular markers and correlative experimental patterning results. We center the present essay on a recapitulative glance at the classic theory of concentric mammalian allo‐, meso‐, and neocortex domains. In its simplest terms, this theory postulates a central neocortical island (6 layers) separated by a surrounding mesocortical ring (4–5 layers) from a peripheral allocortical ring (3 layers). These territories show additional partition into regional or areal subdivisions. There are also borderline amygdalar, claustral, and septal areas of the pallium, nuclear in structure. There has been little effort so far to contemplate the full concentric ring model in current “cortex patterning” models. In this essay, we recapitulate the ring idea in mammals (mouse) and consider a potential causal patterning scenario using topologic models. Finally, we briefly explore how far this theory may apply to pallium models proposed recently for sauropsids.

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Section: Some Molecular Markersmentioning

confidence: 99%

Concentric ring topology of mammalian cortical sectors and relevance for patterning studies

Puelles

Alonso

Garcı́a-Calero

et al. 2019

J of Comparative Neurology

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158

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“…Floating 110-mm-thick vibratome sections were used for in situ hybridization according to the protocol described by Ferran et al (2015b). The sections were washed afterward, and the hybridized product was detected immunochemically with antidigoxigenin coupled to alkaline phosphatase (Roche Diagnostics, Mannheim, Germany).…”

Section: In Situ Hybridizationmentioning

confidence: 99%

Selective early expression of the orphan nuclear receptor Nr4a2 identifies the claustrum homolog in the avian mesopallium: Impact on sauropsidian/mammalian pallium comparisons

Puelles

Ayad

Alonso

et al. 2015

J of Comparative Neurology

184

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The transcription factor Nr4a2 was recently revealed as a very early developmental marker of the claustrum (CL) proper in the mouse. The earliest claustral primordium was identified superficially, dorsal to the olfactory cortex, and was subsequently covered by the Nr4a2-negative cells of the insular cortex. Some tangentially migrating claustral derivatives (subplate cells and some endopiriform elements) also expressed this marker. The present study employs the same genetic marker to explore the presence of a comparable pallial division in chicken in which, in principle, the same pallial sectors exist as in mammals. We were indeed able to delineate an early-developing Nr4a2-positive mantle domain at the expected topologic position within the developing chicken lateral pallium. In the chicken as well as in the turtle (from data in the literature), the earliest postmitotic lateropallial cells likewise express Nr4a2 and occupy a corticoid superficial stratum of the mesopallium, which is clearly comparable in spatial and chronological profile to the mouse CL. Other cells produced in this pallial sector include various tangentially migrating Nr4a2-labeled derivatives as well as Nr4a2-negative and Nr4a2-positive local deeper subpopulations that partially interdigitate, forming mesopallial core and shell populations. We hold that the deep avian and reptilian mesopallial formation developing under the superficial corticoid CL homolog represents a field homolog of the insula, although additional studies are required to underpin this hypothesis.

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“…To generate Cck Cre :Tcf7l2 fl/fl strain, in which the knockout of Tcf7l2 is induced in the thalamic area perinatally (Allen Brain Atlas, 2011), Tcf7l2 tm1a/+ animals were first crossed with flippase-expressing mice (JAX stock #009086; (Farley et al, 2000)), and then with (Walther and Gruss, 1991). Chromogenic in situ hybridisation of cryosections was performed as described elsewhere (Ferran et al, 2015b;Puelles et al, 2016). The stained sections were washed in xylene and mounted using EuKitt (Cat.…”

Section: Methodsmentioning

confidence: 99%

TCF7L2 regulates postmitotic differentiation programs and excitability patterns in the thalamus

Lipiec

Koziński

Zajkowski

et al. 2019

Preprint

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StatementThe study describes a role of TCF7L2 in neuronal differentiation of thalamic glutamatergic neurons at two developmental stages, highlighting its involvement in the postnatal establishment of critical thalamic electrophysiological features. AbstractNeuronal phenotypes are controlled by terminal selector transcription factors in invertebrates, but few examples of such regulators have been provided in vertebrates. TCF7L2 has been identified as a regulator of efferent outgrowth in the thalamus and habenula. We used a complete and conditional knockout of Tcf7l2 in mice to investigate the hypothesis that TCF7L2 plays a dual role in thalamic neuron differentiation and functions as a terminal selector. Connectivity and cell clustering was disrupted in the thalamo-habenular region in Tcf7l2 -/embryos. The expression of subregional thalamic and habenular transcription factors was lost and region-specific cell migration and axon guidance genes were downregulated. In mice with postnatal Tcf7l2 knockout, the induction of genes that confer terminal electrophysiological features of thalamic neurons was impaired. Many of these genes proved to be TCF7L2 direct targets. The role of TCF7L2 in thalamic terminal selection was functionally confirmed by impaired firing modes in thalamic neurons in the mutant mice.These data corroborate the existence of master regulators in the vertebrate brain that maintain regional transcriptional network, control stage-specific genetic programs and induce terminal selection.of the mice colony, and Kacper Posyniak for assistance with sample staining and acquisition.

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Exploring Brain Genoarchitecture by Single and Double Chromogenic In Situ Hybridization (ISH) and Immunohistochemistry (IHC) on Cryostat, Paraffin, or Floating Sections

Cited by 26 publications

References 9 publications

Concentric ring topology of mammalian cortical sectors and relevance for patterning studies

Concentric ring topology of mammalian cortical sectors and relevance for patterning studies

Selective early expression of the orphan nuclear receptor Nr4a2 identifies the claustrum homolog in the avian mesopallium: Impact on sauropsidian/mammalian pallium comparisons

TCF7L2 regulates postmitotic differentiation programs and excitability patterns in the thalamus

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