Prenatal effects of alprazolam treatment on the immature cerebellum of rats

• Published in: International journal of developmental neuroscience 2022-12, Vol.82 (8), p.727-735
• Main Authors: Mascarenhas, Fernanda Naves Araújo do Prado, Silva, Natália Ferreira, Menezes‐Reis, Lorena Tannús, Vieira, Lucélia Gonçalves, Hirano, Líria Queiroz Luz, Botelho, Françoise Vasconcelos, Ribeiro, Daniele Lisboa, Zanon, Renata Graciele
• Format: Article
• Language: English
• Subjects: benzodiazepines; cerebellum; development; glia; nervous system; neuron
• ISSN: 0736-5748; 1873-474X
• DOI: 10.1002/jdn.10222

The use of benzodiazepines (BZDs) during pregnancy, especially alprazolam, is common and its impact on the fetal neural tissue is not known. In this sense, the present study aimed to investigate the effects of prenatal treatment with alprazolam on the cerebellum of Wistar rat pups. Thirty animals (24 females and six males, CEUA protocol 014/17) were separated into pairs for copulation. Females were divided into three groups: Control (CT), treatment 1 (T1, 1.25 mg per animal), and treatment 2, which is an overdose (T2, 30 mg per animal). Alprazolam was administered 10 days before copulation and throughout pregnancy. We evaluated the number and weight of pups and the macroscopic changes in the brain. Eight neonates (n = 8) from each group were used in the following analyses: Cellular and chromatin density, gliosis, synaptic density, inflammation, and oxidative stress. The results showed no significant differences regarding the number of pups, body weight, and macroscopic changes. The morphological study focused on the external granular layer (EGL) that is presented only in the immature cerebellum. Here, we detected more cells after alprazolam treatment; the T2 group showed large nuclei and some pyknotic nuclei; also, both treated groups presented an increase in the euchromatin density compared with the control. The molecular and biochemical analyses used the total protein extract of the entire cerebellum and showed an increased expression of Iba‐1 and NF‐κBp65 but without indication of inflammation or degeneration in the T1 group. Overdose of alprazolam presented an increased level of oxidative degradation of lipids. The treatment with alprazolam during pregnancy involved cellular and molecular changes in the immature cerebellum. The cerebellum of rats that had intrauterine contact with alprazolam showed increased cells, probably microglia (elevated iba‐1 and NF‐kb compared with control), while overdose seems to be harmful showing lipid peroxidation.