Hypoxia Alters miRNAs Levels Involved in Non-Mendelian Inheritance of Autism Spectrum Disorder in Mice

Ecma Güvenilir; Zeynep Yilmaz Şükranli; Mai R. S. Abusalim; Aslı Okan Oflamaz; Züleyha Doğanyiğit; Kemal Erdem Başaran; Mehmet Dolanbay; Minoo Rassoulzadegan; Serpil Taheri

doi:10.3791/68435

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Method Article

Hypoxia Alters miRNAs Levels Involved in Non-Mendelian Inheritance of Autism Spectrum Disorder in Mice

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DOI:

10.3791/68435

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July 11th, 2025

Ecma Güvenilir¹ , Zeynep Yilmaz Şükranli²^,³ , Mai R. S. Abusalim¹^,² , Aslı Okan Oflamaz⁴ , Züleyha Doğanyiğit⁴ , Kemal Erdem Başaran²^,⁵ , Mehmet Dolanbay⁶ , Minoo Rassoulzadegan²^,⁷^,⁸ , Serpil Taheri²^,³

¹Gevher Nesibe Genome and Stem Cell Institute, Erciyes University, ²Betül Ziya Eren Genome and Stem Cell (GENKÖK) Center, Erciyes University, ³Department of Medical Biology, Erciyes University, ⁴Department of Histology and Embryology, Yozgat Bozok University, ⁵Department of Physiology, Erciyes University, ⁶Department of Obstetrics and Gynecology, Faculty of Medicine, Erciyes University, ⁷University of Nice Sophia Antipolis, ⁸Department of Medical Genetics, Erciyes University

Corresponding Authors: Serpil Taheri <serpiltaheri@hotmail.com>

Summary

Here, we present a protocol to induce hypoxia in neonatal mice by delivering them in controlled environments with varying oxygen concentrations. Mouse models subjected to oxygen variations at birth have durably varied the expression of miRNAs involved in the induction of behavioral phenotypic changes such as autism.

Abstract

Hypoxia can occur to varying degrees for different reasons during the perinatal period, affecting development. We aim to examine the consequences of mild to severe neonatal hypoxia in mice, focusing on its potential link to neurodegenerative fetal brain dysfunctions related to autism spectrum disorder (ASD).

We induced hypoxia in neonatal mice by delivering them in controlled environments with varying oxygen concentrations. The results showed elevated levels of HIF-1α (a marker of hypoxia) in the cerebral cortex and hippocampus, as well as increased neurodegeneration in these regions. Moreover, we observed altered expression of ASD-related miRNAs (miR-19a-3p, miR-361-5p, miR-150-5p, miR-126-3p, and miR-499a-5p) in hypoxic groups, consistent with previous results in human and animal models of ASD. These molecular alterations were accompanied by behavioral changes such as decreased movement, speed, social interaction, and repetitive behaviors, as well as an increased tendency to exhibit corner-seeking behavior in mice subjected to hypoxic conditions. Although behavioral and molecular changes were detectable in the 12% O₂ group (mild hypoxia), the most pronounced changes were observed in the more severe hypoxia groups, at 10% O₂ and 8% O₂.

Overall, the results suggest that neonatal hypoxia can induce lasting molecular changes associated with neurodegenerative diseases with behavioral consequences. Moreover, even in normal birth, exposure to hypoxia during early development can impair memory and learning. The results highlight the need for early detection and intervention in newborns exposed to hypoxia to prevent long-term neurological problems. miRNA levels are regulated during development, and their modifications in tissues, especially in germ cells, constitute a risk factor for the next generation. The analyses provide a basis for studying the regulatory pathways that affect functional and behavioral changes under hypoxia.

Introduction

Defined as a state of oxygen deficiency, hypoxia can elicit unique cellular and molecular responses that are critical to understanding its role in various pathologies. İt can disrupt developmental sequences, leading to molecular, cellular, and neuronal alterations and injuries. Furthermore, early exposure to hypoxia interrupts central nervous system (CNS) development, impairing axonal connectivity and synapse development¹.

Additionally, although hypoxia has been shown to have adverse effects on CNS development, it may also play a critical physiological role under specific conditions, such as during embryogenesis....