Immunohistochemical characterization of postnatal changes in cerebellar cortical cytoarchitectures in ferrets

Abstract

We immunohistochemically characterized postnatal changes in cerebellar cortical cytoarchitectures in ferrets using markers for cerebellar cortical neurons and glial cells. Although 10 lobules of the vermis were already observed on postnatal day (PD) 4, Purkinje cells were still arrayed into two to three layers. Purkinje cells were aligned in a monolayer by PD 10 and formed mature shapes on PD 42 by developing their dendritic arbors. Parvalbumin immunostaining revealed relatively slower maturation of Purkinje cells in the Lobule X cortex than in other lobules. Basket and stellate cells emerged in the molecular layer on PDs 21 and 42, respectively. Rosette‐like arranged glutamate decarboxylase 65 and 67‐positive puncta were observed in the inner granular layer (IGL) on PD 21. Proliferating cell nuclear antigen immunostaining appeared in the outer zone of the external granular layer (EGL) containing progenitors of granular neurons on PDs 4–21. Bergmann glial processes extending vertically through the molecular layer and EGL were visible with GFAP immunostaining on PD 10 and thereafter. Their somata, aligned in the Purkinje cell layer, showed immunopositivity to Sox2 already on PD 4 and subsequently to S100 protein on PD 10. Sox2‐positive cells were found sparsely in the IGL. Few of them were NeuN positive on PD 90, predicting the possibility of adult neurogenesis. These immunohistochemical results revealed that ferrets underwent cerebellar cortical histogenesis during their postnatal life in sequences. Relatively slow development or maturation of the ferret cerebellum was revealed by the timing of the monolayer alignment and morphological maturation of Purkinje cells.

Neonatal valproic acid exposure produces altered gyrification related to increased parvalbumin-immunopositive neuron density with thickened sulcal floors

“…(1:1,000; IBL, Gunma, Japan) [53], and mouse antibodies to GFAP (1:1,000, G 3893, Sigma-Aldrich) [54] and parvalbumin (PV; 1:500; PV235, Swant, Switzerland) [52,54]. Anti-PV was used as a marker for bRG-derived neurons in ferret cortex on PD 20, as reported previously [52].…”

Granule Cells Constitute One of the Major Neuronal Subtypes in the Molecular Layer of the Posterior Cerebellum

“…1 A , middle). Therefore, it has been used to label MLIs in various animal species ( Ohshima et al, 1991 ; Kadowaki et al, 1993 ; Fortin et al, 1998 ; Kalinichenko and Pushchin, 2008 ; Kamiya and Sawada, 2021 ).…”

Differential staining patterns of immunohistochemical markers for neurogenesis staging in the premature cerebellum of ferrets and mice

“…Postnatal ages of 20 days in ferrets and 10 days in mice are identical stages of cerebellar corticohistogenesis, with EGL expanding the most. 7 Animals were perfused with 4% paraformaldehyde in phosphate buffer solution (PBS; pH 7.4) under deep anesthesia with 2% isoflurane gas. The cerebellum was removed, immersed in 30% sucrose in PBS, and embedded in an optimal cutting temperature compound.…”

“…Triple immunofluorescence staining was performed according to our previous procedures. 7 The primary and secondary antibodies used in this study are listed in Tables S1 and S2. Since immunohistochemical staining patterns for various neuronal and glial markers did not differ among vermal lobules in the developing cerebellum of the ferrets, 7 the present study examined lobule VI (as a representative lobule) in the deep portion of the primary fissure.…”