The Conflicting Role of Caffeine Supplementation on Hyperoxia-Induced Injury on the Cerebellar Granular Cell Neurogenesis of Newborn Rats

Abstract

Preterm birth disrupts cerebellar development, which may be mediated by systemic oxidative stress that damages neuronal developmental stages. Impaired cerebellar neurogenesis affects several downstream targets important for cognition, emotion, and speech. In this study, we demonstrate that oxidative stress induced with high oxygen (80%) for three or five postnatal days (P3/P5) could significantly damage neurogenesis and proliferative capacity of granular cell precursor and Purkinje cells in rat pups. Reversal of cellular neuronal damage after recovery to room air (P15) was augmented by treatment with caffeine. However, downstream transcripts important for migration and differentiation of postmitotic granular cells were irreversibly reduced by hyperoxia, without rescue by caffeine. Protective effects of caffeine in the cerebellum were limited to neuronal survival but failed to restore important transcript signatures.

Prenatal and postnatal development of the rat cerebellar granule cells following maternal administration of thyme and tramadol

“…TEM, × 10000 DISCUSSION The current study aimed to examine the prenatal and postnatal effect of thyme co administration with tramadol on the cerebellar granule cells of albino rats. The cerebellar granule cells were chosen as a target of study because their unique maturation from mitotic granule cell precursors (GCPs) to postmitotic GCs, and their radial migration from EGL along to the formation of the IGL (28). LM examination of the developing cerebellum of the control and thyme groups revealed that in gestational day 13, the cerebellum was formed of a primordium that covered by the ventricular zone of the cerebellar.…”

Protective Effect of Dexmedetomidine against Hyperoxia-Damaged Cerebellar Neurodevelopment in the Juvenile Rat

“…Wistar rats were term-paired (Janvier Labs, Le Genest-Saint-Isle, France) and kept in single cages under standardized conditions (constant 12 h/12 h light-dark cycle, room temperature, 60% relative humidity, and ad libitum to food and water). In all of our animal experiments [ 28 , 43 , 44 , 45 ], newborn pups were kept with a lactating mother for the duration of the experiment. The animal procedures were assessed and approved by the local animal welfare authorities (LAGeSo, approval number G-0145/13 and G-139/18) and complied with institutional guidelines and ARRIVE guidelines.…”

“…Using an i.p. injection of ketamine (100 mg/kg), xylazine (20 mg/kg), and acepromazine (3 mg/kg), the pups were anaesthetized at P7, P9, P11, and P14 and subsequently transcardially perfused, as previously described [ 28 , 43 , 44 , 45 ]. The entire cerebellar tissue was removed immediately after decapitation and opening of the skull.…”

“…As previously described for the extraction of RNA [ 28 , 46 ], the total RNA from whole snap-frozen cerebellum of each animal was homogenized in peqGOLD RNAPure™ (PEQLAB Biotechnologie, Erlangen, Germany), according to the manufacturer’s instructions. After DNase treatment, 2 µg of RNA was reverse transcribed.…”

“…Paraffin-embedded whole cerebellums were cut into 5 µm-thick serial slices and subsequently mounted on SuperFrost Plus-coated slides (Menzel, Braunschweig, Germany) with three consecutive slices each, as previously described [ 28 , 46 ]. After deparaffinization in Roti-Histol (Carl Roth, Karlsruhe, Germany), twice for 10 min each and rehydration in descending ethanol concentration for 3 min each, heat-induced antigen retrieval was applied to the paraffin sections in citrate buffer (pH 6.0) in a microwave oven for 10 min at 600 W. To block the non-specific binding sites, the sections were incubated with appropriate blocking solutions for 1 h at room temperature (calbindin/DAPI: 3% bovine serum albumin (BSA), 0.05% TW-20 and 0.1% Triton X-100 in PBS; Pax6/PCNA/DAPI: 3% BSA and 0.2% Triton X-100 in PBS).…”