Evaluation of sensomotor development, behavioral reactions and cognitive functions of the second generation of rats with hyperhomocysteinemia

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Abstract

The adverse maternal exposure during pregnancy leads to developmental disorders in the offspring that can be passed on to later generations. Epigenetic regulation of DNA transcription may mediate inherited metabolic diseases. An increase in homocysteine concentration in the blood is associated with epigenetic modifications of the genome, which can alter the fetal brain’s development program and cause cognitive impairment. The aim of our work was to identify changes in sensomotor development, behavioral reactions and cognitive functions of offspring of second generation rats (HcyF2) of hyperhomocysteinemia. Our results indicate that unconditioned reflexes and physical parameters are delayed in HcyF2 rats. In “open field”, HcyF2 rats showed higher levels of anxiety and decreased exploratory and motor activity, while coordination of movements studied in “rotarod” test was not impaired. Decreased limb muscle strength was shown in the “grip strength” test. Additionally, HcyF2 rats demonstrated an impaired learning and longterm memory in the Morris water maze. Biochemical analysis revealed an imbalance in the antioxidant systems, which was attributed to decreased activity of glutathione peroxidases and H2S synthesis enzymes. It was suggested that elevated homocysteine levels during pregnancy may result in epigenetic modifications of the genome, which can impact the metabolism of offspring and be inherited by future generations.

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O. V. Yakovleva

IFM&B, Kazan (Volga Region) Federal University

Author for correspondence.
Email: a-olay@yandex.ru

Department of Human and Animal Physiology

Russian Federation, Kazan

V. V. Skripnikova

IFM&B, Kazan (Volga Region) Federal University

Email: a-olay@yandex.ru

Department of Human and Animal Physiology

Russian Federation, Kazan

A. V. Yakovlev

IFM&B, Kazan (Volga Region) Federal University

Email: a-olay@yandex.ru

Department of Human and Animal Physiology

Russian Federation, Kazan

G. F. Sitdikova

IFM&B, Kazan (Volga Region) Federal University

Email: a-olay@yandex.ru

Department of Human and Animal Physiology

Russian Federation, Kazan

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2. Fig. 1. The homocysteine levels, litter size and weight, and weight gain in the second generation rats born from dams with HHcy. (a) – the scheme shows the protocol for obtaining second-generation offspring, (б) – homocysteine level in animals born from females with HHcy in the first (HcyF1) and second generation (HcyF2). Percentage of animals with plasma homocysteine concentrations within the normal range (0–9 µM, white sector), mild HHcy (10–20 µM, light gray sector), moderate HHcy (21–50 µM, dark gray sector). (в) – Litter size and total weight (shaded) in control females (white square), in females with HHcy (gray square). (г) – Birth weight of rats in control (white square) and HcyF2 groups (gray square). Rectangle – 25–75%, square inside – average value, central line – median, “whiskers” – minimum/maximum values. (д) – dynamics of weight gain in rat pups of the control (white square) and HcyF2 groups (gray square). * p < 0.05 compared to the control group.

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3. Fig. 2. Changes in motor and exploratory activity, level of anxiety in the “open field” test in rats in the control and HcyF2 groups. Number of head elevations (a), vertical stands (б), explored holes “head dips” (в), crossed squares (г), acts of grooming (д) and defecation (е). P8, P16, P26 – age periods. White bar – control group, gray bar – HcyF2 group. * p < 0.05 relative to the control group.

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4. Fig. 3. Distance (a) and time (б) spent on a rotating cylinder in the “rotarod” test and time spent on the grid in the “grip strength” test (в) in rats of the сontrol (white bars) and HcyF2 (gray bars) groups at different ages. P8, P14, P16, P18, P26 – age periods. Rectangle – 25–75%, square inside – average value, central line – median, “whiskers” – minimum/maximum values. * p < 0.05 compared to the control group.

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5. Fig. 4. Evaluation of learning and memory in HcyF2 rats in the Morris water maze test. (a) – changes in platform search time during training in the Morris water maze in both control (white circles) and HcyF2 (gray squares) rats, (б) – examples of the trajectories of platform search by rats from the control and HcyF2 groups, (в) – platform search time 1 and 24 hours after training. * p < 0.05 relative to control.

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6. Fig. 5. Levels of sulfides and oxidative stress in the brain of rats, in control and HcyF2 groups. The concentration of HSions (a), rate of HSproduction (б), level of malondialdehyde (MDA) (в), activity of glutathione peroxidases (GPx) (г) in the brain of rats of the control group (white columns), HcyF2 (gray columns). Rectangle – 25–75%, square inside – average value, central line – median, “whiskers” – minimum/maximum values. * p < 0.05 compared to the control group.

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